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rd3 International Plant Physiology Congress

Challenges and Strategies in Plant Biology Research

December 11-14, 2015Convention Centre, JNU

New Delhi, India

Indian Society for Plant PhysiologyJawaharlal Nehru University

National Institute of Plant Genome ResearchNew Delhi, India

1958

IPPCongress2015

Published by:Indian Society for Plant PhysiologyF-4, A Block, NASC ComplexIARI, New Delhi-110012

Publication Committee

Dr. R.K. Sairam

Dr. Ajay Arora

Dr. Sanjay Kumar

Dr. Rita Sharma

Dr. Viswanathan Chinnusamy

Dr. Suresh Nair

Dr. Sneh L. Singla-Pareek

Dr. Pramod Kumar

Dr. N. Raghuram

Dr. Alok K. Sinha

Dr. Rabi N. Sahoo

The views expressed in this publication by the authors are their own and these do not necessarily reflect those of the organizers

Cover page designed by Prof. Sanjay Kapoor, Department of Plant Molecular Biology, University of Delhi, South Campus, New Delhi.

Printed at Cambridge Printing Works, B-85, Naraina Industrial Area, Phase-II, New Delhi-110028, Tel. 011-25891262

OUR SPONSORS

MESSAGE

The President of India, Shri Pranab Mukherjee, is happy to know that the Indian Society for Plant Physiology in

association with Jawaharlal Nehru University and National Institute of Plant Genome Research, New Delhi is rdorganizing the 3 International Plant Physiology Congress on “Challenges and Strategies in Plant Biology

Research” from December 11-14, 2015.

The President extends his warm greetings and felicitations to the organizers and participants and sends his best wishes for the success of the event.

Deputy Press Secretary to the President

Tel. (O): 91-11-23793528 (Direct), 91-11-23015321 Extn. 4442, Fax : 91-11-23010252Email : [email protected]

Shri Pranab Mukherjee

President of IndiaRashtrapati Bhavan

New Delhi-110004

November 09, 2015

MESSAGE

rdI am immensely pleased to know that the “3 International Plant Physiology Congress: Challenges and

Strategies in Plant Biology Research” is being organized by the Indian Society for Plant Physiology in

association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome

Research (NIPGR), New Delhi.

The theme of the International Congress is quite appropriate to the present scenario of changing climate. Fast

changing global climate is posing tougher challenges to the crop scientists in terms of several abiotic stresses.

In future, it is going to be the synchronized efforts of the Breeders, Crop Physiologists, Agronomists and

Molecular Biologists to work on a common platform to find out different strategies to increased crop production.

Understanding the basic physiological processes and the possible interventions at different steps are further to

be explored at molecular level.

I am sure that the congress would provide a good opportunity to the youth to expose themselves to various

research areas.

I wish the event a grand success.

( )

199-Q, Krishi Bhawan, New Delhi-110001, Tel : 91-11-23782343, Fax : 91-11-23074190

Minister of State of Agriculture and Farmers Walfare

Government of India, Krishi Bhawan New Delhi-110001

November 20, 2015

Sanjeev Kumar Balyan

MESSAGE

rdI am glad to know that the “3 International Plant Physiology Congress: Challenges and Strategies in

Plant Biology Research” is being organized by the Indian Society for Plant Physiology in association with

Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR),

New Delhi.

We have seen time and again as to how science can be effectively put to work to raise agricultural productivity.

In India, the "Green Revolution" ushered in an era of food sufficiency and rural prosperity. The "Green

Revolution" was a success due to the introduction of improved seeds, irrigation and plant protection

measures combine with a visionary and positive policy support, liberal public funding for agricultural research

and development, and dedicated work of farmer.

Although technological interventions can provide a new hope for Indian agriculture, there are two factors that

are likely to be crucial determinants on whether or not we will be able to ultimately use this technological

prowess to gain actual benefits. The first is the issue of public concerns on safety, especially with respect to

health and environment. The second is the restrictive role of IPR in actually denying access of these

technologies to the developing countries. I am sure that appropriate deliberations will be held among other

things, on both these issues, during the Congress.

I wish the organizers of this Congress all success in meeting their objectives.

(Ashutosh Sharma)

Ashutosh Sharma

SecretaryGovernment of India

Ministry of Science and TechnologyDepartment of Science and Technology

November 18, 2015

Technology Bhavan, New Mehrauli Road, New Delhi - 110016Tel: 91-11-26511439/26510068, Fax: 91-11-26863847, Email: [email protected], website: www.dst.gov.in

MESSAGE

I am happy to know that the Indian Society for Plant Physiology in association with Jawaharlal Nehru

University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is rdorganizing the “3 International Plant Physiology Congress: Challenges and Strategies in Plant Biology

Research” from December 11-14 at, JNU, New Delhi, India.

stThe proposed International Congress has great relevance for the 21 century because for feeding the

burgeoning population of India and for attaining food security mission oriented efforts for augmenting food

productivity and its sustainability through physiological, genetic and biotechnological approaches are critical.

Plant Physiologists have a key role to play by providing deeper understanding of the physiological processes

and their effective manipulation for improving the productivity of crops.

I hope that this International Congress will address such important issues and come out with new directions for

researchers for tailoring productive plant types and that would help break the yield barrier and thereby ensure

sustained food and nutrition security.

I wish the Congress all success.

(S. Ayyappan)

Secretary and Director General Government of India

Department of Agricultural Research & Education and

Indian Council of Agricultural ResearchMinistry of Agriculture and Farmers Welfare

Krishi Bhavan, New Delhi-110001

October 27, 2015

Tel.: 91-11-23382629; 23386711; Fax: 91-11-23384773; Email: [email protected]

S. Ayyappan

MESSAGE

rdIt is a matter of immense pleasure to know that the “3 International Plant Physiology Congress:

Challenges and Strategies in Plant Biology Research” is being organized by the Indian Society for Plant

Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant

Genome Research (NIPGR), New Delhi.

The theme of the conference “Challenges and Strategies in Plant Biology Research” is quiet appropriate

and calls for immediate attention by the Plant Physiologists and other scientists across the nation. Plant

Physiologists along with breeders and molecular biologists form a cohesive group that is entrusted with the

responsibility of improving the productivity of our present day crop plants keeping in view of the depleting

natural resources and constantly changing environment. An understanding of the mechanisms underlying the

physiological processes, the effective manipulation of these processes to improve the quality and quantity of

the produce and tailoring of productive eco-specific plant types is the need of the hour.

I hope this International Congress will provide a good forum to discuss some of these issues and evolve future

strategies that will enable us to deal with the problem of food and nutritional security at the national level.

I wish this congress a grand success.

(J.S. Sandhu)

Deputy Director General (Crop Science)Indian Council of Agricultural Research

Krishi Bhawan, Dr. Rajendra Prasad RoadNew Delhi-110001

November 16, 2015

Jeet Singh Sandhu

Ph: (Off.) 23382545 Fax : 23097003 E-mail: [email protected]

MESSAGE

It is a matter of great pleasure that Indian Society for Plant Physiology in association with Jawaharlal Nehru

University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is rdorganizing the “3 International Plant Physiology Congress: Challenges and Strategies in Plant Biology

Research” from December 11-14, 2015 at JNU, New Delhi, India.

The congress will provide a good opportunity to the Plant Physiologists/Biologists to review the recent progress

made in the various emerging areas of Plant Physiology and to focus on some of the pertinent issues

concerning plant productivity in a changing scenario of climatic regime, dwindling natural resources and fast

growing human population.

Plant Physiology, has and will continue to play an important role in agricultural research and development. With

the technological advances in the field of crop breeding and molecular biology, there is a need for integrated

approach wherein knowledge of Plant Physiology in the areas of photosynthesis, abiotic and biotic stresses

tolerance, nutrient use efficiency, post harvest physiology and so on, can be applied to address the key

questions pertaining to improved crop productivity in harmony with the changing environment.

I am confident that the scientific deliberations of the congress will provide the right direction for the future

strategies in overcoming the challenges faced by the agricultural sector in order to provide food and nutritional

security at the international level.

I wish the congress a great success.

(S.K. Sopory)

Vice ChancellorJawaharlal Nehru University

New Delhi 110067&

PresidentIndian Society for Plant Physiology

November 16, 2015

Sudhir K. Sopory

MESSAGE

I am glad to know that the Indian Society for Plant Physiology in association with Jawaharlal Nehru University rd(JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is organizing the “3

International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” from

December 11-14, 2015 at JNU, New Delhi, India.

A rapid growth in new knowledge being generated in diverse areas of Plant Biology is apparent. It is now time to

consolidate the same. Thus, the theme of the Congress “Challenges and Strategies in Plant Biology Research”

is quite appropriate and reflects the current approach. Plant Physiologists have played apivotal role in

deciphering functional aspects of plants to help improve the productivity of our crops. This is even more

relevant today in view of the fast depleting natural resources and climate change. A deeper and precise

understanding of the mechanisms involved in regulation of diverse physiological processes, the effective

manipulation of such processes to improve the quality and quantity of the produce and connecting the same to

societal benefitsneed immediate attention. The Congress will provide an excellent platform to discuss some of

these issues and plan future strategies that will help evolve sustainable food and nutritional security,not only at

the national level but also at global scale.

I understand that a large number of researchers would attend this mega scientific event and feel confident that it

would be a successful and rewarding experience.

My good wishes to the Congress.

(Akhilesh K. Tyagi)

DirectorNational Institute of Plant Genome Research

Aruna Asaf Ali MargNew Delhi-110067

November 16, 2015

Akhilesh K. Tyagi

MESSAGE

It is my great pleasure to learn that the Indian Society for Plant Physiology in association Jawaharlal Nehru

University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is rdorganizing the “3 International Plant Physiology Congress: Challenges and Strategies in Plant

Biology Research” from December 11-14, 2015 at JNU, New Delhi, India.

Several issues that are confronting the Indian agriculture today has to find a solution through the application of

Plant Physiology such as crop tolerance towards drought, heat, salinity and nutrient stresses, greater source-

sink relationship, higher micro-nutrient content in the food grain, better shelf life of vegetable and fruits apart

from increase in vase life of flowers. Organizing this conference at this point of time will explore new

knowledge and approaches in solving some of the chronic issues. I am of the opinion that Plant Physiology,

Crop Improvement and Molecular Biology are to be integrated in a manner that we are in a position to address

some of the issues in a much more precise manner than what we have been doing in the past.

I wish the Congress a grand success.

(Trilochan Mohapatra)

DirectorICAR-Indian Agricultural Research Institute

New Delhi-110012

th29 October, 2015

Trilochan Mohapatra

INDIAN SOCIETY FOR PLANT PHYSIOLOGY

The Indian Society for Plant Physiology (ISPP), established in 1958, is one of the oldest academic societies in

India. It owes its origin to the vision and zeal of eminent plant scientists Prof. Boshi Sen, Prof. R.D. Asana, Prof.

J.J. Chinoy and Prof. K.K. Nanda and the financial support of Government of India. Since then, the society has

been serving the cause of research and education in the field of Plant Physiology, allied disciplines and

agriculture. The society is based at National Agriculture Science Centre of Indian Council of Agricultural

Research (ICAR), New Delhi. The society is a member of Global Plant Council, a coalition of plant and crops

societies from across the globe. The main focus of the society is to bring plant scientists together to work

synergistically towards solving the pressing problems facing humankind and to generate a critical mass to

provide a strong platform for deliberations on important issues of agricultural research, education and

extension and presenting views of the scientific community as policy inputs to planners and decision makers at

national/ international levels. To achieve this, the society organizes and supports national and international

conferences, seminars, symposia, workshops and brainstorming sessions on important issues of plant biology

and agricultural sciences. It also publishes journal and proceedings for advancement of agricultural research

and development. The official publication of the Society, the Indian Journal of Plant Physiology was launched

in the year 1958. Since then this journal is being published regularly and is highly acclaimed for its scientific

content and indexed worldwide. ISPP has instituted various awards to recognize and promote scientists for

their excellent research contribution in the areas of plant sciences and agriculture. The society bestowed

fellowships to >250 eminent researchers during the last fifty years and invites applications annually during the

month of July/August every year for the following awards/fellowship.

Memorial lecture awards

: S.K. Sinha Memorial Lecture

K.K. Nanda Memorial Lecture

G.V. Joshi Memorial Lecture

Recognition awards

: J.J. Chinoy Gold Medal

J.C. Bose Gold Medal

Young Scientist awards (upto 40 years)

: R.D Asana Gold Medal

R.H. Dastur Gold Medal

Best paper award

(Published in IJPP)

: G.S. Sirohi Award

Fellowships Fellowship of ISSP (FISSP)

1958

F-4, A Block, NASC Complex, New Delhi-110012 www.ispponline.org

IPPCongress2015

3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India

CONTENT

S.No. Title Page

Scientific Program

1. ISPP Memorial Award Lectures 1-3

2. Plenary, Lead and Oral Presentations 4-107

3. Young Scientist Presentations 109-114

4. Poster Presentations 115-357

Session A : Abiotic Stress Adaptation and Management (PA001 to PA135) 115-182

Session B : Signaling and Phototransduction (PB136 to PB165) 183-198

Session C : Genomics, Epigenomics and Regulation Biology (PC166 to PC217) 199-224

Session D : Plant Metabolism and Metabolomics (PD218 to PD240) 225-236

Session E : Biotic Stress Management and Interactions (PE241 to PE287) 237-260

Session F : Plant Nutrition Physiology and Genetics (PF288 to PF322) 261-278

Session G : Plant Growth Regulators and Applications (PG323 to PG370) 279-302

Session H : Photosynthesis (PH371 to PH388) 303-311

Session I : Genomics-assisted Plant Improvement (PI389 to PI425) 312-330

Session J : Flowering and Post Harvest Physiology (PJ426 to PJ432) 331-334

Session K : Computational and Systems Biology (PK433 to PK452) 335-344

Session L : Climate Change and Agriculture (PL453 to PL462) 345-349

Session M : Reproductive Biology (PM463 to PM478) 350-357

5. Author Index 359-370

biotic Stres

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ORGANIZATION

CHAIR : Sudhir K. Sopory, Vice-Chancellor, JNU

CONVENORS : Ashwani Pareek, JNU & Madan Pal, IARI

CHIEF PATRONS

Ashutosh Sharma K. VijayRaghavan S. Ayyappan

Secretary, DST, New Delhi Secretary, DBT, New Delhi Secretary, DARE & DG, ICAR, New Delhi

PATRONS

Asis Datta, NIPGR, New Delhi T. Mohapatra, Director, IARI, New Delhi M.S. Swaminathan, MSSRF, Chennai

Anil Grover, UDSC, New Delhi J.S. Sandhu, DDG, ICAR, New Delhi A.K. Tyagi, Director, NIPGR, New Delhi

J.P. Khurana, UDSC, New Delhi D.K. Sharma, Director, CSSRI, Karnal

INTERNATIONAL STEERING COMMITTEE

Hans Bohnert, University of Illinois, USA Andy Pereira, Virginia Bioinfor. Instt. USA Rakesh K. Singh, IRRI, Philippines

O.P. Dhanker, UMass Amherst, USA Himadri B. Pakrasi, I-CARES, WUStL, USA Paul J. Thornalley, Univ. of Warwick, UK

Govindjee, University of Illinois (UC), USA K.G. Raghothama, Purdue Agriculture, USA Neil Turner, UWA, Australia

Abdelbagi Ismail, IRRI, Philippines V.R. Reddy, USDA-ARS, Baltimore, USA Elizabeth Vierling, UMass Amherst, USA

Jez Joseph, WUStL, USA Julian Schroeder, UCSD, USA Jian-Kang Zhu, Purdue University, USA

Autar K. Mattoo, USDA-ARS, USA Vijay Sharma, Monsanto, St. Louis, USA Tanguy Lafarge, FARCID, France

K.H.M. Siddique, UWA, Australia Mohan Singh, Univ. of Melbourne, Australia K. Rajareddy, Mississippi, USA

NATIONAL ORGANIZING COMMITTEE

A.K. Tyagi, NIPGR, New Delhi P.K. Aggarwal, New Delhi K.C. Bansal, NBPBR, New Delhi

K.S. Charak, DBT, New Delhi Bharat B. Chattoo, MSU Baroda M.B. Chetti, ICAR, New Delhi

Renu Khanna-Chopra, IARI, New Delhi Ranjan Das, AAU, Jorhat P.S. Deshmukh, IARI, New Delhi

M.C. Ghildiyal, IARI, New Delhi Ganesh Iyer, RRC, Mumbai James Jacob, RRII, Kottayam

B.L. Kakralya, SKNU, Jobner M. Kar, OUAT, Bhubaneshwar P. Anand Kumar, ANGRAU, Hyderabad

M. Maheshwari, CRIDA, Hyderabad A.N.L. Majumder, BI, Kolkata A.S. Nandwal, CCSHAU, Hisar

K.K. Narayanan, Metahelix Bangalore A. Narayanan, Coimbatore Ajay K. Parida, MSSRF, Chennai

Deepak Pental, UDSC, New Delhi A.S. Raghavendra, Hyderabad V.S. Reddy, ICGEB, New Delhi

P.C. Sharma, CSSRI, Karnal N.K. Singh, IARI, New Delhi V.P. Singh, DU, Delhi

Ramesh V. Sonti, CCMB, Hyderabad G.C. Srivastava, IARI, New Delhi J.P. Srivastava, BHU, Varanasi

Renu Swarup, DBT, New Delhi Inyatullah Tahir, Srinagar H.S. Talwar, IIMR, Hyderabad

M. Udaykumar, UAS, Bangalore K. Veluthambhi, MKU, Madurai C. Vijayalakshmi, TNAU, Coimbatore

Usha Vijayraghavan, IISc, Bangalore Usha B. Zehr, MAHYCO, Jalna M.S. Sheshshayee, UAS, GKVK, Bangalore

S.R. Voleti, IIRR, Hyderabad K.B. Hebbar, CPCRI, Kasaragod Nataraja N. Karaba, UAS, GKVK, Bangalore

PROGRAM COMMITTEE

A.K. Tyagi, NIPGR, New Delhi M.Z. Abdin, Jamia Hamdard, New Delhi V. Chinnusamy, IARI, New Delhi

Anil Grover, UDSC, New Delhi J.P. Khurana, UDSC, New Delhi Hemant R. Kushwaha, ICGEB, New Delhi

Ashwani Pareek, JNU, New Delhi T.R. Sharma, NRCPB, IARI, New Delhi P. Verma, Bioseed Res. India, Hyderabad

IPPCongress2015


PUBLICATION COMMITTEE

R.K. Sairam, IARI, New Delhi V. Chinnusamy, IARI, New Delhi N. Raghuram, GGSIPU, New Delhi

Dr. Ajay Arora, IARI, New Delhi Suresh Nair, ICGEB, New Delhi Alok K. Sinha, NIPGR, New Delhi

Sanjay Kumar, IHBT, Palampur Sneh L. Singla-Pareek, ICGEB, New Delhi R.N. Sahoo, IARI, New Delhi

Rita Sharma, JNU, New Delhi Pramod Kumar, IARI, New Delhi Madan Pal, IARI, New Delhi

FINANCE COMMITTEE

V.P. Singh, IARI New Delhi Madan Pal, IARI, New Delhi G. Ramarao, Tirupati

V.K. Singh, CISH, Lucknow Alok K. Sinha, NIPGR, New Delhi B.C. Tripathy, JNU, New Delhi

LOGISTICS COMMITTEE

Paramjit Khurana, UDSC, New Delhi Anjali Anand, IARI, New Delhi Sanjay Kapoor, UDSC, New Delhi

Sangeeta Khetarpal, IARI, New Delhi Ishwar Singh, IIMR, New Delhi Manoj K. Sharma, JNU, New Delhi

Lekshmy S., IARI, New Delhi Bhupinder Singh, IARI, New Delhi Sneh L. Singla-Pareek, ICGEB, New Delhi

REGISTRATION COMMITTEE

Ashish K. Nandi, JNU, New Delhi Sanjay Kapoor, UDSC, New Delhi S. Naresh Kumar, IARI, New Delhi

Hemant R. Kushwaha, ICGEB, New Delhi Girdhar Pandey, UDSC, New Delhi Sudhir Kumar, IARI, New Delhi

Manoj K. Sharma, JNU, New Delhi V.P. Singh, IARI, New Delhi Puja Rai, IARI, New Delhi

TRANSPORT COMMITTEE

Ajay Arora, IARI, New Delhi Vijay Paul, IARI, New Delhi Nirala Ramchiary, JNU, New Delhi

J.P. Singh, IARI, New Delhi S.D. Singh, IARI, New Delhi Praveen Verma, NIPGR, New Delhi

ACCOMMODATION COMMITTEE

Ishwar Singh, IARI, New Delhi Ajay Arora, IARI, New Delhi Ranjit R. Kumar, IARI, New Delhi

Rakesh Pandey, IARI, New Delhi R.N. Sahoo, IARI, New Delhi Dharmender Singh, IARI, New Delhi

I.P. Singh, IARI, New Delhi V.P. Singh, IARI, New Delhi

FOOD COMMITTEE

S. Chakraborty, JNU, New Delhi Ajay Arora, IARI, New Delhi Madan Pal, IARI, New Delhi

Neera B. Sarin, JNU, New Delhi I.P. singh, IARI, New Delhi Rana P. Singh, JNU New Delhi

Sneh L. Singla-Pareek, ICGEB, New Delhi J.P. Singh, IARI, New Delhi V.P. Singh, IARI, New Delhi

CULTURAL COMMITTEE

Atul K. Johri, JNU, New Delhi Ashish K. Nandi, JNU, New Delhi Renu Pandey, IARI, New Delhi

Rita Sharma, JNU, New Delhi Sneh L. Singla-Pareek, ICGEB, New Delhi Shashi Meena, IARI, New Delhi

Archana Kumari, IARI, New Delhi

ISPP Memorial Award Lectures

1

IPPCongress2015


Professor S.K. Sinha Memorial Lecture

Physiological and molecular basis of abiotic stress tolerance in wheat

Renu Khanna-Chopra*

Stress Physiology Lab, Water Technology Centre, Indian Agricultural Research Institute,New Delhi-110012, India

*Email: [email protected]

Drought and high temperature stress are major environmental factors limiting wheat productivity worldwide.

Understanding physiological and genetic basis of drought tolerance is important for breeding for drought resistance in

wheat to stabilize productivity in stress environments. Both drought and heat stress enhance monocarpic senescence

in wheat which impacts crop productivity. Drought and heat tolerant wheat cultivars exhibited delayed senescence in

stress environments. Oxidative stress is an important component of both abiotic stresses and senescence. Our

studies have shown that effective antioxidant defense plays an important role in delayed senescence in stress tolerant

wheat cultivars compared to susceptible genotypes. Senescence is also influenced by reproductive sink strength in

wheat. Desinked wheat plants had a slower rate of senescence associated with lower ROS levels, slower nitrogen

mobilization and increased antioxidant defense compared to plants with effective sink. Drought acclimation an important

component of drought tolerance is also associated with co-ordinated antioxidant defense in wheat under field condition.

A population of 206 recombinent inbred lines derived from drought susceptible and drought tolerant wheat cultivars

WL711/C306 was phenotyped for grain yield and related traits under water deficit and irrigated conditions in seven

different environments. Novel QTLs for grain yield and components were identified on chromosome 3BL which need

to be further validated and fine mapped for marker development.

Renu Khanna-Chopra


2

IPPCongress2015


Professor G.V. Joshi Memorial Lecture

A plant physiology route to bioeconomy along the Himlayan altitudes

Sanjay Kumar*

CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, HP, India


A change in altitude accompanies alteration in environmental conditions such as partial pressure of gases including

CO2, temperature, and radiation load. Accordingly, plant species possess mechanisms encompassing genes and

proteins that offer adaptive advantage for the niche on one hand and also can potentially be utilized for economic

activities through technological interventions on the other hand. Our strategy revolved around identifying the plant

processes suitable for industrial use. We discovered that plants exhibited biochemical plasticity to compensate for

low partial pressure of CO2 at high altitude through alteration of CO

2 fixation pathway. The pathway was also linked to

efficient nitrogen assimilation. C3 plants fixed CO2 both through phosphoenolpyruvate carboxylase (PEPCase) and

ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and channelized the part of fixed carbon to capture

the ammonia evolved through metabolic reactions such as photorespiration. The identified mechanism was evaluated

in Arabidopsis through transgenic approach, which showed growth advantage to plants at low nitrogen and improved

carbon fixation. Yet another feature of plant species in the hostile environment of high altitude was modulation of

metabolic processes for quick growth and development along with operation of varied protective mechanisms. Some

of the species possessed protein with unusual properties as well. For example, one of the enzymes, Cu, Zn-superoxide

dismutase (Cu, Zn-SOD) tolerated autoclaving in Potentilla atrosanguinea, a plant that is distributed at altitude

>3500 m. Further, the enzyme catalyzed the reaction even at sub-zero temperature of (-)10°C and also beyond 50°C.

The enzyme imparted tolerance to abiotic stresses through varied processes; by lignification of vascular tissues and

altering the signaling pathway as discerned in transgenic Arabidopsis. The enzyme was engineered by directed

mutation of targeted amino acids to enhance the thermo-stability further. Realizing enormous industrial use of the

enzyme, the technology for its production and purification was transferred to an industry at premium price. While the

identified adaptive mechanism have implications for agriculture industry to develop stress tolerant plants, unique

proteins will find place in suitable industry for societal benefits.

Sanjay Kumar


3

IPPCongress2015


Professor K.K. Nanda Memorial Lecture

Tomato functional genomics in reverse: Mutants as an aid to biofortify

tomato fruits

Rameshwar Prasad Sharma*

Repository of Tomato Genomics Resources, School of Life Sciences, University of Hyderabad,India


During last two decades genome sequencing of several crop species has uncovered vast repertoire of gene sequences,

yet the functions of many genes remains to be determined. One of best way to understand the function of a gene is

to examine how the mutations alter the function of the gene and its effect on metabolic or morphological phenotype of

the plant. Currently available reverse genetics tools along with genome-wide mutagenesis and gene-indexed mutant

collections have revolutionized the way in which gene functions are studied in plants. In tomato we have created

EMS-mutagenized populations to investigate gene function both by forward and reverse genetic approaches. EMS-

mutagenized populations were used to isolate novel mutants of tomato using TILLING (Targeted Induced Local

Lesions in Genome). Using the natural accessions and mutants we examined accumulation of carotenoids and folate

in tomato fruits and influence of mutation/SNP on the primary metabolome of tomato fruits. The metabolite data

sorting allowed us to identify the natural accessions with biochemical extremes in the carotenoids and folate levels in

tomato fruits. Our results indicated that genes contributing to folate biosynthesis in plants are recalcitrant and show

little polymorphism in the gene sequences. The mutations in the diverse pathways regulating light signalling, leaf

architecture and ethylene biosynthesis affected the levels of carotenoids and folate in fruits. Our results indicate that

the levels of folate and carotenoids in tomato fruits are regulated by a complex intertwining of the cellular metabolic

pathways.

R.P. Sharma

4

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Day 1: December 11, 2015

Plenary Session-I

Regulation of membrane transport by the CBL-CIPK signaling network

Sheng Luan*

Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA

*Correspondence: [email protected]

Almost all signal transduction processes in plants involve Ca2+ that serve as a vital second messenger. Understanding

how Ca2+ mediates the cellular responses triggered by myriad environmental signals is one of the most important

goals for plant biologists in years to come. Recent studies uncovered a new family of Ca2+ sensors (CBLs) that target

a family of protein kinases (CIPKs), establishing a novel paradigm for Ca2+ signaling in plants. The CBL calcium

sensors and CIPK protein kinases are encoded by two multigene families. In Arabidopsis (with the smallest genome

among flower plants), at least 10 CBLs and 26 CIPKs are identified. Each CBL interacts with a selected repertoire of

CIPKs and each CIPK interacts with one or more CBLs. Some CBLs have common CIPK targets and some CIPKs

share CBL regulatory proteins. Such specific and overlapping scheme in the CBL-CIPK interaction suggests both

specific and redundant functions among the members of the two gene families. Genetic analysis has begun to reveal

the function of individual CBLs and CIPKs. Available results indicate that CBL-CIPK interactions form an extensive

network that functions in a number of signaling pathways including plant responses to abiotic stress, nutrition status,

and abscisic acid (ABA). In addition, more recent work identified protein phosphatases that down-regulate the pathways

activated by CBL-CIPK network and other kinases. In particular, ion channels serve as targets for such regulations in

which kinases and phosphatases form specific pairs. Examples and mechanisms of ion channel regulation by CBL-

CIPK network will be presented.

Sheng Luan

5

IPPCongress2015



Plenary Session-I

Lessons from the good and the evil in plant-microbe interactions

Heribert Hirt*

King Abdullah University of Sciences and Technology, Thuwal, Saudi Arabia and Institute of PlantSciences of Paris Saclay, France


Plants are permanently challenged by a number of adverse environmental factors including a large variety of microbes

and the innate immune system revealed to play a key role in regulating plant growth and defence. Apart from studying

typical plant pathogens, we are interested in extreme cases of plant-microbe interactions, such as Salmonella

typhimurium, one of the major causes of food poisoning and death in humans. S. typhimurium is often found in soil

samples and can infect and propagate both in animals and plants. S. typhimurium was found to apply different

strategies to infect plants, some are specific to plants and others also suppress the human innate immune system.

However, the ability of a variety of plants to cope with stress also depends on their association with rhizosphere

microbes, but the mechanisms of the beneficial interaction are poorly understood. In the DARWIN21 project (http://

www.darwin21.net), the biodiversity of plant root microbes and the mechanisms of enhancing the tolerance of plants

to heat, salt and drought stress are investigated. In short, endophytic microbes are isolated from pioneer plants in

different deserts of the world. In a high throughput screening, we select bacteria for plant growth (PGP) and/or stress

tolerance promotion (STP). Different biochemical and molecular approaches are used to study the mechanisms of

PGP and STP activity of selected strains. Greenhouse and field trials with a number of candidates showed enhanced

stress tolerance of crop plants, confirming that beneficial endophytes offer an alternative way to do sustainable

agriculture in arid regions of the world.

Heribert Hirt

6

IPPCongress2015



Plenary Session-I

Functional analysis of cryptochrome 2 (OsCRY2) and a HY5 ortholog

in regulating rice development

Jitendra P. Khurana*

Interdisciplinary Centre for Plant Genomics & Department of Plant Molecular Biology, University ofDelhi South Campus, New Delhi 110021, India


Light plays a pivotal role in regulating plant development all through its life cycle. The photomorphogenically active

light is perceived by many different sensory receptors, including red/far-red sensing phytochromes, blue/UV-A sensing

cryptochromes and phototropins. In recent years, our laboratory has been focusing on the functional characterization

of cryptochromes from Brassica (Cry1 and CRY2) and rice (CRY2) and an ortholog of Arabidopsis HY5 from rice,

which acts as a central integrator downstream to all the major sensory photoreceptors. The transcript levels of rice

CRY2 gene are not affected by light but it encodes a protein that is rapidly degraded on exposure to blue light. The

OsCRY2 gene when over-expressed in rice caused rapid development of seedlings and the adult plants were semi-

dwarf. Even when grown in dark, the coleoptiles of OsCRY2ox young seedlings were short and displayed early leaf

emergence. Unexpectedly, both the wild type and OsCRY2ox transgenics flowered at the same time. However, when

OsCRY2 gene was over-expressed in Arabidopsis, it caused hyper-photo-morphogenesis, including early flowering.

The HY5 ortholog in rice, which is expected to work downstream to Cry2 as well, however, did not alter flowering time,

when over-expressed both in rice and Arabidopsis but it did complement hy5 mutant of Arabidopsis and also caused

strong reduction in plant height in rice. These data provide evidence that sensory photoreceptors like Cry2 and its

downstream signaling components like Hy5 can be exploited to alter photomorphogenic development, including

control of flowering time and plant height.

Jitendra P. Khurana

7

IPPCongress2015



Concurrent Session A-I

Genes and germplasm for climate resilient agriculture

K.C. Bansal*

ICAR-National Bureau of Plant Genetic Resources, New Delhi-110012, India


Abiotic stresses adversely affect crop productivity worldwide. To develop abiotic stress tolerant and climate resilient

crop varieties, identification of key genes and germplasm lines which can serve as trait donor is critical. We focused

on cloning and functional validation of candidate genes, transcriptomics to identify novel genes and gene networks,

and large-scale phenotyping to identify donor germplasm lines for abiotic stress tolerance in important crops. We

cloned and validated promoters (LEA1) and genes coding for transcription factors (DREB1s/CBFs, Zinc finger proteins),

signalling components (ERA1) and effector genes (osmotin, LEA4, APX, genes for osmolyte synthesis). Deployment

of these genes and promoters for transgenic crops development is underway in various laboratories in the country.

Further, we employed NGS and computational biology approach to unravel stress responses of rice (a C3 plant with

high water requirement) and sorghum (a C4 plant with low water requirement). Comparative transcriptome of drought

sensitive rice cv. IR64 and drought tolerant cv. Nagina 22 and pathway analyses revealed potential role of α-linolenic

acid metabolic pathway as a novel mechanism of drought tolerance of Nagina 22. Utilisation of the large untapped

gene pool is the key for climate resilient agriculture. Since heat stress threatens wheat, we phenotyped the 18,653

wheat accessions for heat tolerance and identified genotypes with very high heat tolerance as donors for development

of heat tolerant wheat varieties. The genes and germplasm identified in these studies will not only help unravel the

molecular mechanisms but also to develop climate resilient crops with improved yield.

K.C. Bansal

8

IPPCongress2015




Enhancement of nitrogen and carbon metabolism in plants through

genetic manipulation of ornithine

Subhash Minocha1*, Rajtilak Majumdar1, Boubker Barchi1, Swathi Turlapati1,2 andRakesh Minocha2

1Department of Biological Sciences, University of New Hampshire, Durham, NH 03824, USA2USDA Forest Service, NRS, Durham, NH 03824, USA


After the twenty proteinogenic amino acids, ornithine perhaps occupies the most critical position among the non-

proteinogenic amino acids in plants. It is an intermediate in the inter-conversions of glutamate and arginine on one

hand, and the production of proline, polyamines, GABA and several alkaloids on the other; all products of great

significance to plants. In a study involving constitutive and inducible genetic engineering of ornithine catabolism in

Arabidopsis thaliana and Populus nigra x maximowiczii, and analyses of substrate flux, qPCR, transcriptomics and

metabolomics, we found that ornithine, whose concentration in cells is rather miniscule (vs. other amino acids), not

only regulates polyamine biosynthesis but also plays an important role in regulating the flux of nitrogen from glutamate

to arginine, proline and GABA. In fact, the metabolic effects of altering Glu-Orn-Arg-Pro-putrescine flux via ornithine

manipulation go far beyond the biosynthesis of polyamines and the associated amino acids. We hypothesize that

ornithine plays a critical role in regulating its own biosynthesis as well as the biosynthesis of glutamate (the primary

nitrogen assimilation product) in the cells. Thus the overconsumption of ornithine leads to increased assimilation of

both nitrogen and carbon, and potentially increased biomass production. Understanding regulation of ornithine

metabolism should improve metabolic engineering of plants for stress tolerance (via manipulation of proline, GABA,

and polyamines), nutritional improvement of food and feed crops (contents of important amino acids like arginine and

citrulline), enhance nitrogen assimilation (reducing environmental pollution from nitrogen fertilizer), and carbon

sequestration for increased biomass production for bioenergy.

Subhash C. Minocha

9

IPPCongress2015




Toward the identification in poplar of a possible involvement of multi-

step phosphorelay partners in drought perception

Lucie Bertheau3, Inès Djeghdir1, François Héricourt1, Françoise Chefdor1, Mélanie Larcher1,Domenico Morabito1, Christiane Depierreux1, Daniel Auguin1, Emilien Foureau2, VincentCourdavault2, Audrey Oudin2, Gaëlle Glévarec2, Franck Brignolas1 and Sabine Carpin1*

1Laboratoire de Biologie des Ligneux et des Grandes Cultures, Université d’Orléans, UPRES EA1207, rue de Chartres, BP 6759, 45067 Orléans Cedex 02, FranceArbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), INRA, USC1318,45067 Orléans, France2Laboratoire Biomolécules et Biotechnologies Végétales (BBV), Université François Rabelais deTours, EA 2106, 31 avenue Monge, 37200 Tours, France3Laboratoire Processus de Transferts et d’Echanges dans l’Environnement (PROTEE), Universitéde Toulon, EA 3819, 83957 La Garde Cedex, France


The molecular basis of plant drought perception and the resulting cellular osmotic stress are still unknown. In yeast,

the HOG pathway controlled by the multi-step phosphorelay system (MSP) SLN1-YPD1-SSK1, is the best characterized

osmosensing pathway, leading to yeast adaptation on high-osmolarity and high-salt media due to glycerol accumulation.

In Populus, we identified MSP proteins such as a Histidine-aspartate Kinase, HK1, homologous to SLN1, Histidine-

containing Phosphotransfer proteins, HPt1 to HPt10, and 9 Response Regulators (RR). By combining several

approaches including yeast two-hybrid and bimolecular fluorescence complementation assays and co-expression

analysis of mRNAs encoding studied proteins, we showed the existence of a partner network linked to HK1, the

putative poplar osmosensor. This network involves HK1, HPt2, 7 and 9 and six transcription factors belonging to the

RR-B family, RR12, 13, 14, 16, 18 and 19. Even if the “osmosensor” function of HK proteins such as AHK1 in

Arabidopsis is still not clearly attested, a role in dehydration avoidance is well established. Thus, in poplar, the HK1

MSP could be involved in dehydration-sensing mechanisms and plant water status control. To go further in this study

and better understand the role of this HK1 MSP in drought tolerance, we are now investigating the HK1 function by a

loss of function approach based on the RNAi strategy.

Sabine Carpin

10

IPPCongress2015




Abiotic stress regulates expression of Galctinol synthase genes in

rice through intron retention and nonsense-mediated decay (NMD) of

mRNA

Sritama Mukherjee, Sonali Sengupta, Abhishek Mukherjee and Arun Lahiri Majumder*

Division of Plant Biology, Bose Institute (Centenary Building), Kolkata-700054, India


Galactinol synthase (GolS), the first committed enzyme in Raffinose Family Oligosaccharide (RFO) synthesis pathway,

produces galactinol from UDP-galactose and inositol. The two isoforms of Oryza sativa GolS (OsGolS) gene are

located in chromosomes 3(OsGolS1) and 7(OsGolS2). The genes are interspersed by conserved introns with each

intron harboring a premature termination codon (PTC). We identified a non-canonical regulation mechanism controlling

the splicing and maturation of both the OsGolS isoforms in photosynthetic tissue. Splicing and maturation of both

OsGolS transcripts are linked to abiotic stress conditions. During abiotic stress, the premature and mature transcripts

of both isoforms accumulate in a rhythmic manner for very small time windows interrupted by phases of complete

absence of either. Reporter gene assay using GolS promoters under different abiotic stresses does not reflect this

accumulation profile, suggesting that this regulation occurs post-transcriptionally. We suggest that there is a surveillance

mechanism triggering the degradation of the premature transcript preventing its accumulation in the cell. This mechanism

fits the paradigm of PTC-induced nonsense-mediated decay (NMD), a less known event in plants. The detection and

degradation of prematurely terminating transcripts coupled to repressed splicing can be described as a form of

regulated unproductive splicing and translation (RUST). This regulatory circuit may have contributed to the adaptation

of plants to stress and phloem loading as suggested by the biological role of galactinol in green tissues.

Arun Lahiri Majumder

11

IPPCongress2015




From ‘Meta-OMICS’ to mutation breeding: Improving salinity and

drought tolerance in rice

Rajeev K. Bahuguna1, Ray Singh Rathore1, Priyanka Das1, Sneh L. Singla Pareek2 andAshwani Pareek1*

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal NehruUniversity, New Delhi-110067, India2Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology,Aruna Asaf Ali Road, New Delhi-110067, India


Rice is a staple food crop for more then half of the world population. An estimated demand for rice would increase at

least by 50 percent in the next 25 years to feed the growing population. Abiotic stresses such as salinity, drought and

heat are major constraints for global rice production in the current and more so in the future climate. Taking clues from

naturally salinity tolerant rice genotypes, we are trying to understand the molecular basis of stress tolerance using an

OMICS-based approach. This work has lead to the identification and functional validation of key stress responsive

genes, which may prove to be a suitable candidate for translational research. Taking an alternative approach of

mutagenesis, we have generated several mutant lines through gamma-irradiation and screened them at seedling

stage for different abiotic stress tolerance. Three screened mutant lines showed significant potential to survive under

severe heat, drought and salinity stress at seedling stage. These mutant lines were further characterized for their

mechanism of stress tolerance at reproductive and grain filling stage. Plants were grown in 15 L pots in the net house

and phenotyping for heat, drought and salinity stress was done across the growth stages. Besides agronomic traits,

key physiological traits such as photosynthesis, physiological water use efficiency, stomatal conductance, spikelet

fertility, pollen germination, K+/Na+ ratio, spikelet fertility and source-sink regulation at grain filling were studied. These

mutant lines could be used for improving rice production and varietal improvement for future climate.

Ashwani Pareek

12

IPPCongress2015




Altered Cu homeostasis regulates drought response in rice

Saurabh Raghuvanshi1*, Sonia Chhillar Balyan1, Mukesh Kumar1, Roseeta Devi Mutum1,Utkarsh Raghuvanshi1 and Saloni Mathur2

1Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road,New Delhi-110021, India2National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India


It is important to understand the molecular basis of natural drought tolerance that evolved in tolerant rice cultivars/

varieties. We conducted in-depth comparative analysis of drought tolerant and sensitive rice cultivars to explore the

molecular basis of drought tolerance. Our study discovered a multi-tiered drought-induced network, involving

transporters, TFs, miRNAs, Cu-proteins and metabolic enzymes, in the leaves that leads to enhanced ROS accumulation

and stomatal closure in the tolerant cultivar. The hallmark was the down-regulation of Cu transporters in the tolerant

rice cultivars (but not in sensitive one) during drought, leading to fall in internal copper levels. This kick-starts the

‘copper economy mode’ via OsSPL9 wherein a group of miRNAs (osa-miR159f, osa-miR1871, osa-miR398b, osa-

miR408-3p, osa-miR2878-5p, osa-miR528-5p and osa-miR397a), referred to as DTA (Drought Trait Associated)

miRNAs, are up-regulated. These DTA miRNAs target several genes coding for copper proteins such as plantacyanins,

laccases and Cu-Zn SODs. Interestingly, even though internal Cu levels fall, the level of plastocyanin remains stable

indicating re-mobilization of internal Cu to stabilize plastocyanin and the photosynthetic electron transport. On the

other hand, down-regulation of Cu-Zn SODs in the flag leaves of the tolerant cultivars raises ROS levels ultimately

leading to enhanced stomatal closure. Physiological advantage of high DTA miRNA levels was demonstrated by

overexpressing MIR408 in the drought sensitive rice cultivar leading to significant enhancement in vegetative growth

and tolerance to water deficit conditions. In summary, differential regulation of copper transporters during drought

defines atleast one way wherein tolerant rice cultivar modulates its metabolism to better endure drought.

Saurabh Raghuvanshi

13

IPPCongress2015



Concurrent Session B-I

SHW1 Interacts with HY5 and COP1, and promotes COP1-mediated

degradation of HY5 during Arabidopsis seedling development

Anjil Kumar Srivastava, Dhirodatta Senapati and Sudip Chattopadhyay*

Department of Biotechnology, National Institute of Technology, Durgapur-713209, India


Light acts as one of the most influential environmental factors for plant growth and development. Arabidopsis seedlings

follow two distinct developmental patterns in the presence and absence of light. The dark grown seedlings have long

hypocotyl with apical hooks, and with small and closed cotyledons. The light grown seedlings have short hypocotyl

with open and expanded cotyledons. Downstream to photoreceptors, multiple positive and negative regulators have

been identified that are involved in Arabidopsis seedling development. SHW1 works as a negative regulator of

photomorphogenesis. SHW1 and COP1, an ubiquitin ligase, genetically interact in an additive manner to suppress

photomorphogenesis. HY5 is a bZIP transcription factor that works as a positive regulator of photomorphogenesis at

various wavelengths of light, and is degraded by COP1 in dark. SHW1 genetically and physically interacts with both

COP1 and HY5. The genetic studies reveal that SHW1 and HY5 work in an antagonistic manner to regulate

photomorphogenic growth. Additional mutation of SHW1 in hy5 mutant background is able to suppress the gravitropic

root growth defect of hy5 mutants. Moreover, SHW1 has been shown to promote COP1-mediated degradation of

HY5 through enhanced ubiquitylation in the darkness. The interplay of these light-signaling pathway regulators and

their concerted role in Arabidopsis seedling development will be discussed.

Sudip Chattopadhyay

14

IPPCongress2015




Light signalling and MAP kinase cascade: A crosstalk

Alok Krishna Sinha*

National Institute of Plant Genome Research, New Delhi-110067, India


Mitogen activated protein kinase (MAPK/MPK) cascade are universally conserved cascade among eukaryotes and

are known to regulate several stress responses and developmental processes. They usually consisit of three tier

components, MAPK, MAPK kinas (MAPKK/MAP2K/MKK) and MAPKK kinase (MAPKKK/MAP3K/MKKK) connected

to each other with an event of phosphorylation. Light signal transduction pathways have extensively been studied in

plants and are known to control several developmental processes. However, a direct connection of MAPK and light

signaling pathways was not reported. Here, we present a MAPK module, consisting of MKK3-MPK6 that is activated

by blue light in a MYC2 dependent manner. MYC2 is a transcription factor that negatively regulates blue light signaling

during Arabidopsis seedling development. MPK6 physically interacts and phosphorylates MYC2, and is phosphorylated

by MKK3. Furthermore, MYC2 binds to MPK6 promoter and regulates its expression in a feedback regulatory mechanism

in blue light signaling. The mutational and physiological studies illustrate the function of MKK3-MPK6-MYC2 module

in Arabidopsis seedling development and provide a new mechanistic view of photomorphogenesis.

Alok K. Sinha

15

IPPCongress2015




CBL-interacting protein kinase, CIPK21, regulates osmotic and salt

stress responses in Arabidopsis

Girdhar K. Pandey1*, Poonam Kanwar1, Amarjeet Singh1, Leonie Steinhorst5, Amita Pandey1,Akhlilesh K. Yadav1, Indu Tokas1, Sibaji Sanyal1, Boem-Gi Kim2, Sung-Chul Lee3,Yong-Hwa Cheong4, Jörg Kudla5 and Sheng Luan6

1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India2Department of Molecular physiology and Biochemistry, National Institute of AgriculturalBiotechnology, Suwon, Korea3Deparment of Life Science, HeukSeok-Dong, Dongjak-Gu, Seoul, Korea4Department of Bio-Environmental Science, Sunchon National Univ., Suncheon, Jeonnam, Korea5Molekulargenetik und Zellbiologie der Pflanzen Institut für Biologie und Biotechnologie der PflanzenUniversität Münster Schlossplatz 4 48149, Münster, Germany6Department of Plant and Microbial Biology, Univ. of California Berkeley, Berkeley, California, USA


The role of calcium mediated signaling has been intensively studied in abiotic stress responses in plants. Calcineurin

B-like proteins (CBLs) and CBL-interacting protein kinases (CIPKs) constitute a complex signaling network in diverse

stress responses. Osmotic stress imposed by soil salinity and drought is a major abiotic stress that impedes plant

growth and development and has been shown to involve calcium-signaling processes. In this study, we report the

functional analysis of the Arabidopsis kinase CIPK21 that is ubiquitously expressed in plant tissues and upregulated

by multiple abiotic stress conditions. A loss-of-function mutant of CIPK21 revealed to be hypersensitive to grow on

salt and osmotic stress conditions. The calcium sensors, CBL2 and CBL3, were found to physically interact with

CIPK21 and target this kinase to the vacuolar membrane. Moreover, preferential localization of CIPK21 to the vacuolar

membrane was detected under salt stress conditions when co-expressed with CBL2/CBL3. These finding suggests

that CIPK21 regulate responses to salt stress condition inli Arabidopsis.

Girdhar K. Pandey

16

IPPCongress2015




Engineering ABA receptors to enhance WUE and drought tolerance of

rice

V.V. Santosh Kumar, Shashank K. Yadav, Sanya Shrivastava, Rakesh K. Verma, VishakhaSharma and Viswanathan Chinnusamy*

Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India


Abscisic Acid (ABA) plays crucial roles in development and stress responses of plants. The START domain proteins

PYRABACTIN RESISTANCE1 (PYR1)/PYR1-like (PYL)/Regulatory Components of ABA Receptors (RCARs) have

been identified as bona fide ABA receptors (ABARs) from Arabidopsis. Towards understanding structural and functional

diversity of ABAR family members from rice, we identified 12 PYR/PYL homolog ABARs from rice. Homology modeling

of ABARs cloned from Nagina 22 revealed that 10 of them may function as receptors in core-ABA signalling. Docking

studies were carried out with ABA, ABA metabolite, agonists and antagonists illustrated differential affinity of different

ABARs to these legends. Expression analysis of ABARs revealed differential tissue specific and stress responses,

suggesting their non-redundant role in stress response and development. Transgenic rice lines with constitutive/

stress responsive over expression and RNAi silencing of ABARs are being developed. Rice transgenics expressing

AtRD29A::OsABAR6 showed hypersensitivity to ABA in germination as compared with non-transformed (NT) wild

type and RNAi lines. This suggests that ABAR6 conferred enhanced ABA perception and thus function as ABA

receptor. The RD29A:ABAR6 transgenic rice lines are more drought tolerant than NT plants in terms of maintenance

of RWC, membrane stability index, chlorophyll content, survival and recovery growth in pot culture under greenhouse

conditions. Water utilization experiment revealed that transgenic plants used about 40% less water per a day as

compared to wild type plants. Results from this study suggest that ABARs have potential to improve WUE and

drought tolerance of rice.

Viswanathan Chinnusamy

17

IPPCongress2015




Tetraspanin family of proteins: New regulators of development and

abiotic stress response in rice

Surekha Katiyar-Agarwal1*, M. Balaji1 and Manu Agarwal2

1Department of Plant Molecular Biology, University of Delhi, South Campus, New Delhi-110021,India2Department of Botany, University of Delhi, Delhi-110007, India

*Correspondence: [email protected], [email protected]

Tetraspanins belong to a super family of highly conserved membrane proteins found in all multicellular and few

unicellular organisms. Tetraspanin proteins (TETs) are comprised of four transmembrane domains, two extracellular

loops with conserved cysteine residues, and two cytoplasmic tails. The dynamic nature of TETs to interact with

multiple partners and form ‘tetraspanin-enriched micro domains’, and their preferential targeting to membranes

emphasizes their participation in signal recognition/perception, and coordination of intracellular signaling cascades in

cells. In animals, tetraspanins play crucial role in multitude of cellular processes, including cell proliferation, adhesion,

fusion, movement, interaction and intracellular trafficking. Unlike animals, the biological function of TETs in plants has

not been well explored. In Arabidopsis, tetraspanins are known to be associated with development processes such

as leaf morphogenesis, root and floral organ formation. The present study was designed to gather comprehensive

information on 15 rice tetraspanin members (OsTETs) with respect to their genomic organization, chromosomal

distribution, phylogeny and domain structure, and expression pattern. Detailed analyses of OsTETs revealed that

these proteins consist of signature ‘GCCK/R’ motif as found in Arabidopsis. Expression profiling studies advocated

their role in rice development, including flag leaf senescence and defining plants response to several abiotic stress

and nutrient deprivation conditions. Subcellular localization of eight OsTETs suggested that they reside on plasma

membrane. Transgenic rice plants for altered expression of OsTET5 have been generated and their preliminary

phenotypic analyses supported role of OsTET5 in abiotic stress tolerance. Detailed characterization of transgenic rice

lines is being carried out and will be presented.

Surekha Katiyar-Agarwal

18

IPPCongress2015




Phosphorylation-dependent regulation of G-protein cycle during nodule

formation in soybean

Swarup Roy Choudhury and Sona Pandey*

Donald Danforth Plant Science Center, 975 N. Warson Road, St. Louis, MO 63132, USA


Signaling pathways mediated by heterotrimeric G-protein complex comprising Ga, Gb and Gg subunits and their

regulatory RGS (Regulator of G-protein Signaling) protein are conserved in all eukaryotes. We demonstrate that the

specific subunits of Gá, Gb and Gg proteins of soybean heterotrimeric G-protein complex regulate signaling during

nodulation. We also show the role of nod factor receptor 1 (NFR1)-mediated phosphorylation in regulation of G-

protein cycle during nodulation in soybean. Our data establish that during nodulation the G-protein cycle is regulated

by the activity of RGS proteins. Lower or higher expression of RGS proteins results in fewer or more nodules,

respectively. NFR1 interacts with RGS proteins and phosphorylates them. Analysis of phosphorylated RGS protein

identifies specific amino acids which when phosphorylated; result in significantly higher GTPase accelerating activity.

These data point to a phosphorylation-based regulation of G-protein signaling during nodule development. We propose

that active NFR1 receptors phosphorylate and activate RGS proteins, which help maintain the Ga proteins in their

inactive, trimeric conformation, resulting in successful nodule development. Alternatively, RGS proteins might also

have a direct role in regulating nodulation because overexpression of their phosphomimic version leads to partial

restoration of nodule formation in nod49 mutant.

Sona Pandey

19

IPPCongress2015



Concurrent Session C

Functional genomics of thermal tolerance in wheat

Paramjit Khurana*

Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India


Climate change and abiotic stress affects agriculture and crop production adversely. Wheat is a temperate crop and

prone to various abiotic stresses. Tolerance to heat stress is a complex phenomenon and controlled by multiple

genes. Heat tolerance related gene transcripts were identified based on their putative functions and validated by

cDNA macroarray and northern/RT-PCR analysis in wheat. A common response to high temperature stress is the

synthesis of Heat Shock Proteins (HSPs). HSP encoding genes are found to be under the control of Heat Shock

Factors (HSF), transcription factors that regulate the expression of HSP genes. One such gene, a chloroplastic small

heat shock protein (sHSP26) was found essential for tolerance to heat stress. Transgenic Arabidopsis plants over-

expressing wheat HSP26 were more tolerant to high temperature induced damage to the photosynthetic machinery,

and the antisense transgenic plants were found to be severely sensitive to heat stress. Similarly, a new heat shock

factor (HSF) possesses the characteristic domains of class A type plant HSFs was identified. Functional genomics

approaches indicated plants overexpressing TaHsfA2d possess higher tolerance towards high temperatures and a

higher yield and biomass under constant heat stress conditions. Analysis of putative target genes showed higher and

constitutive expression of several abiotic stress responsive genes in transgenic Arabidopsis plants over-expressing

TaHsfA2d. TaHsfA2d can also functionally compliment the T-DNA insertion mutants of AtHsfA2. These observations

suggest that discovery of novel genes may be useful in molecular breeding of crop plants, especially wheat, to

improve yield under abiotic stress conditions.

Paramjit Khurana

20

IPPCongress2015




Genomics-assisted mapping of QTLs and genes for salt tolerance in

Indian rice (Oryza sativa L.) germplasm

Nagendra K. Singh1*, Sushma Tiwari1, Sefali Mishra1, Balwant Singh1, S.V. Amitha Mithra1,Vandna Rai1, Ashok K. Singh2, Vinod Kumar2 and S.L. Krishnamurthy3

1National Research Centre on Plant Biotechnology, Pusa Campus, New Delhi-110012, India2Central Soil Salinity Research Institute, Karnal–132001, India3Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India


Soil salinity is a major constraint to rice production in large inland and coastal areas of rice cultivation around the

world. Modern high yielding rice varieties are particularly sensitive to high salt stress. There are salt tolerant landraces

and traditional varieties of rice but with limited information on QTLs and genes for their salt tolerance. Salt tolerance

is generally quantitatively inherited because of its control by multiple genes located in different genomic regions. With

the advent of DNA markers it is possible to genetically map these quantitative trait loci (QTLs) using different types of

mapping populations. However, this requires genotyping of entire population using large number of polymorphic

markers to create a dense molecular genetic map. Bulked segregant analysis (BSA) allows rapid identification of

markers linked to the trait, but it has been applied mainly to map simply inherited traits. Here we describe a method for

rapid identification of genomic regions (QTLs) for seedling and reproductive stage salt tolerance in rice using two

different mapping populations of recombinant inbred lines (RILs) using a bulked segregant analysis (BSA) approach.

We found that the number of RILs with extreme phenotype to be pooled for BSA was thirty each to get the maximum

heterogeneity in the non-associated regions of the genome. The parents and bulks were genotyped using a specially

designed 50K SNP chip to identify genomic regions showing homogeneity of alleles for polymorphic SNPs in the

bulk(s). The method was applied to ‘CSR11/MI48’ RILs segregating for reproductive stage salt tolerance. Genotyping

of the parents and RIL bulks, made on the basis of salt sensitivity index for grain yield, revealed 6,068 polymorphic

SNPs of which 22 genomic regions (QTLs) showed contrasting allelic homogeneity in the two bulks. The method was

validated in another RIL population of ‘CSR27/MI48’ used earlier for mapping QTLs for salt tolerance using low-

density SSR markers. BSA using 50K SNP chip in the CSR27 populations revealed 5,035 polymorphic SNPs and 34

QTLs showing allelic homogeneity in the two bulks. The BSA not only confirmed the location of QTLs mapped earlier

in this population but also identified additional QTLs because of high density SNP genotyping. BSA with high-density

SNP chip provides a rapid approach to scan the whole genome for mapping QTLs for complex agronomic traits in

rice. We have also identified novel sources of salt tolerance in Indian wild rice germplasm collected from different eco-

geographical regions of India.

Nagendra K. Singh

21

IPPCongress2015




Sequence polymorphism in the Waxy locus and its relationship with

apparent amylose content of endosperm starch in cultivars of rice

(Oryza sativa L.) from Northeast India

Nikhil K. Chrungoo* and Ayam Gangarani Devi

Molecular Plant Physiology Laboratory, Department of Botany, North Eastern Hill University, Shillong,Meghalaya, India

*Correspondence: [email protected]; [email protected]

The Northeast India, being at the confluence of Indo-Malayan and Indo-Chinese biogeographic realms, represents

one of the world “biodiversity hotspots”. With a rich diversity of cultivars, the rice gene pool of the region shows

marked variations in various quality traits including aroma and starch quality. Since starch quality is primarily determined

by its apparent amylose content and that the Waxy locus controls much of the variation in apparent amylose content,

the linkage between sequence polymorphism in the Waxy locus and the apparent amylose content assumes

significance. Our results reveal that based on combination of “G/T” polymorphism in 5’leader intron splice site, length

polymorphism in exon 2 and “A/C” polymorphism in exon 6, the Waxy locus of rice can have four allelic forms viz.

“Int1T-Ex2(23bp)2-Ex6C”, “Int1T-Ex2(23bp)

1-Ex6A”, “Int1G-Ex2(23bp)

1-Ex6C” and “Int1G-Ex2(23bp)

1-Ex6A” which

correlate specifically with “waxy”, “very low”, “low/intermediate” and “high amylo-” cultivars respectively. Phylogenetic

tree on alignment matrix of nucleotide sequences of the p-SINEI element of the Waxy locus clustered the upland and

japonica cultivars separately from the lowland and indica cultivars thereby establishing the role of transposons in the

Waxy locus in differentiation of upland/japonica and lowland/indica cultivars.

Nikhil K. Chrungoo

22

IPPCongress2015




Orthoulogues of blast resistance gene Pi54 interacts at different

domains of Avr-Pi54 gene of Magnaporthe oryzae

T.R. Sharma*, B.N. Devanna and Soham Rey

National Research Center on Plant Biotechnology, Indian Agricultural Research Institute,New Delhi-110012, India


Decoding of whole genome sequence of Rice (Oryza sativa) and its one of the important pathogens Magnaporthe

oryzae have provided an excellent opportunity to understand molecular basis of host –pathogen interaction. We

identified, cloned and characterized a major gene (Pi54) for blast resistance from rice using map based cloning

approach. Its orthologue Pi54 of was also cloned from wild species of rice Oryza officinalis and functionally validated

in different genetic backgrounds of rice. Functional complementation of the gene has confirmed its stable and high-

level of resistance expression against the geographically diverse strains of M. oryzae. To understand the molecular

basis of interaction between Pi54 and Avr –Pi54 genes we sequenced the whole genome of an Avirulent strain of M.

oryzae and cloned Avr-Pi54. The docking experiments involving four Pi54 R proteins and an AVR-Pi54 protein revealed

that all the resistance proteins could interact with AVR counterpart, whereas the susceptible Pi54tp protein does not

show any interaction with AVR-Pi54. This finding is not unexpected as there is a large deletion at NBS region of

Pi54tp protein. The protein molecular docking results of this study also revealed that the Pi54 of protein interacted

with AVR-Pi54 only through its NBS domain. Whereas the other Pi54 protein interacted through the CC-NBS-LRR

regions and Pi54rh interacted through the NBS-LRR regions. This differential preference of AVR protein for binding

domains among the Pi54 proteins might be attributed to its different resistance mechanisms against M. oryzae.

Among the resistant Pi54 proteins, a stress inducible protein binding domain, STI1 is present only in Pi54 of and

absent in other two resistant proteins. Different results obtained on cloning and characterization of Pi54 orthologues

will be discussed in this presentation.

T.R. Sharma

23

IPPCongress2015




A multigene cluster underpins a large effect QTL for rice yield under

drought

Ajay Kohli*

Plant Molecular Biology Laboratory, Genetics and Biotechnology Division, International RiceResearch Institute, DAPO 7777, Metro Manila, Philippines


Limited QTLs have been cloned for the underlying cardinal genes. For drought tolerance in plants two QTLs have

been dissected for the cardinal gene. As opposed to such single gene QTLs, which are mostly limited to minor effects

on the traits, large effect QTLs have now been cloned for rice yield under drought. Our efforts at the dissection of one

such large-effect QTL has revealed a gene cluster that is essential for the functionality of QTL qDTY12.1. The

multigene cluster is unique in its functionality in a manner analogous to the regulon-like genetic architecture. A

transcription factor and its co-localized target genes work in concert to affect the trait of drought tolerance. Along with

this novel aspect, it is also the first time that a QTL from a susceptible variety has been successfully dissected at the

molecular level. These results suggest that complex traits may be best addressed through complex loci rather than a

single master switch.

Ajay Kohli

24

IPPCongress2015




Next generation genomics: Connecting genes and (epi)genomic

variations to abiotic stress response/adaptation

Mukesh Jain*



The availability of high-throughput sequencing technologies provide opportunity to unveil the genetic/epigenetic basis

of abiotic stress response and adaptation. Rice is an important crop accounting for food security of over half the world

population. Water-deficit and salinity are the major abiotic factors that affect rice productivity worldwide. Rice germplasm

exhibit variability in their response to abiotic stresses, which may be attributed to genetic and epigenetic variations.

Although several stress-responsive genes have been identified and a few regulatory networks have been proposed,

tolerance/response mechanisms still remain unclear to a larger extent. We performed whole-genome, transcriptome

and bisulphite sequencing of three rice cultivars with contrasting responses to drought and salinity stress. We identified

thousands of high-quality SNPs and InDels among these cultivars. One-fourth of the SNPs and InDels were detected

in genic regions and about 3.5% of the total SNPs resulted in non-synonymous changes. DNA polymorphisms present

in the differentially expressed genes within the known quantitative trait loci were also identified. RNA-seq analysis

identified several novel stress-responsive transcripts/isoforms. We discovered extensive DNA methylation at single-

base resolution in rice cultivars, identified the sequence context and extent of methylation at each site. Numerous

differentially methylated regions (DMRs) among different cultivars were identified and many of them were found

associated with differential gene expression. The transposon-associated DMRs were found coupled to the transcript

abundance of the transposon and/or proximal protein-coding gene(s). These results provide insights into the regulatory

mechanisms underlying abiotic stress response/adaptation and functional markers/ target genes for stress tolerance.

Mukesh Jain

25

IPPCongress2015



Concurrent Session A-II

Elucidation of the molecular machinery components associated with

adaptation of rice plants to heat stress

Anil Grover*

Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road,Dhaula Kuan, New Delhi-110021, India


Previous work from our laboratory has shown that the transcriptional profile of rice (Oryza sativa L.) is significantly

modulated within 10 min of heat stress (HS_10): analysis of differentially expressed genes (DEGs) revealed that

1,556 genes were rapidly up-regulated (>twofold, P<0.05) at HS_10 as compared to 600 down-regulated genes. The

enrichment of GO terms protein kinase activity/protein serine kinase activity, response to heat and reactive oxygen

species in up-regulated genes after HS_10 signifies the role of signal transduction events and reactive oxygen species

during early heat stress. The transcriptional profile shows that the response of rice to heat stress involves signal

perception and transduction, activation/synthesis of heat shock factors, genomic and proteomic alterations, ROS

metabolism and a host of other proteins associated with biochemical, cellular and physiological processes and ‘unknown’

functions. The synthesis of heat shock proteins (Hsps) appears an important response to heat stress. Genome-wide

gene families of rice Hsp20, Hsp40, Hsp70 and Hsp100 as well as heat shock factors (Hsfs) were analysed. Rice

ClpB/Hsp100 family proteins are constituted of three isoforms localized in three different cell compartments namely

cytoplasm (ClpB-c), chloroplast (ClpB-p) and mitochondria (ClpB-m). Our present focus is to analyse the role of ClpB/

Hsp100 proteins in governing heat stress response in Arabidopsis and rice plants.

Anil Grover

26

IPPCongress2015




Development of physiological and biochemical markers for monitoring

forest health in a changing climate

Rakesh Minocha1*, Stephanie Long1, Swathi Turlapati2,1 and Subhash C. Minocha2

1USDA Forest Service, NRS, Durham, NH 03824, USA2Department of Biological Sciences, University of New Hampshire, Durham, NH 03824, USA


Continuous input of pollutants from atmospheric deposition can move ecosystems towards soil Al mobilization/toxicity,

nutrient depletion, heavy metal toxicity, and finally nitrogen saturation. A major goal of our research is to determine

the usefulness of selected polyamines, amino acids, and phytochelatins as early metabolic markers for assessing

forest productivity using asymptomatic forest stands. This type of early evaluation is beneficial in developing risk

assessment and stress remediation strategies. Our objective is to determine the usefulness of polyamines, particularly

putrescine, amino acids (e.g. arginine, Proline and GABA), phytochelatins (inducible by heavy metal toxicity) as foliar

indicators of abiotic stress in visually asymptomatic conifer and hardwood trees. This presentation will be a review of

several studies conducted over two decades in collaboration with scientists from various institutions using hardwood,

conifer, and mixed wood study sites across Northern New England, USA over two decades. These data show a

strong correlation between soil nutrient, these metabolites, and chlorophyll indicating that they can potentially be

used as reliable and easily quantifiable markers of abiotic stress in visually healthy trees. Most of these select metabolites

increased in response to exposure to increased levels of pollutants. In conjunction with these changes, changes in

soil amino acids and microbial population diversity and functions were also observed. In some cases, such as Harvard

Forest, MA, USA and Bear Brook Watershed, ME, USA, long-term ecosystem research (LTER) study sites these

changes accompanied lower site productivity in red pine and red spruce stands, respectively.

Rakesh Minocha

27

IPPCongress2015




Tailoring rice plants for higher yield via genetic manipulation for ion

partitioning, cellular detoxification and cytokinin metabolism

Khirod K. Sahoo1, Brijesh K. Gupta1, Rohit Joshi1, Amit K. Tripathi1, Ashwani Pareek2 andSneh Lata Singla-Pareek1*

1Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology,Aruna Asaf Ali Marg, New Delhi-110067, India2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal NehruUniversity, New Delhi-110067, India


To meet the rising food demand, there is an urgent need to boost crop production. Thus, we need to accelerate our

understanding of reduction in crop yield occurring due to various climatic vagaries. Tremendous progress has been

made over the past years in defining the molecular circuitry underlying cross talk between various plant hormones,

defence related machineries, development associated proteins and environmental response networks. Here, we have

attempted to bridge the yield gap in rice for sustainable food production by genetic manipulation of either stress related

genes or genes participating in hormone homeostasis. Taking clues from the stress tolerance capacity of plants

overexpressing either the glyoxalase (Gly) pathway for enhanced cellular detoxification or sodium proton antiporters

(NHX as well as SOS1), we developed transgenic riceplants expressing two different stack combinations of stress

associated genes viz. GlyI, GlyII along with NHX1 or with SOS1. Evaluation of these multigenic transgenic plants via

assessment of various morphological, physiological and agronomical parameters indicated improved tolerance to abiotic

stress and enhanced yield. Simultaneously, we also raised transgenic rice plants with RNAi based knockdown of one of

the key genes of cytokinin metabolism i.e., Cytokinin oxidase-2. These transgenic plants showed better yield under both

non-stress and stress conditions. Considering the complexity of stress response, our results suggest that a combinatorial

approach targeting diverse pathways is better than the ‘single gene approach’ to tame the deleterious effects of

environmental extremes of high salinity and water deficit under field conditions.

Sneh Lata Singla-Pareek

28

IPPCongress2015




Transcriptome analysis under temperature stress in seabuckthorn, an

important high altitude medicinal plant

Saurabh Chaudhary1, Rajesh Ghangal2, Ankit Jain3, Ashish Kumar1 andPrakash C. Sharma1*

1University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi, India2Department of Biotechnology, Manav Rachna International University, Faridabad, India3Centre of Excellence in Genomics, ICRISAT, Patancheru, Hyderabad, India


Seabuckthorn (Hippophae rhamnoides L.), a high altitude shrub, is known for its immense medicinal, nutritional and

ecological value. The remarkable adaptation of seabuckthorn to harsh environments exhibited by tolerance to abiotic

stresses specifically extreme temperatures has recently attracted attention of researchers as a non model important

plant species. However, little is known about gene regulation and signalling pathways involved in extreme temperature

(low and high) stress responses in this species. We analysed expressed sequence tags (ESTs) data followed by

whole transcriptome sequencing from leaf and root tissues of sea buckthorn to explore the plant responses at molecular

level under extreme temperatures. Moreover, to a gain deep insight into the molecular mechanism under cold and

freeze stress we performed digital gene expression analysis using DeepSAGE analysis. Initially, 3412 ESTs were

generated and assembled into 1665 unigenes, followed by assembly of 86,253,874 high quality NGS short reads into

88,297 putative unigenes. Next, from DeepSAGE data, 11,922 differentially expressed genes (DEGs) including 6539

up regulated and 5383 down regulated genes were identified under cold and freeze stress. Among various DEGs, 38

genes were validated in response to cold and freeze stress, using qRT-PCR. Recently, we have identified and

validated 11 more genes, found to be differentially expressed under heat, cold and freeze stress simultaneously,

suggesting a crosstalk between different stress pathways. The seabuckthorn transcriptome data have also been

used to mine microsatellite markers for diversity analysis and transcription factors. The ample amount of data generated

is under further analysis for gene discovery, development of functional molecular markers and other by products.

Prakash C. Sharma

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IPPCongress2015




Development of salt tolerant varieties following conventional, physi-

ological and molecular approaches

Parbodh C. Sharma1*, S.L. Krishnamurthy1, Neeraj Kulshreshtha2, Jogendra Singh1,Preeti Pundir1 and Dinesh Kumar Sharma1

1Central Soil Salinity Research Institute, Karnal-132001, Haryana India2Sugarcane Breeding Institute RRS, Karnal-132001, Haryana, India


Among abiotic stresses, salt stress is the most complicated problem posing a major challenge. Around 1000 m ha

globally and 6.73 m ha in India are affected by salinity. Central Soil Salinity Research Institute (CSSRI) has developed

15 salt tolerant varieties of different crops; 7 of rice (CSR10, CSR13, CSR23, CSR27, CSR Basmati 30, CSR36 and

CSR43), 4 of wheat (KRL1-4, KRL 19, KRL210 and KRL213), 3 of Indian mustard (CS52, CS54 and CS56) and one

in chickpea (Karnal Chana 1) through conventional breeding approaches to increase the productivity of salt affected

soils for resource poor farmers. Physiological and biochemical approaches have been used to identify different

mechanisms of salt tolerance and used for identification of salt tolerant and sensitive lines. The molecular approaches

are also being used to develop salt tolerant rice varieties. Introgression of Saltol QTL to two mega high yielding rice

varieties namely, Pusa 44 and Sarjoo 52 carried out through Marker Assisted Selection Back Cross (MABC) breeding.

The varieties namely, Pusa 44 and Sarjoo 52 were selected as recurrent parents while FL478 was used as a donor

parent and produce F1 seeds. True F

1s were selected by the Saltol marker RM8094. Further backcrossing with

respective recurrent parent and selfing was performed to produce seeds up to BC3F3 generation. In every back cross

generation, recombinant and foreground selection was carried out. Desirable plants were selected based upon the

(genotyping) marker data and through stringent phenotyping as similar to recurrent parent. Background selection and

DUS characterization of NILs is in progress.

Prabodh C. Sharma

30

IPPCongress2015




Introgression of physiological traits to save water while improving the

productivity in rice

M.S. Sheshshayee1*, M.D. Prathibha1, M.V. Mohan Kumar1, B.R. Raju1, H.G. JalendraKumar1, M.P. Rajanna2 and M. Udayakumar1

1Department of Crop Physiology, UAS, GKVK, Bangalore-560065, India2ZARS, VC Farm, Mandya-571401, Karnataka, India


Saving water and sustaining productivity under water limited conditions are the most important challenges. Global

research strongly suggests an analytical breeding approach in lieu of the empirical breeding adopted presently. Thus,

improving plant traits associated with maintenance of tissue water relations and metabolism have paramount

significance. While scientists equivocally agree on the role of root traits in maintenance of water relations, there are

debates on the relevance of WUE as a drought adaptive trait. We provide strong experimental evidences that

emphatically prove the relevance of WUE as a potential trait for crop improvement under water limited environments.

Further, introgression of several of such drought adaptive traits seems to be essential for a comprehensive improvement

in productivity under water-limited conditions. We adopted conventional as well as an elaborate marker assisted

backcross breeding (MABC) approaches to introgress WUE and water relation traits into a single elite genetic

background. The progeny developed by these approaches led to an increase of yield by more than 15% combined

with an ability to save at least 60% of irrigation water. The results of these investigations emphatically demonstrated

the relevance of improving physiological traits by breeding.

M.S. Sheshshayee

31

IPPCongress2015



Concurrent Session B-II

Signaling in plant immunity

Dierk Scheel*

Leibniz Institute of Plant Biochemistry, Department of Stress and Developmental Biology, Weinberg3, D-06108 Halle (Saale), Germany


Plants detect potential pathogens in their environment via pathogen-associated molecular patterns (PAMPs) that are

recognized by plant plasma membrane receptors. Typical PAMPs include the bacterial flagellin-derived flg22 peptide,

the elf18 peptide of the bacterial elongation factor EF-Tu, bacterial peptidoglycans and lipopolysaccharides, as well

as fungal chitin oligomers and glucan fragments from oomycetes. PAMP-binding to their receptors initiates complex

signaling networks that activate a multi-component defense response and thereby establish PAMP-triggered immunity.

One of the earliest detectable responses after PAMP perception is the activation of ion channels at the plasma

membrane. Using a transgenic Arabidopsis line with the calcium reporter, aequorin, increases in cytosolic calcium

levels are detected after PAMP application. To identify signaling network components, seeds of aequorin-expressing

lines were mutagenized and the population screened for mutants with changed calcium elevation (CCE) in response

to different PAMPs. Among several receptor complex components, a lectin S-domain receptor kinase was identified,

which mediates lipopolysaccharide sensing in Arabidopsis thaliana. MAPK cascades are essential for controlling

defense responses. The elements that prevent erroneous signaling crosstalk may include expression patterns of the

MAPK cascade components, the presence of pathway-specific protein complexes or the MAPK substrate diversity.

Different strategies have been employed to isolate MAPK interacting proteins. Several VQ-motif containing proteins

are MAPK substrates and regulate immune responses.

Dierk Scheel

32

IPPCongress2015




Natural variation and network modelling approaches to identify novel

regulatory genes affecting plant development and water use

Ulrike Bechtold*, John Ferguson and Phil Mullineaux

Plant Molecular Physiology Lab, University of Essex, School of Biological Sciences, Colchester,Essex, United Kingdom


The improvement of plant productivity while reducing water use is an urgent objective to sustain or improve plant

production while conserving natural resources. To identify and exploit genes that promote yield in water limiting

environments is an important challenge in plant biology and requires genes that affect lifetime traits such as water

use. In their natural environment, plants may experience continuous or episodic drought stress, and we aim to

understand how different types of drought stress impact on plant growth. In order to do this we obtained a quantitative

measure of the natural genetic variation of yield and water use in 46 Arabidopsis thaliana accessions. Water use,

developmental and physiological traits were assessed during episodic- and continuous drought experiments to evaluate

if proxy traits for productivity and water use can be identified. Simultaneously we are using high-resolution time series

transcriptome profiling coupled with Bayesian modeling to infer regulatory networks linked to early events of an

episodic drought stress in order to identify key regulatory genes. Using these approaches we identified AGAMOUS-

LIKE 22 (AGL22) as key gene in a regulatory transcription factor network. AGL22 is involved in the transition from

vegetative state to flowering, and loss of AGL22 expression affected photosynthesis and water use providing a link

between early changes in primary metabolism and the subsequent initiation of responses to stress that govern plant

productivity. The combination of genomic, transcriptomic and systems biology approaches holds great promise to

increase the efficiency of identifying novel candidate genes under different drought stress conditions.

Ulrike Bechtold

33

IPPCongress2015




Deregulated symbiosis receptor kinase hyperactivates spontaneous

nodulation

Maitrayee Dasgupta*, Sudip Saha, Ayan Dutta and Abhishek Bhattacharrya

Department of Biochemistry, Calcutta University, Kolkata, West Bengal, India


Plants restricted to a monophyletic clade of four angiosperm orders is capable of undertaking intracellular root nodule

symbioses with nitrogen-fixing bacteria. A compatible interaction triggers the ‘Symbiosis response pathway’ (Sym

pathway) to initiate the genetic reprogramming required for development of nodule primordia and intracellular

colonization of symbionts in the newly divided cells. Gain of function of ‘Sym pathway’ members like CCaMK, CYCLOPS

or its downstream effectors like HK1 can uncouple nodule organogenesis from bacterial infection and cause

development of empty spontaneous nodules in the absence of the bacterial symbiont. The efficiency of spontaneous

nodulation under constitutive activity of these members was either same or lower than the rhizobia induced nodulation,

which meant that pathways restricting nodule numbers were functional under these conditions. Symbiosis Receptor

Kinase (SYMRK), is an upstream receptor kinase of the ‘Sym pathway’. Our efforts have shown that ligand independent

deregulated activation of the intracellular kinase domain of SYMRK triggered hyperactivation of nodule organogenesis.

The merged clusters of spontaneous nodules indicated that constitutive SYMRK had overcome the control over

nodule numbers or their spatial position and suggested the importance of SYMRK ectodomain in restricting nodule

number. In presence of the symbiont the unrestrained kinase activity of SYMRK supported the root hair curls and

epidermal infection threads but rhizobial colonization in the nodule interior was highly compromised in the absence of

ectodomain of SYMRK. Profuse spontaneous nodulation triggered by constitutive SYMRK would be discussed in the

backdrop of prior arts.

Maitrayee Dasgupta

34

IPPCongress2015




Regulation of plant stress responses and development by the ROS-

Ca2+ signaling network

Kazuyuki Kuchitsu1,2*, Kenji Hashimoto1, Hidetaka Kaya1 and Nobutaka Kitahata1

1Department of Applied Biological Science; 2Imaging Frontier Center, Tokyo University of Science,Noda, Chiba 278-8510, Japan


Reactive oxygen species (ROS) are highly toxic molecules produced during photosynthesis and aerobic respiration.

However, ROS are enzymatically produced in a highly regulated manner spatially and temporally. ROS play key roles

in regulating a broad range of physiological processes, such as growth and development including tip growth of root

hairs and pollen tubes, defense responses against biotic and abiotic stresses as well as cell wall metabolism. Respiratory

burst oxidase homologues (Rboh) have been identified as ROS-producing NADPH oxidases, which act as key signalling

nodes integrating multiple signal transduction pathways in plants. Rbohs are synergistically activated by Ca2+ binding

to the N-terminal EF-hand motifs and phosphorylation by several families of protein kinases. We have identified

several proteins that interact with the N-terminal cytosolic region of Rbohs in vivo and regulate their ROS-producing

activity. Tip growth and ROS accumulation in pollen tubes were severely impaired in the rbohH rbohJ double mutant.

Point mutations in the EF-hand motifs impaired Ca2+-induced ROS production and complementation of the double

mutant phenotype, indicating that Ca2+-activated ROS production is essential for the proper tip growth. The ROS-Ca2+

positive feedback regulation seems to be conserved not only in polarized cell growth but also various signaling

processes in plants. We recently identified and characterized two Rbohs from an emerging model liverwort Marchantia

polymorpha. Evolutionary aspects and molecular regulatory mechanisms of Rboh/ROS-mediated signaling will be

discussed. Furthermore, novel chemical screening led us to identify potential compounds that affect Rboh-mediated

signaling events including plant defense responses positively and negatively.

Kazuyuki Kuchitsu

35

IPPCongress2015




Cytosine DNA methyltransferases and components of PRC1 complex

regulate growth, development and abiotic stress tolerance in the moss,

physcomitrella patens

Meenu Kapoor*, Vimala Parihar and Deepshikha Arya

Guru Gobind Singh Indraprastha University, New Delhi, India


Liverworts, hornworts and mosses form a paraphyletic group among bryophytes that diverged early from the lineage

that gave rise to flowering plants. Among these, mosses have had a tremendous impact on different ecosystems and

climatic conditions during the pre-cambrian era that paved way for colonization of land by animals. Physcomitrella

patens occupies a key evolutionary position among bryophytes linking the green algae and the vascular plants. Its

genomic study provides insight into the adaptations to extreme conditions that eventually lead to colonization of land

by plants. The green yeast™, as it is sometimes called, exhibits a high rate of homologous recombination. This,

together with the ease of transformation and high regeneration capacity has resulted in emergence of P. patens as a

model plant for reverse genetics approaches for gene functions analysis. Exploiting this potential, we have generated

stable gene knockout mutants for cytosine DNA methyltransferases, PpCMT and PpDNMT2 and the PRC1 component,

PpLHP1. Whole genome transcriptome analysis has revealed the role of epigenetic modifications, such as DNA

methylation, in regulating genes/biological pathways affecting resilience and abiotic stress tolerance. We provide

genetic evidence for regulation of cell growth, cell wall loosening and gravitropic response by the PpCMT/PpLHP1

complex and for PpLHP1 affecting transition from juvenile to adult leafy gametophore stage. Protein-protein interaction

studies reveal other molecular components involved in these pathways. We also show that the DNA/RNA

methyltransferase, PpDNMT2, regulates salt and osmotic stress tolerance by affecting biogenesis/stability of specific

transfer RNA molecules.

Meenu Kapoor

36

IPPCongress2015




Ethylene receptors and molecular mechanism of ethylene sensitivity

in tomato

Ajay Arora*

Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India

*Correspondance: [email protected]; [email protected]

The past two decades have been rewarding in terms of deciphering the ethylene signal transduction and functional

validation of the ethylene receptor and downstream genes involved in the cascade. Our knowledge of ethylene

receptors and its signal transduction pathway provides us a robust platform where we can think of manipulating and

regulating ethylene sensitivity by the use of genetic engineering and validating through transgenics. Recently, major

progresses have been made in understanding of the molecular mechanism regulating ethylene biosynthesis, perception

and signal transduction pathways. In important ornamental plant species, we aim to identify homologues to genes

involved in ethylene signaling, which previously have been isolated in model plants. Key genes are identified and their

regulation is studied in the postharvest context. An alteration of ethylene action is a valuable target for the genetic

engineering of crops. Manipulation of ethylene biosynthesis or perception allows us to modulate these processes and

thereby create plants with more robust and desirable traits, giving us a glimpse into the role of ethylene in the plant.

We isolated, characterized and transformed the gladiolus ethylene receptor and developed the tomato transgenics

that confer reduced sensitivity to ethylene. The possible strategies for altering the ethylene sensitivity of tomato

plants using ethylene receptor genes will be discussed.

Ajay Arora

37

IPPCongress2015



Concurrent Session D

Metabolic engineering of pathways for enhancing oil and seed yield in

Camelina sativa

Om Parkash Dhankher1*, Sudesh Chhikara1, Hesham Abdullah1, Kenny Ablordeppey1,Parisa Akbari1 and Danny Schnell2

1Stockbridge School of Agriculture, University of Massachusetts, Amherst, MA 01003, USA2Department of Biochemistry and Molecular Biology, Univ. of Massachusetts, Amherst,MA 01003, USA


Liquid transportation fuels based on plant seed oils have tremendous potential as an alternative to petroleum-derived

fuels. Camelina sativa, a member of brassicaseae, has been proposed as an ideal non-food crop for biofuel production.

Increasing seed oil yield by manipulation of the triacylglycerol (TAG) biosynthesis pathway and by increasing the flux

of carbon from increased photosynthesis to TAG synthesis will make this crop highly profitable. In seeds, TAG is

synthesized from glycerol-3-P and fatty acids through the sequential activities of the glycerol-3-P acyltransferase,

lysophosphatidic acid acyl transferase and diacylglycerol acyltransferase. Overexpression of enzymes that catalyze

the synthesis of glycerol backbone and conjugation of fatty acids to glycerol backbone appear to be a promising

target for increasing TAG accumulation. Therefore, to increase the oil contents and seed yield in C. sativa, we

targeted the over expression of several genes involved in fatty acids synthesis and acylation of fatty acids in the

downstream process for TAG synthesis. Over expression of genes involved in TAG synthesis resulted in a significant

increase in seed and oil yield. Since C. sativa is fairly a new oil seed crop and information regarding genes that

regulate the fatty acids and TAG biosynthesis remain poorly understood. It is not known how these genes contribute

to the overall accumulation of TAG in the seed oil and what are the rate limiting steps in the biosynthesis of these

molecules during seed development. Therefore, to identify the bottlenecks for further improving the seed and oil yield

in C. sativa seeds, a comprehensive transcriptome and metabolome analysis was performed in wild type and transgenic

C. sativa over expressing TAG related genes. Both transcriptome and metabolome profiling revealed several genes/

gene networks controlling the synthesis of key metabolite for TAG biosynthesis and storage during seed development.

Efforts are underway to combine the expression of these key genes using gene-stacking approaches for further

enhancing the oil and seed yield.

O.P Dhankher

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IPPCongress2015




Terpenoid sub-class based discrimination between MVA and DOXP

pathways of isoprenogenesis in plants: Exceptional results to

conceptual revision

Rajender Singh Sangwan*

1Center of Innovative and Applied Bioprocessing (A National Institute under DBT, Govt. of India),C-127, Phase-8, Industrial Area, S.A.S. Nagar, Mohali-160071, Punjab, India


Biosynthesis of terpenoids in plants differs from that in animals by its recruitment of two independent routes of

isoprenogenesis: the classical cytosolic mevalonate (MV) pathway and the recently discovered plastid localized deoxy-

xylulose phosphate (DOXP) pathway. Redundancy of routes to generate isopentenyl pyrophosphate (IPP) and its

isomer (dimethyl allyl pyrophosphate, DMAPP) invokes the issue of their relative significance. Early studies have

indicated that sesqui-, tri- and polyterpenes are synthesized through MVA pathway while biosynthetic pathways for of

mono-, di- and tetraterpenes recruit DOXP pathway generated IPP and DMAPP. Very recently, exceptions to such

strict discrimination, based on sub-class of terpenoids, with respect to recruit of isoprenogenic route have appeared

including observations from our on studies on triterpene ancestry specialized metabolites (withanolides) of

Ashwagandha (Withania somnifera). Our studies have revealed that withanolides, despite being of triterpene origin,

are synthesized with the involvement of substantial participation of DOXP pathway. Similarly, even differentially role

of MVA and DOXP pathways has been encountered by some researchers for same the sub-class of terpenoids

produced in different parts of the plant. While an article cites this phenomenon as “exception is the rule”, we have

hypothesized a new conceptual line of biosynthetic load-driven model of relative recruitment of DOXP and MVA

pathways for biosynthesis of isoprenoids (Plant Cell Reports 31: 1889-1897, 2012). An updated synthesis of this

subject would be presented at the conference along with its implications in inter-organelle coordination of metabolic

events and its applications in metabolic engineering for enhanced production of terpenoidal secondary metabolites.

R.S. Sangwan

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IPPCongress2015




Enhancement of therapeutic molecules in medicinal plants using

metabolic engineering approach

M.Z. Abdin*

Centre for Transgenic Plant Development, Department of Biotechnology, Faculty of Science, JamiaHamdard, Hamdard Nagar, New Delhi-110062, India


Plants synthesize a vast array of secondary plant metabolites through a network of complex metabolic pathways that

are regulated by endogenous factors and environmental conditions. In 1988 the NAPRALERT data base contains

more than 88000 natural compounds, most of which were derived from plants. Though these compounds are believed

to be non essential for the plant to live, they play an important role in the interaction of the plant with its environment

ensuring the survival in its ecosystem. The plant secondary metabolites however, are of immense economical

importance due to their use as flavouring agents, fragrances, insecticides, pesticides, dyes and drugs. Of all the

drugs used in western medicine, about 25% are derived from medicinal plants, either as pure compounds or as

natural synthons. The drugs derived from the medicinal plants include morphine, codeine, paclitaxel, vinblastin,

vincristine, scopolamine, atropine, pilocarpine, physostigmine, digoxin, artemisinin etc. Best example of natural synthon

is the steroid skelton, which is the basis of large series of drugs, such as contraconceptives and corticosteroids. The

concentration of these compounds in medicinal plants is however, very low, for example the concentration of artemisinin

in Artemisia annua L. is @1.0 %, while the concentration of curcumin in Curcuma longa is @6.0%, limiting their

commercial exploitation. The metabolic engineering approach of secondary metabolic pathways therefore, has been

used by us and other researchers to enhance the concentration these therapeutic molecules in medicinal plants so as

to improve their commercial utilization.

M.Z. Abdin

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Secondary plant product pathway engineering for improved crop

productivity and human health

Prabodh Kumar Trivedi*

CSIR-National Botanical Research Institute, Rana Prata Marg, Lucknow-226001, India


Plants have developed various approaches to magnetize pollinators, shielding themselves against UV-light and for

their defence. One of the strategies adopted by plants for these purposes is based on synthesis of secondary

metabolites, including flavonoids. Flavonoids participate in number of processes in plants as well as are beneficial for

human health. Biosynthesis and accumulation of these secondary plant products is species-specific and under tight

spatial and temporal regulation of gene expression. Therefore, number of food products consumed by human is

deficient in these phytoceuticals. Due to diversity of secondary transformations and complexity of regulation of this

pathway, single gene strategies have not been much successful in enhancing the flavonoid biosynthesis in target

plants. We used transcription factors, carried out gene pyramiding as well as used artificial miRNAs to engineer

flavonoid biosynthesis in plants. Analysis of tobacco and tomato transgenic lines revealed that use of specific

transcription factors can lead to flux availability for phenylpropanoid pathway in general and flavonol biosynthesis in

particular. The transgenic lines with enhanced flavonols developed resistance against the insect pests. Transgenic

tobacco lines co-expressing MYB transcription factor and isoflavone synthase gene accumulated substantial amount

of isoflavonoid being at the highest levels that could be engineered in non-leguminous plants. In addition, flavonoid

rich tobacco and tomato showed value addition in relation to improvement in bone health in animals. Our results

establish an efficient strategy for successful pathway engineering of flavonoids which can be used for improving

medicinal value as well as insect resistance in plants.

P.K. Trivedi

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IPPCongress2015




Enhancing nutritional value of oilseed Brassica juncea through genetic

manipulation of glucosinolate biosynthesis and transport

Naveen K. Bisht* and Rehna Augustine

National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Glucosinolates are Capparales-specific secondary metabolites that have immense potential in human health and

agriculture. Brassica juncea, a globally important oilseed crop, contains relatively high amount of seed glucosinolates

ranging from 80-120 μmol/g dry weight (DW). One of the major breeding objectives in oilseed Brassicas is to improve

the seed-meal quality through the development of low seed glucosinolate lines, as high amounts of certain seed

glucosinolates are known to be anti-nutritional and reduce the meal-palatability. Unlike for Arabidopsis thaliana, our

knowledge about the complex genetical and biochemical regulation of glucosinolate biosynthesis in Brassica crops is

sparse. Our laboratory have identified multiple homologs of key glucosinolate pathway genes (MAM1, CYP83A1,

AOP2), and its transcriptional regulators (MYB28) from B. juncea and assessed the effects of allopolyploidization on

divergence of gene sequence, structure, expression and function. Subsequently we found that, targeted silencing of

BjuMYB28, a key transcriptional regulator, provided significant reduction of the anti-nutritional aliphatic glucosinolates

fractions (ca. 11.26 μmol/g DW of seed), without altering the desirable non-aliphatic glucosinolate pool. The low

glucosinolate trait was stable in subsequent generations of the transgenic lines with no visible off-target effects on

plant growth and development, when tested under containment field conditions. Enrichment of glucoraphanin, a

glucosinolate known for its potent anti-carcinogenic properties, is another area of ongoing research in our laboratory.

Besides, a part of my presentation will discuss selective manipulation of glucosinolate transport processes, which will

be a favourable strategy to engineer the low glucosinolate trait in oilseeds without compromising the fitness of plants.

Naveen K. Bisht

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IPPCongress2015




Changes in metabolites and defense related enzymes in groundnut

during leaf spot diseases

Mahesh Kumar Mahatma1*, Sujit Kumar Bish1, Lokesh Kumar Thawait1, Kuldeep SinghJadon1, Nilesh Khatediya1, Khyati J. Rathod2 and B.A. Golakiya2

1ICAR-Directorate of Groundnut Research, Post Box No. 5, Junagadh-362001, Gujarat, India2Department of Biotechnology, Junagadh Agricultural University, Junagadh-362001, Gujarat, India


Leaf spots are major diseases affecting groundnut productivity worldwide. Early and late leaf spot diseases are

caused by two distinct, but closely related fungi. Late leaf spot (LLS) is generally more aggressive and difficult to

control than early leaf spot. In the present investigation LLS disease resistant (AK-265 , ICGV-86590, FDRS-10,

GPBD-4 and CS-186) and susceptible genotypes (JL-24, JAL-42, GG 20, GG-7 and TG37A) were sown in the pots

during Kharif 2014 and leaf samples were taken from control and infected plants at 45 and 90 days after sowing.

Sugars and phenol were separated and analyse by Ion chromatography and LC-MS/MS, respectively. Glucose and

sucrose content increased in resistant genotypes whereas decreased in susceptible genotypes after infection. These

results suggested that the sugars might be utilized by the pathogen for growth and development. Inositol content was

increased in all genotypes after infection. Higher constitutive and induced level of salicylic acid was observed in

resistant genotypes. Resistant genotypes had about 2-3 times higher constitutive level of salicylic acid. Moreover,

resistant genotypes also had appreciably higher content of p-coumaric acid than that of susceptible genotypes at

infected and uninfected stages. Peroxidase and polyphenols oxidase activities were also higher in resistant genotypes.

Thus, results revealed that higher sugars in resistant genotypes may provide higher amount of substrate for

phenylpropanoid pathway which leads to synthesis of higher content of salicylic acid and coumaric acid and thus

confers disease resistance in groundnut

M. K. Mahatma


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IPPCongress2015


Plenary Session-II

Rice genome for forward and reverse genetics

Akhilesh K. Tyagi1*, S.K. Parida1, S. Das1, N. Malick1, S. Kapoor2, A.K. Singh3 and P. Agarwal1

1National Institute of Plant Genome Research, New Delhi-110067, India2Department of Plant Molecular Biology, Univ. of Delhi, South Campus, New Delhi-110021, India3Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India

*Correspondence: akhilesh @genomeindia.org

We have sequenced genome of rice as part of IRGSP ten years back. In the post-genomic era, significant progress

has been made in transcriptomics and gene discovery by way of forward and reverse genetic approaches. Efforts are

on to fill the gap between the genome and the phenotype. This may lead to regular practice of genomics-assisted

molecular breeding. One of the primary aims of ongoing investigations in our group is to understand gene function,

phylogeny and regulatory networks. Several general conclusions of great relevance emerging from these studies

include: i. new genes and taxon-specific clades can be identified by using molecular model based comparative

genomics, ii. signatures of co-evolution in interacting proteins are discernible, iii. duplicated genes have undergone

neo-/sub-/pseudo-functionalization, iv. highly significant number of genes is differentially expressed in spatial/temporal

manner and in response to stress, and v. such analysis provides useful input for selecting target genes/promoters for

functional genomics. At the same time investigations on defining the role of seed-specific genes by reverse genetics

and identification of QTLs/genes by genetic and association mapping as well as by haplotyping have been undertaken.

As we integrate molecular biology to genetic enhancement activity and overcome practical impediments in taking the

knowledge from lab to land, we hope to reap the benefit of genomics research for plant improvement.

Akhilesh K. Tyagi


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IPPCongress2015


Plenary Session-II

Natural variation in plant circadian clocks

C. Robertson McClung1*, Richard M. Amasino2, Brent E. Ewers3, Kathleen Greenham1,

Victor Hermand1, Ping Lou1, Todd C. Mockler4, Cynthia Weinig3,5, Scott T. Woody2 and

Qiguang Xie1,6

1Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA2Department of Biochemistry, University of Wisconsin, Madison, Wisconsin, USA3Department of Botany, University of Wyoming, Laramie, Wyoming, USA4Donald Danforth Plant Science Center, Saint Louis, Missouri, USA5Department of Molecular Biology, University of Wyoming, Laramie, Wyoming, USA6Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei NormalUniversity, Shijiazhuang, Hebei, China


We have developed a high-throughput motion estimation system to measure circadian parameters by leaf movement.

Analysis of a nested association mapping (NAM) panel from seven Recombinant Inbred Line (RIL) populations of

Arabidopsis thaliana, supplemented with accessions from the 1001 Genomes Project, has identified multiple Quantitative

Trait Loci (QTL) linked to known clock genes, as well as at least one novel QTL. Among natural populations of

Mimulus guttatus and elite cultivars of soybean (Glycine max), we observe a strong latitudinal cline in circadian period

in both species. This suggests that the clock contributes to performance in both natural and cultivated conditions.

This is consistent with our growing understanding of the roles of the clock in the regulation of flowering and in the

temporal coordination of growth and development, and in abiotic and biotic stress responses. In Brassica rapa, we

have mapped QTL for circadian period, water use efficiency (WUE), shade avoidance, and other morphometric traits.

There is striking colocalization of QTL for circadian period with QTL for shade avoidance and WUE. We have identified

GIGANTEA (GI) as a major QTL responsible for natural variation in circadian period and have identified the causal

nucleotide polymorphism responsible. This same nucleotide polymorphism underlies variation in hypocotyl elongation,

cold and salt tolerance. Thus, natural variants of circadian clock genes such as GI offer attractive targets for the

development of crops with enhanced stress tolerance and improved yield.

C. Robertson McClung


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IPPCongress2015


Plenary Session-II

Functional relevance of stress specific transcription factors bZIP60,

MYC57 and NAC1 from Eleucine coracona in improving stress tolerance

in rice

K.C. Babitha, S.V. Ramu, H.V. Ramegouda, Nataraja N. Karaba and M. Udayakumar*

Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus,Bangalore-560065, Karnataka, India


Moisture stress induced decrease in cell turgor affects several metabolic processes and amongst these, protein

synthesis and protein turnover are the most challenged. Major stress induced constraints are translation processes

and protein folding and stability. Stress specific transcription factor (TFs) might play a pivotal role in regulating the

functional genes to sustain these processes. With this back ground, from the stress specific transcriptome of finger

millet we cloned and characterized three stress specific TFs EcNAC1, EcMYC57, EcbZIP60 involved in regulating the

expression of genes associated with protein folding and stability. These TFs are upregulated under diverse stress

conditions. Overexpression of these TFs in tobacco improved tolerance to diverse abiotic stresses. The EcbZIP60

transformants showed tolerance to DTT and tunicamycin induced ER stress by regulating expression of relevant

genes involved in protein folding. The EcMYC57 and EcNAC1 improved tolerance predominantly by upregulating the

several dehydrins. Transgenic plants co-expressing of EcNAC1, EcMYC57, EcbZIP60 showed substantially improved

tolerance to diverse abiotic stresses expressing ER stress responsive genes like chaperones (BiP, CRT, PDIL),

dehydrins (LEA14, rd29A, rd29B), ABA signaling (PP2C). These results not only signify the relevance of genes

associated with protein folding and stability in improving the stress adaptation but also provide evidences that the

simultaneous expression of relevant genes has specific advantage over single gene transgenics. Based on this proof

of concept the rice transgenic plants developed co-expressing EcNAC1, EcMYC57, EcbZIP60 showed improved

tolerance to ER stress besides salinity and dehydration. Further these transgenics maintained higher spikelet fertility

and increased productivity under semi irrigated aerobic conditions.

M. Udayakumar


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IPPCongress2015


Concurrent Session E

Marker assisted breeding for disease / pest resistance

Ramesh V. Sonti*

CSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500007, Telangana, India


Marker assisted selection (MAS) is a very effective tool for crop improvement that involves use of variations in DNA

sequences as flag posts to make selections of varieties that have new and advantageous combinations of genes.

MAS can be applied for crop improvement when the trait of interest is present within the gene pool/extended gene

pool of the crop of interest. I will present our collaborative work with the Indian Institute of Rice Research, Hyderabad,

on development of a rice variety that is resistant to bacterial blight as a case study on how MAS can be gainfully used

for crop improvement. This variety, called Improved Samba Mahsuri (ISM), is in commercial cultivation and is being

appreciated by farmers in rice growing regions where fine grained varieties are cultivated and bacterial blight is a

production constraint. I will present recent results which indicate that ISM exhibits tolerance to another bacterial

disease called leaf streak; a disease which is gaining in importance and for which very few sources of resistance are

available. Furthermore, we have developed an EMS mutagenized population of the elite rice variety Samba Mahsuri

that we have used to isolate new mutants that are resistant to bacterial blight and other mutants that exhibit enhanced

tolerance to yellow stem borer, a serious insect pest of rice. I will discuss our plans for further characterization of

these mutants and how this work can be used to extend the scope of MAS.

Ramesh V. Sonti


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IPPCongress2015



Maize defense signaling in response to aphid infestation

Joe Louis1,2*, Saumik Basu1 and Suresh Varsani1

1Department of Entomology; 2Department of Biochemistry, University of Nebraska-Lincoln, Lincoln,NE 68583, USA


Maize, one of the world’s significant cereal crops, suffers severe yield losses due to insect infestation. Corn leaf aphid

[CLA; Rhopalosiphum maidis (Fitch)] is an economically important pest of several monocot crops, including maize. In

addition to extensive crop damage, CLA also acts as a vector for viruses that cause devastating diseases in maize.

The maize genotype, Mp708, developed by classical plant breeding confers enhanced resistance to CLA compared

to B73 and Tx601 maize inbred lines. Feeding by CLA triggers the rapid accumulation of the Maize insect resistance1

(Mir1) transcripts, which encodes a cysteine protease. Since Maize insect resistance1-Cysteine Protease (Mir1-CP)

is expressed in the vascular tissues, we examined the effect of vascular sap exudates by including it in the aphid diet.

The number of aphids recovered from diet containing vascular sap collected from uninfested plants was comparable

between Mp708 and Tx601 genotypes. However, if plants were infested with CLA prior to sap collection, Mp708

vascular sap significantly reduced the number of aphids compared to Tx601, suggesting that CLA-feeding induced

accumulation of toxic factors, presumably Mir1-dependent factors, contribute to defense against aphids. Furthermore,

our results demonstrate that recombinant Mir1-CP provides direct toxicity to CLA. Finally, we recently identified that

foliar feeding by CLA rapidly sends defensive signal(s) to the roots that trigger belowground accumulation of the Mir1,

signifying a potential role of long-distance signaling in maize defense against the phloem-feeding insects.

Joe Louis


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IPPCongress2015



Targeting of chitinase and cathepsin L genes of Helicoverpa armigera

by RNAi confers insect resistance in tomato

Mamta and Manchikatla Venkat Rajam*

Polyamine, Transgenic and RNAi Laboratory, Department of Genetics, University of Delhi SouthCampus, Benito Juarez Road, New Delhi-110021, India


Helicoverpa armigera, a polyphagous and notorious insect pest causes huge yield losses in many crop plants, and

various insect control strategies met with limited success. Hence, novel alternate strategies are required to combat

insect pests. RNA interference (RNAi), a sequence-specific gene silencing mediated by dsRNA emerged as a novel

tool for insect control, where host plants are engineered to produce dsRNAs against vital genes of insect pests. In the

present study, we have identified chitinase (HaCHI) and cathepsin L (HaCL) genes as potential targets for control of

H. armigera as they are involved in insect molting, metamorphosis and survival. The off-target free partial sequences

were selected from CDS and 3’UTR regions of HaCHI and HaCL genes respectively, and they were isolated and

used for preparation of hair-pin RNAi constructs. Several RNAi tomato lines were generated by Agrobacterium-

mediated transformation with these constructs. Continuous feeding of insect larvae on leaves of RNAi tomato lines

reduced the larval growth, development and caused mortality. Various developmental deformities occurred in pupa

and adults developed from larvae fed on leaves of RNAi lines. Reduction in fertility/fecundity was also observed in

adults as compared to the control adults. The q-PCR analysis showed reduction in target gene transcripts in larvae

fed on RNAi lines, and this could be correlated with the observed RNAi effects on insects. These results demonstrate

that the chitinase and cathepsin L are essential for insect growth and development, and RNAi holds a great potential

for effective management of H. armigera.

M.V. Rajam


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IPPCongress2015



Rep mediated transcription of geminivirus genome : Bringing in a

new perspective

Nirbhay Kushwaha, Mansi Bhardwaj and Supriya Chakraborty*

School of life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Geminiviruses are a group of plant viruses with ssDNA genome, which replicate in the nucleus of the infected cell.

Replication initiator protein (Rep) encoded by a geminivirus is an early protein that is indispensible for viral replication.

Although, multitasking roles of Rep in replication has been elucidated, information about its role in efficient gene

expression is lacking. Chilli leaf curl virus (ChiLCV) is a monopartite geminivirus that can replicate in permissive host

Nicotiana benthamiana. For efficient and orderly transcription from viral genomic DNA, recruitment of machineries

responsible for histone post-translational modifications (PTMs) on viral promoter is essential. The present study

evidently proves that deposition of histone 2B monoubiquitination (ubH2B) promotes tri-methylation of histone 3 at

lysine 4 (H3K4me3) on ChiLCV promoter. We isolated homologues of two major components of monoubiquitination

machinery, UBIQUITIN CONJUGATING ENZYME UBC 2 (NbUBC2) and HISTONE MONOUBIQUITINATION1

(NbHUB1) from N. benthamiana. TRV based virus induced gene silencing of NbUBC2 and NbHUB1 has indicated

their crucial roles in ChiLCV pathogenesis. Rep redirects NbUBC2 from the cytoplasm to the nucleusand interacts

with NbHUB1 and NbUBC2 in the nucleus. Furthermore, NbUBC2 and NbHUB1 silencing reduced deposition of

H3K4me3 and ubH2B on host chromatin and down-regulated expression of host genes. Together, the current study

reveals that ChiLCV Rep protein binds to the viral genome and recruits two host proteins NbUBC2 and NbHUB1.

These proteins further help in monoubiquitination of histones 2B, and accumulation of H3K4me3 on viral mini-

chromosomes, and in turn enhance transcription of the viral genome.

Supriya Chakraborty


50

IPPCongress2015



Secreted effectors of necrotrophic fungi: Quest for possible host

targets

Praveen Verma*

Plant Immunity Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Marg,New Delhi-110067, India


Recent studies on genomics of both plant and phytopathogens are beginning to unravel how these organisms

manipulate host immunity to cause diseases. Plants recognize pathogens through strategies involving both conserved

and variable pathogen elicitor, and pathogen manipulate the defense response through secretion of virulence effector

molecules. However, only a few studies have explored for necrotrophic fungal pathogen effectors. Chickpea-Ascochyta

rabiei system provides an excellent model for studying necrotrophy. We have sequenced 34.6 megabase draft genome

sequence of Ascochyta rabiei and predicted 10,596 protein-coding genes that encode large and diverse inventories

of secretory proteins, carbohydrate-active enzymes, transporters and primary and secondary metabolism enzymes,

reflecting its necrotrophic lifestyle. Comprehensive analysis predicted a set of 758 secretory proteins with different

conserved motifs. Further analysis of the secretome revealed a high abundance of PEXEL-like effector candidates.

We characterized a novel PEXEL-like effector, PEC25, and found that this effector is indispensable for A. rabiei

virulence. PEC25 translocate into the host nucleus during infection, which is critical for its functionality. To identify the

host target, we explored the candidate genes of major QTLs of chickpea against A. rabiei resistance for PEC-25

interaction. Interestingly, we showed that PEC-25 is interacting with transcription factor present in QTLAR1

in the

nucleus of chickpea. Our research will give new insights to elucidate the necrotrophic mode of infection through

effectors and their targets.

Praveen Verma


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IPPCongress2015



Plant-pathogen interaction with respect to plant parasitic nematodes:

A sophisticated dialogue

Amar Kumar1*, Neha Shukla1, Pritam Kaur1, Rachita Yadav2, Arun Jagannath1, Shailendra

Goel1 and Manu Agarwal1 and Ramneek Gupta2

1Department of Botany, University of Delhi, Delhi-110007, India2Center for Biological Sequence Analysis, Technical University of Denmark, Denmark


Plants are continuously exposed to a wide range of pathogens, including plant parasitic nematodes. Being sessile,

plants have developed elaborate innate defense mechanisms that allow them to recognize potential invading pathogens

and to initiate successful defenses. I will initially present our current state of understanding of plant pathogen interactions

followed by our own research on plant-nematode interactions. Plant endoparasitic nematodes (cyst and root-knot

nematodes) are major pests of crop plants. Potato cyst (Globodera species) and root-knot (Meloidogyne species) are

obligate biotrophs and takes between 3-6 weeks to complete their life cycle in the host roots. Roots are invaded by

infective juveniles (J2) and develop feeding sites with highly modified host cells. Formation of feeding cells is activated

by nematode secretions, which cause dramatic morphological and physiological changes in root cells and converts

them into nutrient sinks. Earlier, we have cloned and characterized a tomato gene (Hero) that confers resistance

against Globodera species by disrupting initiation and development of feeding cells in roots. Recently, we have taken

genome-wide approaches by transcriptome and miRNAome analyses to investigate genes and regulatory miRNAs

from tomato that are involved in disease development during root-knot nematode infection. We have shown that a set

of genes (e.g., WRKY transcription factor, Cyclin D3-1, Putative Hydrophobic protein OSR8, Auxin Efflux carrier) and

miRNAs (e.g., mi156, miR164, miR319) from tomato are up- and down-regulated at various stages of disease

development and these are likely to play a crucial role in determining the outcome of plant-nematode interaction.

Amar Kumar


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IPPCongress2015


Concurrent Session F

Molecular response of plants to phosphate deficiency

K.G. Raghothama1*, V.K. Nagarajan1 and A. Jain2

1Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette,IN-47907, USA2National Research Centre on Plant Biotechnology, Lal Bahadur Shastri Building, Pusa Campus,New Delhi-110012, India


Phosphorus, a key nutrient regulating metabolism and energy balance impacts a multitude of plant processes. Many

of them are regulated at transcriptional and post transcriptional levels to enable plants to acquire, mobilize and

recycle phosphate. Systemic and local responses of root system allow the plant to acquire phosphate from the

rhizosphere. Cross talk with sugar, phytohormones and other nutrient signaling pathways is a part of phosphate

starvation response mechanism. Pi deficiency modulates expression of Pi starvation-responsive (PSR) genes that

are coordinately regulated by several transcription factors (TFs). Our research is focused on dissecting transcriptional

regulation of phosphate starvation responses in Arabidopsis. Studies show that low Pi stress triggers expression of

several transcription factors including Myb62, WRKY75, Zat6, HHO2 and ERF, E3 ligase among others. Analysis of

these transcription factors using gene knockout and overexpression plants reveal a diverse role for them in regulating

root system architecture, interaction with sugar, phosphate acquisition and utilization. Further Myb62 and ERF are

also involved in GA mediated root responses to phosphate starvation. Studies revealed that subset of Pi starvation-

induced (PSI) responses mediated by HHO2 is under regulation of PHR1 which is considered to be a major regulator

of PSI. Transgenic plants with altered expression of transcription factors exhibit interesting root and shoot phenotypes,

sensitivity to sugar and GA highlighting the complexity of plant response to phosphate deficiency. In spite of recent

advances questions related to sensing and signaling, post transcriptional regulation, hormone and sugar interactions

still need to be answered.

K.G. Raghothama


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IPPCongress2015



Genome-wide expression of transcriptomes under iron and zinc

stresses revealed candidate genes for kernel iron and zinc

accumulation in maize (Zea mays L.)

Mallana Gowdra Mallikarjuna1, Nepolean Thirunavukkarasu1, Rinku Sharma1,

Kaliyugam Shiriga1, Shikha Mittal1, Firoz Hossain1 and Hari Shanker Gupta2*

1Maize Research Lab, Division of Genetics, Indian Agricultural Res. Inst., New Delhi-110012, India2Borlaug Institute for South Asia, CG Block, NASC Complex, New Delhi-110012, India


Deficiency of iron (Fe) and zinc (Zn) causes micronutrient-malnutrition or hidden-hunger affecting millions of people

globally including sub-Saharan Africa, South Asia and Latin America where, maize is one of the major sources of food

and feed. Among several available approaches, genetic-biofortification is a sustainable and cost-effective approach

in addressing hidden-hunger. Identification of key genes involved in Fe and Zn homeostasis is an important prerequisite

to breed micronutrient-rich hybrids. A transcriptome assay was performed, with ~18,000 transcripts distributed across

the maize genome, in a SKV-616 inbred after exposing the seedlings to three Fe and Zn stress treatments (+Fe -Zn,

-Fe+Zn, -Fe -Zn) along with control (+Fe+Zn). A total of 306 and 715 genes expressed in root and shoot, respectively

were found common across stress treatments, suggesting presence of common transporters controlling both minerals

in root and shoot. Differential expression of genes under stress treatments revealed that Zn association in many

pathways was more pronounced over Fe pathways and the physiological demand for these minerals in shoot was

much higher than in root. Significant expression of genes involved in mugineic acid pathway, metal-transporters, and

phyto-hormone metabolisms revealed the role of those pathways in understanding the physiological and molecular

mechanisms of Fe and Zn homeostasis. Co-mapping of DEGs with known QTLs for kernel Fe and Zn concentration

identified several candidate genes such as OPT, metal-binding-protein, nramp3 and NAS. The candidate genes

identified in the present investigation could be used for breeding kernel Fe- and Zn-rich maize hybrids.

H. S. Gupta


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IPPCongress2015



Identification of genomic regions and genes for nitrogen use efficiency

in rice (Oryza sativa L.)

C.N. Neeraja*, S.R. Voleti, D. Subrahmanyam, K. Surekha, I. Subhakara Rao and B. Srikanth

Indian Institute of Rice Research, Hyderabad, Telangana, India


Nitrogen use efficiency in rice crop is crucial in climate change for reduction of nitrous oxide emission resulting from

excess nitrogen fertilizer application and also in reduction of cost of cultivation. To identify genomic regions and

genes for NUE in rice, various strategies viz., QTL and association mapping, micro array, real time PCR, transcriptome

and micro RNA analyses were applied to germplasm and biparental mapping populations. Around 700 genotypes

were screened under low and recommended nitrogen levels in field and wide genetic variability was observed. A set

of genotypes was subjected to association mapping with 50 SSR markers and ~100 traits derived from field and

hydroponics screening and 26 genomic regions found to be associated with more than 10 traits. Differential expression

of PAL, OsSPL 14, Ammonium transporter, Glutamine synthetase and Nitrate Reductase was observed through

qRT-PCR. Using genotypes with differential response under low N, applying Next Generation Sequencing strategies,

several up and down regulated genes were analyzed in root and shoot tissues of two genotypes using microarray

analysis; panicles of two genotypes were identified on HiSeq 2000 using 2x100 bp pair-end sequencing and using

whole genome mRNA sequencing. The identified differentially expressed genes are being validated in the mapping

populations and being introgressed into popular varieties.

C.N. Neeraja


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IPPCongress2015



Comparative functional genomics of GPCRs and G-proteins: A new

match-making service?

Nandula Raghuram1*, Navjyoti Chakraborty1, Kostya Kanyuka2, Ravi Pathak1, Devapriya

Choudhur3 and Richard Hooley4

1Molecular Biology and Functional Genomics Laboratory, University School of Biotechnology, GuruGobind Singh Indraprastha University, Sector 16C, Dwarka, New Delhi-110078, India2Department of Plant Biology and Crop Science, Rothamsted Research, West Common, Harpenden,Hertfordshire, UK3School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India4Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, UK


Plant G-proteins and canonical G-protein coupled receptor(s) are best represented by Arabidopsis GPA1 and GCR1.

But their partnership in plant signalling became controversial due to the competing all-or-none hypotheses. We used

comparative functional genomics approach to analyse the whole transcriptomes of GPA1/GCR1 single and double

mutants of Arabidopsis (gpa1-5, gcr1-5 and gpa1-5gcr1-5). By comparing the transcriptomes of the single mutants,

we found that 30% of all GCR1-responsive genes and 57% of all GCR1-regulated processes were similar to those of

GPA1, though there were also 246 DEGs and 43 processes that did not overlap with those of GPA1. Similarly,

pairwise comparison of the differentially expressed genes (DEGs) from the double mutant with each single mutant

revealed commonality of 115 or 33% of the 350 GCR1-regulated genes and 214 or 54% of the 394 GPA1 regulated

genes respectively. When the double mutant was included in the comparison, though the common genes constituted

only 6.2% of the 1,031 DEGs found in all 3 mutants, they represented 34% of all the 160 processes, indicating their

co-regulation. This is by far the most compelling evidence, not only in favour of the GCR1-GPA1 partnership, but also

in favour of its possible coexistence with other alternative partnerships, contrary to all-or-none hypotheses. The

alternative partnerships may involve other GCRs, RGS1 or hitherto unknown GPAs. Our identification of the genes/

processes that could be regulated by such combinatorial partnerships provides a functional basis for match-making

between different components of G-protein signalling for the first time.

Nandula Raghuram


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Foxtail millet: A climate change resilient model C4 crop with rich genetic

and genomic resources for genetic improvement of cereals and

bioenergy grasses

Manoj Prasad*



Foxtail millet (Setaria italica L.) is recently regarded as a tractable experimental model crop for studying functional

genomics of millets and bioenergy grasses. Further its potential abiotic stress tolerance has invited much research

attention and we attempted to generate genomic resources and to decipher the molecular mechanism of stress

tolerance. Though our efforts has generated considerable knowledge on structural and functional genomics of foxtail

millet, the release of genome sequence significantly accelerated the genomic research to a new dimension, as

evidenced by large-scale development of genomic resources, development of web-based database, genome-wide

association studies, genome-wide functional genomics and standardization of molecular protocols specific to foxtail

millet. Our research contributions in both the functional and structural genomic aspects of foxtail millet are remarkable

and our efforts have now resulted in the transformation of this orphan crop into a nutri-crop with rich genomic and

genetic resources. These resources would promisingly assist in crop improvement programs involving millets, non-

millets and bioenergy grass species, thus ultimately addressing global food security.

Manoj Prasad


57

IPPCongress2015



Comparative transcriptome and co-expression network analysis of two

Brassica juncea cultivars in response to nitrate treatments

Parul Goel1,2, Rohit Chauhan1, Mohit Kumar Swarnkar1, Shivalika Pathania1, Vandna

Chawla1, Vishal Acharya1, Ravi Shankar1 and Anil Kumar Singh1,2*

1Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061(HP), India2Academy of Scientific and Innovative Research, New Delhi-110012, India


Brassica juncea is an economically important oilseed crop of India. The Nitrogen use efficiency (NUE) of this crop is

lower than other crops. We employed RNA-seq approach to understand transcriptional response of two contrasting

genotypes Pusa Bold (high NUE) and Pusa Jaikisan (low NUE) of B. juncea in terms of nitrogen use efficiency under

various nitrate treatments. A total of 42 RNA-seq libraries were sequenced using Illumina GAIIx that generated

446,580,418 paired-end reads which were utilized for de novo assembly following quality filtering. A total of 46,556

transcripts were generated with N50 1,393bp. A total of 20,884, 11,327 and 12,575 transcripts were annotated under

GO, EC and KEGG, respectively. Our study showed that nitrate as a signal regulates expression of several genes

involved in processes like signaling, nitrogen metabolism, carbohydrate metabolism, photosynthesis, secondary

metabolite production etc. Moreover, genes involved in nitrate transport and assimilation were found to be induced

more rapidly in Pusa Bold as compared to Pusa Jaikisan after nitrate treatment. Weighted gene co-expression network

analysis of RNA-seq data of both the varieties was performed to explore their differential behavior in response to

nitrate. From our network analysis, conserved modules in both varieties that respond to nitrate treatment were identified.

Based on the comparison of shared genes, a HUB transcription factor mTERF was found to be differentially connected

in both datasets in response to nitrate treatment. Through this study, we made an attempt to understand the molecular

regulation of nitrate response in Brassica juncea.

Anil Kumar Singh


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Concurrent Session G

Structural biology of pre- and post-receptor regulation of auxin

hormone responses

Joseph Jez*, Lucia Strader, Corey Westall, Ashley Sherp and David Korasick

Department of Biology, Washington University, St. Louis, MO, USA


Plants respond to developmental cues and abiotic and biotic stresses by controlling both the level and activity of

various hormones, including auxins, jasmonates and salicyclic acid. Recent structural biology studies provide new

insights on control mechanism before and after binding of auxin and jasmonates to their respective receptors. Pre-

receptor action by acyl acid amido synthetases of the GH3 protein family modulate hormone levels by amino acid

conjugation. The first crystal structures of benzoate-specific Arabidopsis thaliana AtGH3.12/PBS3 and jasmonic acid

(JA)-specific AtGH3.11/JAR1, combined with biochemical analysis, define features for the conjugation of amino acids

to diverse acyl acid substrates and highlight the importance of conformational changes in the C-terminal domain for

catalysis. These results demonstrate how a highly adaptable three-dimensional scaffold is used for the evolution of

promiscuous activity across an enzyme family for modulation of plant signaling molecules. A second molecular control

mechanism in auxin responses involves protein-protein interactions mediated by the Phox/Bem1p (PB1) domains of

Auxin Response Factors (ARF) and Aux/IAA co-repressors to regulate transcriptional events modulated by the

phytohormone auxin. These structures reveal a two-pronged interaction and evolutionarily conservation of this core

recognition feature, along with co-varying interface sequences allows for versatile PB1-mediated interactions in auxin

signaling. Combined structural, functional, and sequence analysis provides new molecular insights that inform future

inquiry into plant auxin responses.

Joseph Jez


59

IPPCongress2015



NPR1-dependent and –independent salicylic acid signaling in

Arabidopsis

Ashis K. Nandi*

School of Life Sciences, Jawaharlal Nehru University, New Delhi-1110067, India


Amongst the plant hormones, salicylic acid (SA) plays the most significant role in defending biotrophic and hemibiotrophic

pathogen. Plants accumulate SA upon pathogen accumulation, which is required for activation of defense. SA activates

a transactivator protein NPR1 (Non-expressor of PR1), the master regulator of SA signalling. Several suppressors of

npr1 mutation suggest the presence of NPR1-independet pathways for defense signaling, which remain elusive.

Through screening of EMS mutagenized Arabidopsis plants, we identified cdd1 (constitutive defence without growth

defect 1) as a suppressor of npr1. In contrast to other npr1 suppressors, cdd1 is not compromised for growth and

development. Further, through transcriptome analysis of cdd1, we identified and characterized a novel zinc-finger

domain containing protein ZFD1 that positively regulates defense signalling. Pathogens or SA induce ZFD1 expression.

Through screening of Arabidopsis cDNA library in a yeast-2-hybrid assay, we identified a large numbers of interactors

that provide mechanistic support for ZFD1 function. The epistatic interaction studies with npr1 mutant suggest that

ZFD1 influences both NPR1-dependent and –independent pathways of SA signaling.

Ashis K. Nandi


60

IPPCongress2015



Biotechnological method of production of orchid tubers containing

biologically active substances for biomedical applications

Lavr Kryukov* and Aleksandr Shirokov

Laboratory of Innovative Production Technologies of Plants, Lobachevsky State University, NizhnyNovgorod Region, Russian Federation, Russia


At present, there is almost no reliable data on the biochemical composition of tuberoids. According to the literature, it is

only known that they contain starch (2.7%), mucilage (48%), glycosides, polysaccharides, dextrin, minerals, essential

oils, proteins, etc. The contents of the components are not exactly specified, and it is impossible to calculate their

percentage. However, this information is important, as this plant material could be of great interest to the pharmaceutical

and parapharmaceutical industry, because it contains polysaccharides that have biologically active properties. We

studied Orchis traunsteineri So3 and Orchis militaris L. from the collection of the botanical garden of UNN. To obtain the

tubers, plants were cultivated for 2 months on the artificial agar medium without hormones with 12-hour photoperiod at

18°C and at 60% relative humidity. For the initial material we used protocorms, which were cultured to the stage of the

seedling. Root-tubers of orchids contain highly vacuolated aquifer parenchyma cells. We extracted mucoid substance

from these cells, dried them, and analyzed them using Shimadzu gas chromatography-mass spectrometer. The analysis

showed that the mucilage contains the variety of secondary metabolites, among which we identified sericic acid (inhibition

of yeast alpha-glucosidase), lupeol (possesses anti-tumor properties) and betulin (antioxidant and anti-inflammatory

properties, immunomodulator). These valuable substances with a strong biological activity were discovered by us for the

first time in this group of species. Our further research focuses on the selection of a line of orchid clones containing the

largest number of biologically active substances for the biomedical applications.

Lavr Kryukov


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IPPCongress2015



AOS networking in seed system biology

Subhash Chandra Naithani*

Seed Biology Laboratory, School of Life Sciences, Pt. Ravishankar Shukla University, Raipur-492010,Chhattisgarh, India


Ever expanding but fragmentary literature on the functions of AOS in the critical regulation of various aspects of seeds

and seedlings demands comprehensive understanding of AOS networking in a consolidated manner appreciating seed

system biology. Dual function of AOS is now established. Depending upon the physiological concentrations of AOS they

may play an active role in either the regulation of signal transduction or cellular damage. Our study revealed that the low

concentrations of AOS regulate maturation drying of seed development, germination and seedling establishment. Whereas,

excess amounts of AOS are central to widely acknowledged “free radical theory” for explaining the mechanism of ageing

in several forest tree seeds. Understanding of AOS functions becomes more complicated due to its various biologically

active forms such as superoxide, H2O

2 and hydroxyl radical with significant variation in their shelf-life and oxidation

potential. Acquisition of desiccation tolerance, one of the remarkable traits of seeds to survive vicissitudes of environment

is induced by ABA mediated AOS that up regulates Lea and AOS detoxifying genes during maturation drying of seed

development of Azadirachta indica. Similarly, AOS signalling coordinate various phases of germination of A. indica

seeds by regulating the antioxidant enzymes and cell cycle proteins. In germinated pea seeds, AOS have been assigned

a new role in the cell proliferation and differentiation in stem cell niche through controlling the cellular redox state thus

important in seedling establishment. Our work is an attempt towards understanding and unifying the tapestry of AOS

network in seed system biology.

S.C. Naithani


62

IPPCongress2015



Relationship of flag leaf photosynthesis with stomatal density, dry

matter partitioning and yield in wheat (Triticum aestivum)

Kushal Kumar Baruah*, Ashmita Bharali and Gulshan Jha

Department of Environmental Science, Tezpur Central Univ., Napaam-784028, Assam, India


Changing atmospheric levels of carbon dioxide (CO2) and the increasing demands for food of a growing world population

are testing the photosynthetic adaptability of a crop and natural ecosystems. A field experiment was conducted during

2013-2014 and 2014-2015 to investigate the efficiency of 07 wheat cultivars [CBW-38(V1), WH-283(V2), WH-1080(V3),

WH-1021(V4), WH-711(V5), WH-542(V6) and PBW-343(V7)] for carbon dioxide fixation, dry matter partitioning and

yield. Differences among the varieties in terms of their photosynthetic efficiency and biomass partitioning ability were

clearly observed. Stomatal frequency of the flag leaf showed a good correlation with photosynthesis at panicle initiation

stage. Average number of stomata observed by Scanning Electron Microscopy ranged from 61-111 mm-2 of leaf area of

the wheat cultivars. The internal to ambient CO2 mole fraction ratio also differed amongst the varieties which may be

attributed to the differences in CO2 requirement of mesophyll cells. Higher stomatal density and flag leaf photosynthesis

in the variety WH 1021 contributed to efficient translocation of assimilates to the developing sink and recorded higher dry

weight of the kernels at harvest. The pooled analysis of data for both the years of experimentation have revealed that

cultivar WH 1021 is superior over other cultivars in terms of photosynthesis and grain yield, and may be beneficial in a

future climate change agriculture. Keywords: leaf gas exchange, stomatal density, yield, wheat, cultivar.

K.K. Baruah


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IPPCongress2015



Enhancing stress tolerance by Silicon in rice

S.R. Voleti1*, D. Subrahmanyam1, P.R. Rao1 and A.P. PadmaKumari2

1Plant Physiology Section 2Entomology Section, Indian Institute of Rice Research, Rajendranagar,Hyderabad-500030, Telengana, India


The beneficial effects of silicon in plant nutrition particularly abiotic and biotic stress tolerance in rice was studied and

reviewed. Lower solubility, uptake, redistribution within the plant organs and polymerization of silicon on the leaf surface

are proposed to be important in rice crop. At national level, in collaboration with IICT, compounds solubilizing silica were

developed and examined their efficacy under field situation and against various stresses. Enhanced solubility and

chemical profile of molecular complexes and sandwich arrangement model was used to explain the mechanism at the

soil-plant interface. From yellow stem borer infected plants, wearing larval mandibles in incisor region due to feeding on

stems was recorded. Histological evidences suggesting rupturing of peritrophic membrane, vacuolation, disintegration

of columnar cells and discharging of cellular contents into the lumen due to abrasion of midgut epithelium using SEM,

and deposition of silicon in cell wall in the form of thick dumb-bell shaped bodies using Electron dispersive X-ray

spectroscopic analysis of stem samples of rice varieties indicated 11-18% accumulation. Enhanced photosynthesis and

water relation characteristics are some of the physiological characteristics that were recorded on silicon application. The

beneficial effects studied across the country in rice growing region under AICRIP revealed that, problematic soils are

benefitted with application. At International level transporter mediated uptake of soluble silica, fine mapping of genes

using low silicon rice mutant, sequencing and identification of aminioacid region and their position are some of the

important milestones in this emerging area of silicon research.

S.R. Voleti


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Plenary Session-III

Methylglyoxal and glyoxalase pathway: markers for plant stress

tolerance

Charanpreet Kaur1,2, Ananda Mustafiz1, Ajit Ghosh1, Shweta Sharma1, Khirod Sahoo1,Ashwani Pareek2, Sneh L. Singla-Pareek1 and S.K. Sopory1,2*

1Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology,New Delhi-110067, India2School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Methylglyoxal (MG), a potent cytotoxic α-oxoaldehyde is generated as a byproduct of glycolysis and increases to toxic

levels under stress and pathological conditions in plants and animals. The ubiquitously present glyoxalase pathway,

comprising glyoxalase I (GLYI) and glyoxalase II (GLYII) genes, is required for the detoxification of methylglyoxal. We

have shown earlier that, glyoxalase pathway is induced in response to various abiotic stress conditions in plants and

over-expression of GLYI and/or GLYII genes impart tolerance towards different stress conditions by keeping MG levels

in control, thus indicating their essential role in stress adaptation in plants. Lately, we have shown that the plant kingdom

possesses multiple isoforms of GLYI and GLYII which have probably evolved to adopt specialized roles, be it in different

organelles, under different developmental stages or in response to different stimuli. A unique Ni2+-dependent GLYI, a

nucleus-localized Zn2+-stimulated GLYI and a glutathione-stimulated GLYII have been characterized in rice. Our studies

also indicate that MG plays a role in stress signaling in plants, inducing large-scale changes in plant signaling transcriptome

and also altering the expression of various genes involved in stress and defense response. Thus, in view of general

inducibility of both MG and glyoxalase pathway under stress, these can be proposed as possible biomarkers in plant

stress response.

S.K. Sopory


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IPPCongress2015


Plenary Session-III

Insights into plant and algal metabolism through metabolic modelling

Mark Poolman1*, Kailash Adhikari1 and Dipali Singh1

1Department of Biology and Medical Science, Oxford Brookes University, Oxford, U.K.


In this presentation, some of the fundamental principles of the structural analysis of metabolic networks will be

introduced, with particular reference to the application of linear programming to such networks. This will be followed

be recent examples of the application of these techniques to the metabolism of Arabidopsis, identifying potential

pathways which could circumvent the effect of genetic knockouts which might otherwise be predicted to be lethal, and

to the metabolism of the diatom, Phaeodactylum tricornutum, to identify novel potential variants of photorespiration,

and roles for enzymes whose purpose is currently unclear. This work is funded as part of the “AccliPhot” project, EU

grant number PITN-GA-2012-316427, http://accliphot.eu.

Mark Poolman


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IPPCongress2015


Concurrent Session H

Discovery and creation of genetic variation to enhance the yield

potential of wheat

Martin A.J. Parry*

Lancaster Environment Centre, Lancaster University, Lancaster, UK


There is an urgent need to develop integrated and sustainable approaches that will significantly increase both crop

production per unit land area and resource use efficiency. Compelling evidence from free-air CO2 enrichment

experiments demonstrates that increasing photosynthesis increases yields, provided that other constraints do not

become limiting. The efficiency with which the sun’s energy can be captured as wheat biomass seldom exceeds 2%

and averages less than 1%; improving this conversion efficiency is widely recognized as the new frontier for crop

improvement. Our research has focused on overcoming the limitations of the CO2-fixing enzyme Rubisco, either

through mining natural diversity or by genetic engineering. We have surveyed a large panel of commercial wheat

varieties to assess the extent of natural variation in photosynthesis in relation to yield. Significant heritable natural

variation in CO2 assimilation was identified. Since photosynthesis is co-limited in the canopy by the kinetics of Rubisco

and the regeneration rate of RuBP, increasing RuBP regeneration by overexpressing rate limiting enzymes involved

in RuBP regeneration is predicted to increase photosynthesis. We have genetically modified UK and CIMMYT wheat

cultivars to increase RuBP regeneration and preliminary results support the predictions of improved photosynthesis

at high CO2. Understanding and exploiting natural diversity and creating novel germplasm based upon exploratory

research provide a powerful combination to achieve the challenging and pertinent goal.

Martin A.J. Parry


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Photorespiration: An adaptive response of plants of oxidative stress

Agepati S. Raghavendra*, Bobba Sunil, Vetcha Aswani, Ramesh B. Bapatla andMohd. Khan Fareed

Department of Plant Sciences, School of Life Sciences, University of Hyderabad,Hyderabad-500046, India

*Correspondence: [email protected], [email protected]

The process of photorespiration achieves two important goals of metabolizing 2-phosphoglycollate, a toxic metabolite,

back into the Calvin cycle, and limiting the loss of carbon. The protection by photorespiration of photosynthesis

against photoinhibition is well known. We emphasize that photorespiratory reactions contribute to the redox-homeostasis

and are part of the adaptive responses of plants to abiotic stress. Photorespiratory reactions can dissipate excess

reducing equivalents and energy either directly (by utilizing ATP, NAD(P)H and reduced ferredoxin) or indirectly (by

sustaining photosynthetic cyclic electron flow, alternative oxidase (AOX) and providing an internal CO2 pool). Further,

photorespiratory H2O

2 could be a signal to modulate relevant gene expression. Our concept of photorespiration as an

important mechanism to meet the challenge of abiotic stress, is based on multiple evidences. Even mild disturbances

in redox status created by deficiency in ascorbate, alternative oxidase or chloroplastic NADP-malate dehydrogenase,

cause increases in photorespiratory components, such as catalase, P-protein of glycine decarboxylase complex

(GDC) and glycine. In a reversal of roles, any deficiency in photorespiratory capacity, results in oxidative stress.

Plants exposed to mild or severe oxidative stress initiate multiple mechanisms to dissipate excess energy and to

prevent ROS accumulation. Photorespiration as an important component of such flexible systems, to optimize

photosynthesis, and to protect against oxidative stress. Photorespiration integrates the function of chloroplasts,

peroxisomes, mitochondria and cytoplasm. Therefore, the strong interaction between the cellular redox status and

photorespiration is not surprising, but demonstrates an interesting network of metabolic systems, including

photorespiration, to prevent oxidative damage.

A.S. Raghavendra


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Genetic manipulation of Uroporphyrinogen III methyltransferase

(Upm1) modulates nitrogen and carbon assimilation in Arabidopsis

thaliana

Sampurna Garai and Baishnab C. Tripathy*



The biosynthesis of siroheme, an Fe-containing modified tetrapyrrole, is one of the common links between C and N

assimilation. Siroheme, the prosthetic group of nitrite reductase (NiR), is synthesized from uroporphyrinogen III, an

intermediate of biosynthetic pathway of chlorophyll and bacteriochlorophyll that are essential for photosynthetic carbon

assimilation of eukaryotic and prokaryotic primary producers. The first enzyme in siroheme biosynthesis pathway that

diverts uroporphyrinogen III from chlorophyll biosynthesis to siroheme is methyltransferase that catalyzes two

methylation reactions of the uroporphyrinogen III to form dihydrosirohydrochlorin that undergoes oxidation to form

sirohydrochlorin which is acted upon by sirohydrochlorin ferrochelatase to generate siroheme. The plastidic

uroporphyrinogen III methyl transferase coded by a nuclear AtUpm1 [At5g40850] was used to raise transgenic sense

and antisense plants to modulate siroheme biosynthesis. Overexpression of AtUpm1 resulted in higher NiR gene and

protein expression and enzymatic activities in sense plants than that of WT. It co-modulated nitrate reductase (NR)

gene and protein expression, and increased NR activity. Enhanced NiR and NR activities in overexpressors increased

their protein content. Higher protein and chlorophyll in sense plants contributed to increased photosynthetic electron

transport, carbon assimilation and plant biomass. AtUpm1 overexpression in sense plants protected them from nitrogen

deficiency. In contrast antisense plants had lower NiR, NR activities, protein content, carbon assimilation and biomass

than the WT and were prone to N starvation. Results demonstrate the co-regulation of two different but inter-dependent

biological processes of C and N assimilation by AtUpm1.

B.C. Tripathy


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IPPCongress2015



BBX32 controls a network of high light-induced transcription

regulators, photosynthesis and dynamic acclimation in mature leaves

Philip Mullineaux1*, Matthews Jack1, Chernukhin Igor1, Jenkins Dafyd2, Wild David2,Bechtold Ulrike1

, Lawson Tracy1, Penfold Christopher2, Ruben Alvarez-Fernandez1 andBeynon James2

1University of Essex, Colchester, UK2University of Warwick, UK


When low light (LL) - grown plants are exposed to high light (HL) for several days their mature leaves can substantially

increase their photosynthetic capacity. This is termed dynamic acclimation and is associated with increased plant

productivity. We hypothesised that nuclear-encoded genes coding for transcription regulators (TRs) that respond

rapidly and transiently to HL could control the establishment of dynamic acclimation. To find such regulatory genes, a

highly replicated temporally clustered transcriptomics dataset was generated from the leaf 7 of Arabidopsis plants

exposed for up to 6h HL. The analysis revealed 3844 genes that were responsive to HL. These genes included a

significant enrichment of those involved in protection against stress and others that were associated with longer term

acclimation. HL-responsive TR genes were selected for Variational Bayesian State Space Modelling (VBSSM), which

defined a 15 gene regulatory network (GRN) centred on B-BOX TRANSCRIPTION (CO) FACTOR32 (BBX32). BBX32

was found to be a negative regulator of photosynthesis in LL-grown plants and dynamic acclimation to daily repeated

exposure to HL. A major, but not exclusive contribution to BBX32 regulation of dynamic acclimation was through its

interaction with LONG HYPOCOTYL5 (HY5), a member of the TR GRN. Our studies indicate that BBX32 is at the

centre of a complex temporal regulation of HL-responsive cellular processes underlying its TR GRN.

Philip Mullineaux


70

IPPCongress2015



Engineering a C4 rice: An update from the international C4 rice

consortium

Anindya Bandyopadhyay*

C4 Rice Center, International Rice Research Institute, Los Baños, Laguna, Philippines


The C4 rice project is a large collaborative program involving sixteen separate organizations world-wide (The C4

Consortium), led by IRRI. The aim of the International C4 rice consortium is to produce a large and sustainable

increase in the productivity of rice in all ecosystems. To achieve this, the engineering of a C4 photosynthetic pathway

into rice (a C3 crop) is underway. C4 cereals typically have grain yields 50% greater than those of C3 crops like rice

and wheat. This is because C4 plants have a more efficient photosynthetic pathway that leads to higher photosynthetic

rates and improved nitrogen and water use efficiencies. The carbon concentrating mechanism of C4 allows CO2 to be

fixed at much lower leaf internal CO2 concentrations. This allows stomatal conductance to be reduced and this lowers

the rate of transpiration and doubles water use efficiency. Most of the C4 biochemical pathway genes, along with

relevant transporters, have been transgenically introduced into rice so far. These genes are now pyramided and

brought into a similar background as a breeding stack. Initial analyses of pyramided lines show signs of a C4 biochemical

pathway. Gene discovery for Kranz anatomy and plastid development is also ongoing with interesting transgenic

gene candidates under molecular verification. Progress has also been made in the areas such as development of

gene stacking technologies for targeted stacking of C4 genes and elucidating the underlying mechanism of stomatal

conductance and water use efficiency.

Anindya Bandyopadhyay


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IPPCongress2015



The dynamic role of cell wall polysaccharides in sustaining sugar

homeostasis during leaf senescence of Arabidopsis thaliana: Induction

of cell wall hydrolases and cellular sugar reprogramming

Basanti Biswal*

CSIR, School of Life Sciences, Sambalpur University, Jyotivihar-768019, Odisha, India


The polysaccharides bound to cell wall are the richest source of organic carbon in plant body and play a critical role

in cellular sugar metabolism and homeostasis during plant development and senescence. We have demonstrated

that senescence induced decline in photosynthesis in Arabidopsis leaves causes loss of sugars, which acts as a

signal for induction of many senescence associated genes (SAGs) that activate several catabolic events to provide

sugars from other sources, primarily cellular macromolecules, to complete energy dependent senescence program.

Expression of SAGs follows a temporal pattern, some expressed early and some expressed late. These genes initiate

and execute an organized and time dependent senescence program. One of the late expressive SAGs codes for bita-

glucosidase responsible for breakdown of cell wall polysaccharides. In addition, other wall bound hydrolases including

bita-glucanase and bita-galactosidase are activated late during senescence, which suggests that breakdown of cell

wall polysaccharides is the terminal event of catabolic network that provides sugars to energy starving cells for

completion of senescence program. That the loss of photosynthetic production of sugar as a signal for induction of

cell wall bound hydrolases is confirmed with the similar response of the hydrolases during loss of photosynthesis in

green leaves of Arabidopsis experiencing abiotic stress. Finally, a model is proposed to explain the nature of signaling

involved in cellular sugar reprogramming during plant senescence.

Basanti Biswal


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IPPCongress2015



Photosynthetic efficiencies among Indian peanut cultivars evaluated

A.L. Singh1*, R.N. Nakar2, K. Chakraborty1, Nisha Goswami1 and K.A. Kalariya3

1ICAR-Directorate of Groundnut Research, PB 5, Junagadh-362001, Gujarat, India2Presently at Department of Botany, Seth PT College, Godhra-389001, Gujarat, India3Presently at ICAR-Directorate of Medicinal and Aromatic Plants Research, Boriavi, Anand-387310,Gujarat, India


Peanut is an important legume and oilseed crop of tropical world mainly grown as rainfed in semi-arid regions, and India

has the largest area and released cultivars. To know how physiologically efficient our peanut cultivars are?, the

photosynthesis (PN), transpiration (E), stomatal conductance (g

s), chlorophyll fluorescence, SCMR and yields were

studied in field grown 186 cultivars during rainy season where large variability was observed. Addition or deduction of

standard deviation from the average value of these parameters categorized 27 cultivars as high PN with >23.9 μ mol m-2s-

1 and 30 cultivars as low PN with <15.7 μ mol m-2s-1, 35

cultivars as high E with >8.9 and 24 as low E with <4.7 m mol m-2s-

1, 35 cultivars as high gs with >0.33 m sec-1 and 28 as low g

s with <0.17, 32 cultivars as high WUE >3.81 and 36 as low

WUE <2.31. These parameters accounted for the yield potential and 33 cultivars with >225 g m-2 pod were high yielder,

26 as low yielder <133 g m-2 pod and 127 cultivars medium with 133-225 g m-2 pod. Positive correlation between, PN and

gs, P

N and E, P

N and number of pods, SCMR and pod yield, SCMR and WUE, haulm yield and plant height, haulm yield

and pods, plant height and pods, and a negative correlations between SCMR and plant height, SCMR and E, and E with

WUE were found, however there was no any correlation of PN with yield.

A.L. Singh


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Concurrent Session I

Male germline development and function in flowering plants

Mohan B. Singh*, Etsuko Itabashi and Prem L. Bhalla

Plant Molecular Biology and Biotechnology Laboratory, Faculty of Veterinary and AgriculturalSciences, The University of Melbourne, Victoria 3010, Australia


The life cycle of flowering plants alternates between the diploid sporophyte and haploid gametophyte phases, with

the latter producing gametes required for sexual reproduction. Male gametophyte development encompasses meiotic

division of microspore mother cells giving rise to four haploid microspores. The asymmetric division of the haploid

microspore into two daughter cells that produces a small generative cell from the larger vegetative cell initiates the

male germline. The generative cell eventually undergoes mitotic division to form two sperm cells required to fuse with

two female gametes (egg and central cell) initiating development of the embryo and endosperm, respectively. The

molecular basis for the diversification of the cell fates of generative and vegetative cells is a central issue in plant

reproductive biology. This diversification is underpinned by the vastly distinct genetic and epigenetic programming of

these two pollen cells. The vegetative cell gene expression program emphasizes cell-cell signalling and cell wall

organization that might be required for cell-cell communication and pollen tube elongation from pollination to fertilization,

whereas the GC transcriptional program is skewed towards DNA/RNA metabolism, histone modification, and cell

cycle progression, indicating that gene expression regulation, changes in the epigenetic state and mitotic cell division

are associated with the generative cell function. This talk will highlight molecular regulatory pathways controlling male

germ line development and function in the flowering plants.

Mohan B. Singh


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Genomic assisted improvement of rice

A.K. Singh*, S. Gopala Krishnan, P.K. Bhowmick, R.K. Ellur, B. Haritha, M. Nagarajan andK.K. Vinod

Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India


Exemplary grain and cooking quality characteristics along with the pleasant aroma is the uniqueness of Basmati rice.

Genetic improvement of Basmati rice has led to the development of varieties with high yield and exceptional cooking

quality traits. These varieties have revolutionized the livelihood of the farmers and increased the annual forex earnings

to the tune of ~US $ 5 billion through export of Basmati rice. However, Basmati rice was highly susceptible to various

biotic stresses such as bacterial blight (BB), blast, sheath blight (ShB), brown plant hopper (BPH) and abiotic stresses

such as salinity. Molecular breeding has been successfully utilized in development of several MAS derived varieties

such as Improved Pusa Basmati 1 and Pusa 1592 for resistance to BB; Pusa 6 (Pusa1612) and Pusa Basmati 1609

for blast resistance. Further, several monogenic and pyramided NILs with resistance to BB (xa13, Xa21, Xa33,

Xa38), blast (Pi54, Piz5, Pi9, Pi1, Pita, Pib, Pi5), sheath blight (qSBR11-1), BPH (Bph18, Bph20 and Bph21), Bakanae

(qBK1), salinity stress (Saltol) and herbicide tolerance in the genetic background of popular Basmati rice varieties

such as Pusa Basmati 1, Pusa Basmati 1121, Pusa Basmati 6 and Pusa Basmati 1509 are being developed. QTLs for

drought tolerance such as qDTY1.1, qDTY2.1, qDTY2.2 and qDTY3.1 are being incorporated into the genetic

background of Pusa Basmati 1 and Pusa 44. Furthermore, pro-vitamin A trait has been incorporated into the mega

rice variety Swarna.Genomic tools have been of great importance to discover novel QTLs for yield and yield related

traits, grain and cooking quality traits, development of heterotic pools as well asassessing genetic purity.

A.K. Singh


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A novel mutagenesis design to revolutionize genetics, genomics and

breeding in maize and beyond

Gurmukh S. Johal1*, Brian Dilkes2, Rajdeep S. Khangura1 and Norman Best3

1Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana, USA2Department of Biochemistry, Purdue University, West Lafayette, Indiana, USA3Department of Horticulture, and Landscape Architecture, Purdue University, West Lafayette,Indiana, USA


The success of any genetic or breeding program depends on having adequate diversity in the germplasm. This

genetic diversity can be of natural origin or generated deliberately by mutagenesis. Various reagents exist to intentionally

induce mutations in plants, but alkylating agents such as ethyl methanesulfonate (EMS) are among the most frequently

used. Unfortunately, many mutagenesis methods, pollen mutagenesis of maize in particular, are highly inefficient due

to a relatively low number of transmitted mutations. As a result, many promising uses of mutagenesis are not feasible

or cost effective. We have developed a technique for stacking mutations to increase the mutation density per individual.

This increases tremendously the efficiency of independent mutant alleles, thereby opening up multiple applications

impossible until now. The process utilizes a multigenerational design to maximize independent changes in the genome

and allows screening of subtle differences in mutant phenotypes. In addition, it results in a vast immortalized mutant

seed library that can be utilized again and again for multiple independent purposes and benefits. A key benefit is that

it maximizes the frequency of polymorphisms of interest for enhancing elite breeding material.

Gurmukh S. Johal


76

IPPCongress2015



Ectopic expression of Arabidopsis thaliana HB17 gene in corn leads

to improved sink potential

Vijay K. Sharma*, Manju Manjunath, Tom Slewinski and Elena Rice

Monsanto Company, 700 Chesterfield Parkway West, Chesterfield, MO 63017 USA


As a result of the large scale screening of candidate genes in transgenic corn, we identified an Arabidopsis thaliana

gene HB17, a member of homeodomain-leucine zipper II (HD-Zip II) family of the plant transcriptional factors, which

affects plant growth and leads to increase in ear size at silking. When expressed in corn, AtHB17 lacks the repression

domain due to the corn specific splicing mechanism and loses the ability to bind the co-repressors and affect transcription

of the target genes. The protein still can form homo-dimers as well as heterodimers with corn endogenous HD-Zip II

proteins and bind to the target DNA sequences due to the presence of the functional leucine-zipper and DNA-binding

domains. We propose that AtHB17 expressed in corn mediates physiological effects through dominant-negative

mechanism by attenuating transcriptional repression activity of endogenous corn HD-Zip II proteins. We hypothesize

that modulation of the activity of HD-ZIP II proteins leads to modulation of corn plant’s growth responses to environmental

and developmental signals and, ultimately, to increased ear size, thus, providing opportunity for enhanced sink potential

in corn plants. Increased sink potential could be manifested through an increase in kernel weight or kernel number

depending on the environmental conditions.

Vijay K. Sharma


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IPPCongress2015



Developing herbicide tolerant rice plants for effective weed

management

Aakrati Agarwal, Dhirendra Fartyal, Mrinalini Manna, Mohan Murali Achary, Vijay Sheri, BabuRam Verma, Tanushri Kaul and M.K. Reddy*

International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India


Weeds are the most serious biological constraint in agriculture and manual weeding over large areas is not feasible.

In this scenario, introduction of genetically engineered herbicide resistant crop plants will be a better weed management

strategy. We developed transgenic rice plants simultaneously resistant to two broad-spectrum systemic herbicides

i.e., sulfonylurea for pre-emergent and glyphosate for post-emergent application. Sulfonylurea is a very strong inhibitor

of acetolactate synthase and glyphosate is a strong inhibitor of EPSP synthase the important plant enzymes involved

in amino acid synthesis. We followed herbicide target site modification strategy by introducing selective amino acid

substitutions in rice EPSP synthase and acetolactate synthase. These multisite-compensating mutations drastically

decreased affinity for herbicide binding without detectable fitness cost. These mutant copies of rice EPSP synthase

and acetolactate synthase encoding genes were cloned under constitutive promoters and linked together on a single

plant transformation vector for simultaneously transformation into rice. These transgenic rice lines showed significant

tolerance to glyphosate and sulfonylurea. Systematic rotation of glyphosate and sulfonylurea application in the rice

fields is expected to prevent the development of herbicide tolerant weeds. We have also developed another strategy

to control the weeds by application of phosphite. Plants cannot metabolize phosphite, as they can only utilize phosphate

for their growth. We have developed transgenic rice plants overexpressing phosphite dehydrogenase that converts

phosphite into phosphate. Phosphite can act as a fertilizer for the transgenic plants whereas, for weeds it acts like a

herbicide and act as herbicide for the weeds.

M.K. Reddy


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Identification and functional characterization of the regulators of

reproductive development in rice

Kapoor Sanjay*, Chawla Mrinalini and Nayar Saraswati

Department of Plant Molecular Biology, Univ. of Delhi South Campus, New Delhi-110021, India


Reproductive development in plants is a dynamic process that involves complex interplay of regulatory networks that are

under the influence of genetic as well as extrinsic cues. Our group has been involved in the identification and

characterization of both genic and epigenetic components that affect development of the pollen and the seed. Our

studies on the rice model have been instrumental in the identification of sets of genes that could potentially play key roles

during sporophytic to gametophytic transition in pollen mother cells, pollen maturation, embryo differentiation and grain

filling. Male gametophyte development has been shown to be a heat-sensitive process in a variety of plant species and

is one of the major determinants of yield in many crops. We have identified components of RNAi and ubiquitin-mediated

protein degradation machinery that mimic pollen heat phenotypes, with reduced starch accumulation and loss of viability.

We have also identified and carried out functional delineation a seed-specific MADS box transcription factor, OsMADS29,

that regulates embryo development and grain filling by affecting hormone homeostasis during seed development in rice.

Rice MADS29 has also been found to interact with several seed-expressed transcription factors and its capability to

localize in the nucleus is affected by these interactions. These and other leads could thus contribute genetic targets and

tools for augmenting yield as well as nutritive value of the seeds.

Sanjay Kapoor


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IPPCongress2015



Towards integrated product development

Amit Das1, Sresty Tavva1, Ratna Kumria1, Priyanka Bhyri1, Bindu Andreuzza1, Hajime Sakai2,Thomas W Greene3 and Amitabh Mohanty1*

1DuPont Pioneer, DuPont Knowledge Centre, Turkapally, Hyderabad-500078, Telangana, India2E.I. duPont Nemours and Company, DuPont Experimental Station, Wilmington, Delaware 19898,USA3DuPont Pioneer, 7000 NW 62nd Ave, Johnson, IA, USA


The global population is estimated to reach 9 billion by the year 2050. Most of this population growth is occurring in

the region of low agricultural productivity. With the ever increasing population, there is a need to produce more food

to feed the growing population. Since agricultural productivity has not kept in pace with the population increase and

therefore, there is a strong need to use cutting edge technology for improving agricultural productivity, especially for

important traits like nitrogen use efficiency (NUE) and drought tolerance (DRT). We have employed forward genetics,

reverse genetics, as well as new breeding technologies, such as CRISPR/Cas9-based guided genome editing to

address some of the key issues. Recent advances in development of tools for precision genome engineering, such as

site directed nucleases (SDN) have opened up a plethora of opportunities for plant breeding and genetics. These new

breeding technologies (NBTs) allow us to perform targeted and precise modification of the genome, thereby imparting

ability to design and develop a new generation of crops with improved traits. Application of these new breeding

technologies has potential to result in significant gains in the area of crop improvement. In this presentation we will

present a holistic view of our integrated approach towards product development.

Amitabh Mohanty


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Concurrent Session J

Unraveling key regulators of floral transition at the shoot apical

meristem of soybean

Prem L. Bhalla*, Chui E. Wong, Chol-Hee Jung, Lim Chee Liew and Mohan B. Singh

Plant Molecular Biology and Biotechnology Laboratory, Faculty of Veterinary and AgricultureSciences, The University of Melbourne, Parkville, Victoria 3010, Australia


Flowering process controls crop productivity. Understanding of the floral transition is essential in ensuring future food

security. Soybean [Glycine max (L.) Merr.], one of the world’s most important crops, is responsible for 55% of the

worldwide oilseed production and a significant source of dietary proteins. Further, soybean is significant in the world

agriculture due to its capacity to fix atmospheric nitrogen. Soybean is a photoperiod sensitive crop whose floral

transition is triggered by exposure to short-day conditions. Our knowledge of the molecular control of flower initiation

in this economically important legume species is limited. We used an integrated experimental and bioinformatic

approach to addresses this gap in our knowledge. Our comparative genomics study revealed complete repertoire of

flowering regulatory genes in the soybean genome. Further, RNA-Seq analysis of shoot apical meristem and leaf

undergoing floral transition revealed major reprogramming events in leaf and the shoot apical meristem that point

toward hormones gibberellins and cytokinin as key regulators in systemic flowering signal(s) production in leaves.

Our data also revealed an extensive reprogramming of genes associated with the epigenetic chromatin modifications

and RNAi gene silencing in the shoot apical meristem during floral transition. Our results highlight the use of a

genomics-based approach in improving our understanding of this important developmental process and provided a

framework to investigate further the molecular network underlying the transition to flowering in soybean.

Prem L. Bhalla


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Vernalization and epigenetic regulation in Arabidopsis

Christopher Helliwell1*, Masumi Robertson1, Jean Finnegan1 and Bob Anderssen2

1CSIRO Agriculture, GPO Box 1600, Canberra, ACT 2601, Australia2Data61, GPO Box 664, Canberra, ACT 2601, Australia


Many plants, including Arabidopsis, respond to extended periods of cold (vernalisation) by initiating the transition from

vegetative to reproductive development. We are interested in this process both because of its biological importance and

as a model for gene regulation by chromatin modification. In Arabidopsis the vernalisation response is conferred by

expression of the FLC gene, a repressor of the flowering pathway. FLC transcription is down regulated in response to

cold and this down-regulation is maintained once plants returned to warm conditions. The maintenance of FLC repression

requires deposition of the chromatin modification H3K27me3 by the polycomb system. I will discuss our understanding

of how FLC is repressed by cold, how this repression is maintained and the wider implications of this for understanding

the role of chromatin modifications in plant development.

Christopher Helliwell


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Physiological and biochemical basis of seed longevity during storage

M.B. Chetti1* and M.K. Meena2

1Assistant Director General (HRD), Education Division, KAB-II, ICAR, New Delhi-110012, India2Assistant Professor, Department of Crop Physiology, UAS, Raichur-584104, Karnataka, India


Great strides in seed packaging and storage have been made under the quality maintenance strategy by using

vapour proof packaging materials which in turn made possible the global distribution of high value and long term

availability of seeds. The factors affecting the storage life of seeds have been identified and characterized for successful

seed storage and accordingly packaging materials are modified considering the storage period. Results of our study

revealed that there was a gradual decline in seed quality parameters during entire storage period of 18 months in

cotton, groundnut, soybean and onion. Not much variation was observed with respect to the vacuum packed bags

from zero to 18 months of storage, irrespective of seeds stored either under cold storage or at room temperature. The

changes in seed quality (germination per cent, mean daily germination, root length, shoot length, total seedling

length, seedling dry weight, mobilization efficiency and vigour index) were attributed to the changes in moisture

content. Comparing the HDPE and gunny treatments under room temperature and cold storage, lower values were

observed in the latter, which is attributed to the high moisture in seeds as the cold storage maintains high humidity.

The electrical conductivity was significantly lower in seeds stored in vacuum sealed containers followed by polythene

bags and was higher in gunny bags and cloth bags throughout the storage period. Higher electrical conductivity

values recorded in seeds stored in gunny bags and cloth may be due to higher level of seed deterioration on account

of age induced membrane damage of various cell and cell organelles or degradation or disruption of cell membranes

leading to subsequent loss of membrane integrity. Our investigations revealed that α-amylase enzyme decreased

with an increased storage period. High density polythene, gunny and cloth bags stored seeds recorded higher

α-amylase activity compared to vacuum packed seeds followed by cloth bags. Decreased α-amylase activity with an

increase in storage period may be due to reduction of free sugars and amino acids. Lipase and protease activity were

found to be slightly less in vacuum packed bags compared to aluminum, HDPE, gunny and cloth bags. There was

higher lipase activity in gunny bags followed by cloth bags. There was increase in the activity of these enzymes with

advancement in storage period. The investigation has thrown light on several aspects that need to be worked out in

detail in future.

M.B. Chetti


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IPPCongress2015



How and why of flower senescence – Understanding from models to

ornamentals

Inayatullah Tahir* and Syed Sabhi Ahmad

Plant Physiology and Biochemistry Research Laboratory, Department of Botany, University ofKashmir, Srinagar-190006, India


Flower senescence represents the last stage of flower development ultimately resulting in the death of the floral parts.

It is regulated by an array of external and internal cues coordinated at the gene level which bear the hallmark of

programmed cell death (PCD). Petals provide an excellent model system for deciphering senescence as they have a

finite lifespan regulated by tight developmental control. While flowers serve an essential role in sexual reproduction,

the maintenance of petals is costly in terms of energy and water loss. PCD in flowers is generally triggered, once

pollination takes place and the plant doesn’t require the floral organs. This results in the breakdown of macromolecules/

organelles and remobilization of their products to developing tissues. Flower senescence is chiefly regulated by a

complex interplay between various growth regulators like ethylene, abscissic acid, auxins, gibberellins, cytokinins,

jasmonic acid, brassinosteroids and polyamines. Ethylene plays main role in some flowers while as in others, it has a

little or no role in the execution of flower senescence. Proteolytic cleavage is thought to play a pivotal role in the

regulation of senescence in some flowers, being one of the earliest signs of gene change resulting in raised enzyme

activity and decline in the protein levels. These changes cause membrane fluidity, increased lipid peroxidation and

decreased activity of antioxidant enzymes, ultimately leading to flower death. Our study tries to gain an insight into

the phenomenon of flower senescence by looking from models to ornamentals.

Inayatullah Tahir


84

IPPCongress2015



Metabolic dominance of spikelets in rice panicle is determined by

genetic expression of starch synthesizing enzymes

Binay Panda and Pravat Kumar Mohapatra*

School of Life Sciences, Sambalpur University, Sambalpur-768019, Odisha, India


Spikelets are the individual flower units of the complex racemose type panicle of rice. Each spikelet produces one

seeded fruit caryopsis at maturity; the success of occurrence depends on completion of starch synthesis in the

endosperm. The spikelets located on the apical part of panicle dominate metabolically and produce well-filled grains

in contrast to the inferior basal spikelets producing poorly-filled grains unsuitable for human consumption. Starch

comprises 80% of the final dry weight of rice grain. Poor sink activity in the inferior spikelets underscores genetic

potential for starch synthesis to the detriment of grain filling which downgrades grain weight and quality. In the recent

years, rice breeders have increased spikelet number of panicle without increment of size resulting in a change over of

spikelet arrangement from lax to compact. Compact arrangement compromised with apical dominance and shortage

of space between spikelets discouraged starch filling of the extra-spikelets accommodated. The discrimination in

starch filling capacity of the inferior spikelets have been measured by estimation of activity of three major starch

synthesizing enzymes, such as, sucrose synthase, AGPase and starch synthase and their expression as seen in RT-

PCR and qRT-PCR studies. Genetic expression of various isoforms of the enzymes was higher in the lax- compared

to compact-panicle rice cultivars and apical verses the basal spikelet. The metabolic dominance of a rice spikelet

depends on the expression of the genes for different isoforms of starch synthesizing enzymes, but the expression of

isoform three of sucrose synthase could be more specific than the others.

P.K. Mohapatra


85

IPPCongress2015



Understanding environmental and physiological basis for low

productivity of Alphonso mango under coastal rainfed ecosystem of

Konkan region in India

Murad M. Burondkar*, Sanjay Bhave, Mahesh Kulkarni and Arun Mane

Dr. B.S. Konkan Krishi Vidyapeeth, Dapoli, Ratnagiri, Maharashtra, India


Konkan region on the west coast of India, is one of the biggest (0.18 million ha) mango (cv. Alphonso), growing belts

in India. However, inspite of several technological interventions, very low mango orchard efficiency (2.5-3.0 t/ha), is

considered as major long standing constraint. Hence, present study was aimed at establishing the association between

the climatic factors prevailing during 3 different seasons (monsoon, winter and summer) and physiological attributes

such xylem sap flow, leaf water potential and rate of photosynthesis, recorded fortnightly during on 30 years old

Alphonso mango trees, grown on red lateritic soils. Results indicated very close association between weather parameters

and physiological response of mango during all the three seasons. Rate of photosynthesis was maximum (8.91 μmol

CO2 m-2s-1) only during October to November and was positively favoured by available bright sun light above light

saturation point (approx. 500 μmol m-2s-1), maximum xylem sap flow and availability of leaf moisture content for the

corresponding period. The rate of photosynthesis during monsoon (June-September) was bare minimum, ranging

from 2.0-2.5 μmol CO2 m-2s-1, due to very poor sunlight (35-40 μmol CO

2 m-2s-1 i.e. almost ten times below light

saturation point. After November, the rate of photosynthesis gradually decreased towards summer on account of

lower xylem sap flow and leaf water potential (-1.9 bar) due to soil moisture stress and low humidity.

Murad M. Burondkar


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Plenary Session-IV

Investigations on cold acclimation induced changes in photosynthesis

in Arabidopsis: Chlorophyll a fluorescence emission measurements

Kumud B. Mishra1 and Govindjee2*

1Global Change Research Centre, Academy of Sciences of the Czech Republic, v. v. i, Belidla 986/4a, 603 00, Brno, Czech Republic2Department of Plant Biology, Department of Biochemistry and Center for Biophysics & QuantitativeBiology, University of Illinois at Urbana-Champaign, Urbana, IL USA


In spite of extensive research on diverse plant species displaying a broad range of cold tolerance, the strategy and

dynamic processes involving the onset of cold acclimation induced cold tolerance and regulatory steps controlling re-

adjustment of physiological homeostasis at low temperatures are inadequately studied and remain an area for future

research. Further, most of the studies on cold tolerance have been carried out on plants grown in static environment,

e.g., inside a growth chamber. In nature, plants are adapted to dynamic environment with varying light and temperature

throughout their growth cycle, and this dynamic environment must have an important role on the development of

tolerance of plants to these environmental stimuli including low temperatures. Since chlorophyll (Chl) a fluorescence

emission has already been established as a powerful, and non-invasive tool for measuring various steps in

photosynthesis, we have used it in this research to study cold acclimation induced changes in the model plant

Arabidopsis (A.) thaliana. We shall present and discuss our results on Chl a emission and relate them to the modulation

of photosynthesis in leaves from non-acclimated and cold acclimated natural accession of A. thaliana C24 (native of

Pourtgal), during linear cooling from room temperature (220C) to -1.5°C.

Govindjee


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IPPCongress2015


Plenary Session-IV

Ionome to genome: Gene discovery in aid of plant nutrition, human

health and environmental remediation

Mary Lou Guerinot*

Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA


Micronutrient deficiencies commonly limit plant growth and crop yields. If the mechanisms of uptake, distribution and

regulation of micronutrients were clearly understood, it might be feasible to engineer plants better able to grow in soils

now considered marginal and to increase crop biomass on soils now in cultivation. Furthermore, as most people rely

on plants as their dietary source of micronutrients, planlits that serve as better sources of essential nutrients would

improve human health. We combine genetics, high throughput elemental analysis via ICP-MS and high resolution

imaging via synchrotron X-ray fluorescence to identify and characterize genes involved in metal uptake, distribution

and storage. Most of our work has been focused on the essential micronutrient iron. We are characterizing several

Arabidopsis mutants that have the potential to improve seed iron content. One of these, Ig14, has a similar metal

content to wild type when grown on normal soil, but thrives on alkaline soil, accumulating significantly more iron in its

shoot and seeds. Another mutant, 93699, accumulates significantly more iron than wild type and is very sensitive to

exogenous iron supply. The mutation in a third mutant has been mapped to an essential bHLH gene we call URI that

controls many of the iron-regulated genes in Arabidopsis. We are also taking similar approaches to determine how

arsenic, a non-threshold, Class 1 human carcinogen, accumulates in rice, a staple food for over half the world’s

population. We have been screening a large collection of rice cultivars and have identified several QTLs for arsenic.

Mary Lou Guerinot


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Plenary Session-IV

Capitalizing on genetic variation for functional genomics of

quantitative traits in rice

T. Mohapatra*

ICAR-Indian Agricultural Research Institute, New Delhi-110012, India


Genetic variaion is essential for evolution. It is the key to the development of improved cultivars through selection of the

best. Besides, genetic analysis of any trait requires variation. Variation in natural populations of cultivated and even wild

forms are first utilized for the purpose. Induced genetic variation often supplements the natural genetic variation while

dissecting target traits. Majority of the traits of agricultural importance are quantitative in nature. Understanding variation

in such traits requires different approaches in comparison to Mendelain traits. In recent years, a combination of genetic,

genomic and statistical tools/approches is being applied to make sense of the variation in complex traits and to infer on

how the genome functions. Rice is the staple food for more than half of the world’s population, especially those living in

Asia. However, the world population is increasing at a fast pace, which might reach nine billion mark by 2050. The

population of India is also estimated to be around 1.6 billion by that time. To feed this huge population, it will be required

to increase production and productivity of rice. This is going to be highly challenging in view of the changing climate that

would accentuate the problems due to abiotic and abiotic stresses. As the economic condition improves along with rising

purchasing power, there will be greater demand for better quality of the produce. Fortunately, rich genetic diversity exists

in rice. More than 100,000 germplasm lines including wild relatives have been collected and conserved in the National

Gene Bank of the NBPGR, New Delhi. Subsets of this germplasm have been evaluated for a range of traits including,

yield, tolerance to abiotic and biotic stresses and nutritional quality at different institutions in the country. Besides, more

than 20,000 EMS mutagenised lines of the upland cultivar Nagina22 have also been developed through national

network funded by the Department of Biotechnology, Govt. of India. Laboratory and field evaluation has led to identification

of mutants with novel phenotypes. We have made use of these genetic resources to understand rice genome function.

The traits that we have targeted include yield components, drought tolerance, salinity tolerance, cold tolerance, anaerobic

germination and grain iron content. Whole genome re-sequencing of contrasting genotypes has revealed huge DNA

sequence level variation with genome scale distribution. Transcriptome sequencing has led to identification of genes

showing differential expression in contrasting lines suggesting thereby involvement of different pathways. Inclusion of

EMS mutants in the study has helped identification of newer alleles of known genes. Integration of genome-wide approaches

has provided opportunity to understand variation in complex traits such as tolerance to drought and salinity. The results

of the above studies will be discussed.

T. Mohapatra


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Concurrent Session K

A day and a night in the life of a unicellular diazotrophic cyanobacterium

Himadri B. Pakrasi*

Laboratory of Himadri Pakrasi, Washington University, International Center for Advanced RenewableEnergy and Sustainability, St. Louis, MO 63130, USA


Cyanobacteria are unique among prokaryotes for their ability to perform oxygenic photosynthesis and for their circadian

lifestyle. Unicellular cyanobacteria like Cyanothece are fascinating organisms that actively use both phases of the

diurnal cycle to separate and carry out antagonistic metabolic processes. The intracellular environment oscillates

between aerobic and anaerobic states during the day-night period, allowing oxygen-sensitive processes like nitrogen

fixation to occur at night while photosynthetic oxygen evolution takes place during the day. This diurnal periodicity

gives a highly dynamic profile to these organisms, which can be captured at the genome, transcriptome, proteome

and ultrastructural levels. Whole genome analysis of six Cyanothece strains revealed that this group of unicellular

nitrogen fixing cyanobacteria specialize in metabolic compartmentalization and energy storage, concomitantly

accumulating metabolic products in inclusion bodies that are later mobilized as part of a robust diurnal cycle.

Transcriptomic analyses showed that the existence of these incompatible processes in the same single cell depend

on tightly synchronized expression programs involving ~30% of genes in the genome, a phenomenon that also

extends to a large extent at the protein level. To examine the proteome of Cyanothece, we used a high-throughput

accurate mass and time (AMT) tag approach and identified a total of 3,616 proteins with high confidence, which is

approximately 68% of the predicted proteins based on the sequenced genome. We have also examined the dynamic

profiles of all of these proteins during a diurnal period. Three-dimensional tomographic reconstructions showed that

the thylakoid membranes in Cyanothece form a dense and complex network that extends throughout the entire cell.

In particular, the organization of these membranes and various intracellular bodies profoundly change between day

and night periods. Together, these studies have provided a comprehensive picture of how a physiologically relevant

diurnal light-dark cycle influences the metabolic behavior of a diazotrophic photosynthetic bacterium.

Himadri B. Pakrasi


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IPPCongress2015



Accelerating product pipeline through computational biology:

deciphering role of Arabidopsis HB17 in maize

Saritha V. Kuriakose1*, Naveen Silvester1, Anuradha Mahindra1, Kyle Skottke2,

Elizabeth Gohara2 and Abha Khandelwal2

1Monsanto Research Centre, Vasanth’s Business Park, Bellary Rd, Hebbal, Bangalore-560092,Karnataka, India2Monsanto Company, 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA


As a result of the large scale screening of candidate genes in transgenic corn, we identified an Arabidopsis thalianagene HB17, a member of homeodomain-leucine zipper II (HD-Zip II) family of the plant transcriptional factors, which

affects plant growth. Transformation of maize with an ATHB17 expression construct leads to the expression of

ATHB17Δ113, a truncated protein lacking the first 113 amino acids which encodes a significant portion of the repression

domain. Because ATHB17Δ113 lacks the repression domain, the protein cannot directly affect the transcription of its

target genes. ATHB17Δ113 can homodimerize, form heterodimers with maize endogenous HD-Zip II proteins, and

bind to target DNA sequences; thus, ATHB17Δ113 may interfere with HD-Zip II mediated transcriptional activity via a

dominant negative mechanism. We used computational approaches to provide first level evidence that maize HD-Zip

II proteins function as transcriptional repressors and that ATHB17Ä113 relieves this HD-Zip II mediated transcriptional

repression activity. This was later confirmed in targeted experiments. Expression of ATHB17Δ113 in maize leads to

increased ear size at silking and, therefore, may enhance sink potential. We hypothesize that this phenotype could be

a result of modulation of endogenous HD-Zip II pathways in maize.

Saritha V. Kuriakose


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IPPCongress2015



CCPM: a web platform for the data capture, analysis and integration of

plant metabolomics

Indira Ghosh1*, Abhijit Mitra2, Vikram Pudi2 and Nita Parekh2

1School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067,India2Center for Computational Natural Sciences and Bioinformatics, International Institute of InformationTechnology, Hyderabad-500032, India


Plant metabolomics represents a global quantitative assessment of metabolites within a cell sample in specific plant

tissue, assisted by advances in high-throughput instrumentation has emerged as a powerful source for the high

throughput phytochemical analysis of plants and rapid generation of large volumes of high quality data, signifying

different chemicals, in quantity & quality. A collaborative platform has been developed to addresses the unique

interdisciplinary character of metabolomics; an integrated and effective tool, for web-based data repository with analysis

& visualization platform for plant metabolomics known as CCPM 3.4 (Computational Core of Plant Metabolomics).

This is a fully functional, state of art web application, addressing the issues of storage, processing, analysis and

sharing of data and results of metabolomics experiments. With a python web framework (web2py) backend and open

source analysis programs embedded, CCPM allows rapid prototyping, easy maintenance and role-based access

control without compromising on security, scalability and efficiency through a user friendly bootstrap-based GUI. It

provides the pipeline for end-to-end analysis of LC/GC-MS data involving raw data capture, data pre-processing,

putative metabolite characterization (linking to METLIN and GOLM), and a wide range of statistical analysis, with

option for customization of parameters from the web interface, modular architecture allows input of externally

preprocessed data in all popular formats, directly to avail the analysis tools. It creates metadata on the details of

sample, experimental conditions and factors. Other features include user friendly tools for on the fly data analysis and

publication quality image download for experimental metabolomics of plant data.

Indira Ghosh


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IPPCongress2015



Wheat phenomics through hyper-spectral remote sensing for water

deficit stress

R.N. Sahoo1*, Bappa Das1, Gopal Krishna1, Rahul Tiwari2, Rakesh Kumar2 and

C. Viswanathan2

1Division of Agricultural Physics, 2Division of Plant Physiology, Indian Agricultural Research Institute,New Delhi-110012, India


Potential of hyper-spectral remote sensing has been explored for non-invasive assessment of water stress for wheat

crop. Hyper-spectral data (350 to 2500 nm) and relative water content (RWC) of 10 different genotypes of wheat crop

were measured at different stress level. Comparative evaluation of newly developed spectral index with existing

water sensitive spectral indices and band depth analysis at selected wavelengths was done with respect to leaf water

status, and spectral based prediction models were developed. Then, the proposed ratio of spectral indices (RSI) and

normalized difference spectral indices (NDSI) based on the leaf reflectance spectra were obtained in all possible

combinations within 350–2500 nm, and their correlation with RWC were quantified to identify the best indices. The

best spectral indices for estimating RWC in wheat were found to be, proposed RSI (R1391

, R1830

) and NDSI (R1391

, R1830

)

with R2 of 0.86 and 0.81, respectively. Hyper-spectral canopy reflectance (350–2500 nm) data recorded at various

stress levels were used to develop a partial least squares regression (PLS) model followed by multiple linear regression

(MLR) to calculate plant RWC. The model R2 and RMSE was found to be 0.96 and 0.88 and 3.88% and 7.16% for

calibration, and 0.91 to 0.88 and 6.52% to 7.54% for validation, respectively. Based on differential spectral response

of genotypes at different water deficit stress levels, genotypes were classified into stress tolerant and susceptible

types. The methodology developed would be further used in high throughput phenomics of different crops for drought

stress.

R.N. Sahoo


93

IPPCongress2015



Mechanistic investigation of floral transition in plants using

computational systems biology

Gitanjali Yadav*

Computational Biology Laboratory, National Institute of Plant Genome Research, Aruna Asaf AliMarg, New Delhi-110067, India


Floral transition represents one of the most critical events in the life of a plant, and refers to the progression from

vegetative to reproductive growth. It is known to be determined not by a single gene but by a highly complex gene

network, in which LEAFY (LFY), a transcription factor with protein binding function that expresses widely in both

vegetative and reproductive tissues, plays an important role as a switch that triggers flower formation by interacting

and coordinating between several other genes. We have investigated the floral transition event through a graph-

theoretical approach at a genome-wide network level in Arabidopsis thaliana by superimposing mRNA expression

profiles and gene ontologies of the known floral genes with the available floral interactome. The regulatory network

provides insights into how LFY, as a master regulator, positively or negatively synchronizes the level or activities of

other genes in the general physiology of flowering. A dynamic analysis of network topology led to the identification of

key regulatory modules of genes during flowering, apart from characterization of relationships among genes in different

floral modules. Since LFY stands at the very center of the network of flower development, even minute modifications

in its expression/interactions may contribute to the appearance of floral structures in evolution. In summary, this work

massively expands the known floral network of Arabidopsis thaliana besides offering new perceptions regarding the

principal mechanism of flowering and molecular basis of diverse floral morphologies.

Gitanjali Yadav


94

IPPCongress2015



Comparative analysis of glutaredoxins (GRXs) in monocot and dicot

species

Hemant Ritturaj Kushwaha1*, Minesh Jethva1 and Saurabh Yadav2

1Synthetic Biology and Biofuels Group, International Centre for Genetic Engineering andBiotechnology, New Delhi-110067, India2Department of Biotechnology, Hemwati Nandan Bahuguna Garhwal University, Srinagar Garhwal,Uttarakhand-246174, India


Environmental variations often lead to the production of reactive oxidative species (ROS), thus causing oxidative

stress in plants. One of the enzymatic antioxidant systems, Glutaredoxins (GRXs) has been characterized for its role

in redox homeostasis. GRXs are the small thermo-stable thiol based oxidoreductases of thioredoxin (TRX) superfamily.

GRXs are involved in the maintenance of cellular redox homeostasis in bacteria, yeast and plant and mammalian

cells. Glutaredoxin proteins perform the glutathionylation and de-glutathionylation reactions. Broadly they have been

classified into two types based on the redox active motif: Monothiols (CGFS) and dithiols (CXXC). CC-type GRXs are

specific to land plants and participates in the more complex functions in the plants. GRXs are widely conserved in

both prokaryotes and eukaryotes and are known to have many roles in addition to oxidative stress. In this study, we

have identified and characterized GRXs in nineteen monocot and dicot species. We have analyzed domain architecture

in their protein sequences, in order to establish their evolutionary relationship. Further, using phylogenetic tree, we

have analyzed the conserved features in monothiol and dithiol sequences in both monocots and dicots. The localization

of GRXs on various chromosomes of monocots and dicots highlights their probable expansion in the various plant

genomes. Thus, this analysis provides novel insights to the functions of GRX proteins in various monocots and dicot

species.

Hemant R. Kushwaha


95

IPPCongress2015


Concurrent Session L

Role of transpiration in regulating leaf temperature in wheat:

Physiological basis of contrasting genotypic responses

S.R.W.M.C.J.K. Ranawana1,2,5, H, Bramley4, J.A. Palta1,3 and K.H.M. Siddique2*

1School of Plant Biology, The University of Western Australia, Perth, WA 6009, Australia2The UWA Institute of Agriculture, The University of Western Australia, Perth, WA 6009, Australia3CSIRO Agriculture, Private Bag No. 5, Wembley, WA 6913, Australia4Plant Breeding Institute, Faculty of Agriculture and Environment, The University of Sydney, 12656Newell Highway, Narrabri NSW 2390, Australia5Department of Export Agriculture, Faculty of Animal Science and Export Agriculture, Uva WellassaUniversity, Badulla 90000, Sri Lanka


The projected increase in global temperature may drastically reduce wheat yields without adaptation strategies. The

role of transpiration in dissipating heat in wheat leaves through evaporative cooling was assessed in relation to

changes in vapour pressure deficit (VPD), air temperature and soil water under controlled environmental conditions.

A reductionist approach was taken starting with evaluating a morpho-physiologically diverse group of 20 wheat

genotypes under two temperature regimes for total water use, leaf temperature and biomass production. Selected

contrasting genotypes were then evaluated under controlled VPD conditions with varying levels of soil water to

identify the physiological basis of the contrasting responses. Greater water use and its effects on leaf cooling contributed

to greater biomass production under high temperature, which depended on genotype. Genotypes differed in their leaf

temperature regulation that did not always correlate with transpiration response to VPD, indicating other heat dissipation

mechanisms are involved in leaf cooling in addition to transpiration. Two contrasting transpiration responses to VPD

were identified: (1) Transpiration increased gradually at slower rate with VPD while stomatal conductance was

unresponsive, and (2) transpiration responded to VPD in two phases separated by a breakpoint (segmented linear

response) where transpiration responds little to VPD after the breakpoint. Genotypic variation in transpiration to VPD

response could be used to develop new wheat cultivars targeting specific environments. The first response pattern

may be better suited to rain-fed agricultural systems, where yield depends on stored water. The second pattern may

be better suited for agricultural systems with intermittent in-season rainfall.

K.H.M. Siddique


96

IPPCongress2015



Functional mechanisms and strategies for improving drought tolerance

in rainfed crops

M. Maheswari*

Division of Crop Sciences, Central Research Institute for Dryland Agriculture, Santoshnagar,Saidabad PO, Hyderabad-500059, India


Developing tolerant genotypes is crucial for stabilizing crop yields under drought stress conditions as it is one of the

most important abiotic stresses affecting crop productivity. Maize and pigeon pea genotypes were evaluated for

water deficit toler-ance across seasons under well watered and water stressed conditions to identify interactions

amongst various tolerance traits and grain yield as well as their association with SSR markers. Significant positive

correlations were elucidated amongst the water relations traits with yield as well as with a few SSR markers under

water stressed conditions indicating the importance of functional mechanisms of intrinsic tolerance and cumulative

traits for drought tolerance in rainfed maize. The importance of recovery patterns in identifying tolerant genotypes

was highlighted using high through put phenomics. In pigeon pea genotypes, water deficit impacted biomass

accumulation, partitioning while it enhanced the flower drop and decreased flower to pod conversion and molecular

genetic diversity based on SSR markers associated with drought tolerance revealed useful information for genetic

enhancement of drought tolerance. The cross talk of different abiotic stresses is mediated through anti-oxidative

metabolism and differential expression of AOX isozymes seemed to be involved in stress tolerance in rainfed crops

like pearl millet. Similarly differential expression especially of transcription factors and regulatory genes is a key

response in crop plants under climatic stresses. The differential expression analysis of SnRK2, a plant specific protein

kinase family involved in stress signalling indicated that this family of genes has a functional role in stress tolerance.

Integrated approaches utilizing the phenomics, functional genomics and molecular breeding are essential for enhancing

climate resilience in rainfed crops.

M. Maheswari


97

IPPCongress2015



Climate information, crop modeling and impacts on global food security

Vangimalla R. Reddy* and Shardendu K. Singh

Research Leader, USDA-ARS Crop Systems and Global Change Laboratory, Beltsville, MD, USA


Climate change is already affecting the natural resources such as terrestrial vegetation, animal husbandry, and

fisheries that societies depend on to provide food, fiber, fuel, several industrial products, and recreational services.

These carbon-based products and fresh water supply come from the thin living skin covering the earth’s land surface

called the terrestrial ecosystem. Within the next fifty years, the human population is projected to double, and economic

buying power for carbon-based products could triple. As there are no more unexplored frontiers, this increased

demand from terrestrial ecosystem will have to be met with the existing natural resource base and in the face of a

changing climate and extreme weather events. In addition, regional increases in soil erosion and atmospheric pollution

could also have negative impacts on crop productivity and the natural resource base of the planet. With existing

scientific knowledge it is impossible to accurately predict how these changes in the global climate may change the

productivity of various crops worldwide and overall productivity of the terrestrial ecosystem. One way to deal with the

complexity of the system, and its impact on crop productivity, is to develop and use mechanistic, process level

computer simulation models, both at the field level and at the ecosystem level. This presentation outlines some

examples of the development and use of the crop models for various applications to increase crop productivity and to

mitigate the harmful effects of adverse environmental variables on natural resources, both in the current and in the

future changing environment.

Vangimalla R. Reddy


98

IPPCongress2015



Elevated CO2 ameliorates drought and high temperature stress effects

in rice

Madan Pal*, Ashish K. Chaturvedi and Divya Shah



Apart from rising CO2, high temperature and drought stress are important climate change factors, which are predicted

to increase in frequency and magnitude under changing climate. These factors often interact with each other under

field conditions and their resulting impacts may be different compared to individual exposure. This work is focused on

physiological impact of combinations of these factors compared to independent exposure. The experiments were

conducted to analyse the role of elevated CO2 (eCO

2, 550±34 μmol mol–1) in combination with drought and high

temperature stress on growth, gaseous exchange, carbon metabolism, grain yield and quality attributes in Nerica L-

44 and Pusa 1121 rice genotypes, raised inside FACE and open top chambers. Rate of photosynthesis, non-structural

carbohydrates and ADPGase and starch synthase enzyme activity was stimulated under eCO2, decreased under

drought and high temperature stress and reduction was less under combined exposure of eCO2 with drought or high

temperature. Similar response was recorded for dry matter production and grain yield. Chalkiness in grains was more

under eCO2 individually and further enhanced under combined exposure of drought and high temperature. Grain

protein concentration decreased under eCO2 individually

but reduction was less under combined exposure of drought

and high temperature with eCO2. Amylose concentration in grains was less under combined exposure of drought and

high temperature with eCO2 compared to exposure to eCO

2 individually. These findings suggest that negative impacts

of drought and high temperature stress on grain yield and quality in rice can be lowered if combined with eCO2

exposure.

Madan Pal


99

IPPCongress2015



Rice adaptation pathways in response to heat stress at anthesis

Tanguy Lafarge*, C. Julia, N. Ahmadi and M. Dingkuhn

French Agricultural Research Centre for International Development, France


Coping with heat stress involves different options that account for the ability of plants to escape (early anthesis time),

avoid (panicle cooling through transpiration) or tolerate (presence of genes of interest) heat at flowering. First, variability

in the time of day of anthesis was correlated with the mean of climatic variables calculated for the 7-day period before

flowering, over four distinct seasons and four contrasted varieties. The best predictive variables (negative correlations)

were Tmin and VPD, with high values of both being associated with early times. Second, depending on conditions,

panicle temperature varied between 9°C below and 2°C above air temperature at 2 m. A significant positive correlation

was obtained between spikelet sterility rate and maximum panicle temperature at flowering, whereas no correlation

was obtained with air temperature. By extrapolation, this correlation predicted minimal sterility with a panicle temperature

of 3°C, and 50% sterility with a panicle temperature of 33-34°C. Third, a genome-wide association study of the

sterility rate of 167 traditional and modern varieties (grown for six consecutive days at 37°C between 8 am and 2 pm

at anthesis) detected 91 significant associations grouped into 12 independent regions located on eight chromosomes.

The highest heat tolerance was detected for N22, an aus variety from India, and Peh Kuh, a traditional indica variety

from Taiwan. Avoidance and tolerance appear as main genetic improvement pathways, whereas sensitivity of time of

day of anthesis to air temperature and humidity seems already effective within the rice crop.

Tanguy Lafarge


100

IPPCongress2015



Implications of climate change on plantation sector with special

reference to coconut

K.B. Hebbar*, Ravi Bhat and P. Chowdappa

Plant Physiology, Biochemistry & Post Harvest Technology, ICAR-Central Plantation CropsResearch Institute, Kasaragod-671124, Kerala, India


Plantation crops have a key role in the socio-economic development and transition from a subsistence agrarian

economy to market oriented commercial cultivation due to their trade significance. Cultivation of plantation crops in

India also has a rich diversity and varied history with each crop having its own distinct historical and economic context

of development. The major plantation crops in the country include coconut, arecanut, oil palm, cashew, tea, coffee,

rubber and several spice crops. Amongst these coconut is the predominant crop which provides sustenance to more

than 10 million people and contributes Rs. 83000 million annually to the Gross Domestic Product (GDP) of the

country. Most of these crops are grown either in hilly areas or coastal tracks where the threat of climate change is

more. The impending climate change will affect coconut plantation through higher temperatures, elevated CO2

concentration, precipitation changes, increased weeds, pests, and disease pressure. As per the latest IPCC report

rainfall becomes sporadic and temperature would rise. Coconut requires large volume of water (50 to 100 litre per

day) for the production of highly productive canopy and with water limitation the canopy reduces and yield may

reduce as high as 50% of irrigated gardens. In order to adopt the crop to climate change various soil, water and crop

management techniques have been evolved at ICAR-CPCRI. The cropping and farming system models developed at

the Institute involving grasses, vegetables, fruit trees, and animal component showed encouraging results in the

farmer’s fields.

K.B. Hebbar


101

IPPCongress2015



Staygreen QTL3B is associated with improved grain and stover yield

under postrainy sorghum growing conditions

H.S. Talwar1*, S.P. Deshpande2, Shiwesh Kumar1 and V. Vadez2

1Indian Institute of Millets Research, Rajindranagar, Greater Hyderabad-500030, Telangana, India2International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Patancheru, GreaterHyderabad-502324, Telangana, India


Postflowering drought is the major production constraint that adversely affects the crop growth and grain yield. Stay-

green has been described as the best characterized trait contributing to the adaptation of sorghum to terminal drought

conditions. The objective of this study was to assess the contributions of identified QTLs to grain and stover productivity

under control and water-stressed conditions. Using B 35 as the donor for staygreen QTLs, introgression lines (ILs)

were generated in two genetic background (S 35 and R 16). These lines along with local checks, recurrent parent and

staygreen donor were evaluated for two years during the postrainy season and at four locations in India. A total of 42

introgressed lines (ILS) targeting stay-green QTL Stg1, Stg3A, Stg3B and Stg4 in R 16 genetic background, and 56

ILs targeting Stg1, Stg2, Stg3A, Stg3B, Stg4 and StgC in S 35 genetic background were tested under irrigated and

terminal water-stressed (WS) treatments. Staygreen ILs had improvement in green leaf area retention at maturity

(GLAM) in both R 16 and S 35 genetic backgrounds under both WW and WS conditions, and more so under WS

condition. R16 was more responsive to the staygreen QTL introgression than S 35 in terms of GLAM, suggesting that

effect of individual staygreen QTL introgression interacted with genetic backgrounds. Stg3B was the key staygreen

QTL in enhancing the GLAM, particularly under WS conditions. Our results, established the superiority of stg3B in

enhancing the Gy and Tdm accumulation over other individual QTLs or in combination under both the genetic

backgrounds, particularly under WS conditions.

H.S. Talwar


102

IPPCongress2015


Concurrent Session M

Control of meiotic gene expression in plants

I. Siddiqi*, S. Andreuzza, A. Singh, J. Davda, D.K. Singh, S. Pandey, Ramesha, A.V.

Pardha Saradhi, A. Nalli, K.F. Max, S. Mahesh, S. Gharpure, V. Vijaybhaskar, V. Subbiah,

V. Vinya, K. Bannerjee, S. Surendra, K. Bethoju and Y. Kavya

Centre for Cellular and Molecular Biology (CCMB), Hyderabad, Andra Pradesh, India


Meiosis produces haploid cells essential for sexual reproduction. Meiosis is a critical event in sexual reproduction.

During meiosis, chromosomes recombine and segregate twice consecutively to produce haploid daughter cells,

which differentiate into gametes. In humans, errors in meiosis are the leading causes of congenital birth defects. In

plants, bypassing the meiotic program can lead to production of clonal seeds that retain hybrid traits that otherwise

segregate. Thus, understanding the controls of meiosis has major implications for both health and crop improvement.

In yeast, entry into meiosis activates transcription factors which trigger a transcriptional cascade that results in sequential

co-expression of early, middle and late meiotic genes. However, these factors are not conserved, and the factors and

regulatory mechanisms that ensure proper meiotic gene expression in multicellular eukaryotes are poorly understood.

We have identified and characterized higher plant genes that control meiotic gene expression at different levels and

through differential action in the two sexes. ACTIN RELATED PROTEIN6, a component of the SWR1 chromatin

remodeling complex controls meiotic gene expression in the female lineage through loading of the histone variant

H2A.Z. DUET/MMD1 controls expression of middle meiotic genes in the male lineage, and CDM1 an RNA binding

protein controls levels of meiotic RNAs in the male lineage. Our results point to an integration of sex-specific gene

expression programmes within a common meiotic programme leading to sexual dimorphism in plant meiosis and

gametogenesis.

Imran Siddiqi


103

IPPCongress2015



Novel phenotyping technique for reproductive-stage salinity tolerance

in rice

Mostafa Ahmadizadeh1,2, Naireen A. Vispo1, Cecilia Diana O. Calapit-Palao1,3, Iris Dawn

Pangaan1 and Rakesh K. Singh1*

1Plant Breeding, Genetics, and Biotechnology Division, International Rice Research Institute,Philippines2Department of Plant breeding, Sari Agriculture Science and Natural Resources University, Sari,Mazandaran, Iran3Institute of Biological Sciences, University of Philippines, Los Baños, Laguna, Philippines


Rice responds differentially at different stages of development. The seedling and reproductive stages are the most

sensitive growth stages with very weak association, suggesting that they are regulated by different processes and

sets of genes/QTLs. There are hardly any studies exist on reproductive-stage salinity tolerance mainly because of the

lack of reliable reproductive-stage-specific phenotyping techniques and incomplete knowledge of the stage-specific

mechanisms of salinity tolerance. Two major challenges for screening exclusively for the reproductive stage are, 1)

how to stress plants at the reproductive stage without stressing them at the seedling or late vegetative stage; and 2)

how to impose the stress on different genotypes or mapping populations at equivalent growth stages of development

because of the variability in the progression of growth rate through developmental stages. We standardise a

methodology that allows salt translocation to the reproductive organs such as the flag leaf (the largest source for the

sink) and panicle as quickly as possible just at the initiation of booting when the genotypes are at the same stage of

tissue/organ development rather depending on age of the plant. Extra leaves were clipped, leaving only a minimum

number of leaves needed by the rice plant that will not significantly affect grain yield and yield components. Salt stress

equivalent to EC 10 dSm-1 was imposed on rice plants with trimmed leaves starting from boot leaf emergence up to 20

days in an experiment. Stage-specific effect of salt stress was verified by observing salt-sensitive and salt-tolerant

genotypes. Details will be presented.

Rakesh K. Singh


104

IPPCongress2015



Integrated genomics-assisted breeding strategy for rapid quantitative

dissection of complex traits in crop plants

Swarup Kumar Parida1*, Alice Kujur1, Deepak Bajaj1, Shouvik Das1, Shailesh Tripathi2,

Hari D. Upadhyaya3 and Akhilesh K. Tyagi1

1National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India2Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India3International Crops Research Institute for the Semi-Arid Tropics, Patancheru-502324, Telangana,India


Development of genetically-tailored superior high-yielding stress tolerant (climate resilient) crop varieties is essential

to improve the productivity and sustainability of global agriculture. However, most of these stress tolerance and yield-

component traits targeted for crop improvement in current plant genomics and breeding research are complex and

quantitative in nature, and multiplicatively governed by many genes/QTLs. The intervention of multiple modern structural,

functional and comparative genomics approaches in classical/advanced breeding strategies i.e., “Genomics-assisted

Breeding (GAB)” offers unprecedented opportunities to identify functionally relevant novel trait-associated major/

minor genes (QTLs) and elite rare alleles for precise molecular dissection of complex traits. This approach is also

effective in studying the natural allelic interactions of favorable loci influencing trait variation in diverse crop genotypes

that have adapted to varied agro-climatic conditions. More recently, the significance of GAB for dissecting complex

genetic architecture of quantitative traits by implementing an integrated approach of association and QTL/eQTL

mapping, associative transcriptomics and whole genome selective sweep-scans, has truly been realized. Essentially,

the GAB expedites selection of desirable plants/recombinants with superior combinations of favorable alleles at

multiple loci (gene/QTL pyramiding) specifically for trait genetic enhancement studies in crop plants via marker-

assisted selection (MAS) and genomic/haplotype selection. The advent of various modern genomics and breeding

tools, including sequence-based informative molecular markers, high-throughput marker genotyping assays and

methods of rapid trait-specific gene/allele identification and their introgression into genotypes, have provided tremendous

opportunities and flexibility to researchers for adopting diverse integrated approaches in genomics-assisted crop

improvement program. This will greatly enrich our understanding of rapid quantitative dissection of complex traits for

crop genetic enhancement through translational genomics (GAB and transgenics).

Swarup K. Parida


105

IPPCongress2015



Molecular regulation of reproductive cold tolerance in chickpea

genotypes

Harsh Nayyar1* and Kamal Dev Sharma2

1Department of Botany, Panjab University, Chandigarh-160014, India2Department of Agricultural Biotechnology, Himachal Pradesh Krishi Vishva Vidyalaya,Palampur-176062, H.P., India


Chickpea is a cool-season food legume, which is sensitive to chilling temperatures (<20/10C), especially at its reproductive

stage, leading to abortion of flowers to substantially reduce the podding potential. The underlying molecular mechanisms

governing cold tolerance in reproductive components of chickpea are not known, which are imperative to be examined

to address cold tolerance in this legume. In the present study, we investigated various physiological, biochemical and

molecular mechanisms in leaves and anthers in chickpea genotypes contrasting for cold sensitivity. The findings revealed

that maintenance of a stable sucrose metabolism in anthers is a vital mechanism affecting pollen development during

cold stress in cold-tolerant genotypes. Investigations on the regulation of expression of differentially expressed genes in

anthers of cold-tolerant genotypes under cold stress revealed two important features that govern cold tolerance in

anthers i) differential expression of less number of genes and ii) up-regulation of majority of the differentially expressed

genes. The main categories of genes governing cold tolerance in anthers were carbohydrate/triacylglycerol metabolism,

signal transduction, pollen development and transport. Almost all the genes in these categories were up-regulated.

Regulation of gene expression suggests that chickpea anthers use a dual cold tolerance mechanism wherein anthers

sustain their development under cold by enhancing triacylglycerol and carbohydrate metabolism while pollen grains

maintain normal development by regulating pollen development genes.

Harsh Nayyar


106

IPPCongress2015



Anthesis, anther dehiscence, pollen viability, stigma receptivity and

pre, post-pollination biochemical changes in three varieties of mango

Kodthalu S. Shivashankara*, Tapas K. Roy and Gouribidanur A. Geetha

ICAR- Indian Institute of Horticultural Research, Bengaluru, India


The study was carried out in order to evaluate the floral biology in terms of anthesis, anther dehiscence, pollen

viability and in-vitro stigma receptivity as well as pre-, post-pollination biochemical changes in stigma of mango cvs.

Alphonso, Totapuri and Amarapali, differing in fruit set intensity. Higher anthesis and anther dehiscence were observed

before 8:00 AM and between 11:00 AM to 12:00, respectively in all the cultivars. As the temperature increased,

anther dehiscence occurred early which wascoincided with higher anthesis resulting in higher fruit set. Higher stigma

receptivity was observed within 3 hrs of anthesis in all the cultivars. The pH of the stigma was neutral during higher

anthesis, anther dehiscence and also with higher stigma receptivity, and later on the pH was changed to acidic in all

the cultivars. The stigmatic exudate was analysed to determine its components with a view to understanding their role

in pollination and fertilization inmango.Free sugars (Sucrose, Galactose, Glucose and Fructose), free amino acids

(Serine, Citrulline, Alanine, Proline, Phenylalanine and Glutamic acid) and hormones (Abscisic acid, Salicylic acid

and Jasmonic acid) analysed using LCMS were consistently predominant and increased after anthesis. Among all the

cultivars, Amarapali was observed to be good in terms of floral biological observations as well in biochemical parameters

resulting in higher fruit set.

K.S. Shivasankara


107

IPPCongress2015



Can one-sided sterility be regarded as a contrivance for sexual

variability in Cleome viscosa?

Veenu Kaul* and Shveta Saroop

Department of Botany, University of Jammu, Jammu-180006, J&K, India


Cleome viscosa L. belonging to family Cleomaceae is reportedly hermaphrodite. However, investigators have found

the species to be quite complex in so far as its sexual system is concerned. Almost all floral features at structural and

functional levels exhibit variation which quite likely leads to a flux in the breeding system of the species. The primary

sex expression on an individual plant differs tremendously. Overall, six flower types differentiate on an individual

plant. These can broadly be characterized as hermaphrodite, staminate (with rudimentary pistil), pistillate (with

rudimentary stamens), male (with no pistil), hermaphrodite with staminodes and staminate with staminodes. The

average production of these flower types varies from plant to plant and so does the frequency of the plants bearing

them. Majority of plants are, however, and romonoecious with some exhibiting functional monoecy. This restriction is

imposed due to the structural arrangements of sex organs while functional ones include stigma receptivity, pollen

germination, ovular and pollen sterility. For-example female sterility is imposed by the non-functionality of ovules and

male sterility due to pollen non-viability of shrivelled anthers. Interestingly, the one-sided sterility in context to the

flower sex expression varies among plants subjected to simple manipulations of defoliation, deflowering and fruit

removal. Results suggest major shift in resource investment towards the hermaphrodites further cluing towards the

complex game of resource allocation influencing this sterility.

Veenu Kaul

ent Session M

Young Scientist Session

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IPPCongress2015


YSS01 (IPPC0359)

Plants actively avoid state transitions upon changes in light intensity: Role of light-harvesting complex

II protein dephosphorylation in high light

Nageswara Rao Mekala*, Mikko Tikkanen and Eva Mari Aro

Department of Biochemistry, University of Turku Finland, Sirkkalankatu 8a B10, Turku-20520, Finland

*Presenting author: [email protected]

Photosystem II (PSII) core and light-harvesting complex II (LHCII) proteins in plant chloroplasts undergo reversiblephosphorylation upon changes in light intensity (being under control of redox-regulated STN7 and STN8 kinases andTAP38/PPH1 and PSII core phosphatases). Shift of plants from growth light to high light results in an increase of PSIIcore phosphorylation, whereas LHCII phosphorylation concomitantly decreases. Exactly the opposite takes place whenplants are shifted to lower light intensity. Despite distinct changes occurring in thylakoid protein phosphorylation uponlight intensity changes, the excitation balance between PSII and photosystem I remains unchanged. This differs drasticallyfrom the canonical-state transition model induced by artificial states 1 and 2 lights that concomitantly either dephosphorylateor phosphorylate, respectively, both the PSII core and LHCII phosphoproteins. Analysis of the kinase and phosphatasemutants revealed that TAP38/PPH1 phosphatase is crucial in preventing state transition upon increase in light intensity.Indeed, tap38/pph1 mutant revealed strong concomitant phosphorylation of both the PSII core and LHCII proteins upontransfer to high light, thus resembling the wild type under state 2 light. Coordinated function of thylakoid protein kinasesand phosphatases is shown to secure balanced excitation energy for both photosystems by preventing state transitionsupon changes in light intensity. Moreover, PROTON GRADIENT REGULATION5 (PGR5) is required for proper regulationof thylakoid protein kinases and phosphatases, and the pgr5 mutant mimics phenotypes of tap38/pph1. This shows thatthere is a close cooperation between the redox- and proton gradient-dependent regulatory mechanisms for proper functionof the photosynthetic machinery.

YSS02 (IPPC0269)

The role of inositol pyrophosphates in the perception of the plant defense hormone jasmonate

Debabrata Laha* and Gabriel Schaaf

Plant Physiology, ZMBP University of Tuebingen Germany, Auf der Morgenstelle 32, Tuebingen-72076, Germany


Diphosphorylated inositol polyphosphates, also referred to as inositol pyrophosphates, are important signaling moleculesthat regulate critical cellular activities in many eukaryotic organisms. In mammals and fungi, two distinct classes ofinositol phosphate kinases mediate their biosynthesis: KCS1/IP6K- and VIP1/PPIP5K-like proteins. Recent studies in ourlab show that PPIP5K homologs are widely distributed in plants and that Arabidopsis VIH1 and VIH2 are functionalPPIP5K enzymes. We will report a specific induction of the inositol pyrophosphate InsP8 by jasmonate and will provideevidence that steady-state and jasmonate-induced pools of InsP8 in Arabidopsis seedlings depend on VIH2. We willfurther report a role of VIH2 in regulating jasmonate related defenses by potentiating jasmonate perception. Using insilico docking experiments and radioligand-binding based reconstitution assays we can show high affinity binding ofinositol pyrophosphates to the F-box protein COI1–JAZ jasmonate co-receptor complex and will provide evidence thatcoincidence detection of jasmonate and InsP8 by COI1–JAZ is a critical component in jasmonate-regulated defenses.


110

IPPCongress2015


YSS03 (IPPC0398)

Transcriptomic insights into the plant structure and function: A parasitic plant and a unicellular alga

Aashish Ranjan1*, Daniel Chitwood2, Brad Townsley3, Moran Farhi3, Yasunori Ichihashi4 and Neelima Sinha3

1National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India; 2Donald Danforth Plant Science Center,Saint Louis, USA; 3University of California, Davis, USA; 4RIKEN Center for Sustainable Resource Science, Japan


Recent advances in the omics approaches, such as next generation sequencing in combination with genomics andtranscriptomics, have facilitated the global insights into the genome and transcriptome to investigate the plant structureand function. Parasitic plants, which penetrate and establish vascular connections through specialized organs calledhaustoria to steal nutrients and water from host plants, are one of the most destructive agricultural pests. In order tounderstand the genetic basis of plant parasitism, we de novo assembled the transcriptome of an obligate stem plantparasite Cuscuta pentagona (Dodder) using RNAseq reads from various stages i.e. seed, seedling, vegetative stem,prehaustoria, haustoria and flowers. Subsequent gene expression analysis and dissection of transcriptional dynamicsacross the stages identified key genes and gene categories, such as plant defense and transporter genes, involved in theprocess of plant parasitism. The study is being used to identify potential gene targets for use in controlling infestation ofcrops by parasitic plants. In a parallel project, transcriptomics deciphered the molecular and genetic basis of patterning inone of the largest unicellular coenocytic alga, Caulerpa taxifolia, with distinct functional pseudo-organs. The study revealeda global, apical-basal pattern of the transcript distribution across the algal body. Interestingly, transcripts associated withspecific cell compartments localize to organs with related functionalities. The results not only provided an intracellularatlas of transcript localization, but also demonstrated the contribution of transcript partitioning to morphology, independentfrom multicellularity, in plants.

YSS04 (IPPC0720)

Physiological, metabolic, and transcriptional analysis of Arabidopsis to gain insight into plant responses

to climate extremes

Gaurav Zinta1,2*, Hamada AbdElgawad1, Ivan A. Janssens1, Gerrit T.S. Beemster1 and Han Asard1

1Department of Biology, University of Antwerp, Belgium; 2Shanghai Center for Plant Stress Biology, SIBS, CAS, China


Ongoing climate change involves co-occurring changes in various environmental factors, such as atmospheric CO2,

temperature and precipitation. Extreme heat and drought limit plant growth and yield, whereas elevated CO2 alone is

beneficial. Current climate change predictions indicate that climate extremes are not only projected to become morefrequent and intense, but also to last longer. Molecular and physiological mechanisms underlying plant responses toclimate extreme conditions under ambient and elevated CO

2 are still poorly understood and little attention has been given

to the temporal variation of plant stress responses, and on the transgenerational effects induced by climate extremes. Weused Arabidopsis thaliana as a model system to understand plant responses to climate extremes. We investigated plantstress responses from whole-plant level, to the level of metabolites, enzymes, transcriptome and epi-genome. Mainfindings of this work are: 1) Elevated CO

2 mitigates the impact of combined heat and drought, which could be explained

by the higher up-regulation of antioxidant defense system (improved ROS scavenging) as well as by reducedphotorespiration (less ROS formation) under high CO

2, 2) Temporal metabolic profiling of plants exposed to combined

heat and drought showed that short- and long-term stress exposure altered the plant primary metabolism differentially,and 3) Multigenerational exposure (for four generations) of Arabidopsis to heat stress resulted in more tolerant off springs.The descendants of the stressed plants showed a primed transcriptional state and greater up-regulation of defenserelated metabolites. Here, we expect the imperative role of epigenetic mechanisms in transgenerational stress memory ofplants.


111

IPPCongress2015


YSS05 (IPPC0091)

Saltol QTL localized OsGATA transcription factor: An important candidate gene contributing salt tolerance

in transgenic rice

Kamlesh Kant Nutan1*, Sneh Lata Singla-Pareek2 and Ashwani Pareek1

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


Plant being sessile faces adverse environmental condition like drought, salinity, flooding and high and low temperature,which affects plant yield. Among them salinity is a major contributor of plant yield loss by abiotic stresses. One-third of theworld’s population consumes rice as a staple food, but this crop is highly sensitive to salinity stress. In our study, we havestudied differential expression of transcription factor genes localized within Saltol QTL of rice under salinity stress. SaltolQTL are present on short arm of chromosome number 1, accounts for more than 60% of the salt tolerance. On the basisof differential expression of 13 transcriptions factor genes (localized in Saltol) in contrasting cultivars of rice IR64 (sensitive)and pokkali (tolerant) under salinity stress, we have chosen GATA transcription factor. Constitutive overexpression ofGATA transcription factor in transgenic rice leads to higher biomass accumulation as compared to wild type (WT) plants.Under salinity stress, transgenic rice seedling overexpressing GATA transcription factor shows higher salinity tolerance,high chlorophyll content and less ion leakage as compared to WT counterparts. Furthermore, at the cellular level, lesserdamage of sub cellular organelles mainly chloroplast was observed in transgenic plants as compared to wild type andRNAi knock out plants. Osmolytes and other metabolites accumulation which help the plant in osmotic stress tolerancehave been found to be higher in transgenic rice as compared to wild type. Our results suggest that GATA transcriptionfactor may play a pivotal role in salinity stress tolerance in rice.

YSS06 (IPPC0171)

Decoding the transcriptome and small RNAome to reveal genotype/developmental stage-specific

molecular barcodes during drought and salinity stress in chickpea

Rohini Garg*



Drought and salinity are the major factors that limit chickpea production worldwide. In response to various stresses,plants evoke some common and stress-specific signature pathways that enable plants to discriminate between differentstimuli and respond in unique ways. In the present study, we identified stress associated transcriptome of differentchickpea genotypes under drought and salinity stress. RNA-seq of the root tissues of drought and salinity related genotypeswere carried out under control and stress conditions at vegetative and/or reproductive development stages. Deepsequencing identified thousands of novel gene loci and alternatively spliced mRNA isoforms. A comparative analysis ofthe transcriptomes revealed divergent gene expression profiles in different chickpea genotypes at different developmentalstages. A total of 79.1% and 84.8% transcripts exhibited genotype- and developmental stage-specific differential expressionunder drought stress and salinity stress, respectively. Overall, a significant fraction (~47%) of the transcription factorencoding genes showed differential expression under stress conditions. The key enzymes involved in metabolic pathways,such as carbohydrate metabolism, photosynthesis, lipid metabolism, generation of precursor metabolites and energy,post-translational protein modification, nucleic acid metabolism, cell redox homeostasis, cell wall component biogenesisand trehalose biosynthesis, were affected by drought and/or salinity stresses. Interestingly, different isoforms showedspecificity of expression patterns across the chickpea genotypes and/or developmental stages as illustrated by the AP2-EREBP family members. We also identified genotype-specific and salinity regulated miRNAs in salt-sensitive and salt-tolerant genotypes of chickpea. These findings provide basis to dissect the gene regulatory network involved in droughtand/or salinity stress response.


112

IPPCongress2015


YSS07 (IPPC0651)

Marker assisted recurrent selection (MARS) for drought tolerance using physiological and agronomic

traits in bread wheat (Triticum aestivum L. em. Thell.)

Harikrishna1*, N.K. Singh2, A.R. Rao3, Parveen Chhuneja4, P.C. Mishra5, S.C. Misra6, Niveditha Saini1, Kavitha Bhisth1,Arun Kumar1, Amitha C.R. Mithra2, Priyanka Vijay1, P.K. Singh1, Neelu Jain1, G.P. Singh1 and K.V. Prabhu7

1Division of Genetics, IARI, New Delhi-110012, India; 2National Research Centre on Plant Biotechnology, IARI, New Delhi-110012,India; 3Indian Agricultural Statistics Research Institute, New Delhi -110012, India; 4Punjab Agricultural University, Ludhiana, Punjab,India ; 5Jawaharlal Nehru Krishi Vishwa Vidyalaya, Powerkheda, India; 6Agharkar Research Institute, Pune, India; 7Joint DirectorResearch IARI New Delhi-110012, India*Present author: [email protected]

The term drought carries networks of complex phenomenon. Because of its complex nature in terms of networks ofpathways involved, complex inheritance and difficulties in phenotyping, breeding for drought tolerance is difficult. Aneffort was made to combine QTLs for drought adaptive traits using biparental population HI1500 X DBW 43; both parentscarrying positive alleles for drought tolerance. Among 25 different traits phenotyped, NDVI at boot and grain filling stagecanopy temperature at grain filling and milky stage, Leaf area index(LAI), grain weight per spike, thousand grain weight,biomass were showing significant correlation and regression values with yield under drought. These parents were diversebased on polymorphic 120 SSRs and 802 SNPs using 9K wheat SNP bead chip array. Based on validated QTLs informationand multi-location (four location two year) testing of base populations 25 best individuals were selected for inter mating atF6 generation. Carefully chosen pairwise inter crosses (100) were attempted in order to combine QTLs using limitedpopulation size. A total of 17 loci were targeted and validation of molecular marker SSRs/QTLs present on1B,1D,2D,2A,2B,2D,3A,3B,4A,5A,5B,6A, and 7B was done. Based on validated markers, wide range of inter crossedprogenies in F5 generation with 4-12 positive alleles were obtained in first round of recombination out of 17 targeted locisegregating in the population. Genotypes which contains more number of QTLs out performed over checks in multi-location testing (four locations). These individuals will be subjected for further testing in co-ordinated trails at nationallevel.

YSS08 (IPPC0195)

OsGLYI-8 is a nucleus-localized glyoxalase I required for methylglyoxal metabolism in rice

Charanpreet Kaur1*, Sneh Singla-Pareek2 and Sudhir Sopory2

1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

The glyoxalase system generates D-lactate from methylglyoxal (MG), a cytotoxic byproduct of glycolysis which canotherwise increase to toxic levels under pathological conditions and form irreversible adducts with proteins and nucleicacids. The first enzyme of the glyoxalase system, glyoxalase I (GLYI), is usually a cytosolic metalloprotein that requireseither Ni2+ or Zn2+ for its enzymatic activity. Plants have a variety of diverse GLYI isoforms that can play a crucial roleduring drought and high salinity. However, the exact subcellular localization, mechanism and physiological relevance ofseveral of these diverse GLYI isoforms remain to be determined. Here, we analyzed a Zn2+-stimulated OsGLYI-8 fromrice that has a so far unique nuclear localization and exhibits unusual biphasic steady-state kinetics. Further, loss of itshomologue (AtGLYI2) in Arabidopsis leads to severe developmental defects on exposure to even low MG concentrations,much like yeast GLYI mutant which is also sensitive towards low MG concentrations. Our results thus, provide firstevidence for MG metabolism in the nucleus and indicate towards a critical role of OsGLYI8 in plants.


113

IPPCongress2015


YSS09 (IPPC0772)

CBL-interacting protein kinase, CIPK9, differentially interacts and phosphorylates its upstream regulators,

Ca2+ sensor CBLs in Arabidopsis

Akhilesh K. Yadav*, Saroj K. Jha, Sibaji K. Sanyal and Girdhar K. Pandey

Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi-110021, India


Potassium is essential macronutrient required for various cellular processes and its deficiency leads to yield penalty incrops. Arabidopsis genome encodes 10 calcineur in B-like (CBLs) calcium sensors, which differentially interacts with26CBL-interaction protein kinases (CIPKs). These CBL-CIPK complexes regulate various developmental, biotic, andabiotic as well as nutrient deficiency signaling processes. One of the CIPK family members, CIPK9 was identified as acritical regulator of K+ deficiency. In this study, we are discussing the detail biochemical regulation of CIPK9 activity, andphosphorylation of its interacting CBLs. CIPK9 specifically interact with CBL1, CBL2, CBL3 and CBL9 in yeast two-hybridassay. The full length CIPK9 differentially phosphorylates its interacting CBLs in in-vitro kinase assay, hence specificCBLs act as physiological substrate of CIPK9.Autophosphorylation of CIPK9 is enormously increased specifically inpresence of CBL2 or CBL3 while unaltered in the presence of CBL1 or CBL9. In contrast, CIPK9ÄR (CIPK9 withoutregulatory region i.e. having only kinase domain) also phosphorylates its interacting CBLs butitspreferential phosphorylationof CBLs is lost. Moreover, we found that mutated T178 to D178 residue in the activation loop of CIPK9 enhances kinaseactivity while T178 to A178 mutation reduces the level of autophosphorylation of CIPK9. However, in presence of CBL2orCBL3, autophosphorylation of CIPK9, CIPK9T178D and CIPK9T178Aas well as transphosphorylation of CBL2 or CBL3increases dramatically in in vitro kinase assay. Our study suggests that CBL1, 2, 3 and 9 interact differentially withCIPK9.Upon interaction CIPK9 can also differentially phosphorylate the CBLs and this differential phosphorylation of CBLs alsoinfluences the autophosphorylation as well as substrate phosphorylation activity of CIPK9 in a Ca2+ dependent manner inin-vitro kinase assay.

YSS10 (IPPC0327)

Physiological and biochemical plasticity of Lepidium latifolium L. suggests it as Sleeper Weed in Western

Himalayas under climate change scenario

Tarandeep Kaur*, Hilal A. Bhat and Dhiraj Vyas

Biodiversity and Applied Botany Division, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu Tawi-180001, India


Unlike the aggressive invasive character of Lepidium latifolium in riparian plains of North America, its growth is restrictedin Ladakh Himalayas. In order to understand its future spread under climate change scenario in ecologically sensitiveregion of Western Himalayas, native populations of Lepidium latifolium in different microclimates were studied forphysiological and biochemical plasticity. Different populations from sparsely populated site (SPS; 2807 m asl; 34.54 N,76.13 E), moderately populated site (MPS; 4139 m asl; 33.20N, 78.64E) and densely populated site (DPS; 3164 m asl;34.15N, 77.57E) were studied for their fluorescence, photosynthetic and biochemical characteristics. Results showedthat the DPS had higher photosynthetic accumulation and transpiration rate despite lower VpdL and higher relativehumidity. This suggests the regulation of its leaf temperature by evaporative cooling. Apart from its nutritive value, higheramounts of Sinigrin obtained in DPS might play an important role in water transport under environmental stress. Thequantum efficiency of PSII photochemistry (F

v/F

m, NPQ, PSII) and light curve at various PPFD’s suggests better light

harvesting potential and light compensation point at DPS than the other two sites. Concomitantly, plants at SPS hadsignificantly higher lipid peroxidation, suggesting a stressful environment, and higher induction of antioxidative enzymes.High light intensities at MPS are managed by specialized contrive of carotenoid pigments and PsBs gene product. It issuggested that L. latifolium is present as ‘sleeper weed’ that has inherent biochemical plasticity involving multiple processesand its potential spread is linked to site-specific micro-environment, whereby, it would prefer flat valley bottoms withalluvial fills having high water availability under changed climate.


114

IPPCongress2015


YSS11 (IPPC0162)

Ambient ionization mass spectrometry imaging of rohitukine, a chromone anticancer alkaloid, during

seed and seedling development in Dysoxylum binectariferum Hook.f (Meliaceae)

P. Mohana Kumara1*, Amitava Srimany1, G. Ravikanth2, R. Uma Shaanker2 and T. Pradeep1

1DST Unit on Nanoscience and Thematic Unit of Excellence, Department of Chemistry, IITM, Chennai- 600036, India2School of Ecology and Conservation, Deprt. of Crop Physiology, University of Agricultural Sciences, GKVK, Bengaluru-560065, India


Chromone alkaloids are a unique group of compounds containing a flavonoid or a noreugenin chromone componentlinked with a ring containing one or more nitrogen atoms. These are sparsely distributed in plants. Rohitukine, a prominentchromone alkaloid, has been reported from only four species, belonging to the families, Meliaceae and Rubiaceae.Among them the Dysoxylum binectariferum (Meliaceae) is reported to accumulate the highest amount of rohitukine. Inrecent years, chromone alkaloid, such as rohitukine has gained wide attention because of their phrmacological activities.In fact, flavopiridol, a derivative of rohitukine, has been approved as an anticancer drug. The biosynthesis of rohitukine isnot yet elucidated. In this study, we examine the spatial and temporal distribution of chromone alkaloids during variousstages of seed and seedling development in D. binectariferum using desorption electrospray ionization mass spectrometryimaging (DESI MSI). Rohitukine (m/z 306.2) accumulation increased from early seed development to seed maturitystage. The spatial distribution of rohitukine was largely restricted to the cotyledonary tissue followed by the embryo andleast in the seed coat. In seedling, root and leaves contained the high amount of rohitukine compared to the stem andtwigs. Besides rohitukine, rohitukine acetate (m/z 348.2) and glycosylated rohitukine (m/z 468.2) were also detected,both through mass fragmentation and exact mass analysis. All of these molecules showed the tissue specific distributionof metabolites in seed, stem, root and leaves. In summary, DESI MS results indicated that spatial and temporal pattern ofdistribution of metabolites. It would be interesting to examine the underlying genetic basis for such a pattern.

YSS12 (IPPC0761)

Biochemical characterization of an Arabidopsis thaliana cyclophilin AtCyp19-3

Gundeep Kaur1*, Supreet Singh1, Harpreet Singh1, Mrinalini Chawla2, Tanima Dutta1, Harsimran Kaur1, Kyle Bender3,W.A. Snedden3, Sanjay Kapoor2, Ashwani Pareek4 and Prabhjeet Singh1

1Guru Nanak Dev University, India2Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India3Queen’s University, Kingston, Canada4Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Cyclophilins are ubiquitous proteins and constitute a multigene family in higher organisms. Several cyclophilins showpeptidyl-prolyl cis-trans isomerase (PPIase) activity that catalyzes cis to trans isomerization, the rate limiting step inprotein folding. Although Arabidopsis genome is predicted to contain 35 cyclophilin genes, biochemical characterization,imperative for understanding their cellular functions, has been reported only for few of these proteins. The present studyreports that Arabidopsis cyclophilin, AtCyp19-3, is an enzymatically active protein and possesses PPIase activity that isspecifically inhibited by CsA, with an inhibition constant of 18.75 nM. The catalytic efficiency of AtCyp19-3 for the substrateN-succinyl-ala-ala-pro-phe-p-nitroanilidine is 4.88 x 106 mol-1 s-1. Gel overlay and bimolecular fluorescence complementationassays provided evidence that AtCyp19-3 interacts with calmodulin in vitro and in vivo. The interaction of this protein withcalmodulin is Ca2+- dependent. Further, the PPIase activity of AtCyp19-3 appears to be redox regulated and in silicomodelling studies suggested that these mechanisms may be distinct in different members of this family, signifying specificityin their roles.

Session A : Abiotic Stress Adaptation and Management

115

IPPCongress2015


PA001 (IPPC0010)

Mitigating effects of paclobutrazole on morpho-physiological, biochemical and antioxidant enzyme

activities of mungbean [Vigna radiata (L.) Wilczek] under flooding stress

A. Hemantaranjan* and D.K. Yadav

Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, India


An extensive investigation was conducted during kharif (rainy season) on the role of paclobutrazol (PBZ) on morpho-physiological, biochemical and antioxidant enzyme activity in flooding tolerant (HUM-16) and susceptible (HUM-12)mungbean (Vigna radiata L.) genotypes at pre-flowering stage. Paclobutrazol was applied both as seed soaking treatment(ST) and foliar spray (FS) (at pre-flowering stage). The flooding stress was carefully imposed at pre-flowering stage up to7 days. Observations recorded revealed that paclobutrazol @ 10 ppm (ST) + 10 ppm (FS) significantly increased numberof leaves per plant, fresh and dry weights of shoot and root, relative water content (RWC) and nitrate reductase activity(NRA) followed by antioxidant enzyme activity, viz., superoxide dismutase (SOD), ascorbate peroxidase (APX) and alcoholdehydrogenase (ADH) while, reduced plant height per plant and leaf area per plant in both the genotypes were also notedin PBZ treated plants under stress. Control plants without PBZ registered lesser significant values on the aforementionedparameters under flooding stress. Hence, findings elucidated that PBZ treatments positively influenced the aforementionedparameters. Nevertheless, the threshold concentration of PBZ @10 ppm ST + 10 ppm FS played significant role inmitigating the flooding stress by enhancing number of leaves per plant, fresh and dry weights of shoot and root; leafrelative water content, NRA, SOD, APX and ADH activities. Interestingly, the advantageous threshold level of PBZ asseed treatment combined with foliar spray treatment in this experiment was evaluated to be an economic source havingconsiderably high potential as compared to other plant growth regulators and data for which shall be further discussed.

PA002 (IPPC0075)

Abiotic stress tolerance in transgenic potato with reduced expression of PSII manganese stabilizing

protein

Chandrama Prakash Upadhyaya1* and Se Won Park2

1Department of Biotechnology, Dr. Hari Singh Gour Central University, Sagar-470003, Madhya Pradesh, India2Department of Molecular Biotechnology, Konkuk University, Seoul, Korea*Presenting author: [email protected]

Manganese stabilizing protein (MSP) is an important component of the Photosystem II (PSII) oxygen evolving complex.We have reported the transgenic potato plants with reduced expression of MSP (named as MSP-As) and the response ofMSP-As plants towards salinity, heavy metal and osmotic stresses. MSP-As plants treated with NaCl, ZnCl2 or mannitolsolution showed significant level of tolerance under all the stress conditions. Specific enzyme activities of major ROS-scavenging enzymes were found significantly higher in MSP-As plants than the control plants. MSP-As plants accumulatedincreased levels of proline and low molecular weight metabolites such as ascorbate and α-tocopherol, which indicatedthat these plants were much more resistant to stress compared to the corresponding control plants. The primaryphotochemical efficiencies and the OJIP kinetics analyses further confirmed that MSP-As plants were in better optimalhealth under stress compared to the control plants. Although the exact reason behind the increased stress tolerance instressed MSP-As plants is unclear, our results strongly indicate the role of MSP, which unknown function in abiotic stresstolerance.


116

IPPCongress2015


PA003 (IPPC0085)

Comparative performance of wheat genotypes adjusting by sowing windows and moisture regimes in

Eastern Indo-Gangatic Plains

S.K. Dwivedi1*, Santosh Kumar1, S.K. Singh1, J.K. Mishra1 and S. Singh2

1Division of Crop Research, ICAR Research Complex for Eastern Region, Patna, Bihar-800014, India2International Rice Research Institute (IRRI), Manila, Philippines


Understanding of physiological mechanisms that enable plants to adapt to water deficit stress and maintain growth andproductivity during stress period could help in screening and selection of tolerant genotypes. Keeping these views inmind, an experiment was conducted at ICAR Research Complex for Eastern Region, Patna to study the comparativeperformance of five wheat genotypes (HD2987, K7903, HI1563, HD2824 and HD2967) grown under irrigated (at CRI) andcompletely rainfed conditions adjusted by sowing windows (15 Nov. & 30 Nov.). Study revealed that least impact ofsowing dates was observed among genotypes in terms of physiological traits and yield attributes. The average yieldreduction from irrigated to rainfed condition was 31 percent. Under rainfed condition, the higher yield was observed withgenotype K7903 (1.9 t ha-1) followed by HD 2987 (1.8 t ha-1) as compared to other genotypes. Moreover, the physiologicaltraits (relative water content; RWC, membrane stability index; MSI, total chlorophyll content and photosynthetic rate)were declined while proline content was improved under rainfed condition across the genotypes at both the developmentalstages (anthesis and A+20 days). However, the genotype HD 2987 followed by K7903 were able to main high RWC(67.59 and 64.84%; 67.73 and 61.86%), and photosynthetic rate (15.70 and 10.73 μmol m-2 s-1; 17.76 and 10.74 μmolm-2 s-1) as compared to other genotypes at anthesis and post-anthesis stage, respectively. High RWC and photosyntheticrate is an important indicator to drought tolerance. This study will boost wheat production under completely rainfedcondition.

PA004 (IPPC0090)

Development of a robust screening protocol and, identification and characterization of novel heat tolerant

rice mutants

Priyanka Das1*, Ray Singh Rathor1, Sneh Singla-Pareek2 and Ashwani Pareek1

1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


Global warming is expected to have negative effect on plant growth due to the damaging effect of high temperature onplant development and yield. The increasing threat of high temperature might lead to catastrophic loss of productivity ofvital food crops like rice. Therefore, it is necessary to develop heat tolerant rice plants by transgenic or mutation breedingapproaches to overcome this problem. Mutation breeding is accepted as a powerful technique in the present era to obtainstress tolerant rice plants. We have developed a robust screening protocol and obtained three heat tolerant mutant plantsfrom M6 mutant population of gamma-irradiated rice. High temperature tolerance is determined at first round of screeningby phenotypic performance of mutant lines as compared to wild type after heat treatment and recovery. As indicators ofheat tolerance at seedling stage, parameters such as leaf colour, root length, shoot length, fresh weight, leaf tip burningand ‘leaf tip kink’ were taken. In the second round of screening, physiological and biochemical analysis (lipid peroxidation,electrolyte leakage study and antioxidant enzyme assays) were carried out to authenticate the tolerance of mutantstowards heat stress. Third round of screening of the heat tolerant mutant (confirmed at seedling stage) plants was carriedout at flowering (during anthesis) stage following heat treatment and subsequent physiological and biochemicalcharacterization. Our results showed that the putative heat tolerant rice lines are better in comparison to WT plants interms of better phenotype, better enzymatic activities, better photosynthesis and higher yield.


117

IPPCongress2015


PA005 (IPPC0100)

Transgenic Arabidopsis thaliana over-expressing MuSMT1, a gene encoding sterol methyl transferase1

from Macrotyloma uniflorum improves tolerance to abiotic and biotic stresses

Shikha Masand* and Sudesh Kumar Yadav

Plant Metabolic Engineering Laboratory, Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology,Palampur-176061 (HP), India


Sterol methyl transferase1 (SMT1), a rate regulating enzyme catalyzes the production of 24-ethyl sterols. TransgenicArabidopsis over-expressing MuSMT1 encoding SMT1 enzyme protein was isolated from Macrotyloma uniflorum and thebeneficial effect of enhancing 24-ethyl sterol biosynthesis was studied. Transgenic Arabidopsis showed tolerance againstmultiple abiotic stresses such as high temperature, low temperature, drought, salinity, and oxidative stresses, as observedby retention of RWC, chlorophyll content and reduced MDA and H

2O

2 levels compared to WT plants. Transgenic Arabidopsis

also showed significant resistance to Botrytis cinerea, a necrotrophic fungus due to enhanced accumulation of 24-ethylsterols. Paclobutrazol, a potent inhibitor of sterol biosynthetic pathway, severely impaired plant growth by reducing 24-ethyl sterols production. The impaired growth of WT plants was rescued by the exogenous application of stigmasterol,suggested the requirement of 24-ethyl sterol for growth and development. Taken together, this study has for the first timedocumented the importance of 24-ethyl sterols accumulation in providing biotic and abiotic stress tolerance by MuSMT1

over-expression.

PA006 (IPPC0113)

Elucidating molecular markers for the loss and re-establishment of desiccation tolerance in pea (Pisum

sativum) seeds

Balram Sahu* and Subhash Naithani

Seed Biology Lab., SoS in Life Sciences, Pt. Ravishankar Shukla University, Raipur-492010, India


Most outstanding feature of dry seeds necessary for ex-situ storage is desiccation tolerance (DT). DT is the ability of anorganism to withstand removal of its intracellular liquid water, then resume normal metabolism after rehydration and tosurvive in this state for ecologically significant period. The DT which is lost in the germinated seeds can be re-induced bythe treatment of PEG, osmotic stress agent etc. Our study offers insight on the role of AOS, antioxidative enzymes anddesiccation proteins [LEA and HSP] as a biomarker in pea seeds during loss and re-establishment of DT. The respirationcapacity of germinated pea seeds declined with prolonged exposure of dehydration using silica gel. The loss of DT in thegerminated pea seeds was negatively correlated with the accumulation AOS [superoxide and H

2O

2] measured in situ and

in vivo. The accumulation of AOS was promoted due to impairment of antioxidative capacity [quantitative, isoenzyme andgene expression analysis of SOD, CAT and APX in the germinated pea seeds]. The pattern of desiccation toleranceprotein [LEA and HSP’s] was similar to antioxidative enzyme. The re-establishment of desiccation tolerance in thegerminated pea seeds was accompanied by reorientation of AOS, antioxdative system and DT-proteins thus confirmingtheir vital role in the DT. The biomarkers of DT such as AOS, antioxidative enzymes and DT-proteins is discussed in lossand re-establishment of DT in the germinated pea seeds a model system developed for understanding the less knownpathways of DT.


118

IPPCongress2015


PA007 (IPPC0118)

Moisture stress tolerance in lentil (Lens culinaris Medik.) genotypes in relation to physiological, biochemical

parameters and 13C discrimination

B.K. Mishra1, J.P. Srivastava1* and J.P. Lal2

1Department of Plant Physiology, 2Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras HinduUniversity, Varanasi-221005, India


Three microsperma (HUL-57, KLS-218, NDL-1) and five macrosperma (DPL-15, DPL-62, IPL-81, IPL-406 and L-4076)lentil genotypes were sown in field under irrigated and rainfed conditions. Drought sensitivity indices indicated microspermatypes to be more tolerant to drought than macrosperma types. Pod formation stage was the most sensitive stage todrought. Stress reduced durations between different phenophases, total dry matter production and seed yield, but reductionwas of lesser magnitude in drought tolerant genotypes. Efficient remobilization of assimilates and nitrogenous substancesfrom vegetative parts to seeds, especially during later phase of crop growth, was observed in drought tolerant genotypes.In drought-stressed plants reduction in seed yield was mainly due to reduction in pods per plant and fruiting nodes perplant. Drought tolerant genotypes maintained higher relative water content (RWC), higher amounts of proline and solublesugars, and lower amounts of malondialdehyde and H

2O

2 in leaves under stress. Drought tolerant genotypes exhibited

lower 13C discrimination (Δ13C) and in normal plants Δ13C was associated negatively with seed yield, dry matter productionand specific leaf weight (SLW) of plants under drought. Efficient remobilization of assimilates and nitrogenous substancesfrom vegetative parts to seeds during later period of crop growth, maintenance of higher leaf relative water content andaccumulation of osmolytes (proline and soluble sugars) were the parameters associated with moisture stress tolerance inlentil. It is suggested that Δ13C technique may be used to identify drought tolerant and susceptible lentil genotypes.Nevertheless, SLW under stress condition may be taken as a reliable alternative of Δ13C.

PA008 (IPPC0121)

Trichoderma-colonization induced morpho-physiological and molecular amendment in drought stressed

rice plant

Veena Pandey1*, Mohammad Wahid Ansari2, Suresh Tula2, Ranjan Kumar Sahoo2, Gurdeep Bains1, Atul Kumar1,Alok Shukla1, Narendra Tuteja2 and Jitendra Kumar1

1Department of Plant Physiology, G.B. Pant University of Agriculture & Technology, Pantnagar-263145, Uttarakhand, India2International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


The present study was carried out to assess the potential of Trichoderma harzianum to induce resistance under droughtstress in two rice genotypes namely Pusa Basmati 1 and Pant Sugandh Dhan 17. Root-dip treatment of rice seedlingswith Trichoderma formulation was given at the time of transplanting i.e. 21 days after sowing. The formulation wasprepared by using Echinocloa frumentacae as substrate and the concentration of viable propagules was found to be 107cfu/g. For determining the dose response, concentrated solution of Trichoderma, containing 10g/L (Dose1), 20g/L (Dose2)and 30g/L (Dose3) of Trichoderma formulation was prepared. Rice seedlings of all the genotypes were dipped in thesethree solutions for 25 min and thereafter planted in pots having sterilized soil. Seedlings dipped in distilled water wereused as control. Drought stress was given at the time of flowering with withdrawn of water for five days. The soil moisturecontent in pots on fifth day was 8±2%. Results showed a significant amendment in the transcript levels of dehydrin andaquaporin genes in a dose responsive manner, as compared to control. Inoculation with Trichoderma induced prominentleaf rolling and reduced the incidence of leaf senescence and tip burn caused by drought stress in rice plant. Thus,providing evidence that T. harzianum colonization in rice plants can be considered as a promising strategy to mitigatedrought stress. This strategy can be explored further so that it can be applied in field to eliminate undesirable consequencesof drought stress.


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PA009 (IPPC0139)

Raising stress tolerant rice for better yield and quality through genetic manipulation of cyclophilin

Suchismita Roy1*, Sneh Singla-Pareek2 and Ashwani Pareek1



Cyclophilins are a set of ubiquitous proteins present in all subcellular compartments, involved in a wide variety of cellularprocesses. Cyclophilin 2 (OsCyp2) is a cytosolic member of the immunophilin family from Oryza and is a 519 bp geneencoding 172 amino acids. qRT-PCR data suggests that transcript abundance of OsCyp2 is regulated under differentstress conditions. Localization studies confirm that OsCyp2 is localized in both cytosol and nucleus, indicating its possibleinteraction with several other proteins. Promoter analysis of this gene demarcates the presence of several stress relatedcis-acting elements, which did show inducibility of the promoter under different stress conditions. Stable overexpressingtransgenic lines of this gene in Oryza sativa imparted tolerance towards multiple abiotic stresses as evidenced by highergermination efficiency, higher root length, more number of lateral roots, higher shoot length, chlorophyll content, and K+/Na+ ratio as well as less calcium accumulation observed in the transgenic plants, under stress conditions. Transgenicplants also showed reduced MDA content and less electrolyte leakage under stress conditions than WT plants, suggestingbetter ion homeostasis in the transgenic plants than WT plants. RNAi lines of OsCyp2 gene validated these results withan observed phenotypic change in the seed morphology, seed weight and yield quality. Thus, it is proposed thatoverexpression of this gene is suitable for genetic engineering for better grain quality, yield and tolerance to cope withunfavourable conditions.

PA010 (IPPC0143)

OsHK5, a histidine kinase of rice, working as multi-stress responsive gene in Saccharomyces cerevisiae

Priyanka Gupta1*, Sneh L. Singla-Pareek2 and Ashwani Pareek1

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, JNU, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


Signal transduction cascades regulate cellular functions under normal and stress conditions. Two component signallingsystem (TCS) is one of the examples of signal cascades which play significant role in signal transduction in both prokaryotesand eukaryotes. TCS members include membrane bound histidine kinase (HK) as receptor and its cognate responseregulator (RR) in cytosol. Plants have histidine phosphotransfer protein (HPT) as adaptor protein along with HK and RR.Perception of an environmental stimulus by membrane bound histidine kinase leads to autophosphorylation of conservedhistidine residues. Signalling proceeds through transfer of the phosphoryl group to histidine residue of Histidinephosphotransfer protein (Hpt), which subsequently transfers phosphoryl group to aspartate residue of response regulatorsmaking a relay of phosphate transfer from His to Asp to His to Asp. In this study, we have investigated the role of ricehistidine kinase, OsHK5 in abiotic stresses along with the rest of the sensory histidine kinases of rice. In silico analysisrevealed that OsHK5 is a close homolog of Arabidopsis cytokinin receptor AHK2, and functions as membrane localisedhomodimer. OsHK5 is highly expressed throughout various developmental stages in rice. Expression of OsHK5 is inducedin response to multiple abiotic stresses. OsHK5 along with the other OsHKs can functionally complement yeast Äsln1temperature sensitive mutant. Yeast transformants carrying OsHK3b and OsHK5 can cope up with salinity, osmoticstress and high temperature. Assessment of growth of yeast transformants carrying OsHK5 in response to plant hormonecytokinin reveals that it may be functioning independent of cytokinin signalling.


120

IPPCongress2015


PA011 (IPPC0167)

Thermal imaging to assess genetic variation in drought adaptation of soybean cultivars

Mahesh Kumar*, Ajay Singh, Susheel Raina, V. Govindasamy, R.L. Choudhary, Jagadish Rane and P.S. Minhas

ICAR-National Institute of Abiotic Stress Management, Malegaon, Baramati-413115, Pune India


Soil moisture deficit is the most important factor limiting soybean yields in drought prone regions. With diminishing shareof fresh water for agriculture this problem is likely to aggravate hence water use efficient varieties are essential to enhanceand stabilize the production of crops like soybean. Though advances in genomics of this crop are remarkable, genescontributing to drought-tolerance are elusive mainly because of lack of precision in phenotypic characterisation of responsesof large set of diverse genotypes. Hence non-invasive tools such as thermal imaging system to understand crop canopytemperature dynamics are recently gaining more attention. However, methods for employing these tools for acquiring theimage, processing and analysis are not standardised. Hence to optimise methods for differentiating responses of soybeanto drought, 32 diverse genotypes with similar phenology were selected from the pool of 174 collections screened inpreliminary experiments. They were raised in field in RBD with 8 replications. More than 2500 thermal image wereacquired at different growth stages of crop to assess the genetic variation in canopy temperature. Simple methods weredeveloped to process the thermal image and analysis was carried out with customised software supplied with instrument.The method could identify superior and high yielding genotypes despite higher canopy temperature and also those whichcan keep their canopy cooler than local adapted cultivars. However, high productivity of NRC-7 despite relatively highercanopy temperature than other genotypes throughout growth period could be partially attributed to high net assimilationrate and quantum yield, indicated by chlorophyll fluorescence parameters.

PA012 (IPPC0169)

Comparative effects of salicylic acid and its derivatives on the water relation, antioxidant defence

mechanism, nitrate reductase, malate dehydrogenase, lipid peroxidation and yield of clusterbean

(Cyamopsis tetragonoloba) under water stress

N.S. Nathawat1,*, V.S. Rathore1, Birbal1, N.D. Yadav1 and R. Bhargav2

1ICAR-Central Arid Zone Research Institute, RRS, Bikaner-334004, Rajasthan, India2ICAR-Central Institute of Arid Horticulture, Bikaner-334006, Rajasthan, India


Drought is an important factor influencing the growth and physiological characteristics of plants. Salicylic acid and itsderivatives are known to affect various physiological and biochemical activities of plants. A field experiment conductedduring rainy (kharif) season of 2014 at CAZRI, RRS, Bikaner, Rajasthan to evaluate the efficacy of salicylic acid (SA),thiosalicylic acid (TSA) and 5-sulfosalicylic acid (5-SSA) on growth, physio-biochemical parameters and yield of clusterbean under water stress condition. The foliar application of SA and its derivatives had 10-23% greater membrane stabilityindex and 13-23% higher NAR than control. The application of SA and its derivatives had 11-40, 14-45, 13-30, 9-22%higher superoxide dismutase (SOD), ascorbate peroxidise (APX), nitrate reductase (NR), malate dehydrogenase (MDH)activity, whereas the level of lipid peroxidation in terms of malondialdehyde (MDA) content was 12-29% lower comparedto water spray control. The MDA content in leaves varied from 8.9 to 12.7 μmol MDA g-1 fr. wt.; being lowest with 1.0 mMSA (29%) over WS. The activities of SOD, APX and NR were highest with 1.0 mM SSA over control. Exogenous applicationof SA and its derivatives brought 10-19 % improvement in seed yield, being highest with 1.0 mM SSA, followed by 1.0 mMSA and 1.0 mM TSA. Application of SA and its derivatives resulted in higher tissue water content; increased activity ofantioxidant enzymes, decreased level of lipid peroxidation and membrane injury and it also protected nitrate reductaseactivity against inhibition under water deficit conditions.


121

IPPCongress2015


PA013 (IPPC0183)

The effect of sowing dates on frost tolerance in faba bean (Vicia faba L.)

Najeeb Alharbi1*, Kedar Adhikari2 and Helen Bramley2

1Plant Breeding Institute, Faculty of Agriculture and Environment, The University of Sydney, 7/48-52 St Hilliers Road AUBURN,Sydney 2144, Australia2The University of Sydney, New South Wales 2006, Australia


Radiation frost is an important abiotic stress affecting grain yield in faba bean. The purpose of this study was to find frosttolerant genotypes and investigate whether frost damage can be managed by adjusting sowing time. A selection of 15diverse faba bean genotypes were evaluated under three different sowing dates (16 April, 7 May and 26 May 2014) withtwo replications in a field condition at Narrabri in New South Wales, Australia. A visual 1-9 scale was used for scoring thefrost damage at vegetative and reproductive stage by examining stem and pod, respectively. There was a highly significantdifference among genotypes for frost tolerance at the vegetative stage, but sowing date and its interaction with genotypewas not significant. There was no significant variation in pod damage among genotypes, however, the sowing time had ahighly significant impact indicating frost can be managed by altering the sowing time. The highest damage was found inthe early sowing date. Moreover, frost damage in stems and pods was not correlated with any of the morphological traits,such as leaf area, number of leaves and internodes length. In conclusion, frost tolerance can be improved at the vegetativestage by including more tolerant genotypes in the breeding program, but there was no variation for tolerance at thereproductive stage. Frost events are not predictable which might be a challenging task if sowing date alone is consideredin frost management. Therefore, more tolerant genotypes need to be identified in the future.

PA014 (IPPC0189)

Multigene engineering bridges the yield gap in rice under abiotic stress conditions

Brijesh Gupta1*, Khirod Sahoo1, Ashwani Pareek2, Sudhir Sopory1 and Sneh Singla-Pareek1

1Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India2School of Life Science, Jawaharlal Nehru University, New Delhi 110067, India


Increasing the yield of crops is an important objective for biotechnologists. Considering the multigenic nature of stressresponse in plants, gene pyramiding or multigene engineering, act as a promising approach to develop stress tolerantplants. Glyoxalase pathway enzymes, viz. glyoxalase I (GlyI) and glyoxalase II (GlyII), are the two enzymes required forglutathione-based cellular detoxification of a metabolically produced cytotoxin methylglyoxal (MG). A sodium protonantiporter contributes towards cellular ion homeostasis by sequestering excess Na+ ions. In the present study, we haveshown that gene pyramiding using GlyI, GlyII and sodium proton antiporter significantly improves tolerance to multipleabiotic stresses in both tobacco and rice. The photosynthetic efficiency of triple (GlyI+GlyII+sodium-proton antiporter)transgenic tobacco and rice lines was significantly higher than double or single gene transgenic plants and they were ableto flower and set more seeds under salinity and drought stress conditions. Further, as compared to wild type, single ordouble gene transgenic plants, the yield penalty was significantly lower in the plants expressing all the three genes undersuch stress conditions. These results suggest that strategies of pyramiding genes to remove the cytotoxic metaboliteslike methylglyoxal (by engineering glyoxalase system) and sequestration of sodium ions (by engineering ion transporters)may significantly improve stress tolerance limits. Future studies may utilize a similar strategy to generate stress tolerantvarieties in other agronomically important crops as well.


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IPPCongress2015


PA015 (IPPC0193)

OsCBSCBS3 is a stress-responsive ATP and/or AMP binding protein involved in abiotic stress adaptation

in rice

Ashish Subba1*, Thilini Ariyadasa1, Charanpreet Kaur2, Ashwani Pareek2 and Sneh Lata Singla-Pareek1

1Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


CBS domain is a small protein module (approx. 60 residues long) present in various classes of proteins in all kingdoms oflife. This domain has been identified to have binding sites for adenosine derivatives, thereby regulating the activity ofassociated proteins or domains. In plants, CBS domain containing proteins are differentially expressed during abioticstress and few of them have been shown to be involved in stress adaptation, but still much is yet to be known about theirphysiological role in the system. Here, in order to understand the role of CBS domain containing proteins, we have clonedand characterized OsCBSCBS3 from rice which encodes a two CBS domain pair-containing protein and is differentiallyinduced in response to abiotic stress conditions in salt-tolerant Pokkali and salt-sensitive IR64 rice cultivars. Our in silicostudies revealed high similarity of OsCBSCBS3 to gamma regulatory subunit of AMP-activated protein kinase from differentspecies. Further, subcellular localization studies indicated cytoplasmic localization of OsCBSCBS3. Notably, OsCBSCBS3was found to possess binding affinity towards AMP and ATP, with higher affinity for AMP than ATP and could alsohydrolyze ATP. Heterologous expression of OsCBSCBS3 in E. coli led to enhanced tolerance of bacterial cells towardssalinity and oxidative stress. Likewise, over-expressing OsCBSCBS3 in plant system also imparted tolerance to transgenictobacco and rice plants towards multiple abiotic stresses. We thus, propose that OsCBSCBS3 acts as sensor of cellularenergy status by binding to AMP and/or ATP and is involved in stress adaptation in plants.

PA016 (IPPC0194)

Physiology and proteome analysis of rice and its mutant under salinity stress

Nita Lakra1*, Khalid Anwar1, Sneh L Singla-Pareek2 and Ashwani Pareek1



Salinity is one of the major environmental constraints that affect the growth and productivity of rice worldwide. Salttolerant mutants are promising varieties for saline soil and they constitute the unique genetic stocks for gene discovery.We have generated several mutant lines by gamma-irradiation of IR64 and screened for their salt tolerance. In order toget better understanding of molecular pathways affected in rice mutant in response to salinity, we have studied theirphysiological characteristics and applied 2D-DIGE approach to explore the proteome changes associated with salt stressresponse. Physiological analysis showed that chlorophyll contents, relative water content and carotenoids were significantlyhigher in mutants under salinity. The level of anti-oxidative enzymes like POD, SOD, APX and CAT showed higher activityin mutant (D100-200) as compared to parent (IR64). Proteomics analysis showed that ~200 protein spots were reproduciblydetected which were sequenced and identified using protein function databases. Gene ontology analysis showed thatthese proteins are involved in metabolism, protein synthesis, photosynthesis and stress response. Some of the proteinswhich showed specifically higher accumulation in mutant as compared to their parent (IR64) were identified as putativeSOD, transcription factors, glycosyl hydrolase, nuclear transport factor, ribosomal proteins, fructose bis-phosphate andRubisco small subunits etc. Few proteins were specifically up-regulated in IR64 and down regulated in leaves tissues ofits mutant such as serine/threonine protein kinases, eukryotic translation initiation factor, OsFBX139 F box protein. Thepossible role of all the identified proteins in salt stress adaptation will be discussed in detail.


123

IPPCongress2015


PA017 (IPPC0198)

Impact of early and delayed transplantation of paddy on physico-chemical, pasting, cooking, textural

and protein characteristics of milled rice

Priyanka Paul1, Amardeep Singh Virdi1*, Parmeet Kaur, Gulshan Mahajan2 and Narpinder Singh1

1Department of Food Science and Technology, Guru Nanak Dev University, Amritsar- 143005, India 2Department of Plant Breeding & Genetics, Punjab Agricultural University, Ludhiana-141004, India


The present study aimed to evaluate the impact of early and delayed transplantation of paddy on physico-chemical,cooking, pasting and protein profiling of milled rice of four cultivars (PR114, PR116, PR118, and PAU201) harvested fromearly (5th June, 15th June) and delayed transplantation (25th June and 5th July) of paddy. At the time of active grain fillingand seed maturation stages, early transplanted paddy experience an average minimum and maximum temperature of32.8 and 22.3°C, against 30.8 and 15°C, respectively, by delayed transplanted paddy. Lightness (L*) value, proteincontent, and lipid content of milled rice was lower while, apparent amylose content, cooked grain hardness, final viscosity,and setback viscosity were higher in the delayed transplanted paddy. Amylose content was positively correlated with finalviscosity, setback viscosity and cooked grain hardness and negatively correlated with breakdown viscosity andadhesiveness of cooked grains. Cooked rice of PAU 201 showed exceptionally higher adhesiveness and lower hardnessas compared to other cultivars, which may be attributed to lower average amylose storage in PAU 201. Protein profilingshowed that the accumulation of prolamin (18 kDa; 16 kDa and 14 kDa polypeptides) was reduced in the milled rice fromPR114, PR116 and PAU 201 upon delaying the transplantation from 5th June to 5th July. The changes in milled rice, whichwere caused by delayed transplanting of paddy, were due to cool night air temperature that affected the accumulation ofstarch, protein and lipids, thereby, influencing the quality of milled rice.

PA018 (IPPC0203)

Exploring the role of AtTAF4b in abiotic stress tolerance

Paomipem Phazang1*, Ulrike Bechtold2, Philip M. Mullineaux2 and Neera Bhalla Sarin1

1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2School of Biological Sciences, University of Essex, Colchester, UK


AtTAF4b is one of the TATA binding protein associated factors constituting the TAFIID, a general transcription factor. InArabidopsis thaliana there are about 21 such TAFs. Recent findings show that each TAF regulates a distinct subset ofgenes in contrast to the earlier belief that they are required for the accurate transcription of all genes. While a large bodyof data is available on TFIID complex in mammalian systems, not much has been elucidated in plants. In this study weventured to investigate the implication of overexpressing the AtTAF4b gene in plants for abiotic stress tolerance. TheAtTAF4b was isolated from Arabidopsis thaliana and was overexpressed in Nicotiana tabacum under the control of CaMV35S promoter. Stable integration of the transgene was confirmed by Southern Hybridization using the gene specificprobe. The expression of the gene at the transcript level was also confirmed by RT-PCR. The transgenic lines of Nicotiana

tabacum overexpressing the gene AtTAF4b showed a significant increase in tolerance to NaCl and mannitol stress ascompared to the untransformed control plants. Although the exact mechanism of how the overexpression of AtTAF4bleads to multiple stress tolerance is not known, our preliminary findings suggest that AtTAF4b may play a role in regulatingthe abiotic stress tolerant genes. Further high through-put studies need to be carried out to have a clear insight into thefunction of the AtTAF4b gene in plants.


124

IPPCongress2015


PA019 (IPPC0204)

Evaluation of management strategies to mitigate adverse effects of waterlogging on productivity and

quality of flue-cured tobacco (Nicotiana tabacum L.)

M. Anuradha1*, D. Damodarreddy2, K. Sivaraju2 and T. Goplakrishnamurthy2

1Plant Physiology, ICAR-Central Tobacco Research Institute, Research Station, Kandukur-522101, India2ICAR-Central Tobacco Research Institute, Rajahmundry-533105, Andhra Pradesh, India


In recent years tobacco crop has been facing a series of cyclonic rains leading to waterlogging and prolonged droughtscoupled with high temperatures. It is predicted that the changes in the climate may lead to extreme weather conditionsincluding floods and droughts or uneven and untimely rains in the future. These changes may result in reduction ofproductivity and quality of tobacco. Though tobacco, can withstand water deficit to a fair degree, it is known to exhibitvulnerability to excess-water or waterlogging. There is need to develop technologies to minimize the negative effects ofwaterlogging on productivity and quality of flue-cured tobacco. In this context, field and pot culture experiments wereconducted to evaluate management strategies (Soil application of KNO

3, Foliar application of KNO

3, Hoagland solution,

Salicylic acid, Putriscine, Kinetin) for their potential to alleviate waterlogging induced adverse effects on tobacco. Thecondition of waterlogging was imposed in both pot and field culture at 60 DAP for 48 hours and after relieving the plantsfrom excess water the plants were treated with different management interventions. Leaves were harvested at maturity,cured and were analyzed for quality. In the pot culture experiment, soil application of KNO

3, foliar spray of kinetin and

putriscine performed better in terms of enabling plants to recover faster. As soil application of KNO3 found better, all the

foliar spray treatments were again tested in combination with soil application of KNO3. Foliar spray of kinetin in combination

with soil application of KNO3 could minimize the adverse effects of waterlogging. These treatments were tested in the field

condition and the results revealed that soil application of KNO3 (5 g on either side of the plant by making holes) in

combination with kinetin spray @ 50 ppm twice at 10 days interval could minimize the adverse impact of excess waterstress on yield and quality to some extent in flue-cured tobacco.

PA020 (IPPC0206)

Role of Glucose Auxin responsive gene GAX2 towards thermotolerance in model plant system Arabidopsis

thaliana

Mohan A. Sharma*, Brihaspati N. Shukla and Ashverya Laxmi

Plant Signaling, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Glucose-Auxin and heat responsive gene GAX2 plays role in thermotolerance. The subcellular localization of GAX2 asdetermined using YFP was found to be in cytosol and nucleus. The expression levels of GAX2 was examined in glucosesignaling mutants and it was found that expression of GAX2 was not dependent on HXK1 dependent and independentsignaling pathway. Overexpression lines of TOR showed increased expression of GAX2 mRNA whereas overexpressionand RNAi lines of KIN10 exhibited decreased and increased GAX2 expression. Data extracted from Genevestigator alsoshowed reduced transcript levels of GAX2 in TOR RNAi lines which further confirmed the plausible involvement of energymediated glucose signaling. Kinetics of GAX2 expression in response to starvation and energy enriched condition revealedthe role of energy signaling in GAX2 regulation. Furthermore, Cis-regulatory elements analysis indicated that 942bppromoter of GAX2 contains several heat shock elements (HSE and CCAAT) and other stress-response elements.Quantitative measurement of chlorophyll content and seedling survival assay revealed stronger thermotolerance in GAX2overexpression lines than wild type. Bio-computational analysis through Genevestigator showed reduced expression ofGAX2 in HsfA1 quadruple mutant (KO-hsfa1a/b/c/d). Besides, overexpression of GAX2 exhibited up-regulation of thoseHsp genes, which are also up-regulated by glucose and HsfA1a, which may lead to stronger thermotolerance. Takentogether, these results suggest that glucose-mediated TOR kinase signaling control GAX2 expression level, which eventuallyregulates thermotolerance response in Arabidopsis seedlings.


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PA021 (IPPC0210)

Pseudomonas putida regulates physiological, biochemical and molecular mechanisms of Cicer arietinum

L. plants for coping with short-term drought stress and recovery

Charu Lata* and Shalini Tiwari

Division of Plant-Microbe Interaction, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India


Drought is one of the most important environmental stress factors that adversely affect plant growth and yield. However,some plant growth promoting rhizobacteria (PGPRs) are known to improve plant health and promote growth duringabiotic stresses. Present study suggested the potential role of Pseudomonas putida MTCC5279 (NBRIRA) in droughtstress amelioration in two contrasting chickpea varieties namely, cv. BG-362 (desi) and cv. BG-1003 (kabuli) under bothin vitro and greenhouse conditions. Drought stress significantly affected various growth parameters, water status, membraneintegrity, osmolyte accumulation, reactive oxygen species (ROS) scavenging ability and stress responsive gene expressionsat all stress durations, which were positively attuned upon application of NBRIRA in both chickpea cultivars. Whencompared to control, NBRIRA inoculation also helped in better recovery of both chickpea types. Quantitative (qRT) PCRanalysis showed differential expression of genes involved in transcription regulation (DREB1A and NAC1), stress response(LEA and DHN), ROS scavenging (CAT, APX, GST), and ethylene biosynthesis (ACO and ACS) in both desi and kabulichickpea subjected to drought stress and recovery with or without NBRIRA. Our results thus indicate that NBRIRA confersdrought tolerance in chickpea by altering various morphological, physiological and biochemical parameters, as well as byregulating differential expression of at least 9 stress responsive genes. To the best of our knowledge, this is the first reporton detailed analysis of plant growth promotion and stress amelioration in one-month-old desi and kabuli chickpea subjectedto short-term drought stress and subsequent recovery in the presence of a PGPR.

PA022 (IPPC0212)

Synchronization of antioxidant defense machinery by facilitation of redox state equilibrium in Naringenin-treated

and Funneliformis mosseae inoculated-Cicer arietinum L. genotypes under salt stress

Priyanka Singla * and Neera Garg

Department of Botany, Panjab University, Chandigarh - 160014, India*Presenting author: [email protected]

Under prolonged salinity, equilibrium between production and scavenging of reactive oxygen species (ROS) is overriddenby oxidative burst in legume plants. Arbuscular mycorrhizal (AM) symbiosis as well as flavonoid application helps plantsto thrive in saline soils by countering stress induced redox imbalance of plants. However, probable cross-talk betweenAM and flavonoid and their relative influence in upholding cellular redox stability in stressed plants is an aspect that hasnot been explored hitherto and thus necessitates its in-depth investigation. Consequently, fully factorial experimentswere designed to investigate the potential role of naringenin (Nar, 4 ìM) and AM (Funneliformis mosseae) in transformingantioxidant network of two salt stressed (0, 40, 60, 80, 100 mM NaCl) Cicer arietinum L. genotypes (PBG 5, DCP 92-3).Despite increase in primary antioxidants, Na+ accumulation-induced ROS build-up was coupled with lowering of ascorbate/dehydroascorbate (AsA/DHA), reduced/oxidised glutathione (GSH/GSSG) and flavonoid content, more significantly inDCP 92-3 than PBG 5. In Nar treated and /or AM plants, increased antioxidants efficiently attenuated oxidative loads, withmaximum redox stability attained by +Nar+AM plants. Improved efficacy was related with elevated recycling of reducedglutathione and ascorbate alongwith improved flavonoid concentration, thereby facilitating higher H

2O

2 scavenging and

subsequently higher O2•- dismutation. However, PBG 5 was more responsive to Nar treatment and/or AM inoculation and

displayed better redox equilibrium than DCP 92-3. Study suggested that in-order to improve chickpea growth and curtailsalt induced oxidative constraints; shifting redox towards more reduced form constitutes an important defensive mechanismadopted by Nar and F. mosseae.


126

IPPCongress2015


PA023 (IPPC0213)

Effect of high temperature stress on physiological attributes, antioxidative enzymes and expression of

heat shock proteins in wheat (Triticum aestivum L.)

Narendra Gupta1*, Aarif Khan2 and Radheshyam Sain1

1Rajasthan Agricultural Research Institute, Durgapura- Jaipur-302018, India2Plant Biotechnology Centre, SK Rajasthan Agricultural University, Bikaner, India

Presenting author: [email protected]

Experiment was conducted to study the effect of heat stress on various parameters in five diverse wheat genotypes atseedling stage. Seeds were sown in plastic pots in laboratory at 25±2°C. For heat treatment, 20 days old seedlings wereexposed to 36°C/42°C for 6 hours. Results showed that the heat stress significantly reduced the dry weight and length ofseedlings in all the genotypes. Chlorophyll content and membrane stability index decreased but proline content increasedin all the genotypes under stress conditions. Among genotypes, Raj 4083 exhibited better heat tolerance, particularly at42°C. Activities of SOD, CAT, APX and POX increased significantly in all the genotypes but per cent enhancement wasmaximum in Raj 4083 and minimum in PBW 502. In SDS-PAGE protein profiling, 15 polypeptides were recovered incontrol plants with slight variations in temperature treated plants of most genotypes. Proteins with molecular weights of85 kDa and 15 kDa were constitutively expressed in all the genotypes but proteins of 45 kDa were expressed only intolerant genotypes. Expression analysis of heat shock proteins, particularly hsp 90 further indicated the existence oftolerance mechanism in Raj 4083. Thus, using a multi parametric approach, Raj 4083 was found most tolerant and PBW502 most susceptible to heat stress. The study also provides evidence that antioxidant enzymes like SOD, CAT, GPX,APX and heat shock proteins like HSP 90 have association with heat tolerance in wheat and thus can be used as efficientmarkers for heat tolerance at seedling stage.

PA024 (IPPC0214)

Development of drought tolerant variety in potato: present status and way forward

Devendra Kumar1*, J.S. Minhas2, V.K. Gupta1, S. Rawal1, Name Singh1 and Brajesh Singh3

1Crop Physiology Biochemistry & PHT, Central Potato Research Institute Campus, Modipuram-250110, Meerut, India2Central Potato Research Station, Jalandhar, India3Central Potato Research Institute, Shimla-171001, India*Presenting author: [email protected]

Potato is highly sensitive to water stress. Current potato varieties have been bred for optimum inputs viz fertilizers,irrigation, temperature, soil type etc. Minor scarcity of any input therefore, offers substantial challenge for sustaining theproductivity. Considering future water scarcity, a programme to breed drought tolerant varieties was started in 2005.Besides field screening of existing varieties, hybrids and exotic clones; carbon isotopes discrimination, proline profile etcwere also considered in the selection of parents. Established protocols were followed for breeding and selection ofdesired tubers traits in subsequent generations. Promising hybrids were evaluated for leaf area pattern, photosynthesisrate and associated parameters, antioxidant defense mechanism, leaf senescence, tuber yield and tuber defects underreduced water regimes (25 % less). Four seasons (2010-11 to 2013-14) data at Modipuram showed that hybrid WS/05-146 had >33.0 t ha-1 mean tuber yield as compared to 27.9 t ha-1 in Kufri Bahar (popular high yielding variety) which isdistinctly higher (>18%) under mild water stress conditions and recorded highest Drought Tolerance Index (DTI) andlowest Drought Susceptibility Index (DSI). Similarly under well watered conditions (control), it had 36.6 t ha-1 (>15%)mean tuber yield as compared to 31.6 t ha-1 in Kufri Bahar. Consequently, WS/05-146 has qualified for multi-location trialsunder All India Co-ordinated Research Project-Potato (ICAR) and will be evaluated during 2015-16 at Bhubaneswar,Deesa, Faizabad, Gwalior, Hisar, Kota, Pune and Raipur. Further, efforts are under way to enhance N economy throughimproved agro-technology.


127

IPPCongress2015


PA025 (IPPC0215)

In vitro drought induced changes in antioxidant systems of Rauwolfia serpentina

Narendra Kumar*, Deepika Sharma, S.C. Shankhdhar and Deepti Shankhdhar

Department of Plant Physiology, College of Basic Sciences and Humanities, GBPUA&T, Pantnagar, Uttarakhand, India


The present study conducted in department of plant physiology, GBPUA&T, Pantnagar to study the in vitro droughtinduced changes in antioxidant systems of (Sarpagandha) Rauwolfia serpentina. It is endangered woody perennial medicinalshrub, belongs to apocynaceae family, having various secondary metabolite as well as enzymatic and non enzymaticantioxidant to prevent various disease like high blood pressure, mental disease, sleeplessness and several other disease.Due to their poor seed germination percentage, tissue culture technique is very useful method for rapid multiplication anddrought induced study. In our experiment the leaf derived callus induction and proliferation of R. serpentina (variety Cim-sheel) observed best in MS media supplemented with 2,4 D (2 mg/l) and BAP (0.5 mg/l). Drought stress was given toproliferated leaf derived callus under different concentration of PEG (1%, 2% and 3%) along with 2,4 D (2 mg/l) and BAP(0.5 mg/l). Proliferation of callus under different concentration of PEG observed in 15, 30 and 45 day after inoculation.Tissue water content was found to be decreased while enzymatic activity of antioxidants i.e. SOD and CAT was enhancedwith increasing PEG treatments in comparison to control. In case of non enzymatic antioxidant like flavonoids contentalso increased with increasing level of drought in different interval of days. So above results indicate that Sarpagandhagood source of antioxidant under drought stress conditions. The use of different concentration of PEG helps to measurethe level of antioxidant and making the drought tolerant genotype through tissue culture technique.

PA026 (IPPC0221)

Thermotolerance in wheat (Triticum aestivum L.) in relation to gibberellic acid

Shivani Nagar1, R. Dhakar2, Neera Singh3, Ajay Arora1 and V.P. Singh1*

1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Agricultural Chemicals, Indian Agricultural Research Institute,New Delhi-110012, India*Presenting author: [email protected]

Most of the wheat growing areas of the world experience water deficit and high temperature stress. Recent report ofIntergovernmental Panel on Climate Change (IPCC 2014) has indicated an increase of 4.80C from pre-industrial temperatureby the end of the century. It will result in exposure of reproductive and ripening phases of wheat to terminal heat stress,leading to severe drop in productivity. In the absence of adaptation and CO

2 fertilization benefits, a one degree celsius

rise in temperature alone could lead to a decrease of six million tonnes of wheat production. Paclobutrazol (PBZ) a plantgrowth regulator is inhibitor of gibberellic acid (GA) biosynthesis. PBZ also confers the thermotolerance to the cropplants. Hence, we hypothesize that lower level of GA should provide thermotolerance. Four wheat cultivars were sown atdifferent dates to create temperature variation at reproductive phase. GA and PBZ were applied exogenously. Heatstress induced lipid peroxidation, membrane deterioration and reactive oxygen species production, and this trend wasreversed by PBZ but not by GA. Activities of antioxidant enzymes like superoxide dismutase (SOD), glutathione reductase(GR), ascorbate peroxidase (APX) and catalase (CAT) were enhanced in all the cultivars. Application of PBZ enhancedactivities of all these enzymes, whereas, GA enhanced the activity of SOD and APX only. We also studied the expressionof heat shock factors (HSFs) and heat shock proteins (HSPs). Application of paclobutrazol enhanced the expression ofHSF A1, HSP 101, HSP 70 and HSP 17.8. Among the cultivars highest expression of HSP’s and HSF’s was observed inC-306. GA enhanced the endogenous level of GA, while PBZ decreased it. Endogenous GA

3 levels in plants showed

negative correlation with MSI, yield/plant, test weight and SOD activity and positive correlation with lipid peroxidation,ascorbate peroxidase and H

2O

2 content.


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PA027 (IPPC0224)

In vitro drought induces oxidative stress and its impact on phenol and flavonoid content in callus of

Withania somnifera

Deepika Sharma, Narendra Kumar, Shailesh Chandra Shankhdhar and Deepti Shankhdhar*

Plant Physiology College of Basic Sciences & Humanities, G.B. Pant University of Agriculture & Technology, Pantnagar-263145,Uttarakhand, India


An important herbal plant Withania somnifera (genotype J 20) was evaluated for its growth, water content, necrosis,phenol and flavonoid changes in callus tissue under PEG induced drought. Leaf derived callus were initiated on Murashigeand Skoog medium supplemented with 1.5 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D), 0.20 mg L-1 kinetin (Kn), andsubcultured on same media containing different concentrations of Poly Ethylene Glycol (1-3%). After 4 weeks of culture,callus growth and water content were determined. The results showed that increasing PEG concentration in the mediumcauses a gradual decrease in growth, water content of callus and create necrosis. However, in between the black necroticcallus, whitish embryogenic mass of cells were observed. The tolerant cells were again proliferated on stress media andused for further study of phenol, flavonoid and MDA content. It was observed that higher concentrations of PEG alongwith longer exposure (up to 45 days) induces oxidative stress in surviving callus and resulted in increased phenolics,flavonoid and malondialdehyde (MDA), which was positively correlated with an increase in the accumulation of PEG.Increasing concentration of phenols and flavonoids in the tissue act as scavengers of free radicals and also prevent theirformation by chelating metals.

PA028 (IPPC0230)

Isolation and characterization of SOS pathway genes from a halophyte, Oryza coarctata

Showkat Ahmad Ganie* and Tapan Kumar Mondal

Division of Genomic Resources, National Bureau of Plant Genetic Resources, IARI Campus, Pusa, New Delhi-110012, India


Salinity is an important abiotic stress which limits the agricultural production world-wide. Rice is regarded as a traditionallysalinity sensitive crop. However, one of its wild relative species, Oryza coarctata, is highly salt tolerant (halophyte) whichgrows in the coastal regions under high saline conditions. It has been demonstrated previously that genes of O. coarctata

can induced salinity tolerance in other crop plants including rice. Thus, we cloned full length sequences of SOS pathwaygenes encoding SOS1, SOS2 and SOS3 from this halophyte. The ORFs were found to be 3353bp, 1370bp and 633bp forSOS1, SOS2 and SOS3 respectively. The deduced amino acid sequences registered cAMP/cGMP binding domain,protein kinase domain and Ca2+ binding domain respectively which showed high degree of homology with correspondinggenes of angiosperm. We further studied three dimensional model of all the genes and quality of predicted models wereanalysed with PROCHECK. Molecular docking of model revealed similar binding pockets for different substances andproducts. All the transcripts were found to salinity stress responsive indicated by qPCR study. Finally, a comparativeanalysis of these gene sequences from susceptible and tolerant glycophtes as well as facultative and obligatory halophytesof individual genotypes of Oryza species revealed interesting differences that might be responsible for differential salinityadaptation.


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PA029 (IPPC0246)

Simultaneous over-expression of Eif4A and Lea2 genes in rice (Oryza sativa cv. PB1): Linking salt and

drought tolerance

Aakrati Agarwal*, Yashwanti Mudgil, K. Chandrasekhar, Subramanyam Chinreddy, Dhirendra Fartyal, Donald Jamesand M.K. Reddy

Plant Molecular Biology, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


Abiotic stress is a huge roadblock in crop-plant productivity worldwide especially in the case of rice (Oryza sativa).Drought and salinity are the most prominent abiotic stresses and are responsible for the major yield loss in rain-fed ricecultivation. Both these stresses are thought to be linked together as they lead ultimately to osmotic stress and the plantresponses in both cases are similar. We developed a strategy to combat against these stresses simultaneously by usingLea2 and Eif4A genes. Lea2 gene is a late embryogenesis abundant protein, which specifically protects plant integrityagainst water loss. Eif4A has a reported role in providing salinity tolerance. To validate their relationship in rice, bothgenes were over-expressed simultaneously in indica rice plant variety PB1. The construct harbouring these two genes inaddition to a plant selectable marker gene hptII and Bar was transformed into rice calli with a virulent Agrobacterium

strain EHA105. The calli were selected in hygromycin B and the presence and stable integration of transgene in putativetransgenic plants was confirmed by PCR for Lea2, Eif4A, Bar and hptII genes and Southern hybridization, which revealedthe copy number in different transgenic events. Transgenic lines having a single and double copy number were selectedfor physiological analysis showing resistance to phosphinothricin. Simulated drought and salinity stress assays revealedan interdependent relationship between the two stresses.

PA030 (IPPC0253)

Cross-genus role of endophytic microorganisms on growth promotion and metal phytoremediation in

an arsenic accumulator terrestrial plant Solanum nigrum

Gairik Mukherjee* and Anindita Seal

Department of Biotechnology, University of Calcutta, Kolkata-700019, West Bengal, India*Presenting author: [email protected]

Endophytes are microorganisms that reside normally inside plant tissues and have been shown to have beneficial impacton the plant. Many of them are known for their growth promoting effects on the host as well as ensure their survival undervarious stress conditions. Among several plants growing in an arsenic contaminated site in West Bengal, a shrub Solanum

nigrum was found to accumulate arsenic in its shoot with a transfer factor greater than 1. When grown under laboratoryconditions, the plant was found to accumulate significant levels of arsenic with a transfer factor greater than 3. Therefore,the plant was chosen to be a suitable terrestrial model for arsenic phytoremediation. Lantana camara is an invasive weedthat can survive environmental extremes. As such, seven arsenic tolerant endophytes were isolated from L. camara

growing in the arsenic contaminated site. All endophytes had phosphate solubilization potential and produced the growthhormone auxin. One of the endophytes could even grow in nitrogen-free medium. All these properties are indicative oftheir probable growth promoting effects. When applied to Solanum nigrum under arsenic treated conditions, the endophyteconsortium of L. camara was found to exhibit growth promoting effects on the plant. The consortium was also found toincrease the leaf chlorophyll content related to growth promotion by enhanced photosynthesis. Besides, application ofthe consortium resulted in enhanced removal of arsenic from the soil and increased root to shoot arsenic translocationsignifying suitability of the system in microbe- assisted phytoremediation of arsenic.


130

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PA031 (IPPC0258)

Impact of high temperature and elevated CO2 in blackgram genotypes (Vigna mungo (L.) Hepper)

H. Vijayaraghavan*, M. Pandiyan and C. Partheeban

Department of Crop Physiology Tamil Nadu Agricultural University Coimbatore-641003, India


Investigation was carried out to screen the black gram genotypes for high temperature tolerance and to evaluate elevatedCO

2-high temperature interaction. A protocol for rapid screening for high temperature was also developed namely, the

temperature induction response (TIR) technique. Evaluation of black gram genotypes for heat tolerance under naturalfield condition indicated that six genotypes, VBG-06-002, VBG-06-005, VBG-06-010, VBG-07-001, VBG-10-008 andCOBG-759 have highest yield components than the rest of the genotypes. These six genotypes fall under the category ofheat tolerant, eight genotypes under moderately heat tolerant and five under the heat susceptible group. TIR at seedlingstage exactly matched with that of field experiment as well as studies conducted under controlled environment chamber,suggesting the usefulness of TIR for screening for heat tolerance. The impact of elevated carbon dioxide and temperatureon select six black gram genotypes was evaluated in the carbon dioxide chamber. The results showed that carbondioxide concentration at 700 ppm under ambient temperature seems to be optimum in producing more number of flowerswith consequent high fertility coefficient. Interestingly the two genotypes, VBG-06-002 and VBG-07-001 were identifiedas highly heat tolerant ones and deserve further research. These genotypes are being forwarded to the breeding programsand for further progeny selection.

PA032 (IPPC0275)

Sorghum improvement for abiotic stress adaptation and climate change resilience in dryland conditions

S.S. Rao1*, H.S. Talwar1, B. Subbarayaudu1, N. Kannababu1, C. Aruna1, K. Hariprasanna1, T.G.N. Rao1, Prabhakar Bhat2,P.V.V. Prasad3 and J.S. Mishra4

1ICAR-Indian Institute of Millets Research, Rajendranagar, Hyderabad-500030, India2Centre on Rabi Sorghum (IIMR), Solapur, Maharashtra, India3Kansas State University, KS, Manhattan, USA4ICAR-ICAR Research Complex for Eastern Region, Patna, Bihar, India


Sorghum (Sorghum bicolor (L.) productivity is reduced due to the occurrence of drought and heat stresses in pre-and postflowering stages. Current research aimed at 1) quantifying the impact of drought (D), high temperature (HT) and combinedstress (D*HT) on sorghum physiological traits and 2) assessing the climate effects on bioenergy sorghum quality. Treatmentsimposed in growth chambers 1) control [Irrigation + Optimum temperature (OT)(32/22 degrees C day and night, resp.]; 2)drought stress [drought for 20 days + OT (32/22 d C)]; 3) HT stress [Irrigation+ HT (38/28 d C)]; and 4) combined stress[drought + HT (38/28 d C)]. Leaf area decreased by 25% in D *HT stress over OT. Average leaf temperature increased by8.0, 5.0, 2.5% resp., in combined, HT and D over OT. Root length and root -shoot ratio significantly increased by 11.0%in combined stress over optimum. Leaf water content decreased by 16.1% and 35.0% respectively, in D and D*HT overOT. Across rainy season plantings, sweet sorghum CSH22SS produced 29.0% more bioethanol than SSV84, whileSSV74 yielded 16.6% more than SSV84. CSH22SS produced higher stalk, grain, sugar and bioethanol yields over SSV84or SSV74. Both CSH22SS and SSV74 were more stable across plantings and showed climate resilience. In post rainyseason, stalk yield decreased by 26-36% in late plantings over early. Based on stalk yield and sugar quality, PAC52093and CSV19SS were stable across plantings and were climate adapted. The implication of these results is discussed forgenetic enhancement of sorghum for climate change resilience.


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PA033 (IPPC0276)

Dehydration-responsive extracellular matrix of chickpea identifies CaFer1, a secretory ferritin that

promotes stress tolerance and growth

Shaista Parveen*, Deepti Bhushan and Niranjan Chakraborty


*Presenting author:[email protected]

Plants strictly regulate iron homeostasis to avoid iron deficiency which can impair key metabolic functions, and ironexcess that produce damaging reactive oxygen. As key components of iron homeostasis, ferritins can store up to 4500Fe (III) atoms in its central cavity and can release the stored iron when the need arises. However, the functions ofphytoferritins, particularly in stress responses, remain poorly understood. In a previous proteomics study, we identified anarray of stress-responsive proteins in the chickpea extracellular matrix that include ferritin. Here we describe the structure,expression and stress-responsive function of the chickpea ferritin, designated CaFer1. Water-deficit conditions triggerCaFer1 expression indicating its dehydration-responsive function, besides its role against Fusarium oxysporum infection.Functional complementation of the yeast frataxin-deficient mutant Deltayfh1 indicates that CaFer1 also functions in oxidativestress. CaFer1 expression increases in response to iron deficiency, and CaFer1-overexpressed seedlings showed alteredexpression of iron-responsive genes and improved growth and development. The mechanism of functions of secretedferritins is well characterized in insects and animals, but no secreted ferritin has yet been identified in plants. Thecompartmentalization of CaFer1 to the extracellular matrix and the secretome, reported here for the first time, indicatesthat its function differs from that of other phytoferritins. CaFer1 interacts with the iron transporter IRT1, suggesting adistinct role in maintenance of iron homeostasis. Our results suggest that CaFer1 plays key role in iron-buffering andstress tolerance.

PA034 (IPPC0277)

Influence of stress mitigating compounds on biophysical parameters and yield attributes under drought

in groundnut (Arachis hypogeal L.)

R.P. Patil1*, M. Harish1, A. Amaregouda1, M.M. Dhanoji1 and M.R. Umesh2

1Department of Crop Physiology, 2Department of Agronomy, University of Agricultural Sciences, Raichur-584133, India


Field experiment was conducted during the year 2014-15 at college of Agriculture, Raichur to investigate the appropriatetechnology to overcome the ill effects of water stress on growth and development of groundnut crop. Water stress plotswere treated with various stress mitigating compounds, significantly higher pod yield was recorded with application oftricontanol @ 2.0 ml/l (28.3 q/ha) followed by KCl @ 2.5% (26.9 q/ha) methanol (2%). Biophysical parameters TR wereshown maximum in tricontanol and minimum diffusive resistance was seen in same treatment. Whereas, among thestressed plot RWC was found more in tricontanol followed by KCl (76.6%). At 45 DAS, Finally yield deciding parameter inphotosynthetic rate was showed significant differences between the treatments. Photosynthetic rate (31.87 μmol CO

2

m-2 s-1) was higher in unstressed plot wherein continuous irrigation without stress. Among the different stress mitigatingcompounds foliar application of tricontanol has recorded significantly higher photosynthetic rate (30.13 μmol CO

2 m-2 s-1)

followed by Nitrobenzene (30.03 μmol CO2 m-2 s-1) Methanol (28.33 μmol CO

2 m-2 s-1) Salicylic acid, Kaolin and lower in

stressed plots. Finally the yield components No. of pods plant-1, Pod weight (g), pod yield (q ha-1) and harvest Index (%)were studied and maximum yield was recorded in the foliar spray of tricontanol (28.3 q ha-1) followed by KCl (26.9 q ha-1)and methanol (6%) (26.6 q ha-1).


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PA035 (IPPC0281)

Screening of cotton (G. hirsutum) genotypes for water stress tolerance underrainfed conditions

D.G. Dalvi*, D.B. Deosarkar and K.S. Baig

Department of Agricultural Botany, Vasantrao Naik Marathwada Agricultural University, College of Agriculture, Parbhani-431402,Maharashtra, India


A field experiment was undertaken during kharif season for the period of eight years (2007-08 to 2014-15) with an objectto screen cotton genotypes for water stress tolerance under protected and unprotected conditions, 155 genotypes ofcotton (G. hirsutum) were screened, out of which the genotypes viz., KH-138, Sahana, NH-615, PH-1009, RCR-102,HBB-101, GTHV-02145,CPD-817, NH-630, WGHH-411, GTHV-0135, BS-30, ARBH-813, NH-635, GBHV-175, PH-1075,CSH-1111, GBHV-162 WDLH-1938, RHC-0717, TSH-04-115, RAN-806, BS-37 and BS-39 were found tolerant to waterstress under rainfed conditions on the basis of physiological parameter i.e. high degree of relative water content (%),proline content (%), leaf area index, specific leaf weight and stress indices viz., dry matter stability index, leaf areastability index, plant height stability index, yield stability index with least (<1.0) drought susceptibility index (S) and chlorophyllstability index. Similarly, these genotypes also recorded highest seed cotton yield (kg/ha) with least reduction (%) in yieldunder rainfed condition. These genotypes may be used for breeding purpose in further development of drought tolerancegenotypes in cotton.

PA036 (IPPC0290)

Association of ascorbic acid with drought tolerance in wheat

Suman Roy1*, Ajay Arora2 and V.P. Singh2

1ICAR-Vivekanand Parvatiya Krishi Anusandhan Sansthan, Almora-263601, Uttarakhand, India2Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India


Wheat is one of the premier cereal crops in India but confronts a severe threat to sustainable production due to waterdeficit stress. Plants have evolved various strategies to overcome the adverse effects of such stresses. One of thestrategies developed by plants to fight such stress includes generation of antioxidants as a response to stress. Evidencesconfirm that ascorbic acid (AA), a non enzymatic antioxidant, is one of the major antioxidant that respond to both bioticand abiotic stresses. The biological activity of AA is defined by its oxidation and subsequent regeneration into the reducedform. However, phenotyping of genotypes on the basis of any antioxidant has not been done till date. In this study, wehave screened 30 wheat genotypes on the basis of endogenous reduced ascorbic acid content under water deficitconditions. A wide genotypic variability for endogenous reduced ascorbic acid content was observed under control andstress condition. Endogenous reduced ascorbic acid content decreased under stress conditions in most of the genotypes.Results of Drought Susceptibility Index (DSI), Relative Water Content (RWC) and Membrane Stability Index (MSI) confirmedthat susceptible genotypes have a lower content of endogenous reduced ascorbic acid as compared to the tolerantgenotypes. In addition lower endogenous reduced ascorbic acid under stress condition was due to restriction of stressinduced hydrogen peroxide levels. Endogenous levels of the other antioxidants may provide a clear picture which can beused as a trait for phenotyping genotypes under water deficit condition.


133

IPPCongress2015


PA037 (IPPC0293)

Overexpression of a stress responsive MuWRKY3 transcription factor leads to enhanced drought tolerance

in groundnut (Arachis hypogaea L.)

K. Kiranmai, A. Nareshkumar, V. Amarnatha Reddy, O. Sudhakar Babu, G. Lokanadha Rao, U. Lokesh, B. Venkateshand Chinta Sudhakar*

Plant Molecular Biology Unit, Department of Botany, Sri Krishnadevaraya University, Anantapur-515003, India


WRKY transcription factors are plant specific transcription factors regulates several developmental events and stressresponses in plants. The WRKY domain is defined by the conserved amino acid sequence named WRKYGQK at its N-terminal end and Zinc finger like motif. WRKY genes often respond to several stress factors which may act as negative orpositive regulators. The aim of present study is to clone, characterize and overexpress WRKY3 gene from horsegram(Macrotyloma uniflorum) a dry-land drought tolerant pulse crop. To this direction, we have successfully amplified MuWRKYgene from horsegram using cDNA with heterologous primer sequences (#EU375350.1). PCR product was purified andcloned into pTZ57R/T vector and sequence confirmed. The nucleotide sequence of the full-length CDS (MuWRKY3)comprised 1476 bp; the deduced protein showed 491 amino acids with an apparent molecular weight of 53.73kD and thetheoretical pI is 6.12 (#KM520390.1). Phylogenetic tree analysis revealed that gene is related to GmWRKY3. Conserveddomains analysis revealed the presence of two conserved WRKY domains and gene belongs to group I WRKY family.Further, MuWRKY3 gene was cloned into plant transformation vector pCAMBIA2301 under the control of CaMV 35Spromoter. Subcellular localization studies in tobacco epidermal cells showed the presence of MuWRKY3 in the nucleus.In planta transformation of groundnut carrying MuWRKY3 was standardized using pCAMBIA2301:MuWRKY3 as binaryvector. Putative groundnut tranformants were screened on a medium containing 200 mg/L kanamycin. T1 plants wereanalyzed for molecular and physiological parameters to confirm transgenic groundnut plants. T2 plants demonstratedincreased activity of antioxidative efficiency and osmolite accumulation over wild plants.

PA038 (IPPC0305)

Identification of high temperature stress-responsive proteins in bread wheat (Triticum aestivum L.)

Divya Mishra*, Shubhendu Shekhar, Subhra Chakraborty and Niranjan Chakraborty



Ambient temperature of earth is drastically increasing due to the effect of global warming. The IPCC report (2014) suggestsan approximate increase of 4-5ºC by the end of 21st century. This elevation in temperature has certainly a negative impacton the growth and metabolism of plant, and crop production. Keeping this in mind, we sought to know how plant respondsto high temperature stress and the underlying mechanism of tolerance. Bread wheat is one of the most important cropspecies, with global annual production currently over 600 million tonnes. It is a winter season crop and hypersensitive tohigh temperature stress. Wheat simulation model predicted that the impact of high temperature stress is more severethan that of dehydration for wheat. Most of the molecular understanding of high temperature stress-responsive cellularadaptation has evolved from transcriptome analyses. However, comparative analysis of high temperature stress-responsiveproteins, particularly proteins in the subcellular fraction, is limiting. We screened the status of thermotolerance of severalvarieties of bread wheat using different physiological and biochemical indexes, and selected relatively tolerant and sensitivecultivars for comparative proteomics analysis. This study would reveal differentially regulated organellar proteins,presumably involved in cellular adaptation and impinge on the molecular mechanism of thermotolerance in wheat. Thefindings may also provide the basis for targeted alteration of metabolic routes for effective engineering strategies in cropspecies for agricultural benefits.


134

IPPCongress2015


PA039 (IPPC0312)

Amelioration of salt stress by exogenous application of salicylic acid and arbuscular mycorrhiza in

Cicer arietinum L. (chickpea) genotypes

Neera Garg and Amrit Bharti *

Department of Botany, Panjab University, Chandigarh-160014, India


Soil salinity is one of the major environmental stresses affecting crop production worldwide. Cicer arietinum L. (chickpea)is the third most important food legume grown in the world and is considered highly sensitive to salt stress. Salicylic acid(SA) is a well known phenolic compound and is extensively studied as a growth regulator, since it plays a key role ineliminating the adverse effects of abiotic stresses. In addition, arbuscular mycorrhizal (AM) fungi have also been reportedto have a great potential to reduce negative effects of salt stress. Therefore, a study was conducted to investigate theimpact of exogenous application of Salicylic acid (0.5 mM) along with arbuscular mycorrhiza on AM colonization, ionicstatus, growth and yield under various concentrations of NaCl (0, 4, 6, 8 ds m-1) in two chickpea genotypes (salt-tolerantPBG-5, salt-sensitive BG-256). Salinity negatively affected plant growth, mycorrhizal colonization, K+/Na+, Ca

2

+/Na+ ratiosleading to decline in plant productivity. AM inoculation significantly improved root and shoot dry weights, nutrient status(P, K), chl a/b ratio, carotenoid, anthocyanin contents, reduced Na+ uptake and increased ultimate yield. SA enhanced %AM colonization and further improved the growth and productivity when given in combination (SA+AM). PBG-5 displayedbetter ability to colonize and ensured better nutrient status and growth, which manifested itself in terms of significantlyhigher seed yield as compared to BG-256. The study concluded that salicylic acid along with arbuscular mycorrhizalinoculation can be applied as a usefull strategy to ameliorate salt stress in chickpea for sustainable agriculture.

PA040 (IPPC0324)

Improved drought resistance in a wheat stay green CMS line is associated with enhanced antioxidant

competence compared to its fertile line under water deficit stress

Renu Khanna-Chopra and Vimal Semwal*

Stress Physiology Lab, Water Technology Centre, Indian Agricultural Research Institute, New Delhi-110012, India


Reproductive sink regulates monocarpic senescence in wheat as desinking delays senescence. Wheat fertile (HW 2041)and its isonuclear male sterile line (CMS) were subjected to post-anthesis water deficit stress (WDS) to understand theassociation between sink strength, oxidative stress, antioxidant defense and drought resistance. CMS plants maintainedbetter water relations and exhibited delayed onset and progression of flag leaf senescence in terms of green leaf area,chlorophyll and protein content than fertile plants. Stay green character in CMS plants under WDS was associated withless ROS generation, lower damage to membranes and better antioxidant defense both in terms of antioxidant enzymeactivities and metabolite content compared to fertile plants. CMS plants also exhibited delay in expression of senescenceassociated genes i.e. WRKY53, serine and cysteine protease (WSP and WCP2), glutamine synthetase (GSI and GSII)and catalase under WDS compared to fertile plants. CMS line exhibited better antioxidant defense in chloroplasts than itsfertile line. In conclusion stay green trait contributed to improved drought resistance (DR) in wheat. Reproductive sinkregulated drought response as CMS wheat plants exhibited stay green character and improved DR coupled with enhancedantioxidant competence than its fertile line under WDS.


135

IPPCongress2015


PA041 (IPPC0325)

Performance of sugarcane varieties under different moisture stress in subtropical zone

Pooja Dhansu1*, A.S. Nandwal1, Mehar Chand2, Ashwani Kumar3 and Anita Kumari4

1Department of Botany & Plant Physiology, CCS Haryana Agricultural University, Hisar-125004, Haryana, India2Regional Research Station, CCS HAU, Karnal-132001, Haryana, India3ICAR-Central Soil Salinity Research Institute, Karnal-132001, Haryana, India4Department of Chemistry & Biochemistry, CCS HAU, Hisar-125004, Haryana, India


A field experiment was conducted at Regional Research Station, CCSHAU, Karnal during 2014-15 with the objective toevaluate the effect of different soil moisture regimes (during pre-monsoon period) on growth and yield of sugarcanevarieties. The experiment consisting of three moisture regimes based on available soil moisture (ASM), i.e., 50 % ASM(control), 40% ASM (mild stress) and 30% ASM (severe stress) in main plot and four commercial varieties, i.e., CoS 767,CoH 128 (mid late) and CoJ 64, Co 0238 (early) in sub plot was laid out in split plot design with three replications. The soilmoisture stress was started after 45 days of planting (DAP). Significant differences were observed among varieties forgermination, growth and yield parameters. Sugarcane variety CoJ64 showed higher germination (64.2%), followed byCoS 767, CoH 128 and lowest in Co 0238. A significant reduction in stalk height (up to 32-43 %), number of tillers (11-24%), millable canes (up to 24%), cane length, cane growth and cane yield were observed in different varieties underwater stress treatment at 30 and 40% ASM as compared to 50% ASM (control). Highest numbers of tillers were recordedin the first week of July. Total chlorophyll content significantly decreased (up to 27%) in stressed plants as compared tocontrol plants. Highest numbers of tillers were recorded in variety CoH 128, followed by CoS 767, CoJ 64 and lowest in Co0238 in the month of July. Highest numbers of millable canes were recorded in CoS 767, followed by CoH 128, Co 0238,and lowest in CoJ 64.

PA042 (IPPC0328)

Differential response of medium duration pigeonpea genotypes to water deficit stress at flowering

N. Jyothi Lakshmi*, M. Vanaja, S.K. Yadav, Amol Patil, P. Sathish, Ch. Ram Prasad, Jainender, K. Salini, Arun K.Shanker, B. Sarkar, M. Maheswari and Ch. Srinivasa Rao

Central Research Institute for Dryland Agriculture, Santhoshnagar-500059, Hyderabad, India


Four medium duration pigeon pea genotypes (Asha, GT-1, Maruti, Vipula) were evaluated at flowering stage in fieldunder well watered and water deficit stress conditions to assess the effect of water-deficit stress and its relief on physiologicaland biochemical parameters including the assay of antioxidant enzymes to ascertain their role in stress tolerance. Waterdeficit reduced the leaf area per plant in all genotypes. The percent reduction in seed yield with water deficit stress was46.2, 57.4, 50.4, 54.3% and drought tolerance efficiency (DTE) was 53.8, 42.6, 49.5, and 45.7 for genotypes Asha, GT-1, Maruthi and Vipula respectively. The drought susceptible index (DSI) was 0.90, 1.11, 0.97 and 1.05 for genotypesAsha, GT-1, Maruthi and Vipula respectively. Genotype Asha was found to be less sensitive to moisture stress conditionfor days to 50% flowering as well as maturity. The genotype Asha was found to be promising for total dry matter productionunder moisture stress (61.8 g/plant) as well as irrigated (115.2 g/plant) conditions. Genotype Asha followed by Marutiwere superior over Vipula and GT-1 in terms of leaf water relations, accumulation of compatible solutes, photosynthesis,WUE, membrane stability index and activity of antioxidant enzyme viz., superoxide dismutase (SOD). Genotype Asharecorded highest grain yield and total dry matter, low reduction in yield due to water stress, high DTE, low DSI, lessreduction in HI under stress. Maintenance of superiority in terms of these physiological and biochemical parameterscoupled with better recovery ability upon stress relief are crucial physiological mechanisms contributing to water deficitstress tolerance in the genotype Asha.


136

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PA043 (IPPC0343)

Over-expression of BrHsf3 helps in multiple stress tolerance in Brassica juncea (Indian mustard)

Subhash Chandra Prasad1*, Philip Mullineaux2, Ulrike Bechtold2 and Neera Bhalla Sarin1

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2University of Essex, UK


Heat-shock transcription factors (HSFs) are known to play an important role in regulation of expression of heat shockproteins in eukaryotes. Plants have a much higher number of HSFs in comparison to other species and each HSFs haveunique functions in response to environmental stress. Earlier studies indicated that HSF3 acts beyond its previouslyreported function. It seems to stand at the interface of abiotic and biotic stress signalling pathways and thus HSF3overexpression confers multiple abiotic and biotic stress resistance. Brassica species are highly sensitive to heat stressat the flowering stage that causes a decline in seed yield. In the present study Brassica juncea (Indian mustard) wasgenetically engineered by overexpressing BrHsf3 (Brassica rapa; Br, Heat shock factor 3) gene and the morphologicaland physiological characters of the transgenic lines were compared with the untransformed control plants and it wasfound that different transgenic lines showed normal morphology and growth similar to the untransformed (UT) controlplants. Leaf disc senescence assays undertaken for two representative transgenic lines showed increased tolerancewhen exposed to NaCl and mannitol stress. Histochemical staining of transgenic lines exposed to heat stress (35°C for12 hr) showed lower accumulation of hydrogen peroxide (H

2O

2) and superoxide (O

2

-) radicals in leaves. The specificenzyme activity of ascorbate peroxidise (APX) was increased in transgenic lines in comparison to UT under heat stresscondition this could be the reason, less ROS accumulating in transgenic lines in comparison to UT.

PA044 (IPPC0348)

Rice nuclear protein OsNAPL6 is an H3/H4 histone chaperone functioning in response and adaptation to

various abiotic stresses

Amit Tripathi1*, Ashwani Pareek2 and Sneh Singla-Pareek1

1Plant Molecular Biology Group, Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna AsafAli Marg, New Delhi-110067, India2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Understanding the mechanistic details of stress response in plants is a prerequisite to develop superior crop varieties thatmay withstand environmental extremes. Focusing on ‘master regulators’ which can drive the expression of a variety ofdownstream target genes is considered to be a significant approach in this endeavour. By altering the DNA accessibility,histone chaperones represent an important class of such ‘master regulators’; however, their physiological role in plantsvis-à-vis abiotic stresses largely remains enigmatic. We have previously found significant differential regulation of a fewhistone chaperones under one or more environmental stress conditions. In the present study, we narrowed down to astress-responsive member of the NAP superfamily of histone chaperones, OsNAPL6, which is expressed at a higherlevel in the salt-tolerant genotype Pokkali under most of the abiotic stress conditions. We found OsNAPL6 to be a nuclear-localized H3/H4 histone chaperone capable of assembling nucleosome-like structure on a relaxed plasmid template.Further, we utilized overexpression and knockdown approach to carry out detailed functional characterization of OsNAPL6with respect to its role in abiotic stresses in rice. We showed that alteration in cellular levels of OsNAPL6 positivelycorrelates with adaptation to various abiotic stresses as measured via analyzing morphological, physiological and agronomictraits. Considering that stress response and adaptation require altered expression of thousands of genes, together, ourresults suggest that the stress-responsive histone chaperone OsNAPL6 may serve a regulatory role in stress physiologyby controlling the expression of stress-related genes, possibly via modulating nucleosome dynamics at the correspondingloci.


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IPPCongress2015


PA045 (IPPC0356)

Effects of water stress on crop physiology of groundnut, aerobic rice, soybean and management options

for mitigation

K.G. Mandal1*, A.K. Thakur1 and K. Kannan2

1ICAR-Indian Institute of Water Management (Formerly Directorate of Water Management) Bhubaneswar-751023, Odisha, India2Regional Research Station, ICAR-Indian Institute of Soil and Water Conservation, Ooty, Tamil Nadu, India


Field experiments were conducted on rice and groundnut, and a pot experiment on soybean to study water stress effectson crop physiology under different management. For groundnut, fv/fm and actual fluorescence efficiency (ÔPS II) showedlower values under water stress compared to well-watered plants; intercepted PAR and pod yield was greater (64.5-74.4%) with paired-row planting on beds than flat-bed (46.3-57.7%); hence, paired-row planting of summer groundnutwith optimum irrigation to its critical growth stages would mitigate crop water stress and enhance WUE by 39%. Waterstress effect on aerobic rice was prominent on cross section of xylem vessels and cortex cells, root mass density, fv/fm,ÖPS II, yield attributes and grain yield than transplanted rice; the varieties viz. ‘Surendra’, ‘Apo’ and ‘Lalat’ showed thehighest yield potential (3.9-4.6 t ha-1) under aerobic conditions. Results on pot experiment on soybean showed physiologicalchanges in soybean seedlings. Due to water stress, relative leaf water content, SPAD values, total chlorophyll, rate ofphotosynthesis, fv/fm, ÔPS II and soluble protein content decreased, whereas proline content and enzyme activities i.e.catalase (CAT), superoxide dismutase (SOD), guaiacol peroxidase (POD) activity and malondialdehyde (MDA) contentincreased in soybean seedlings due to increase in reactive oxygen species (ROS). However, application of methyl jasmonate(MeJA @ 1 mmol L-1) reduced the adverse effects due to ROS and thereby increased rate of photosynthesis, stomatalconductance and soluble protein content. Hence, methyl jasmonate has the potential to mitigate water stress and toenhance the water stress tolerance to soybean seedlings.

PA046 (IPPC0361)

PROTEIN L-ISOASPARTYL METHYLTRANSFERASE confers tolerance to abiotic stress in Arabidopsis

thaliana

Shraboni Ghosh*, Pooja Verma and Manoj Majee

National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

Abiotic stresses usually lead to deterioration of structure and function of proteins. Maintenance of such proteins duringstress condition is vital for survival under stress. PROTEIN L-ISOASPARTYL METHYLTRANSFERASE (PIMT, EC.1.1.77)is a protein repairing enzyme that is known to repair damaged and aged proteins. Here we examine the function of PIMTin Arabidopsis thaliana plants under salinity, dehydration, oxidative, heat and cold stress conditions. Involvement of PIMTin stress tolerance was investigated by analysis of PIMT overexpression and RNAi lines in seedlings as well as in matureplants. During stress conditions elevated PIMT activity was seen which limits the accumulation of abnormal L-isoAspresidues. The RNAi lines exhibited greater sensitivity to the stress conditions whereas constitutive overexpression ofboth cytoplasmic (AtPIMT1) and nuclear (AtPIMT2) PIMT confers tolerance to various stresses. The damage caused byinduced stresses were assayed in terms of malondialdehyde content, chlorophyll estimation and H

2O

2 accumulation. The

overall higher PIMT activity and lower isoaspartyl accumulation leads to improved survival of the transgenic plants. Theresults indicate that PIMT not only functions in seed vigor and longevity but also provides stress tolerance to plants byrepairing isoAsp mediated damaged proteins which in turn limit reactive oxygen species formation.


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IPPCongress2015


PA047 (IPPC0366)

Ectopic expression of a wheat SNARE-binding protein confers drought tolerance in transgenic Arabidopsis

thaliana

Brinderjit Singh1*, Paramjit Khurana1, J.P. Khurana1 and Prabhjeet Singh2

1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India2Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab 143005, India


Drought stress is one of the major factors which limits crop productivity and has been reported to result in upto 50% lossin the average grain yield. The response of plants to drought stress is highly complex and is contributed by multiplegenes. Therefore, identification of genes involved in stress tolerance in crops is a prerequisite for developing stresstolerant plants by transgenic approaches. In the present study, gene encoding for vesicle-associated membrane protein(VAMP)-associated protein (TaVAP) was used to develop transgenic Arabidopsis plants for studying its role in abioticstress tolerance. The VAMP-associated proteins (VAPs) have been proposed to play roles in vesicle trafficking, microtubuleorganization, lipid metabolism, inositol auxotrophy and unfolded protein response. However, the physiological role ofthese genes is still a matter of conjecture in plants. This study demonstrated that VAP is functionally conserved, sinceTaVAP was able to rescue the yeast mutant cells, which were mutated in orthologue of this gene. Our studies provide thefirst evidence that TaVAP play an important role in abiotic stress response in plants. The expression of this gene wasenhanced in wheat seedlings in response to different stress conditions. Furthermore, ectopic expression of TaVAP inArabidopsis conferred significantly higher tolerance to transgenic plants under abiotic stress conditions. Importantly, theoverexpression of TaVAP did not result in any yield penalty, thereby, indicating that this gene is an ideal candidate forenhancing abiotic stress tolerance of crops through transgenic approaches.

PA048 (IPPC0369)

Differentially expressed galactinol synthase gene (s) of chickpea (Cicer arietinum L.) improves seed

vigor and longevity by limiting the hydroxyl radicals during aging

Prafull Salvi* and Manoj Majee

Plant Molecular Biology and biochemistry, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Galactinol synthase (EC: 2.4.1.123) (GolS) catalyzes the rate limiting step in Raffinose Family Oligosaccharide (RFO)biosynthetic pathway. RFOs including galactinol are known as anti-stress agents and suggested to play important role inabiotic stress responses and seed desiccation tolerance. In this study, we have shown galactinol synthase activity as wellas galactinol content is highly concentrated in pod and dry seed and gradually decreased as seed germination progresses.Galactinol synthase activity was also found to be enhanced in response to stresses. In chickpea, galactinol synthaseenzyme is encoded by two divergent genes (CaGolS 1 and CaGolS 2) which produce 5 different types of transcripts.Transcription analysis revealed that the CaGolS1 and CaGolS2 are differentially regulated. Further, all isoforms werebacterially expressed, purified and biochemically characterized. All isoforms exhibit differential enzyme activity. Nativemolecular weight and subunit composition was determined for enzymatically active isoform through gel filtrationchromatography. Confocal microscopy showed that a transiently transfected CaGolS-YFP fusion proteins primarilydistributed to nucleus and membrane in onion epidermal cell. Further we found that seed-specific expression of CaGolS1and 2 in Arabidopsis play a role in maintaining seed vigor and longevity, which was correlated well with increasedgalactinol and raffinose contents and reduced hydroxyl radicals in seed of transgenic lines. Collectively, our work clearlydemonstrates that CaGolS (s) are differentially regulated in chickpea and play important role in enhancement of seedvigor and longevity.


139

IPPCongress2015


PA049 (IPPC0371)

Surviving metabolic arrest: understanding the mechanisms of desiccation tolerance in the resurrection

plant Craterostigma plantagineum

Dinakar Challabathula*

Department of Life Sciences, Central University of Tamil Nadu, Thiruvarur-610101, Tamil Nadu, India


Drought is the most common abiotic stress that is encountered by the plants. Most angiospermic plants are droughtsensitive and do not survive if the relative water content decreases below a certain limit, whereas, the seeds of manyplants can tolerate extreme dehydration. A small group of resurrection plants from Linderniaceae family have developedthe ability to survive severe water loss in vegetative tissues and thus regarded as desiccation tolerant eg. Craterostigma

plantagineum. Transcriptome analysis of C. plantagineum at different dehydration stages identified the genes that areup-regulated/down-regulated during desiccation. Existence of both desiccation tolerant and sensitive plants in Linderniaceae

family provides an advantage for comparative analysis there by to understand the mechanisms of desiccation tolerance.Transcripts encoding group II late embryogenesis abundant (LEA) proteins from C. plantagineum were identified andquantified. While the transcripts encoding LEA-like 11-24 protein are abundantly expressed during desiccation in thedesiccation tolerant plants C. plantagineum and Lindernia brevidens, the transcripts are expressed at a low level in thedesiccation sensitive Lindernia subracemosa. Differential activities of the promoters were observed along with someconservation in Cis elements. While desiccation sensitive plants showed degradation of mRNA of specific genes duringdesiccation, in the desiccation tolerant plants the mRNA for these genes was highly stable. Further, comparative studiesutilising physiological, genetic and biochemical approaches are essential to decipher the underlying mechanisms ofdesiccation tolerance.

PA050 (IPPC0389)

Effects of gamma radiation stress on production of 20-hydroxyecdysone in Sesuvium portulacastrum L.

shoot cultures

Vishwadeep Kapare1*, Ramesh Satdive2, Nutan Malpathak1 and Devanand P. Fulzele2

1Department of Botany, Savitribai Phule Pune University, Pune-411007, India2Plant Biotechnology and Secondary Metabolites Section, NABTD, Bhabha Atomic Research Centre, Mumbai-400085, India


Sesuvium portulacastrum Linn. a member of family Aizoaceae, is naturally growing plant in coastal areas and rich sourceof phytoecdysteroid 20-hydroxyecdysone (20E). Various biological properties including antidepressant, antioxidant,prevention of infectious diseases and cytotoxic activity are associated with 20E. Due to its anabolic activities it is commonlyused by body builders and sportsmen in form of phytoecdysteroid containing herbal extracts as well as OTC drug. In vitro

shoots cultures of S. portulacastrum were irradiated with low doses of 5, 10, 15, 20, 25, 30, 35 and 40 Gy from a cobalt-60 source. Growth kinetics and content of 20E of the in vitro cultures were studied for three generations: M1, M2, and M3.High Performance Liquid Chromatography (HPLC) revealed that gradual increase in gamma doses resulted in significantincrease in product synthesis. Shoot cultures were irradiated at 20 Gy synthesized highest 1.19 mg/gm of 20E, whereasnon irradiated shoot cultures produced 0.35 mg/gm. High doses of gamma rays impaired product synthesis and growthrate of shoot cultures. The present investigation corroborated that in vitro irradiation is an important method for improvementof secondary metabolites at suitable doses of gamma rays.


140

IPPCongress2015


PA051 (IPPC0403)

Physiological trait based breeding: An obligate for future food security under abiotic stress

H.M. Mamrutha1*, Ritika Choudhary1, Dheeraj Rana1, Omprakash Tuteja1, Ravish Chatrath1, Sai Prasad2, D.C. Saxena2,Satish Mishra3, Sonia Sheoran1, Chetan Kumar Choudhary1, Mitchelle Watt4 and Indu Sharma1

1Indian Institute of Wheat and Barley Research, Karnal-132001, Haryana, India; 2Indian Agricultural Research Institute (IARI) RegionalStation, Indore, India; 3Agarkar Research Institute, Pune-411004, India; 4CSIRO, Australia


By the year 2050, the projected global and India’s wheat demand will be 1090 and 140 mt, respectively. This target canbe achieved with annual production growth of 1.7%. There is ceiling in wheat yield under high productive areas, hence theother potential source to increase yield is stressed environment. Worldwide 84 mha of wheat growing area is affected bydrought, and traditional breeding efforts are focused on yield rather than survival, which present a challenge to wheatscientists to develop wheat cultivars that are both productive and adapted to abiotic stresses. Recently physiologicaltraits (PTs) have become a target for plant breeders aiming to improve yield potential in stressed environment. In thepresent study, the different PTs, viz., early vigour, tillering inhibition, deep rooting system and alternate dwarfing genesfor deep sowing under drought were studied. A set of wheat genotypes were validated using novel field screening methodsin three different locations in India and Australia. Morphological markers and germplasm source for these traits havebeen identified. The morphological marker includes third leaf width at seedling stage, as a proxy for early vigour, longcoleoptile length for emergence from deeper soil, spike pubiscence for tillering inhibition and canopy temperature at 15days after anthesis for deep root system. The germplasm identified were MW257, Synthetic 9589/Kennedy, CM18/Viguor18with Rht13 and HI 1531 for tillering inhibition, early viguor, long coleoptile length and deep root system, respectively.These traits have been introgressed into Indian wheat genotypes, mapping populations have been developed andidentification of relevant molecular marker is under progress.

PA052 (IPPC0404)

Differential transcript abundance of genes of antioxidative enzymes and redox cues reveal two faces of

hydrogen peroxide-mediated oxidative stress in rice seedling

Chakrabarty Ananya* and Bhattacharjee Soumen

Centre for Advanced Study, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India


Treatments with different titre of H2O

2, both elevated (20 mM) and low (500 μM) during early imbibitional phase of germination

resulted in the generation of differential redox cues in metabolic interface as being investigated in terms of significantchanges in prooxidant/antioxidant ratio, titre of individual reactive oxygen species, efficacy of Ascorbate-Glutathionecycle, activities of enzymatic antioxidants, radical scavenging activities, changes in thiol compounds and expression ofoxidative damage to membrane protein and lipid. All the redox parameters tested, corroborate the finding that treatmentwith elevated concentration of H

2O

2 cause serious loss of redox homeostasis, whereas low titre H

2O

2 treatment not only

restores redox homeostasis but also up-regulate oxidative stress-acclimatory processes as evident from reduction inoxidative damages of newly assembled membrane system. 500 μM H

2O

2 treatment exhibited enhanced activities of

Ascorbate-Glutathione pathway enzymes, which was otherwise inhibited significantly in 20 mM treated seedlings. Acomparison between imbibitional chilling stress-raised experimental rice seedling with 20 mM H

2O

2 treated rice seedling

reveal similar kind of generation of redox cues. Further, H2O

2 treatments in rice seedlings also reveal the dose-dependent

regulation of expression of genes of antioxidative enzymes. Low titre H2O

2 treatment combat oxidative stress by up-

regulation of gene expression of some antioxidative enzymes, particularly of Ascorbate-Glutathione pathway, which inturn offers better capacity of redox regulation and mitigation of oxidative damage of juvenile membrane system. Inconclusion, a dose-dependent regulation of H

2O

2 mediated redox cues, redox regulatory properties and oxidative damages

in rice seedling is suggested.


141

IPPCongress2015


PA053 (IPPC0405)

Efficacy of expression of genes of ascorbate-glutathione cycle and redox regulatory mechanisms under

dehydration stress as the basis of screening land races of some aromatic rice of West Bengal

D. Nivedita* and Bhattacharjee Soumen

Centre for Advanced Study, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India


PEG-induced dehydration stress during early germination causes disruption of redox-homeostasis and oxidative damagein four aromatic land races (Jamainadu, Tulaipanji, Sitabhog and Badshabhog) by enhancing the accumulation of pro-oxidants (DCFDA oxidation, O

2

.- and H2O

2 accumulation), significant reduction of antioxidative defence (total antioxidant

and radical scavenging capacity, total thiol content and activities of antioxidative defence enzymes) and aggravatingprotein oxidation and membrane lipid peroxidation (assessed in terms of free carbonyl content and accumulation ofthiobarbituric acid reactive substances). When compared between the land races, a clear trend in differential redoxregulatory properties in which ROS-antioxidant interaction acts at metabolic interface was observed in the order of theland races Tulaipanji> Sitabhog> Badshabhog> Jamainadu. Moreover, when the efficacy of Ascorbate- Glutathione cyclefor scavenging H

2O

2 generated during dehydration stress were assessed and compared between the land races exposed

to PEG-induced dehydration stress, it also exhibited almost the same trend with the land race Tulaipanji and Jamainaduexhibiting maximum and minimum efficiencies respectively. The land race Tulaipanji and Sitabhog resist dehydrationstress better than the other land races due to its early preparedness to combat oxidative stress by upregulating expressionof genes of some enzymes of Ascorbate-Glutathione cycle along with some other antioxidative enzymes. A model ofredox homeostasis in which ROS- antioxidant (Ascorbate- Glutathione system) acts at metabolic interface for up-regulationof antioxidative gene expression necessary for differential drought stress tolerance among the land races of aromatic riceis suggested.

PA054 (IPPC0407)

Loss of adaptive plasticity of mangroves in degraded ecosystem-a study based on osmolyte accumulation

Krishna Ray1*, Sandip Kumar Basak2, Tapan Sutradhar1, Chandan Mukherjee1, Rajojit Chowdhury1 and Momtaj Begam1

1Department of Botany, Environmental Biotechnology Group, West Bengal State University, Barasat-700126, West Bengal, India2Department of Botany, Kandi Raj College, Kandi, Murshidabad, West Bengal, India


In degraded mangrove forests, it has been well documented that due to various environmental constraints, the ecosystemhomeostasis gets hindered to an extent when it can no longer continue the normal processes of secondary successionand mangrove forests come to the verge of extinction. Mangroves bear a fascinating complex network of morpho-physiological adaptations, but their loss of adaptive plasticity is causing gradual disappearance under degraded ecosystem.Synthesis of nitrogen-rich and energy-intensive organic solutes proved to be strong selective advantage for mangrovesto encounter salt stress, but this adaptation is too expensive in terms of metabolic costs for this group of plants resultinginto inevitable trade-off against its growth. The cost of generating enough organic solutes to achieve full osmotic adjustmentin hypersaline soil exceeds all other energy consuming adaptive processes. It becomes far more challenging for mangroves,when they have to adjust osmotically under degraded soil conditions. Our studies established that nutrient cycling underdegraded mangrove soil is largely impaired and limit the synthesis of nitrogen-rich and reduced carbon based compatiblesolutes. The pioneer colonizer halophytic grasses like Porteresia, Myriostachya, Paspalum, and Sporobolus sp. InSundarban mangrove ecosystem contribute to nutrient cycling through the bacterial population harbouring at therhizosphericregions, hence augmenting the secondary succession and restoration process in the degraded mudflats. These grasseswith their ideal osmotic and strong mechanical adaptivitypave the way for stabilization and stress management of incomingmangrove species under a multitude of abiotic stressesin degraded mangrove ecosystem of Sundarban.


142

IPPCongress2015


PA055 (IPPC0411)

Selection of castor (Ricinus communis L.) germplasm with good root traits for drought tolerance in field

P. Lakshmamma*, Lakshmi Prayaga and K. Alivelu

Department of Plant Physiology, ICAR-Indian Institute of Oilseeds Research, Rajendranagar- 500030, Hyderabad, India


Fourteen germplasm lines with good and two with poor root growth identified by screening in root structures along with avariety (48-1) and hybrid (DCH-519) check were sown in field for two years during late rabi (Nov. 1st week) 2010-11 &2011-12 and water stress was imposed from 30-90 days after sowing (DAS) by withholding irrigation in three replicationsin split plot design. Pooled analysis of data shows significant growth reduction in terms of reduced plant height, leafnumber, stem girth, branches and total dry matter (56%) with water stress. On an average, there was 13% reduction inTDM at harvest, 35% reduction in seed yield and 19% reduction in harvest index (HI) with drought stress. Among thegenotypes studied, RG 1494 and RG 2139 recorded <20% reduction in seed yield and also showed low drought susceptibilityindex (DSI) of <0.5. Primary seed yield reduced significantly with stress, but the reduction was less in secondaries andcompensated in later order spikes in these genotypes. The root growth in terms of root volume (RG 1494-310 cm3, RG2139-557 cm3) and root dry weight (RG 1494-122 g/pl., RG 2139-93 g/pl.) was also more in these genotypes. Chlorophyllcontent measured with SPAD chlorophyll meter (SCMR) and bloom (wax) content increased in stressed plants as anadaptive feature. The genotypes RG 1494, RG 2139 that showed good performance during both years with good root andshoot growth characters can be used in drought breeding programs.

PA056 (IPPC0413)

Differential accumulation and gene expression of various osmolytes in Brassica juncea under temperature

stress

Sudhir Kumar1*, R.K. Sairam1, K.V. Prabhu2 and Akshay Sakhare1

1Division of Plant Physiology, 2Division of Genetics, Indian Agriculture Research Institute, New Delhi-110012, India


A study was carried out to evaluate the role of various osmolytes in temperature stress tolerance in Brassica junceagenotypes. Three dates of sowing were used for temperature stress i.e. D1 (30 Oct), D2(15 Nov) and D3(30 Nov).Different osmolytes were studied. Result showed that Temperature stress due to late sowing (D2 and D3) resulted inincrease in contents of total sugar, proline, glycine-betaine and trehalose as compared to normal (D1) sowing. Amongvarious genotypes CS 52, Progaro and NDR 8801 accumulated highest content of different osmolytes as compared toPusa Agrani, EJ-15 and Pusa Tarak, which showed lesser accumulation under temperature stress. Gene expressionstudied by RT-PCR showed temperature stress induced increase in mRNA transcript of β-pyrroline-5-carboxylatesynthetase (P5CS), betaine aldehyde dehydrogenase (BADH) and trehalose-6 -phosphate synthase (T6PS), which wasagain higher in Proagro and CS 52 compared to Pusa Agrani, EJ 15 and Pusa Tarak. Results suggested that hightemperature induced expression of P5CS, BADH and T6PS in Proagro and CS 52 resulted in greater accumulation ofosmolytes, which could be one of the reasons of their tolerance to temperature stress.


143

IPPCongress2015


PA057 (IPPC0418)

Evaluation of rice varieties and hybrids under high temperature stress

Beena Radha1*, Veena Vigneswaran1, P. Sindhumol1, M.C. Narayankutty1 and S.R. Voleti2

1Department of Plant Physiology, Regional Agricultural Research Station, Pattambi, Kerala Agricultural University, Palakkad-679306,Kerala, India2Department of Plant Physiology, Directorate of Rice Research, Hyderabad-500030, AP, India


A study was conducted to evaluate the performance of rice varieties and hybrids under high temperature stress during2014 at Regional Agricultural Research Station, Pattambi, Kerala Agricultural University. In this study seven rice hybridsand seventeen hybrid derivatives were evaluated at 6-8°C above the normal temperature and ambient condition fromseedling to maturity stage. Biomass accumulation of high temperature exposed plants were higher than plants kept atambient condition. Spikelet fertility of rice hybrids were very low but hybrid derivatives were maintained spikelet fertility assimilar to that of ambient condition. In another study it is revealed that when plants were shifted to high temperature frompanicle initiation to maturity stage, hybrid derivatives also showed reduction in spikelet fertility. From these studies, it canbe concluded that when plants were exposed to high temperature from seedling stage to maturity occurrence of spikeletsterility is very less due to the acclimatization process but hybrids were highly susceptible to high temperature stress thanhybrid derivatives. Spikelet fertility of hybrid derivatives ranged from 78.35% (Lalat) under high temperature and 92.70%(IET 23216) under ambient condition. Spikelet fertility of rice hybrids ranged from 64.52% (IET 24122) under hightemperature and 90.22% (IET 24117) under ambient condition. Photosynthetic rate and spikelet fertility of high yieldingvarieties were maintained on par between ambient and high temperature condition except Lalat and MTU-1010. In ricehybrids, both photosynthetic rate and spikelet fertility significantly reduced under high temperature condition except inIET 21404 and US-312.

PA058 (IPPC0425)

Characterization of delayed senescence traits in chickpea (Cicer arietinum L.) germplasm under drought

stress

Gurumurthy Shekharappa1*, P.S. Basu1, Arora Ajay2, Jagdish Singh1 and N.P. Singh1

1Division of Crop Physiology, Biochemistry & Microbiology, Indian Institute of Pulses Research, Kanpur-208024, India2Division of Plant Physiology, ICAR-Indian Agriculture Research Institute, New Delhi-110012, India


An experiment was conducted with 276 chickpea genotypes of ICRISAT minicore with an objective to screen the genotypeshaving drought induced delayed leaf senescence trait. The physiological parameters such as SPAD chlorophyll meterreading (SCMR), phenology, water use efficiency (WUE), membrane stability index (MSI), leaf Senescence Rate (LSR),greenness index (GI), yield and yield components were studied. We identified thirteen contrasting genotypes belong todelayed senescence and the other thirteen falls in the category of faster senescence among the genotypes studied.Higher and lower reduction in the WUE, MSI and SCMR during stress condition was observed with faster and delayedsenescence genotypes, respectively under moisture deficit stress condition. These contrasting genotypes did not differedin flowering and podding behavior of both the irrigated and stress condition. Delayed senescence genotypes showedlesser reduction in leaf LSR and significantly higher GI compared to genotypes with faster senescence. Genotypes withdelayed senescence showed higher biomass as well as better adaptation under drought. LSR was recorded by visualscoring; the maximum and minimum score was recorded in genotypes ICC 762 and ICC 6279, respectively. The presentstudy revealed that SCMR, WUE, MSI, LSR and GI are good physiological indices under water deficit stress condition forchickpea crop and can be used for screening of chickpea genotypes for drought induced leaf senescence.


144

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PA059 (IPPC0427)

Assessment of terminal heat tolerance ability of wheat genotypes based on physiological traits using

multivariate analysis

Shivani Nagar1*, V.P. Singh1, Ajay Arora1, Neera Singh3, Rajkumar Dhakar2 and Shiv Ramakrishnan1

1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Agricultural Chemicals, Indian Agricultural Research Institute,New Delhi, India-110012, India

*Presenters author: [email protected]

Terminal heat stress is one of the limiting factors in wheat production and it is expected to rise under present climatechange scenario. Present study was conducted to evaluate the performance of 40 wheat genotypes under terminal heatstress conditions based on eight physiological traits. The plants were sown late (i.e., on 5th January) to expose them toterminal heat stress. The genotypes were evaluated using multivariate analysis, viz., Ward’s method of hierarchicalclusters analysis, discriminant analysis and principle component analysis. The genotypes were categorized in to threegroups namely tolerant, intermediate and sensitive. Tolerant genotypes like DBW-14, RAJ-3765, HD-2643 and HALNAperformed physiologically better in terms of higher membrane stability index (MSI), chlorophyll content, photosynthesisrate (P

N), harvest index (HI) under heat stress condition. Genotypes HD-2987, SHAINGHAI-1, HD-2402 and WH-730

were found to be heat sensitive. Physiological traits like MSI, SPAD value, Fv/Fm ratio and PN were found to be most

important contributor in grouping of genotypes, and showed positive correlations (r) of 0.73, 0.47, 0.41 and 0.39 with grainyield, respectively, which were significant at p<0.05. The large genetic diversity was found among the genotypes basedon physiological traits. These genotypes can be utilized in wheat improvement programs for heat tolerance.

PA060 (IPPC0430)

Effect of salicylic acid to mitigate the high temperature stress in Indian mustard [Brassica juncea (L.)

Czern & Coss

B.L. Kakralya* and O.P. Godara

Department of Plant Physiology, S.K.N. College of Agriculture, Jobner, Rajasthan, India


A field experiment was conducted to evaluate the role of salicylic acid in mitigating high temperature stress effects in twogenotypes of Indian mustard viz.RGN-236 and RGN-229. Both the genotypes were exposed to high temperature throughdelayed sowing and three concentrations of salicylic acid viz. 50, 100 and 150 ppm were sprayed at pre anthesis andsiliqua initiation stage. Under delayed sowing, relative water content, photosynthetic rate, chlorophyll stability index,membrane stability index and oil content decreased significantly. Whereas, transpiration rate, stomatal conductance andleaf temperature significantly increased under delayed sowing. On the other hand, application of salicylic acid showedless reduction in growth and above physiological traits. Among three concentration of salicylic acid, 150 ppm showed thebest response and was followed by 100 and 50 ppm. Yield attributes like number of siliqua per plant, number of seed persiliqua, plant height, test weight, seed yield, biological yield, harvest index decreased with delayed sowing under controlbut plants treated with 150 ppm salicylic acid showed significant improvement and less reductions due to high temperaturestress under delayed sowing. Among two genotypes, RGN-229 showed less reductions in growth and yield and can beconsidered as superior in terms of high temperature tolerance over RGN-236.


145

IPPCongress2015


PA061 (IPPC0436)

Differential response of rice cultivars to high night temperature

Sangeeta Khetarpal1, Anjali Anand1*, Archana Yadav2, S.M. Trivedi1, Dinesh Kumar3 and Himanshu Pathak2

1Division of Plant Physiology, 2CESCRA, 3Division of Agronomy, Indian Agricultural Research Institute, New Delhi-110012, India


Global climate change is leading to asymmetric atmospheric warming with reduced temperature differences between dayand night. Minimum night temperatures are increasing at a faster pace than maximum day temperatures and this increaseis expected to be larger in the tropics and subtropics (IPCC, 2013) which correspond to the main rice production area.Twenty rice cultivars were subjected to high night temperature (HNT) from panicle emergence to maturity. During theexposure period and despite some variability, mean minimum temperature increased by about 1.5oC in the polychambersover the mean minimum temperature of 24oC in ambient. Flag leaf night respiration rates increased manifold in all cultivarsaccompanied by reduction in biomass. Based on grain yield heat susceptiblity index Nerica L 44, Nagina 22, Ganga andAbhishek performed better under HNT. Leaf starch content reduced by 14% in genotypes showing better performanceunder HNT compared to 26% in genotypes with high susceptibility index suggesting more loss of stored carbohydrateunder HNT. The reduction in stem starch and increase in reducing sugar content observed in most of the genotypes couldbe due to reduced synthesis of stem starch as a consequence of lower availability of photoassimilates from leaves due torespiratory losses or increased breakdown of starch in stem. Grain filling was adversely affected in the middle stage ofgrain growth that could be another cause for lower yields under HNT in some genotypes. Quality was affected in terms ofdecrease in proportion of high density grains and increase in chalkiness and grain protein content under HNT.

PA062 (IPPC0437)

Oxidative stress and antioxidative defence system in Ricinus communis (L.) under high temperature

stress

Praduman Yadav* and C. Lavanya

Department of Biochemistry, ICAR-Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad-500030, India


The study was undertaken to investigate the effect of high temperature (HT) induced stress and revival on antioxidativeenzymes and metabolites in leaves of the castor (Ricinus communis L.) seedlings of HT susceptible and HT tolerantgenotypes. Seeds of thirty five genotypes were sown in trays having sandy loam soil. Ten days old seedlings wereexposed to threshold high temperature (45±0.5°C) and 30% relative humidity continuously. Time taken to 50% mortalitywas recorded in all the genotypes for screening against high temperature tolerance. Two genotypes were identified as HTsusceptible (DCS-89 and DCS-117) and two genotypes as HT tolerant (DCS-107 and DCS-113). After subjecting theseedlings to HT stress, the seedlings were revived by placing the trays at 25±0.5°C for 24 hrs. Activities of superoxidedismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX) and glutathione reductase (GR)increased in both HT tolerant and susceptible genotypes under stress but increase was found significantly higher intolerant genotypes. On revival antioxidative enzymes viz. SOD and CAT started decreasing, but the activities of POX,APX and GR still continued to increase in all the genotypes. The malondialdehyde (MDA) content H

2O

2 concentration and

lipoxygenase (LOX) activity in leaves of castor seedlings increased at the high temperature but higher percent increasewas observed in susceptible genotypes. However, on revival, LOX activity, MDA and H

2O

2 content decreased as compared

to stressed conditions in all the genotypes.


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IPPCongress2015


PA063 (IPPC0446)

Overexpression of ABA receptor gene OsABAR6 enhances drought tolerance of rice

V.V. Santosh Kumar1*, Shashank K. Yadav1, Sanya Shrivastava1, Rakesh K. Verma1, Suchitra Puskar1, M.V. Rao2 andViswanathan Chinnusamy1

1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, IndiaDepartment of Plant Sciences, Bharathidasan University Tiruchirappalli, Tamil Nadu, India


Abscisic Acid (ABA) plays crucial roles in development and stress responses of plants. ABA receptor (ABAR) family inArabidopsis consists of 14 members. Bioinformatic analyses led to the identification of 12 PYR/PYL homolog ABARsfrom rice. However, the specific function of members of ABARs in developmental and stress responses needs to beunderstood. In present study, OsABAR6 gene from rice was used for functional validation. Expression analysis of ABAR6

in different tissues and stress conditions revealed differential expression patterns. ABAR6 gene was cloned in binaryvector pCAMBIA1300 under AtRD29A promoter and pANDA RNAi vectors. Trangenic rice plants were developed byusing Agrobacterium mediated genetic transformation and confirmed through PCR, qRT-PCR and southern blotting.Rice transgenics expressing AtRD29A::OsABAR6 showed hypersensitivity to ABA in germination as compared withnon-transformed (NT) wild type and RNAi lines. This suggests that ABAR6 conferred enhanced ABA perception andthus function as ABA receptor. Analysis of drought tolerance of RD29A:ABAR6 rice transgenics (T

2) in pot culture under

greenhouse conditions revealed that RD29A:ABAR6 transgenic rice lines are more tolerant than NT plants in terms ofmaintenance of RWC, membrane stability index, chlorophyll content, survival and recovery growth. However, due toleaky expression of AtRD29A::ABAR6 even under control growth conditions, ABAR6 plants showed reduced height andyield. Water utilization experiment revealed that transgenic plants used about 40% less water per a day as compared towild type plants. Results from this study suggest that ABAR6 has potential to improve WUE and drought tolerance of rice.

PA064 (IPPC0449)

Abiotic stress and wood anatomy of some lianas from Western Ghats

Vidya Patil* and N.P. Malpathak

Department of Botany, University of Pune, Pune-411007, India


Lianas constitute an important floristic component of tropical forests, (approximately 25%) where their high abundancehas been used to differentiate tropical from temperate forests. Lianas are vines that begin life on the ground as small self-supporting shrubs and rely on other plants to reach the light-rich environment of the upper canopy competing directly withother trees for sunlight. They adapt themselves for support, climbing and twisting. As thickness increases in lianas, theyshow abnormal secondary growth and the storage organs exhibit anomalous structure. These anomalous structures maybe directly influenced by external environmental conditions. Precise reason behind occurrence of cambial variants is yetnot known, but it is considered that this is due to the effect of physiological or environmental stress. Acclimatized oradaptive structures like presence of wide vessels in xylem helps in high hydraulic conductivity. Abundance of parenchymaand ray parenchyma cells within and between xylem groups provides good flexibility with minimum damage to waterconduits in the stem during climbing and twisted growth. It also limits physical disruption of vascular tissues duringtwisting and bending, and also promotes wound healing after girdling. It might also play a role in photosynthate storage.Present study focused on abiotic stress and their management and adaptations for growth and existence.


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IPPCongress2015


PA065 (IPPC0454)

Analysis of genotypic variability for stem reserve mobilization in rice germplasm with contrasting drought

tolerance

A.S. Sakhare1*, Puja Rai1, Sudhir Kumar1, P.C. Nautyal2 and V. Chinnusamy1

1Division of Plant Physiology, 2Division of Seed Science and Technology, Indian Agricultural Research Institute, New Delhi-110012, India


Non-structural carbohydrates accumulation in stem contributes substantially to grain development under stresses andhence, considered important for yield stabilization in grasses. In order to understand the contribution of non-structuralcarbohydrates remobilization in drought tolerance, we performed an experiment to analyze the genotypic variability forstem reserve mobilization (SRM) in rice germplasm with contrasting drought tolerance. Fourteen rice genotypes withcontrasting drought tolerance were grown in the field and drought stress was imposed by withholding irrigation water. Forestimation of SRM, plant samples were collected at flowering and at maturity stages. The relative change in dry weightand non-structural carbohydrates in stem, sheath, leaf and panicle tissues were analyzed to estimate SRM. Significantvariability was observed among the rice genotypes for non-structural carbohydrates remobilization under drought stress.Under control conditions, reserve mobilization varied from 0 to 53%. Drought stress enhanced reserve mobilization inCrossa, MTU 1010, Vandana, CR-143-2-2 and Nagina 22 with maximum increase in CR-143-2-2. In case of Pusa 44,IR64 and Swarna, mobilization percentage decreased considerably under stress. Physiological and molecular basis ofthe genotypic differences in SRM are being investigated.

PA066 (IPPC0474)

Characterization of a novel zinc finger transcription factor (TaZnF) conferring heat stress tolerance in

plants

Preeti Agarwal* and Paramjit Khurana

Department of Plant Molecular Biology, University of Delhi, New Delhi-110021, India


High temperature is known to affect the crop production worldwide especially wheat. Under high temperature stress, anumber of Transcription factors are activated including HSF’s, MYBs, bZIP in wheat. Recently, we have identified a zincfinger transcription factor in wheat from a suppression subtractive heat stress library. TaZnF belongs to C

3CH

4 type which

constitutes the largest family in the plant kingdom and is reported to play important roles during various abiotic stressconditions. The characteristic feature of TaZnF includes a typical RING domain and a transmembrane region. The clustalalignment studies revealed high degree of conservation of zinc finger domain across various other orthologous members.TaZnF expression was upregulated under high temperature, salt, mannitol and cold stress. Transcriptional activation ofTaZnF was confirmed by trans-activation in yeast. Localization studies revealed the possible localization to the nucleusand the membrane. The Arabidopsis transgenic plants over-expressing TaZnF showed high tolerance to high temperaturestress at 42°C and performed better with respect to their growth and yield. Besides these responses to high temperature,the OE Arabidopsis transgenics showed considerable tolerance to drought and salinity stress. These observationssuggest that TaZnF plays an important role during heat stress. Therefore TaZnF can be of great importance in improvingthermal stress tolerance in plants.


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IPPCongress2015


PA067 (IPPC0480)

Gene pyramiding of antioxidant enzymes leads to abiotic stress tolerance in Brassica juncea

Neelam Prabha Negi1*, Vinay Sharma2 and Neera Bhalla Sarin1

1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Department of Bioscience and Biotechnology, Banasthali University, P.O. Banasthali, Vidyapeeth, Rajasthan-304022, India


Reactive oxygen species (ROS) are generated by biotic and abiotic stresses such as drought, salinity, strong light,extreme temperatures, heavy metals, UV radiation, atmospheric contamination, mechanical wounding, nutrient starvationand pathogen attacks. Antioxidant enzymes play a significant role in eliminating toxic levels of reactive oxygen species(ROS), generated by living cells under stress. The antioxidative defense is initated by SOD, which converts superoxide(O

2

-) radicals to H2O

2. The H

2O

2 that is also potentially harmful is converted to non-toxic water and monodehydroascorbate

by the APX enzyme utilizing ascorbate as the electron donor. In the present study, genes for two different cytosolicantioxidant enzymes, superoxide dismutase (AhCuZnSOD) and ascorbate peroxidase (AhcAPX) isolated from salt tolerantcell lines of Arachis hypogaea were simultaneously over expressed in highly important oilseed crop, Brassica juncea(Indian Mustard) to check the effect on alleviation of abiotic stress. Compared with wild type plants, transgenic plantssurvived under longer period of water deficiency and salinity stress and displayed improved recovery after rehydration.The enhanced levels of antioxidant enzymes in the transgenic plants correlated with higher relative water content, improvedphotosynthetic efficiency, less electrolyte damage, elevated accumulation of compatible osmolytes, less malondialdehydeas well as H

2O

2 accumulation and O

2

- accumulation under stress conditions compared to untransformed wild-type controls.Our analysis showed that an increase in the activity of antioxidant enzymes can significantly enhance salt tolerance intransgenic plants, suggesting an important role of combined over expression of two antioxidant enzymes.

PA068 (IPPC0483)

High temperature tolerance in chickpea (Cicer arietinum L.): Possible involvement of zeaxanthin cycle

pigments

Pramod Kumar*, Sanjay Yadav and Madan Pal

Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India*Presenting author: [email protected]

Chickpea being a cool season crop often experiences abnormally high temperature during reproductive phase. Terminalhigh temperature stress is one of the major constraints for its production. In nature, plants have developed severalmechanisms to combat the high temperature stress. Our approach leads to a consideration of the photosynthetic pigmentsfor high temperature tolerance. So, in order to analyze the heat tolerance in relation to photosynthetic pigments, anexperiment was conducted using 4 contrasting genotypes namely BG 240 and JG 14 (relatively heat tolerant), SBD 377(moderately tolerant) and ICC 1882 (relatively heat sensitive). High temperature was imposed by delaying the sowingdate i.e. normal (18th November) and late sown (18th December). Under late sown condition, high temperature wasexperienced by crop from flowering stage to crop maturity (during this period maximum temperature ranged from 25oC to39.9oC). Under late sown high temperature condition, in general, high temperature tolerant genotypes BG 240 and JG 14maintained higher level of membrane stability index, RWC (%), osmolytes, dry matter partitioning, grain yield and heattolerance index and had higher values of zeaxanthin, quantum yield of PS II (Fv/Fm ratio), non-photochemical quenching(NPQ), photosynthetic rate, level of photosynthetic pigments (chlorophylls and carotenoids) and lower level of violaxanthin,and lipid peroxidation as compared to high temperature sensitive one (ICC 1882). Furthermore, Fv/Fm ratio and NPQshowed positive association with high temperature tolerance which in turn indicated the involvement of zeaxanthin cyclepigments in high temperature tolerance of chickpea.


149

IPPCongress2015


PA069 (IPPC0484)

Plasticity in root-shoot development under water stress: exploring genotypic variation in wheat progenitor

species, Aegilops tauschii and Triticum dicoccoides

Yadhu Suneja1*, Anil Gupta1 and Navtej Singh Bains2

1Department of Biochemistry, 2Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana-141004, Punjab, India


A set of fifty seven Aegilops tauschii (DD genome), twenty six Triticum dicoccoides (AABB genome) accessions, wildwheat progenitors, along with tetraploid durum and hexaploid wheat cultivars were evaluated for their responsiveness towater stress. Observations across stress and non-stress environments allowed elucidation of genotypic variability in thelevel of induced responses for various traits. Though under PEG-induced water deficiency, average root length decreasedfrom 3.82 cm to 3.66 cm for Ae. tauschii and from 5.40 cm to 5.26 cm for T. dicoccoides, several accessions demonstratedroot elongation. The spectrum of variation was wider for increase than decrease in root length and biomass. Thirty threeAe. tauschii accessions and eighteen T. dicoccoides accessions showed an increase in root dry biomass under stressthat ranged from 2% to 295%. Shoot parameters- length and biomass, by and large were suppressed by water deficitstress. Change in root length had a significant negative correlation with change in shoot length (r= -0.519). Change in rootdry weight was significantly negatively associated with change in shoot length (r= -0.242) and change in shoot drybiomass (r= -0.369). Cross talk of root-shoot development with proline dynamics was also investigated. Aegilops tauschiiaccessions- 9803, 9814, 14109, 14191 and T. dicoccoides accessions- 7130, 7056 that showed maximum rootenhancement and minimum shoot retardation under stress are worthy targets of molecular genetic analysis and subsequentmarker-assisted mobilization of appropriate levels of inducibility for root and shoot development in the elite wheatbackgrounds.

PA070 (IPPC0495)

Deep rooted crops under center pivot irrigation: managing water stress

Sangamesh Angadi*, Sultan Begna, Sukhbir Singh and Khulbhushan Grover

Plant and Environmental Sciences, New Mexico State University, 2346 SR 288, Clovis 88101, United States


Agriculture in the Southern Great Plains of the United States perennially suffers from abiotic stresses including water,wind and temperature. Irrigation water from the Ogallala Aquifer converted this “Dust Bowl of the 1930s” into a highlyproductive agriculture region in the country. Center Pivot Irrigation Systems was invented in the region to improve irrigationapplication efficiency and reduce energy costs, which is used extensively. However, continued decline of well outputsand need for crop diversification have led to assess deep rooted alternative crops for the region. Center pivot systemsimulates frequent rainfall events and applies small quantities of water frequently to wet a shallow depth of the soil profile.With the current practice of irrigation deeper soil profiles are less frequently used. A multiyear study using the deep rootedsafflower crop was conducted to assess effects of storing water in the deep soil profile using pre-season irrigation,varying in-season irrigation levels and critical growth stage based irrigation management on safflower stress physiology,seed yield formation and productivity. Safflower responded to water stress with multiple strategies. It ranged frommorphological adjustment of plant size and leaf area to physiological response of stomatal and photosynthesis regulation.Safflower relied on its strong root system to extract moisture from depth. Although the amount of water extracted fromdepth was relatively small, the efficiency of using that water in yield formation was much higher.


150

IPPCongress2015


PA071 (IPPC0496)

Interplay between aldo-keto reductase (AKR) and glyoxalases as an important strategy for methylglyoxal

detoxification during abiotic stress in Jatropha curcas L.

Shalini Mudalkar* and Attipalli Ramachandra Reddy

Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India


Methylglyoxal (MG), a by-product of oxidative stress, is responsible for protein oxidation, DNA damage and lipid peroxidationin plants leading to the generation of advanced glycation end products (AGEs). The accumulation of MG was studied inJatropha curcas under progressive salinity and drought stresses, wherein the levels of MG increased with increasingstress severity. To understand the mechanism of MG detoxification and plant protection in J. curcas under abiotic stress,aldo-keto reductase (JcAKR) and glyoxalases (JcGLY1 and JcGLY2) were cloned, characterised and heterologouslyexpressed in E. coli BL-21 (DE3). MALDI-TOF analysis was performed to confirm the purified proteins, which showedsignificantly high enzyme activities and catalytic efficiencies with methylglyoxal (MG) as substrate. The expression ofJcAKR, JcGLY1 and JcGLY2 determined through qRT-PCR analysis, showed up-regulation in leaf and root tissues underprogressive drought and salinity stresses. To further confirm the protective roles of JcAKR, JcGLY1 and JcGLY2, spottingassays with supplemental PEG (5%), NaCl (200 mM) and MG (5 mM) were performed with E. coli and yeast. The bacteriaand yeast harbouring the respective genes were able to grow in the presence of external stress compared to controls. Inconclusion, JcAKR, JcGLY1 and JcGLY2 act together to protect the plant from the harmful effects of MG under conditionsof abiotic stress.

PA072 (IPPC0498)

Genetic engineering of elite rice cultivars by inducible expression of late embryogenesis abundant gene(s)

from wild genotype, O. nivara improves tolerance towards soil water stress without yield penalty

Ranjeet Kaur*, Soumitra Kumar Sen and Ananta Kumar Ghosh

Indian Institute of Technology, Kharagpur-721302, West Bengal, India


Soil water stress (SWS) is one of the major environmental stress conditions that limits crop yield. Inducible expression ofgene(s) conferring tolerance to SWS is desirable for producing farmer-friendly stress tolerant crop plants. Gene(s) encodinglate embryogenesis abundant (LEA) proteins could be a prospective candidate in this regard. A comparative expressionprofiling of Wsi18, a group 3 LEA gene, performed by means of stage specific genome wide microarray and subsequentreal time qPCR analysis, revealed differential response under SWS conditions across different rice genotypes. Geneticpolymorphism in the coding DNA sequence as well as in the 5' regulatory elements of Wsi18 were looked into betweenwild rice and elite cultivars and it revealed interesting insight into their evolutionary structure-function relationship. Wsi18promoter from Oryza nivara (wild progenitor of cultivated rice) showed enhanced expression of the reporter gusA gene incomparison to its orthologue from IR20. A further attempt was undertaken to functionally ascertain the role of such variantallelomorphic forms to influence the capacity to withstand SWS condition of any stress sensitive elite rice cultivar in termsof their ultimate grain yield via transgenic approach. When driven by the endogenous OnWsi18 promoter, the transgenicIR20 rice plants carrying the OnWsi18 gene showed improved tolerance towards SWS without any yield penalty. Thestudy, to the best of our belief, is expected to contribute to the development of molecular strategies to generate SWStolerant rice lines that can produce well.


151

IPPCongress2015


PA073 (IPPC0511)

Drought stress impact on physiological, biomass and yield parameters- genotypic variability in maize

P. Vagheera, R. Abdul, M. Vanaja*, M. Maheswari, P. Sathish, N. Jyothi Lakshmi, G. Vijaya Kumar, S.K. Yadav andB. Sarkar

Central Research Institute for Dryland Agriculture (CRIDA), Santoshnagar, Hyderabad, India


Five maize (Zea mays L.) genotypes- DTL-3, DTL-4, DTL-4-1, DTL-9 and DTL-11 from CRIDA maize germplasm wereevaluated along with drought tolerant genotype Varun for their response to drought stress at flowering and to identify thetraits contributing to it. The moisture deficit stress was imposed at initiation of anthesis and stress was relieved oncewilting symptoms appeared. The measurements of photosynthesis and transpiration were recorded at stress point andafter stress release and compared with well watered controls. Genotypes DTL-3 and DTL-4-1 recorded higher photosyntheticrate, lower stomatal conductance, lower transpiration rate and higher WUE even under drought stress and stress releasedconditions. With imposition of drought stress, photosynthetic rate reduced from 38% (DTL-3) to 62% (Varun), howeverDTL-3 and DTL-11 recovered to normal when stress was relieved. Drought stress decreased total biomass of maizegenotypes from 6% (DTL-4-1) to 29% (DTL-11) due to decreased stem and cob biomass. Among the genotypes, DTL-4recorded highest per se total biomass and seed yield under both irrigated and stress condition. Drought stress reducedseed yield (7% to 43%) and the reduction was mainly due to decreased seed number (6% to 44%) while change in testweight was non-significant. DTL-4 performed best among the selected genotypes both under well watered and droughtstress condition due to its better ability to partitioning of biomass to shoot and reproductive parts.

PA074 (IPPC0512)

Hydraulic dynamics were associated with expression of major intrinsic proteins (MIPs) in mulberry

(Morusspp L.) subjected to progressive drought stress

Kanubothula Sita Rami Reddy* and Attipalli R. Reddy



Two mulberry genotypes including a drought tolerant (DT), Selection-13 (S13), and a drought susceptible (DS), Kanva-2(K2), were subjected to progressive drought stress to elucidate the hydraulic dynamics in relation to changesinphotosynthetic leaf gas exchange characteristics as well as to understand the expression patterns of major aquaporins.Drought imposition induced severe reduction in photosynthetic performance in both S13 and K2 genotypes after 15 daysof stress (DAS) than their respective controls. However, S13 showed better photosynthetic rate (P

n) and water use

efficiency (WUEi), with minimal reduction in stomatal conductance (g

s) and transpiration (E). These characterizations of

photosynthesis and water use efficiency in turn were associated with greater leaf water potential (øL), sap flow (F), leaf

(KL) and stem (K

S) hydraulic conductance compared toK2 at 15 DAS. Further, S13 showed an upregulation of mRNA

transcripts of MIP isoforms including plasma membrane intrinsic proteins (PIPs) and tonoplast intrinsic proteins (TIPs) inboth leaf as well as in roots after 15 DAS. In conclusion, plant hydraulics and the expression levels of certain MIPs shouldserve as desirable traits in crop improvement programs for drought tolerance.


152

IPPCongress2015


PA075 (IPPC0518)

Proteogenomic landscape of an orphan legume, grasspea, challenged with dehydration stress

Divya Rathi*, Subhra Chakraborty and Niranjan Chakraborty

Plant Stress Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Plants respond and adapt to adverse environmental stresses by altering their cellular metabolism and activating variousdefense machineries. Investigation of genetic resource for stress adaptation, in plants, is crucial not only for betterunderstanding the underlying mechanism, but also as an attractive target for improving stress tolerance. Grasspea, oftenregarded as an ‘orphan’ legume, serves as a model system for rich source of nitrogen beneficial for soil fertility, and is avaluable source of dietary protein. Further, many of its cultivars are capable to withstand a myriad of constraints, makingit a suitable model species to study stress tolerance. As part of proteomics studies on legumes, we reported earlier thesalinity- and cold-responsive proteome of grasspea. In the present study, 3-week-old grasspea seedlings were subjectedto dehydration for 144 h. There were no visible changes in the seedlings till 48 h of dehydration. The leaflets rolled after60 h and displayed chlorosis, and the damage aggravated further during 72-144 h. The physiological changes of stressedseedlings were thereby monitored. The temporal effects of dehydration on grasspea were evaluated at the proteomics aswell as transcriptomics levels using classical 2-DE and RNA-seq, respectively. The 2-DE analysis revealed approximately200 proteins that were dehydration-responsive and were differentially regulated in a qualitative as well as quantitativemanner. Conversely, approximately ten-fold numbers of transcripts were found to be dehydration-responsive. Thiscomparative stress-responsive proteome of grasspea would provide new information that would be used for targetedbreeding approaches for crop improvement.

PA076 (IPPC0522)

Specific leaf weight: Better indicator to screen drought tolerance in cotton genotypes

S.B. Amarshettiwar*, T.H. Rathod, G.R. Shamkuwar and W.K. Shembekar

Department of Agricultural Botany, PGI, Dr. PDKV, Akola-444104, India


Present investigation was conducted during kharif season of 2010 and 2011 in RBD on the experimental field under nonstress and water stress conditions in pot culture. The seeds of 31 cotton genotypes (10 G. arboreum and 21 G. hirsutum)were sown in three replications. Experimental findings are based on two years pooled data under water stress and nonstress conditions. Results indicate significant superiority of G. arboreum over G. hirsutum, which indicates its tolerancetowards water stress on the basis of morpho-physiological and seed cotton yield. Result revealed that Under water stresscondition, five G. arboreum genotypes, viz., AKA-8 (65.03 g/plant), AKDH-1 (62.30 g/plant), AKDH-5 (55-69 g/plant),Garrohill (55.15 g/plant), GAK-423 (54.97 g/plant) and three G. hirsutum genotypes, viz., PKV Hy-2 (58.22 g/plant), PKV-Hy-5 (57.87 g/plant), PKV-Hy-4 (52.43 g/plant) recorded significantly higher seed cotton yield, which were 46.7, 40.54,25.63, 24.41, 24.0, 31.33, 30.54 and 18.27% higher than general mean (44.33 g/plant) of both the species, respectively.At 120 DAS, both under stress and non stress conditions, G. arboreum genotypes showed significantly higher SLW thanG. hirsutum. Specific leaf weight was found to be best indicator for drought tolerance. Among G. arboretum, AKA-8 (1.78g/dm²) Garrohill (1.91 g/dm²), AKDH-1 (1.82 g/dm²), KWA-8 (1.53 g/dm²), HD-110-151 (1.50 g/dm²), AKA-7 (1.49 g/dm²)and among G. hirsutum, PKV-Hy-4 (1.51 g/dm²), PKV Hy-5 (1.34 g/dm²), Yamuna (1.17 g/dm²) and PKV-Hy-2 (1.10 g/dm²) were found more tolerant genotypes for water stress condition. The correlation co-efficient revealed positive correlationsbetween seed cotton yield and SLW (r=0.741),+ and this criteria may be useful for assessing genotypes for droughtstudies.


153

IPPCongress2015


PA077 (IPPC0525)

Screening of cotton (G. hirsutum L.) germplasm for adaptation to drought stress resilient

Jayant Meshram* and J. Annie Sheeba

Crop Production Division, ICAR-Central Institute for Cotton Research, Nagpur-440010, India


Though cotton is drought tolerant crop, drought is a major problem in rain-fed areas. . Leaf reddening and wilting are alsogetting more pronounced in recent years, following drought stress. A field experiment was conducted under rain-fed andirrigated condition at ICAR-CICR, Nagpur during 2014-15 to study drought tolerance traits in twenty cotton genotypes onthe basis of various stress indices for yield and yield attributing characters. The physiological changes induced due towater stress were quantified by withholding irrigation for continuous 25 days to 45 days old cotton plants in July sowncrop. In all twenty genotypes comprising Bt cotton (Mallika), non-Bt cotton (var. Suraj), susceptible and drought tolerantgenotypes were evaluated for leaf area, leaf relative water content, mid-day water potential, epicuticular wax in theleaves, plant height, number of bolls per plant, boll weight (g) and correlated with temperature and volumetric soil moisturecontent. Data revealed that Nagpur 9, F-1226, and SGNR-27 genotypes recorded highest epicuticular wax content andyield attributing characters under both irrigated and rain-fed conditions. While IC-357045 germplasm accession showedreduction in drought tolerance traits and seem to be susceptible to moisture stress. A field phenotyping approach is toinitially identify promising germplasms with high cellular tolerance to drought stress to withstand dry spell for improvingfield level drought tolerance. The differences of root traits in terms of drought tolerance will enable to augment cottonproductivity under water limited environments but combining these traits with high yield potential is required.

PA078 (IPPC0527)

A Class-I TCP protein modulates ABI4-mediated pathways and lipid droplet synthesis to influence

developmental and abiotic stress signaling

Pradipto Mukhopadhyay* and Akhilesh Kumar Tyagi



Abiotic stresses severely affect development of plants and facilitate the production of stress-related factors at the cost ofgrowth-related proteins. A balance between these two classes of proteins determines the stress tolerance limit of theplants. TCP transcription factors form an indispensible group of plant-specific developmental regulators. We identified astress-upregulated Class-I TCP transcription factor in rice, OsTCP19, the expression of which was found to be higher instress-tolerant varieties under dehydration and high salinity. This gene also presents a case of tissue, variety and stressdependent alternative splicing. Overexpression of this gene not only affects the expression auxin, abscisic acid andjasmonate-dependent pathways but also increases the synthesis of lipid droplets and related genes in Arabidopsis. Thetransgenic Arabidopsis plants show many developmental alterations in addition to increased tolerance to abiotic stress.In silico analysis also predicts OsTCP19-mediated similar alteration in gene expression in rice. This protein was furtherfound to interact with OsABI4 and OsULT1, a transcriptional co-regulator, and directly activate a triacyglycerol biosynthesisgene in transient assays. Together, these results suggest OsTCP19 as an important regulator which crosslinksdevelopmental and abiotic stress signaling.


154

IPPCongress2015


PA079 (IPPC0536)

Transcriptome analysis of salt susceptible blackgram (Vigna mungo L. Hepper) under saline conditions

Pooja Bangar1,2*, Ashok Chaudhury1 and K.V. Bhat2

1Bio & Nano Technology, Guru Jambheshwar University of Science and Technology, Gurgaon-122002, India2National Bureau of Plant Genomic Resources, Pusa Campus, New Delhi-110012, India


Among various abiotic stresses, salinity is one of the world’s most serious problems as plant agriculture faces a loss of$12 million annually due to salinity stress worldwide (FAO stat 2006). Blackgram (Vigna mungo L. Hepper, 2n=22) animportant grain legume grown for its protein rich edible dry seeds in the tropical and subtropical regions of the Indiansubcontinent, is adversely affected by salinity stress. We aimed to identify differentially expressed genes (DEGs) and itstranscription factors by comprehensive analysis of transcriptomes of both root and leaf tissues of Blackgram genotypegrown under the control and saline conditions. A total of427 million high quality reads were generatedby employingIlluminamiseq sequencing platform which were assembled (de nova) using CLC Genomics Workbench 7.5.1 at defaultparameters into 591,822 contigs with an average length of 300 bp and N50 value of 369 bp. Functional classificationrevealed 65.83% unique contigs into 20 subcategories of the biological process, 32.77% unique contigs into 14subcategories of molecular functions and 37.80% unique contigs into 11 subcategories of cellular components. Weidentified 5659 candidates salt responsive genes in which 1259 are upregulated and 4400 are down regulated genes(with a RPKM fold change >2 and a cut off of p <0.001) with significantly different expression under salt water and freshwater treatments. The data generated in this study will help in understanding the fundamentals of salt tolerance in blackgramand will provide resources for functional genomics studies and in crop improvement.

PA080 (IPPC0559)

Oxidative stress tolerance correlates with high temperature stress tolerance at plant level in rice

P. Boominathan*, R. Sivakumar and V. Ravichandran

Department of Crop Physiology, Tamil Nadu Agricultural University, Coimbatore–641003, Tamil Nadu, India


Among the many challenges to achieve higher productivity in rice, climate change associated phenomenon especiallyhigh temperature is increasingly becoming an important factor. Most of rice is currently cultivated in the regions wheretemperature is close to optimum or above for growth. Therefore, any further increase in mean temperature would certainlyreduce the rice yield. One of the most common high temperature stress effects at cellular level is the oxidative stresscaused by various reactive oxygen species. An investigation was conducted with eleven genotypes including popularvarieties of Tamil Nadu, landraces and tolerant and susceptible cultivars known for the high temperature tolerance withthe hypothesis that oxidative stress tolerant genotypes would also tolerate abiotic stresses including high temperaturestress. To assess the genetic variability in response to oxidative stress, rice seedlings were treated with a quinine, knownas menadione, to generate reactive oxygen species. The duration of exposure and concentration of menadione optimizedas 2 mM for 24 hrs in rice. The study revealed that the genotypes white Ponni, Moroberekan, Norungan which showedhigher sensitivity to high temperature in the growth chamber at plant level, also exhibited higher percent reduction insurvival during menadione treatment. Thus, developing the oxidative stress tolerant genotypes would eventually helps inidentifying genotypes tolerant to high temperature stress. The tolerant genotypes displayed lower membrane injury thusstrongly supporting the fact that membranes are primary sites for oxidative stress.


155

IPPCongress2015


PA081 (IPPC0568)

Influence of waterlogging on physio-biochemical traits and expression behavior of SOD and ADH genes

in sugarcane genotypes

Radha Jain1*, S.P. Singh1, Amaresh Chandra1, Anshu Singh1, Smita Singh1, R.K. Singh1, A.D. Pathak1, M. Swapna1 andV.K. Srivastava2

1Plant Physiology and Biochemisrty Division, Indian Institute of Sugarcane Research, Lucknow-226002, U.P., IndianUttar Pradesh Council of Sugarcane Research, Shahjahanpur, U.P., India


Waterlogging is one of the serious abiotic stresses affecting sugarcane productivity. It usually occurs due to rise in watertable in almost the entire canal irrigated areas during rainy season (July-September) in North Bihar, Eastern Uttar Pradesh,West Bengal, coastal areas of Orissa, Andhra Pradesh and Kolhapur area of Maharashtra. Cane yield and juice qualitylosses due to waterlogging depend on genotypes, environmental conditions, development stage and inundation duration.In present study, an effort was made to characterize sugarcane genotypes for waterlogging tolerance based on physio-biochemical traits. Three bud setts of twenty four sugarcane genotypes including six commercial cultivars, CoLk 94184,BO 91, CoS 767, CoJ 64, CoS 97264 and UP 9530 were planted at Kharika Block, Indian Institute of Sugarcane Research,Lucknow during spring season under two different conditions; control (non-waterlogged) and waterlogged. In waterloggingtreatment, crop was grown in deep plot which was naturally waterlogged during rainy season. Results obtained indicateddecrease in leaf area, SPAD reading, chlorophyll a, b, carotenoids, specific leaf weight (SLW), nitrate reductase (NR)activity under waterlogged conditions. Almost all the genotypes exhibited aerial rooting due to waterlogging. Genotypesvaried significantly for SLW, SLA, SPAD and NR activity. Quality attributes; oBrix, sucrose and juice purity were decreaseddue to waterlogging; only a few genotypes showed slight increase over control. Waterlogged affected plants exhibitedhigher expression of ADH and SOD genes in leaf, root and stalk; ADH expression was significantly high in roots. Presentfindings help in identifying and characterizing sugarcane genotypes tolerant to waterlogging.

PA082 (IPPC0585)

OsHsfA6a directly regulates a subset of Hsp genes and functions as omphalos for transcriptional regulation

of ClpB/Hsp100 genes in rice (Oryza sativa L.)

Dhruv Lavania*, Anirban Chakraborty, Anuradha Dhingra, Juarez Road, Dhaula and Anil Grover

Department of Plant Molecular Biology, University of Delhi, South Campus, Benito Juarez Road, Dhaula, New Delhi-110021, India


ClpB/ Hsp100 proteins are critically important in governing heat stress tolerance in plants. The expression of OsClpB-C(cytoplasmic isoform) is mainly regulated at the transcriptional level. We have earlier shown two important events in thisprocess: (a) OsHsfA6a regulates OsClpB-C promoter and (b) OsHsfA6a interacts with OsClpB-C and OsHsfB4b proteins.In this study, OsHsfA6a was identified as the only direct transcriptional activator of OsClpB-C and OsClpB-P (plastidialisoform) genes and as direct transcriptional activator of a subset of heat inducible OsHsp genes. The transcriptionalactivity of OsHsfA6a was repressed in reporter gene assays by interactions with OsClpB-C and OsHsfB4b proteins.Protein-protein interaction analysis revealed heteromeric interactions of OsHsfA6a with several class A OsHsfs and thecomplexes were nuclear localized. This is the first study on heteromeric interactions of subclass A6 Hsf member and thehighest number of heteromeric interactions for any plant Hsf to date. The interacting OsHsfs showed differential transcriptexpression kinetics under a time-course regime of heat stress and recovery. We thus infer that the target gene selectivityand activity of OsHsfA6a may be regulated by virtue of these interactions at post-translational level. Our findings suggestthat binding of OsClpB-C with OsHsfA6a may work in a negative feedback-loop manner and OsHsfB4b might function asa repressor protein. We conclude that a supra-complex involving several OsHsfs regulates Hsp promoters under heatstress and recovery in rice.


156

IPPCongress2015


PA083 (IPPC0598)

Better antioxidative defense mechanism of rainfed lowland rice varieties is associated with their tolerant

to submergence

Ashish Kumar Srivastava1*, Akhtar Hussain Khan2, Phool Chand Ram2, Sudhanshu Singh1 and Abdelbagi M. Ismail3

1Plant Breeding Genetics and Biotechnology, International Rice Research Institute, New Delhi-110008, India2Narendra Deva University of Agriculture and Technology, Kumarganj, Faizabad, UP, India3International Rice Research Institute, Los Baños, Philippines*Presenting author: [email protected]

21-days-old seedlings of 4 tolerant (FR13A, Swarna-Sub1, NDR9730018, NDR9930111) and 2 susceptible (Swarna andIR42) lowland rice varieties were submerged for 10 days in outdoor flooded water tank. Same set of lines were plantedwithout imposing submergence to record grain yield under natural conditions. Enzymes of Halliwell-Asada pathway (SOD,APX, GR) were estimated before submergence and 0, 2, 4, 6 and 24th hours after de-submergence, whereas enzymeCAT and antioxidants (AsA and GSH) were estimated before and just after de-submergence. SOD activity increased (3-4 fold in tolerant vs 1-1.5 fold in susceptible) up to 4th hr after de-submergence and subsequently declined till 24th hr.Upon de-submergence successive increase in APX activity was recorded up to 24th hr in all varieties. Tolerant genotypesshowed consistently higher GR activity after de-submergence. Higher CAT activity was recorded in tolerant varieties aftersubmergence (48.8-58.2%) when compared to IR42 and Swarna (28.4 and 38.3%). AsA and GSH contents showedopposite trend after de-submergence, IR42 and Swarna showing 7.6 and 14.7% increase in AsA values and more than80% reduction in GSH values. After submergence, FR13A and Swarna-Sub1 recorded more than 85% survival, whereasIR42 and Swarna showed only 41.5% and 61.5% survival. FR13A and Swarna-Sub1 exhibited only 41.7 and 50% reductionin grain yield, whereas Swarna and IR42 showed 60 and 68% reductions when compared to their yield in natural conditions.It can be thus concluded that better antioxidative defense mechanism of tolerant lines is associated with better survivaland higher grain yield under submergence.

PA084 (IPPC0617)

Identifying plant genetic resources adapted to elevated temperature

Sunil Archak1*, Anuj Singh1, Sarika Mittra2, D.P. Semwal1, P.N. Mathur3, P.K. Aggarwal4 and K.C. Bansal1

1ICAR-National Bureau of Plant Genetic Resources, New Delhi-110012, India2Bioversity International, New Delhi-110012, India3Regional Representative, Central and South Asia, Bioversity International, New Delhi-110012, India4Regional Program Leader South Asia, CCAFS, New Delhi-110012, India


Climate smart agriculture hinges on the cultivars with greater adaptability and resilience. This in turn necessitates the useof a wider range of intra-specific diversity conserved in the genebanks. Estimation of adaptive capacity of genebankaccessions based on passport data and identification of germplasm accessions with excellent adaptation potential canhelp develop climate smart crop varieties. Globally, genebanks are working to bring together the inherent diversity of ex

situ collections and power of climate analogues for enhancing adaptive capacity of food crops to climate change. Weattempted to employ climate tools to identify pre-adapted germplasm among ~39,000 genebank accessions belonging tofive select crops- wheat, pearl millet, chickpea, pigeon pea and sorghum. The methodology comprised geo-referencingand clustering the accessions, climate matching, designating pre-adapted material and collecting germplasm from predictedsites.


157

IPPCongress2015


PA085 (IPPC0621)

Overexpression of AtICE1 gene improves abiotic stress tolerance in transgenic rice

Rakesh Verma1*, Santosh Vinjamuri1, Shashank Yadav1, M.V. Rao2 and Viswanathan Chinnusamy1

1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India2Department of Plant Sciences Bharthidasan University, Tiruchirappalli, Tamil Nadu, India


Abiotic stresses such as drought, soil salinity and freezing, greatly affect plant productivity and quality. Many genes areinvolved in the process of adaptation to adverse environmental conditions. ICE1 (Inducer of CBF Expression 1), anupstream transcription factor, regulates the transcription of CBFs and other genes, and confers freezing tolerance toArabidopsis. Recent studies from different plant species suggest that ICE1 also regulates tolerance to other abioticstresses. Arabidopsis ICE1 gene was isolated by RT-PCR and cloned in a modified pCAMBIA1300 under stress inducibleAtRD29A promoter and transformed into indica rice variety Pusa Sugandh 2 by the Agrobacterium-mediatedtransformation. Cold, drought and salt stress tolerance of transgenic events were compared with the non-transgenicplants in seedling, vegetative and flowering stages. Transgenic plants exhibited enhanced tolerance to these stressesin terms of RWC, chlorophyll and membrane stability. In addition, transgenic rice plants overexpressing AtICE1 exhibitedhigher stomatal density but smaller stomatal size as compared to non-transgenic plants. Further, leaf width and lengthwere significantly lower in transgenic plants. The results suggest that the function of AtICE1 in stress tolerance andstomatal development is conserved in rice and the over-expression of ICE1 can improve tolerance to abiotic stresses inrice.

PA086 (IPPC0625)

Adverse effects of short exposure to high temperature on wheat and mustard and its amelioration through

iron and zinc application

Uday Burman* and Praveen Kumar

Division of Integrated Land Use Management & Farming System, Central Arid Zone Research Institute, Jodhpur-342003, India


Changes in temperature significantly influence plant processes and subsequent crop productivity. Temperature increasedby 3-5 OC when plants are partially covered with <100 μm thick transparent polythene sheet. This resulted in 69.0, 70.7,45.8 and 59.5 % reduction in plant water potential, nitrate reductase (NR) activity, total soluble carbohydrate (TSC) andstarch in mustard. Corresponding values in wheat were 40.0, 39.3, 5.5 and 13.1%, respectively. In a two year study,seeds of both mustard (var. Pusa Jai Kisan) and wheat (var. Raj 3077) were primed with FeSO

4 (10%), Fe EDDHA (0.6%),

ZnSO4 (36%) and Zn EDTA (6.0%) prior to sowing in pots. In another experiment, 0.125, 0.250, 0.50 and 1.00 % iron and

zinc sulphate were foliar sprayed at pre flowering stage in field. Thereafter plants were subjected to high temperatures atflowering stage for seven days in both the experiments. Priming with 0.6% Fe EDDHA was associated with highestincrease in RWC (30.7%), TSC (26.5%) and starch (68.0%) compared to corresponding control in wheat. Similar responsewas observed in mustard also. Foliar spray of 0.125% Zn was associated with less reduction in yield upon exposure tohigh temperature compared to control in both wheat and mustard. High NR activity (21% more than control) in wheat andmaintenance of high membrane stability index in mustard could be responsible for the partial alleviation in yields.


158

IPPCongress2015


PA087 (IPPC0627)

Phenotypic traits as selection tools for screening mango rootstock against salt stress

V.K. Singh* and S. Rajan

Crop Production Division, ICAR-Central Institute for Subtropical Horticulture, Lucknow-226101, India


Mango is highly sensitive to soil salinity and alkalinity and tolerant rootstocks have potential for making mango cultivationpossible in these problem soils. Keeping this in view, poly-embryonic cultivars as rootstock, viz., Bappakai, Goa, Kurukkan,Moovandon, Mylepelian, Nekkare, 13-1, Kitchner, Peach, Starch, and Chandrakaran, Olour, Phillippino and Vellaikulambanwere evaluated under salt stress conditions. Symptoms of salt injury in seedlings appeared within one month and athigher EC (2-4 dS m-1), all plants died except Nekkare, 13-1 and Kurukkan. Salinity factor index revealed that 13-1 hasbetter tolerance to salt stress and can survive at high level of salinity. With increasing salinity levels, highest reduction inroot/shoot ratio was recorded in Goa and lowest in 13-1. Electrolyte leakage showed severe membrane damage in thestressed plants with drastic reduction in chlorophyll fluorescence and PSII photochemistry in majority of the rootstocks.Proline and glycine-betaine content were higher in leaves (4.5–6.0 mg g-1 dry wt.) than roots (1.2–3.5 mg g-1 dry wt.) withelevated salt stress, and results indicated low contribution towards osmotic adjustment. Leaf water potential was drasticallyreduced with greater reduction (-19.78 ψ

w) in Starch and Kitchner, whereas, Kurukkan, 13-1 and Nekkare exhibited least

reduction. Rootstock 13-1 exhibited higher cuticular wax (608.50 μg cm-2) content even in control (normal soil) plants,however, Kurukkan and Nekkare exhibited higher level of leaf wax when they were exposed to salt stress. Thus, thesetraits may be utilized for screening polyembryonic mango cultivars for rootstock purpose against salt stress.

PA088 (IPPC0630)

Effect of salinity on morphological traits of F2 population in rice

P. Pundir*, S.L. Krishnamurthy, P.C. Sharma and D.K. Sharma

Department of Biotechnology, Central Soil Salinity Research Institute, Karnal-132001, India*Presenting author: [email protected]

Salinity causes significant yield reduction in rice. There are substantial opportunities to improve grain yield by usingconventional breeding under stress conditions. The F

2 segregating population (PS 5/CSR10) of rice was studied under

saline stress and non stress conditions at reproductive stage. The objective of this study was to unveil the performanceof same F

2 progenies under both saline and non stress environment (EC ~ 10 dSm-1) and to find out the suitable stress

indices. Significant variability was found in segregating population for grain yield and other traits under both saline andnon-saline condition. There was a mean reduction of 89% in grain yield due to high salt stress (EC~10 dsm-1). Under non-stress environment the significant positive correlation was noticed between grain yield and biomass (r=0.64) followed bygrain yield with plant height (r=0.36) and grain yield with productive tillers (r=0.30). Under salinity weak and insignificantassociation of grain yield was observed with other traits. Grain yield of the F

2 population ranged from 400 to 9333 kg h-1

under normal environment while it reduced to 40-1591 kg ha-1 under salinity. Grain yield and biomass were the affectedtraits most by salinity. Skewness and Kurtosis indicates the presence of complementary gene interaction under bothenvironments thus population can be further improved for selection of salt tolerant lines. Correlation coefficient and biplotanalysis of stress indices indicated that GMP, STI, MP.


159

IPPCongress2015


PA089 (IPPC0641)

Elucidating dominant enzyme mediated oxidative stress tolerance pathway in groundnut under salinity

stress

Koushik Chakraborty*, Sujit Bishi, Kiran Patel, Nisha Goswami, Debarati Bhaduri, A.L. Singh, Pratap V. Zala

Directorate of Groundnut Research, Junagadh Agricultural University, Junagadh-362001, Gujarat


Salinity, a major abiotic factor limits plant growth and productivity globally. Oxidative stress, aroused due to higheraccumulation of reactive oxygen species (ROS), restricts plant metabolism under salt-stress. Previous reports suggestedexistence of more than one mechanism for ROS-detoxification adapted by plants under abiotic stress. Hence, the presentstudy focuses on elucidating major oxidative stress tolerance pathway in groundnut under salinity stress using differentiallysalt-sensitive genotypes. For this, a pot experiment was conducted with 6 genotypes (‘CS 240’, ‘NRCG 357’, ‘TMV 2’,‘Girnar 1’, ‘TPG 41’, ‘Somnath’) and four different levels of salt treatment (T1:0, T2:25, T3:50 and T4:100 mM NaCl).Imposition of salinity treatments resulted in significant stress build-up evident from higher soil ECe value of 3.7 and 5.4 dSm-1 in T3 and T4, respectively and significant drop in leaf water potential (-15.5 MPa in control to -40 MPa under T4).Severe plant mortality was observed at T4 treatment. Determination of ROS level build-up by both spectrophotometricand dye-staining method revealed comparatively higher oxidative stress mediated damage in ‘NRCG 357’ and ‘TMV 2’than ‘Somnath’ and ‘CS 240’. Activity assay of major ROS-detoxifying enzymes (superoxide dismutase, catalase,peroxidase, glutathione reductase, ascorbate peroxidase) and their relative transcript abundance through real-time qPCRtechnique revealed up-regulation of SOD, CAT and POD under salt stress, while no significant induction observed in GRand APX. Thus present study concludes, minimal role of ascorbate-glutathione mediated ROS-detoxification pathwayand major dependence on SOD, CAT, POD pathway for oxidative stress tolerance in groundnut under salinity stress.

PA090 (IPPC0646)

Phenotyping of rice genotype for high temperature stress tolerance at anthesis stage

Sourabh Karwa*, Ashish K. Chaturvedi, Divya Shah, Shashi Meena and Madan Pal Singh



Rising temperature is one of the most important climate factors affecting rice yield .Rice is considered sensitive to hightemperature stress at flowering stage. High temperature can affect rice grain quality. This study reports phenotyping ofthirty-six rice genotype raised in pots under ambient environment and exposed to high temperature stress (>38-40p C)after anthesis till maturity of crop. After ten days exposure to high temperature, spikelets were collected from control andhigh temperature stressed plants and pollen viability and fertility were analyzed using fluorescent microscopy. Thermalimages of the plants were captured using IR thermal image camera and CTD was calculated. Spikelet fertility and panicleyield was calculated after harvest of crop. There was large variability in spikelet fertility among the genotypes, and it wassignificantly correlated with grain yield per plant. Based on the spikelet fertility, ten contrasting genotypes were identified.Tolerant rice genotypes showed higher pollen viability and fertility (65-80%) under high temperature, while in sensitivegenotypes both the traits showed up to 70% reduction. Similarly, tolerant genotypes showed 3.4-5.5p C CTD under hightemperature stress. All the genotypes will be characterized on physiological basis of high temperature tolerance andmost tolerant genotypes will identified for future breeding program of rice crop.


160

IPPCongress2015


PA091 (IPPC0667)

Evaluation of walnut (Juglans regia) genetic diversity in central Himalayan region for moisture stress

and quality

A.K. Trivedi1,2*, S.K. Verma1 and R.R. Arya1

1ICAR–National Bureau of Plant Genetic Resources, Regional Station Bhowali District Nainital-263132, Uttarakhand, India2Present Address: ICAR–Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow-226101, Uttar Pradesh, India*Presenting author: [email protected]

Walnut (Juglans regia L.) is an important temperate nut grown in India. Walnut cultivation is sensitive to abiotic stresses,particularly moisture stress at seedling stage. Dry weather reduces potential productivity of walnuts even in the absenceof soil water deficit. Although, it is one of the major nut fruit grown in Himalayan region but it has remained a low prioritycrop in temperate fruit growing areas of India. Genetic diversity of walnuts available in the Himalayan region, which maypossess valuable stress resistance that helps them to cope with the unfavourable environmental conditions. Hence, inorder to collect available genetic diversity of walnut, germplasm survey and exploration expeditions were conducted incentral Himalayan region. A total of 15 accessions having distinct traits of horticultural importance were collected andconserved in the field gene bank along with four exotic accessions for comparative evaluation. Conserved germplasmhave been evaluated for different morpho-physiological traits, antioxidants and nutrients, and under normal and moisturestress condition at seedling stage. Considerable diversity in vegetative parameters, fruiting traits, antioxidants, nutrientsand response to moisture stress at seedling stage has been found. Moisture stress tolerant accessions have been foundto show higher proline accumulation accompanied by higher activity of antioxidant enzymes. Untapped genetic variabilityavailable in this region may be utilized for cultivation as well as for crop improvement programmes.

PA092 (IPPC0668)

Role of reactive oxygen species (ROS) during the growth of mungbean [Vigna radiata (L.) Wilczek]

seedlings under water stress

Satyajit Das* and Rup Kumar Kar

Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva-Bharati University, Santiniketan, West Bengal, India


Recently a role of ROS in growth and development in plants has been revealed. While working with Vigna radiata seedlingswe observed that mild water stress (simulated by PEG 6000) promoted root growth while inhibiting growth in hypocotyl,which might be controlled by extracellular ROS. Thus, root growth of 3-day-old seedlings was retarded, both under waterstress and non-stress condition, by treatment with CuCl

2, a superoxide scavenger, as well as by ZnCl

2, an inhibitor of

NADPH oxidase (PM-located superoxide generator). However, in case of hypocotyl these treatments were without effect.Treatments with DMTU (H

2O

2 scavenger) and DEDTC (SOD inhibitor), though inhibited normal growth, could not inhibit

stress-induced root growth. Again these treatments did not effect hypocotyls. When seedlings were treated with sodiumbenzoate a OH scavenger, and SHAM, a peroxidase inhibitor, root growth under both stress and non-stress conditionswas inhibited, will the former being more effective. Again these were not effective in hypocotyl. In-gel assay showedhigher activity of NADPH oxidase (RBOH) in root, but not hypocotyl, under PEG-induced stress confirming the involvementof apoplastic superoxide in stress-induced root growth, although data on assessment of extracellular production ofsuperoxide by estimating in bathing medium did not support this. But localization of superoxide and H

2O

2 by staining root

and hypocotyl separately with NBT and TMB, respectively confirmed the involvement of ROS in water stress-inducedgrowth promotion of root whereas these are not involved in growth inhibition of hypocotyl under water stress simulated byPEG 6000.


161

IPPCongress2015


PA093 (IPPC0683)

Identification of promising salt tolerant genotypes using stress indices of morphological and physiological

trait at seedling stage in rice

S.L. Krishnamurthy*, V. Batra, P. Pundir, P.C. Sharma and D.K. Sharma

Division of Crop Improvement, Central Soil Salinity Research Institute, Karnal-132001, Haryana, India


A set of 131 rice accessions was evaluated in two levels namely, non-stress (EC ~ 1.2 dS/m) and saline stress (EC ~ 10dS/m) in hydroponics at seedling stage. Root length and shoot lengths were reduced by 52 and 50%, respectively insaline stress in comparison to non stress conditions. The effect of the increased Na+ concentration in the medium isdetrimental to root length and shoot length as observed by reduction in root length and a concomitant reduction in shootlength. The increased concentration of Na+ leads to augmented Na+/K+ ratio with increasing stress in medium and decreasingexpression of traits. A significant positive correlation (r=0.60) was noticed between stress tolerance index (STI) for rootlength and STI for shoot length. The stress susceptibility index (SSI) for root length was expressed significant positivecorrelation with SSI for shoot length (r=0.43). SSI for K+ content was registered significant negative correlation with STIfor Na+ content (r=-0.43). The three accessions namely, IC 545004, IC 545486 and IC 545215 were found to be the bestperformers adjudged on the morphological and physiological criteria in saline stress situation. These three rice accessionscould be used as a donor parent or for genotypic studies in future breeding programs.

PA094 (IPPC0687)

Development of salt tolerant varieties following conventional, physiological and molecular approaches

Parbodh Sharma1*, S.L. Krishnamurthy1, Neeraj Kulshreshtha2, Jogendra Singh1, Preeti Pundir1 and D.K. Sharma1

1Central Soil Salinity Research Institute, Karnal-132001, Haryana, India2Sugarcane Breeding Institute RRS, Karnal-132001, Haryana, India


Among abiotic stresses, salt stress is the most complicated problem posing a major challenge. Around 1000 m haglobally and 6.73 m ha in India are affected by salinity. Central Soil Salinity Research Institute (CSSRI) has developed 15salt tolerant varieties of different crops; 7 of rice (CSR10, CSR13, CSR23, CSR27, CSR Basmati30, CSR36 and CSR43),4 of wheat (KRL1-4, KRL 19, KRL210 and KRL213), 3 of Indian mustard (CS52, CS54 and CS56) and one in Chick pea(Karnal Chana 1) through conventional breeding approaches to increase the productivity of salt affected soils for resourcepoor farmers. Physiological and biochemical approaches have been used to identify different mechanisms of salt toleranceand used for identification of salt tolerant and sensitive lines. The molecular approaches are also being used to developsalt tolerant rice varieties. Introgression of Saltol QTL to two mega high yielding rice varieties namely, Pusa 44 and Sarjoo52 carried out through Marker Assisted Selection Back Cross (MABC) breeding. The varieties namely, Pusa 44 andSarjoo 52 were selected as recurrent parents while FL478 was used as a donor parent and produce F

1 seeds. True F

1s

were selected by the Saltol marker RM8094. Further backcrossing with respective recurrent parent and selfing wasperformed to produce seeds up to BC3F3 generation. In every back cross generation, recombinant and foregroundselection was carried out. Desirable plants were selected based upon the (genotyping) marker data and through stringentphenotyping as similar to recurrent parent. Background selection and DUS characterization of NILs is in progress.


162

IPPCongress2015


PA095 (IPPC0688)

Rice F-box protein, OsFBK, regulates abiotic stress signaling

Satyam Vergish*, Nitin Jain and J.P. Khurana

Department of Plant Molecular Biology, Interdisciplinary Centre for Plant Genomics, Delhi University, New Delhi-110021, India


The F-box proteins (FBPs) are components of SCF ubiquitin-ligase complexes and are one of largest multi-gene superfamilyin plants containing a conserved N-terminal F-box motif that binds to the SKP, a component of SCF, and a variable C-terminal protein interaction domain that selectively binds target proteins for turn-over by 26S proteasome. There are over900 FBPs in Arabidopsis and rice that are involved in all the facets of plant biology such as stress signalling,photomorphogenesis, hormone signaling, flower development, self incompatibility, biological rhythms, defence and soon. Our lab identified a few F-box proteins in rice that are regulated under abiotic stresses, one of which was OsFBK. Thegene was found to be upregulated under drought, heat and salt stresses and also in Y-leaf and flag leaf. The proteinharbors a canonical F-box domain which interacted with OsSKPs demonstrating that it is a component of a functionalSCF-type E3 ligase. The OsFBK protein showed formation of homodimers and heterodimers with its paralogs in yeasttwo-hybrid assays. Also screening of heat stress rice library showed interaction of OsFBK with some proteins known toplay roles in heat tolerance. The protein was found to localize in the nucleus as well as cytosol. Over-expression linesin Arabidopsis showed role of this gene in heat response and other abiotic stresses. Also, the over-expression linesflowered earlier. The above results indicate involvement of OsFBK in abiotic stress signalling and photoperiodic flowering.

PA096 (IPPC0693)

Assessment of physio-biochemical traits of rice landraces of Goa for salt stress tolerance

Kundan Kumar* and Swapnil Punyapwar

Department of Biological Sciences, BITS Pilani K.K. Birla Goa Campus, Goa- 403726, India*Presenting author: [email protected]

Rice is the major staple food crop for more than half of the world’s population. Salinity stress is the most common andserious problem in rice cultivation mainly in the coastal saline soils of Goa locally known as Khazan land and occupies 33percent of total rice cultivated area. Developing rice cultivars with tolerance to salt with better agronomic and quality traitsacceptable to farmers is a feasible approach to address this problem. Salt tolerance trait available with certain traditionalland races, not exploited so far, has significant relevance in crop improvement. The indigenous rice germplasm of Goa isrich in genetic diversity and valuable gene system for traits associated with salinity tolerance. In the present study, wehave analyzed three rice genotypes of Goa namely Dhave, Ek Kadi and Korgut for their salt tolerance at seedling stage.IR64, a salt sensitive cultivar of Oryza sativa was used as a control. Their physiological parameters viz. root length, shootlength, fresh weight, dry weight, germination percentage were taken into consideration for salt stress tolerance. Biochemicalparameters like salinity induced free proline accumulation, chlorophyll content, lipid peroxidation, catalase activity, totalprotein and elemental (Na+, Ca2+, K+) analysis were considered for salt stress tolerance in all 4 genotypes. Taken togetherour results suggest that Korgut cultivar showed better tolerance to salinity stress among all landraces tested here. Therice genotypes showing salinity stress tolerance will be highly useful in rice breeding for improved rice varieties to surviveunder higher salt stress condition.


163

IPPCongress2015


PA097 (IPPC0699)

Impact of ZnO nanoparticle on water stress and its recovery in Oryza sativa L.

H. Upadhyaya1*, S. Shome1, M.K. Bhattacharya1, S. Tewari2 and S.K. Panda3

1Department of Botany and Biotechnology, 2Department of Physics, Karimganj College Karimganj-788710, Assam, India3Plant Biochemistry and Molecular Biology Laboratory, School of Life Sciences, Assam, University, Silchar-788011, India


Zinc oxide (ZnO) nanoparticles (NP) is being widely used in various research. Plant system being extremely criticalrequires the fundamental understanding on the influence of ZnO-NP on its cellular growth and functions. Our studyevaluates the effect of ZnO-NP on growth and physiological changes in Oryza sativa L., and its ameliorative effect onPEG induced water stress and its recovery in rice. Water stress induced growth reduction in rice seedlings, but the - ZnO-NP alleviatied the adverse effect of water stress on rice seedlings by stimulation of growth parameters and the increasein photosynthetic pigments. It can also be suggested that the water stress recovery may be counteracted by ZnO-NP atlower concentration. Further, detail study on the ZnO-NP effect on physiological events during water stress, andphysiochemical and antioxidative responses of rice seedlings can reveal the exact mechanism underlying the ameliorationof water stress and its recovery in rice. The present study gives an insight on the possible role of zinc transporter proteinand their interaction with ZnO-NP during water stress acclimatization and stress recovery process in rice, which in turnwill help in understanding biointerface on protein–nanoparticles interaction in future. Detail understanding of the regulationof zinc transporter in response to Zn nanoparticles is required to dissect the physiological mechanism regulated by ZnO-NP during water stress and its recovery in rice.

PA098 (IPPC0711)

Validation of Sub1 QTL in CO 43 from submergence tolerance using key physiological traits

R. Samundeswari1*, D. Vijayalakshmi1 and M. Raveendran2

1Department of Crop Physiology, 2Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore-641003, India


In Tamil Nadu, Cauvery delta zone is frequently affected by flash flooding during the monsoon period. Hence, researchwas aimed towards improving submergence/flooding tolerance of popular rice genotypes of Tamil Nadu viz., CO43 byintrogressing Sub1 locus from the tolerant FR13A through marker assisted selection. BC1F1 generation was forwardedup to BC4F1 and selfed progenies of BC2 and BC3 generations were phenotyped for submergence tolerance using keyphysiological traits. Advanced generations of the Sub1 introgressed CO43 lines were subjected to flooding stress 13days in a simulated submergence screening facility and evaluated for traits namely leaf and stem carbohydrates, chlorophyllcontent, chlorophyll fluorescence (Fv/Fm), photosynthetic rates, stomatal conductance and transpiration rates. All theSub1 introgressed lines survived 13 days of complete submergence showing more reserved carbohydrates before andafter submergence, higher chlorophyll contents and chlorophyll fluorescence ratios coupled with high photosyntheticrates. CO43 could not survive 13 days of flooding and was found dead and decayed. Almost all the introgressed lineswere found to be superior to FR13A, the donor parent. Superior progenies of CO43 improved for their submergencetolerance has been developed which will be further tested for their agronomic performance and efforts will be taken forrelease as a smart rice variety to submergence tolerance.


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PA099 (IPPC0714)

Marker assisted introgression of Sub1 locus from FR13A into CO43 for improved submergence tolerance

D. Vijayalakshmi1*, M. Raveendran2, R. Samundeswari and S. Muthulakshmi

1Department of Crop Physiology, 2Central Institute for Cotton Research, Tamil Nadu Agricultural University, Coimbatore-641003,Tamil Nadu, India*Presenting author: [email protected]

The study was aimed at developing submergence tolerant version of popular rice variety of Tamil Nadu viz., CO43,through marker assisted introgression of Sub1 locus from a FR13A. BC1F1 generation was forwarded up to BC4F1 andselfed progenies of BC2 and BC3 generations were genotyped and phenotyped for submergence tolerance. Twenty oneBC3F1 plants were subjected to BGS using 37 SSR markers. The effect of introgression of Sub1 locus in terms oftolerance against submergence was studied in BC2F3 and BC3F3 progenies under simulated submergence condition.CO43 plants were completely affected. All the survived plants in BC2F3 and BC3F3 generations were evaluated for plantheight, carbohydrates, chlorophyll, chlorophyll fluorescence, photosynthetic rate, stomatal conductance and transpirationrates before and after flooding. All the introgressed lines proved to be flood resistant with improved physiological traits.Superior progenies of BC2F3 were selfed to generate BC2F4. BC2F4 progenies were raised in the field along with theparents and evaluated for the morphological traits. These superior Near Isogenic Lines (NIL’s) recorded the plant heightand grain quality similar to that of CO43. Superior NIL’s of CO43 improved for their submergence tolerance have beendeveloped which will be further multiplied and tested under multi-location trails.

PA100 (IPPC0725)

Genetic engineering of RSA and source-sink relationship in rice for improving abiotic stress tolerance

Preeti Kaur*, Pragya Yadav, Santosh Vinjamuri, Shashank Yadav, Rakesh Verma and Viswanathan Chinnusamy



Rice requires about 5000 litres of water to produce 1 kg of grain. Drought stress is a major constrain for rice production inIndia. Phytohormones play significant roles in dehydration tolerance mechanisms of plants. A low level of cytokinin hasbeen shown to promote root growth and drought tolerance in Arabidopsis and tobacco. Genetic and transgenic analysesin different plants revealed that cytokinin is vital for determination of grain number and maintenance of photosynthesis.This study was aimed at simultaneous improvement of root system architecture (RSA) and source-sink balance to enhancedrought tolerance in rice. Rice plants were engineered to down regulate cytokinin levels in root by over-expression ofcytokinin oxidase (CKX) by using root specific promoter (RCc3), and maintain appropriate levels of cytokinin in shoot byengineering drought responsive down-regulation of CKXs expression by RNAi in the shoot. Genes for cytokinin biosynthesis(IPT) and degradation (CKX) were identified from Oryza sativa genome. Selected genes were cloned into planttransformation vector. Transgenic rice of genotypes MTU1010 and IC305692 were developed using these constructsthrough Agrobacterium mediated transformation. Molecular analysis of transformants was carried out using PCR andRT-PCR. Rice transgenics will be further analyzed for altered RSA and source-sink relationship under normal anddrought stress conditions. These transgenics may be useful in developing rice lines with improved RSA and enhancedyield stability under drought stress.


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IPPCongress2015


PA101 (IPPC0741)

Expression analysis of SOS pathway genes under salinity stress in contrasting rice genotypes

T.V. Vineeth1*, Parbodh Sharma1, Ashwani Pareek2, Manjari Mishra2, Priyanka Das2 and S.L. Krishnamurthy2

1Crop Improvement Division, Central Soil Salinity Research Institute, Karnal-132001, Haryana, India2School of Life Science, Jawaharlal Nehru University, New Delhi-110067, India*Presenting author: [email protected]

Salinity is one of the major problems negatively influencing rice productivity. Salinity tolerance is a very complex trait.SOS (Salt Overly Sensitive) Pathway has been established as a major candidate in cellular ion homeostasis and therebysalt tolerance.The present study was carried out to examine the role of SOS pathway in salinity stress tolerance inselected rice genotypes. The experiment was conducted in hydroponics with 8 rice genotypes, viz., Pokkali, VSR 156,CSR 30, Pusa Basmati 1, Pusa 44, CSR 27, CSR 10 and IR 29 subjected to salinity stress (200 mM NaCl) along withcontrol after 12 days of sowing. Imposed salinity treatment lead to significant reduction in relative water content (RWC)and membrane stability index (MSI) in all the genotypes. However, decline in RWC and MSI was significantly less inPokkali, CSR 10 and CSR 27 as compared to other genotypes. In contrast, the sodium (Na) content increased underimposed salinity stress in the shoot tissues of all the genotypes. However, the increase was less in Pokkali, CSR 10 andCSR 27, which also maintained lower sodium/potassium (Na/K) ratio enabling cellular ion homeostasis. Gene expressionanalysis of CSR 10 and IR 29 revealed the presence of a more efficient salt overly sensitive pathway composed of SOS1,SOS2 and SOS3 in CSR 10 leading to a good correlation with the effective salinity stress tolerance of CSR 10 at thephysiological level.

PA102 (IPPC0748)

Analysis of sequence variation in HKT family genes of wheat for their association to salt tolerance

Vijendra Singh*, Amit Kumar Singh, Rakesh Singh, Sundeep Kumar and Jyoti Kumari

ICAR- National Bureau of Plant Genetic Resources, New Delhi-110012, India


Salinity is the major abiotic stress which limits wheat production in various parts of India and around the world. Some ofthe Indian wheat genotypes are known to possess very high level of tolerance to salinity stress. However, the molecularbasis of salt tolerance in these genotypes is not clearly understood. A detailed understanding of molecular mechanismsunderlying salt tolerance in these wheat genotypes will facilitate development of salt tolerant high yielding varieties. In thepresent study, a diverse set of 81 wheat genotypes including salt tolerant and salt sensitive genotypes were evaluatedunder controlled and stressed i.e., two different salinity conditions (4.0 and 5.0 dS m-1) in the net house at NBPGR, NewDelhi. The leaf and shoot tissues from all the genotypes were harvested and analysed for Na/K concentration. Further,HKT (high affinity K transporter) genes have been amplified from wheat genotypes and analysed for the presence ofsequence variations. Finally, nucleotide sequence variations among the HKT genes will be analysed for identification ofSNPs/ Indels for their association with variation in level of Na/ K concentrations in the plant tissues of all the wheatgenotypes.


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PA103 (IPPC0762)

Transgenic expression of cytosolic ascorbate peroxidase gene (apx1) from finger millet confers enhanced

oxidative stress tolerance potential in Arabidopsis thaliana

P. Yadukrishnan1*, Bhawna Negi2 and Sandeep Arora2

1Department of Biological Sciences, Indian Institute of Science Education and Research Bhopal- 462023, India2Department of Molecular Biology and Genetic Engineering, G.B. Pant Univ. of Agriculture and Technology, Pantnagar-263145, India*Presenting author: [email protected]

A drastic increase in reactive oxygen species (ROS) level is associated with most of the biotic and abiotic stresses andattributes to the reduced crop productivity under stress. Halliwell-Asada pathway (Ascorbate-glutathione cycle) is a centralpathway that effectively detoxifies hazardous ROS in plants. Ascorbate peroxidase (APX), an antioxidant enzyme playsa crucial role in this cycle by reducing toxic H

2O

2 to water using ascorbate as an electron donor. In this study, successful

attempts were made to transform Arabidopsis thaliana (ecotype Col-0) with finger millet (Eleusine coracana) cytosolicascorbate peroxidase (apx1) gene, cloned from a stress tolerant variety, PR202. Successfully transformed Arabidopsisseedlings carrying Ecapx1 driven by CaMV35S promoter showed increased net expression of ascorbate peroxidase. Thefunctional validation of the transgenic plants for their ability to tolerate oxidative stress was done by carrying out quantitativeestimation of biochemical parameters such as free proline accumulation, malondialdehyde (MDA) content and H

2O

2

content, under salinity-induced oxidative stress. Higher net accumulation of proline was observed in transformed plantswhen compared to wild types. Significant reduction in accumulation of MDA and H

2O

2 contents was recorded in transgenic

plants as compared to the wild-type plants. The greater accumulation of proline and reduction in the toxic oxidativespecies contents indicate that transgenic Arabidopsis plants expressing apx1 from finger millet are better equipped totolerate oxidative stress which could be well attributed to the presence of enhanced ascorbate peroxidase activity.

PA104 (IPPC0768)

Mitigating effect of salicylic acid on biochemical and antioxidant enzymes in maize (Zea mays L.) genotypes

under low temperature stress

Preeti Singh* and Vijai Pandurangam

Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, U.P., India


An experiment was carried out in field condition to find out sowing effects along with salicylic acid treatments with twoscreened maize genotypes i.e., HUZM-185 (tolerant) and HUZM-80-1 (sensitive) under low temperature stress duringRabi season. Seeds were primed with salicylic acid (SA) @ 20 ppm and 40 ppm along with hydro (distilled water) forovernight and dry seeds as control before sowing in split plot design with three replications having two sowing conditioni.e., normal and delay. Observations were recorded at 20, 40 and 60 (days) after sowing viz., malondialdehyde content(MDA), hydrogen peroxide content (H

2O

2), superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) activities

in both normal and delay sowing. It was found that 20ppm salicylic acid significantly reduced MDA, H2O

2 and enhanced

SOD, CAT and POX activities in both genotypes on normal and delay sowing, but values were higher in delay sowingcondition under low temperature stress. Remaining treatments were also performing better as compared to dry treatmentunder low temperature stress. Tolerant genotype significantly reduced MDA and H

2O

2 production and enhanced antioxidants

(SOD, CAT and POX) levels as compared to sensitive genotype. Hydrogen peroxide is a reactive oxygen species (ROS),which leads to disarrangement of bio-membrane and hampers its selectivity. A higher antioxidant activity in tolerantgenotype minimized ROS production (H

2O

2) and overcomes low temperature stress as compared to sensitive genotype.

The deleterious effect of low temperature stress was managed by enhancing SOD, CAT and POX activities in delaysowing as compare to normal sowing condition.


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PA105 (IPPC0769)

Copper mediated effect on reactive oxygen species generation and antioxidant responses in Oryza sativa

Nisha Kevat* and Prabhat K. Sharma

Department of Botany, Goa University, Panjim-403206, Goa, India


The aim of the present study was to investigate the effect of copper (Cu) on external morphology, photosynthesis,photosynthetic pigment content, reactive oxygen species (ROS) generation and succeeding oxidative damage in riceplants (Oryza sativa L. var. Jaya). Isoforms of GPX (Guiacol peroxidase), CAT (Catalase), SOD (Superoxide dismutase)and SOD activity were also studied. Plants were grown in pots (12 cm diameter) containing vermiculite and irrigated with0.1-3.0 mM CuSO

4 in Hoagland’s solution. SEM images of the leaf surface revealed reduction in stomata and trichome

size in a concentration dependent manner. Quantum efficiency of PSII (Fv/Fm), photochemical quenching (qP), netphotosynthetic rate (A), transpiration rate (E) and stomatal conductance (gs) decreased radically with incriminate in Cutreatment. Chlorphyll a, chlorophyl b, neoxanthine, violaxanthine and antheraxanthin were reduced linearly with increasingCu concentration. Hydrogen peroxide (H

2O

2) and hydroxyl radical (OH•) were effectively generated with an elevated Cu

concentration and induced oxidative stress measured as oxidative damage to lipid peroxidation and protein oxidation.Osmotic stress measured as proline content showed three-fold accumulation at 3 mM Cu concentration. Isozymes ofGPX and SOD were greatly affected due to Cu stress. Pronounced enhancement in the SOD activity was observed tometabolize ROS. The results demonstrated that exposure to elevated Cu concentration resulted in ROS (H

2O

2 and OH•)

generation leading to overall inhibition of photo-phosphorylation, brought morphological changes and induced oxidativestress. Eventually, the alleviated antioxidant responses emerged as major defensive mechanism adopted by rice plantsto moderate the Cu stress.

PA106 (IPPC0770)

Cloning and characterization of salt gene from Pusa Basmati 1

Navdeep Kaur*, Isha Sharma and Pratap Kumar Pati

Division of Biotechnology, Guru Nanak Dev University Amritsar, Amritsar-143005, India


Salinity is one of the major stresses affecting rice production worldwide. It is fairly a complex process regulated by anarray of genes. One such gene is SalT that is located in the Saltol QTL and shows higher expression under salt stress.However, precise mechanism through which it confers tolerance is not well understood. In the present work, attemptswere made to extensively characterize this gene in rice. The expression of SalT was analyzed under different conditionsusing qRT-PCR and was found to be up-regulated in different tolerant varieties as the scale of tolerance increases.Further, it was found to be regulated by different hormones and range of abiotic stresses. SalT gene from rice was clonedand overexpressed in Nicotiana tobaccum for developing transgenic plants overexpressing rice SalT. These transgenicplants were analyzed for increased salt tolerance and for finding putative interactive partner using FLAG-pull down assay.Initial analysis showed that this gene could be regulated through MAPK pathway in a calcium dependent manner. Thegene was further cloned into the protein expression vector and species specific antibody was developed. The antibodywas used for the validation of gene expression at the protein level. The heterologously expressed purified protein wasused for protein structuaral characterization using SAXS and was found to exist in different isoforms, dimeric form beingmost stable. The protein was observed to bind specifically with mannose that was confirmed using ITC and gel retardationassay indicating the probable link between carbohydrate metabolism and SalT.


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PA107 (IPPC0771)

Physiological characterization of mungbean genotypes for salinity stress tolerance

Sunder Singh1*, Neeraj Kumar2 and A.S. Nandwal3

1Department of Botany, Maharshi Dayanand University, Rohtak-124001, Haryana, India2Department of Botany and Plant Physiology, CCS Haryana Agricultural University, Hisar-125004, Haryana, India*Presenting author: [email protected]

Two mungbean genotypes (Asha and Muskan) were evaluated for salinity stress tolerance on the basis of physiologicaltraits viz. plant water status, solute accumulation, ionic composition, reproductive behavior and grain yield. Both thegenotypes were raised in green house and treated with salinity stress of 0, 2.5, 5.0 and 7.5 dS m-1 at flowering stage forten days and analysed for above traits. The water potential (ψ

w) of leaves and osmotic potential (ψ

s) of leaves and

decreased with increase in salinity levels from 0 to 7.5 dS m-1 in both the genotypes. Relative water content of leaves androots reduced, while proline and total soluble sugar (TSS) increased under salinity stress. The Na+/K+ ratio in leaves androots enhanced significantly and highest was estimated in roots. The genotype ‘Asha’ exhibited a low Na+/K+ ratio in aerialplant parts compared to ‘Muskan’. Salinity stress delayed the time of first flowering initiation by 2-4 days and reduced thenumber of flowers per plant. Pollen viability and in vitro pollen germination decreased in both the genotypes due tosalinity. Similarly, the pollen tube length decreased with increasing the level of salinity but did not effect on the number ofovule production. Salinity stress reduced the number of pods, number of seeds and weight of seeds per plant; and seedtest weight. Based upon above physiological traits, the genotype ‘Asha’ was identified relatively tolerant to salinity stressthan ‘Muskan’.

PA108 (IPPC0802)

Effect of water stress on physiological, biochemical, yield and yield contributing components in pigeon

pea (Cajanus cajan L. Millsp.)

Damor Parthvee R1, Ajay V. Narwade2*, Lalit Mahatma3, D.A. Chauhan4 and S. Sree Ganesh5

1,2,5Department of Genetics & Plant Breeding, 3Department of Plant Pathology, 4Main Pulses and Castor Research Station, N.M.College of Agriculture, Navsari Agriculture University, Navsari-396450, Gujarat, India*Presenting Author: [email protected]

A field experiment was conducted during winter (rabi) season of 2013 at Navsari Agricultural University, Navsari, toevaluate the effect of water stress on physiological, biochemical, yield and yield contributing components in pigeon pea(Cajanus cajan L. Millsp.). Four irrigations treatments were given during the experiment, viz., I

0- all the irrigations given at

25, 50 and 75 days after sowing (DAS), I1- Irrigation given at 25 and 50 DAS, I

2- Irrigation given at 25 DAS and I

3- kept as

rainfed. Six varieties of pigeon pea were used, viz., GT-102, Bharboot local, GNP-304, GT-1, AGT-2 and C-11 in split plotdesign with three replications. The response of pigeon pea varieties were observed by measuring various physiological,biochemical, growth, yield and yield attributing parameters. The experimental results indicated that the seed yield ofpigeon pea was significantly lower due to water deficit, due to the reduction in various physiological, biochemical, growth,yield and yield attributing characters. Among the six pigeon pea genotypes, GT-102 recorded significantly highest yieldas compared to other varieties due to highest harvest index, number of pods per plant, number of seeds per pod, 100-test weight, seed yield per plant, leaf area index, leaf area ratio and photosynthetic rate. It also had the highest accumulationof chlorophyll, carotenoid, glycine-betaine, proline and protein. By these results, GT-102 was identified as the droughttolerant variety, followed by C-11, GT-1 and AGT-2, while two the other genotypes, GNP-304 and Bharboot local weredrought susceptible.


169

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PA109 (IPPC0806)

Photosynthesis and yield of chickpea (Cicer arietinum) under interactive effect of elevated CO2, drought

and high temperature

Sadhana Maurya1,2*, Ashish K. Chaturvedi1, Shashi Meena1, Madan Pal1 and A.K. Chopra2

1Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi- 110012, India2Department of Zoology and Environmental Science, Gurukula Kangri Vishwavidyalaya, Haridwar-249404, Uttarakhand, India*Presenting author: [email protected]

India, the largest producer of pulses in the world, shares 14.6% of global pulse production. Chickpea has been knownsensitive to drought and high temperature which causes reductions in yield. On the other hand, CO

2 enrichment studies

revealed beneficial effects of CO2 in terms of photosynthesis and yield in chickpea. However, combination experiments

on elevated CO2, high temperature and drought are meagre. Therefore, an attempt was made to determine the potential

impact of elevated CO2, drought and high temperature alone and in combination of all the factors on photosynthesis and

seed yield of two contrasting chickpea genotypes viz. ICC-4958 and FLIP 90-166 raised under open top chambers. Pot-grown plants grown inside open top chambers were exposed to elevated CO

2 (ambient +250 μl/l), high temperature (>5°C

above ambient) and drought conditions (upto -1.6MPa) throughout the growth season. Elevated CO2 alone significantly

increased photosynthetic rate, plant biomass and seed yield of both the genotypes while high temperature and droughtalone caused significant reduction in photosynthetic rate and contributed to poor vegetative growth with lesser biomassand seed yield compared to control. Elevated CO

2 in combination to drought and high temperature as well showed

ameliorative effects on both the genotypes, however, the response was more prominent in FLIP 90-166. This studyconcludes that elevated CO

2 can alleviate the adverse effect of drought and high temperature stress on photosynthetic C

assimilation and yield in chickpea.

PA110 (IPPC0809)

Evaluation of maize genotypes for cold tolerance

Hari Singh Meena1, Upama Mishra1*, R.N. Gadag1 and H. Pathak2

1Division of Genetics, 2CESCRA, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India


Cold stress is a major environmental factor that limits the agricultural productivity of maize plants. Though maize is amajor crop in kharif season, cultivation during winter season in the Indian sub-continent is assuming importance due tovarious factors. Low temperature often affects plant growth and crop productivity, which causes major loss in maize cropin the northern part of India. Identification of suitable genotypes under northern belt of the country, therefore, assumespriority, to cater to such agro-climatic conditions. About 1300 lines of maize, in various stages of their derivation anddiverse sources were screened against cold stress in field conditions. Observation recorded were days to germination,days of silking, days of anther shedding, stem colour, root colour, anther colour, silk colour, uniformity, vigour, no of cobs,height. Days to germination ranged from 25 days to 41 days after sowing. Days of silking ranged from 84 days to 112days. Extensive variability was found in stem colour, ranging from light yellow to dark green including purple colour.Secondary roots colour ranged from light green to dark green including purple. Silk colours were brown and red. Antherscolour in the maize genotypes ranged from red brown to blue. Based on these observations and taking into account theproductivity of the lines in the experimental field of IARI, 52 lines which performed relatively better under cold stress, wereidentified. These selected lines will be subjected to re-screening in next rabi season and also would be involved ingenerating hybrids specifically suitable for rabi season in the Northern part of India. This strategy is expected to make useof wide flexibility and diversity of maize in terms of adaptation to varied climatic conditions and could serve as additional,economically viable option for the farmers.


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PA111 (IPPC0810)

Fatty acids composition of fenugreek (Trigonella Foenum-graecum L.) seed oil as influenced by water

stress

S.N. Saxena*, R.K. Kakani and S.S. Rathore

Department of Plant Physiology, National Research Centre for Seed Spices, NRCSS, Tabiji Farm-Ajmer 305206, Rajasthan, India


A study was conducted with thirteen fenugreek genotypes to evaluate the effect of water stress on oil content and its fattyacids composition. Oil content under non stress environment was ranging from a minimum of 2.62% in genotype AFg 6 toa maximum of 5.33 in AM 327-3. When stress imposed at midterm growth stage the oil content was increased from aminimum of 3.29% in AFg 6 to a maximum of 5.31 in AM 327-3. Significant genotypic variation was observed in fatty acidscomposition. Linoleic acid (34-54%), linolenic acid (27-46%), stearic acid (0- 5%) and palmitic acid methyl ester (7-13%)were major contributors. Genotypes AFg 3, AFg 5, B2 19, and CL 32- 17 showed increase in lenoleic acid and eitherdecrease or no change in lenolenic acid under stress conditions while genotypes AFg 4, AFg 6, AM 292, AM 327-3, HisarSonali, AL-1-2 and RMt 305 showed decrease in linoleic acid and increase in lenolenic acid content. Stearic acid andpalmitic acid methyl esters either reduced or unaffected under stress conditions. Under non stress genotype AFg 3 gavemaximum seed yield (8.93 g plant-1). Genotype AFg 6 produced highest seed yield (8.99 g plant-1) under stress conditionswhere lenoleic acid drastically reduced from 49.57 to 34.85% and lenolenic acid increased from 31.69 to 37.66%. Othergenotypes viz. AFg 4, AFg 6, Hisar Sonali and RMt 305 yielded good under stress showed similar fatty acid composition.

PA112 (IPPC0823)

Improving salt tolerance of crop plants : prospects and perspectives

Surinder Kumar Sharma*

Department of Crop Improvement, ICAR-Central Soil Salinity Research Institute, Karnal-132001, Haryana, India


More than 800 million hectares of land throughout the world and 6.73 million hectares in India are salt-affected and / oraccompanied by poor quality waters. Despite the increasing severity of these problems and their detrimental effects onagricultural production, programs to improve plant tolerance are in place in few countries, national and internationalresearch organizations. Selection for tolerance in the saline / sodic soils is further complicated as soil heterogeneity isoften accompanied by other individual or multiple stresses like drought, waterlogging, mineral deficiencies or toxicities.Four decades of researches at CSSRI, Karnal involving conventional breeding techniques supplemented by physiologicaland genetic mechanisms helped to develop suitable materials culminating in release of 15 salt tolerant varieties in rice,wheat, Indian mustard and chickpea besides identifying tolerant genetic stocks and donors. Some of these materials alsopossess multiple tolerance to drought, waterlogging or micro-elements enabling better performance and adaptability indiverse stress situations. These efforts were supported by demonstrations in research farms and farmer’s fields,supplemented by production and supply of quality seed promoting their cultivation and spread in many states having saltaffected lands. Adoption of these varieties has made significant contribution to national food production and povertyalleviation, benefiting resource poor and marginal farmers and restoring the marginally productive / barren lands into theproduction chain with additional environmental benefits. This paper describes some of these endeavors, adaptation andspread of such varieties in India and other countries highlighting the need to strengthen such programs globally.


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PA113 (IPPC0836)

Physiological and molecular analysis of root system architecture under drought and osmotic stress

in rice

G.K. Krishna1*, C. Vishwakarma, S. Thomas, P. Yadav, S. Srivastava, S. Yadav, R.K. Verma, V.V. Santosh Kumar,M. Das and V. Chinnusamy

ICAR-Indian Agricultural Research Institute, New Delhi-110012, India


Rice, the staple food for more than half the world population, is the second most important food grain crop. According toFAO, rice requires 1000 litres of water to produce 1 kg grain. Hence, reduced availability of irrigation water significantlyaffect its production. We targeted the avoidance mechanism exhibited by rice ie., the improved root system architecture(RSA) under drought stress. In the present study, 50 genotypes of rice were screened for their performance in RSA traits.Plants were raised in field, soil filled PVC pipes and hydroponic culture. In soil, treatment was given by cycles of droughtstress and recovery at -50 and -10 kPa respectively. Under hydroponics culture, osmotic stress was given at -100 kPa by8% PEG. From screening experiments, three contrasting genotypes were identified for RSA under stress viz. IC 305692(better performing), Nagina22 (moderate) and RCPL 1-3c (poor). Further, to understand the genetic basis for the regulationof this complex trait, 29 candidate genes which were induced in roots under drought were selected from rice oligonucleotidearray database. RT-PCR was performed to study their genotypic and tissue specific gene expression under 8% PEG and400 nM ABA. Five genes were identified by this experiment - homeobox associated leucine zipper, universal stressprotein, OsFBX132, MYB transcription factor and protein phosphatase 2C. The genes were subsequently cloned fromrice, to be used for functional validation in Arabidopsis. The present study will provide light in to the mechanismsregulating root system architecture in rice, especially under drought stress.

PA114 (IPPC0837)

Oxidative stress response in Lycopersicon esculentum mill. exposed to malathion and endosulfan

Manisha Gautam1, Shefali Poonia1* and Purushottam2

1Department of Botany, Deva Nagri College, Meerut-250001, India2Sardar Vallabh Bhai Patel University of Agriculture & Technology, Modipuram, Meerut-250110, India*Presenting author: [email protected]

A study was conducted with two varieties of tomato, viz. Pusa Ruby (PR) and Pusa Early Dwarf (PED). The plants wereexposed to three different concentrations (0.05, 0.15 & 0.25%) of pesticides namely malathion and endosulfan at regularintervals. Proline and ascorbic acid content in leaves were estimated for oxidative stress response. There was a highaccumulation of the two biochemical compounds which was concentration dependent. A substantial percent increasecould be observed in proline content upto 22.22 and 25.21 and ascorbic acid content upto 22.52 and 43.12 with malathionand endosulfan respectively at plant age of 30d in PR. For the same in PED, proline was 12.41 and 19.71 while ascorbicacid was 28.31 and 34.04, which is comparatively less than PR. The two biochemical components exhibited an increasingtrend with the age of the plant which slowed towards late maturity. At 90d, there was a significant decline in the increasingpercent values of ascorbic acid both in PR and PED. Similar was the observation for proline in PR but in PED theincreasing trend was observed. The enhanced accumulation of proline and ascorbic acid may be supportive to both thevarieties of tomato plant. All the values obtained were subjected to statistical analysis and level of significance wasevaluated.


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PA115 (IPPC0843)

Role of advanced unsaturated soil mechanics in understanding soil-root-water interaction

V.K. Gadi1*, A. Garg1, S. Sreedeep1, W.H. Zhou2 and L. Sahoo3

1Department of Civil Engineering, 3Department of Biotechnical Engineering, Indian Institute of Technology, Guwahati-781039, India2Department of Civil and Environmental Engineering, University of Macau, India*Presenting author: [email protected]

Fundamental understanding of soil-root-water interaction is important for analyzing and design of plantation scheme,irrigation scheduling and also in geotechnical infrastructures such as green roof, biofiltration units and vegetated slopes/landfill covers. Unsaturated soil parameters such as soil water retention curve and hydraulic conductivity could be affectedby soil type (particle size distribution), soil density and root growth (root depth and root distribution). The objective of thisstudy is to critically review literature and highlight the short comings in terms of measurement of unsaturated soil parameters.A wide literature focusing on plant biotechnology and geotechnical discipline is reviewed. In most of the cases, parametersare burrowed from literature. On the other hand, literature in geotechnical discipline lacks studies on investigation ofeffects of root growth on unsaturated soil parameters. In addition, in their studies, the boundary condition (i.e., evaporation)at surface of ground is considered to be uniform without considering canopy effects of vegetation. The review suggeststhe importance of integration of advanced concepts of unsaturated soil mechanics with that of plant biotechnology forthorough understanding of soil-root-water interaction.

PA116 (IPPC0850)

Comparative phenotyping under pot and field conditions to decipher component traits of drought tolerance

in rice

Rakesh Verma, Chandapal Vishwakarma*, Sitaram Kushwaha, Suchitra Pushkar, Shashank Yadav, Rahul Tiwari, SantoshVinjamuri and Viswanathan Chinnusamy



Genetic improvement of rice for enhanced WUE and drought tolerance is necessary to meet the food demand underdeclining fresh water availability. Evaluation of germplasm and breeding populations in augmented design for droughttolerance under field conditions often faces limitations due to soil moisture variability across the field and phenologicaldifferences among genotypes. Dehydration tolerance traits are poorly utilized in genetic improvement. Under limited soilconditions as in pots, mechanisms of transpiration minimization and cellular dehydration tolerance help survival of plantsunder drought stress. Hence, a comparative analysis of performance of rice genotypes was carried out for drought stresstolerance under pot and field conditions. Drought stress was imposed at flowering stage. Soil moisture status was quantifiedby tensiometer, soil moisture meter and gravimetric methods, while relative water content was measured to quantify plantwater status. Various component traits such as tissue temperature, excised leaf water loss, chlorophyll stability index,spikelet fertility and yield components were measured. MSAP analysis revealed genotypic differences in drought inducedchanges in DNA methylation levels and methylated sites between drought tolerant rice genotype Nagina 22 and relativelydrought sensitive genotype Swarna. Significant variation in dehydration avoidance and tolerance traits were found amonggenotypes. The results suggest that combined use of both pot and field methods will help greatly to tap the unexploitedgene pool for cellular tolerance traits to enhance the yield of rice under drought stress environments.


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IPPCongress2015


PA117 (IPPC0856)

Evaluation and validation of Saltol QTL for seedling stage salinity tolerance in rice (Oryza sativa L.)

K.T. Ravikiran*, S.L. Krishnamurthy and Parbodh Sharma

Genetics and Plant Breeding, Central Soil Salinity Research Institute, Karnal-132001, Haryana, India*Presenting author: [email protected]

The present investigation was undertaken to evaluate a set of 192 rice genotypes using hydroponics under saline stress(EC ~ 12 dS/m) and non stress (EC ~1.0 dS/m) conditions for morpho-physiological traits. Shoot length, root length andchlorophyll content decreased significantly under salinity as compared to non stress. Vigour score ranged from 3 (tolerant)to 9 (highly susceptible) with 14 and 16 genotypes categorized as tolerant (score 3) and moderately tolerant (score 5),respectively. The genotypes were further assessed for their molecular diversity using 10 SALTOL associated markers. Atotal of 30 alleles were generated at 10 microsatellite loci among the 192 genotypes. Combining morphological andmolecular assessment, four genotypes i.e. CSR IR 34, CST 7-1, Arvattelu and IR 87938-1-1-3-1-2-B were found tolerantwith distinctive allelic pattern from the tolerant check (FL 478). These could be used for further study to confirm theirnovelty and future breeding programme for seedling stage salinity tolerance in rice.

PA118 (IPPC0863)

Insights into various mechanisms adopted by Arbuscular mycorrhizal (AM) symbiosis for sustainable

legumes production under salt and metal(loid)s stresses

Garg Neera*



Chickpea and pigeonpea are sensitive to salt and metal(loid)s stresses. Increased level of stresses negatively influencedgermination, rhizobial symbiosis, photosynthetic efficiency, membrane properties, nutrient and redox homeostasis,osmoregulation; which eventually led to reduced plants growth and reproducibility. In pigeonpea, presence of highconcentration of zinc counteracted the negative effects of Cd, while co-occurrence of Cd-Pb and Cd-NaCl in rhizospheredisplayed synergism in the form of compound negative effects on growth and metabolism. In addition to rhizobial symbiosis,legumes can establish dynamic interaction with arbuscular mycorrhizal (AM) fungi, which is an essential link for effectivephosphorus (P) nutrition, leading to enhanced N

2 fixation. The mechanisms adopted by AM in alleviating salt and metal(loid)s

stress included higher water absorption capacity and nutrient status by increasing root rhizosphere; enhanced nitrogenfixation; improved ionic and redox homeostasis. Besides this, mycorrhization reduced negative impact of metal(loid)sthrough dilution effect and phytochelatins induced vacuolar sequestration. Among different AM species, exotic singleisolates conferred higher salt tolerance to pigeonpea than the native inoculums isolated from saline soils. Moreover,Rhizophagus irregularis (alone or in combination with Funneliformis mosseae) displayed higher effectiveness correlatedwith their more stable viability under salt stress. Although, abiotic stresses reduced AM colonization in both legumespecies; application of exogenous alleviants (flavonoids and silicon) further enhanced salt tolerance in chickpea throughimproved mycorrhization. Individually F. mosseae inoculation outperformed flavonoid and silicon application in impartingsalt stress tolerance, however functional complementarity of F. mosseae with both flavonoids and silicon provided utmostyield and productivity in chickpea.


174

IPPCongress2015


PA119 (IPPC0868)

Screening of rice genotypes for heat tolerance based on chlorophyll fluorescence measurement

N. Veronica1, Desiraju Subrahmanyam2*, S.R. Voleti2, Y Ashoka Rani1, K.L. Narasimha Rao1, M Lal Ahamed1 andP. Prasuna Rani1

1Agricultural College, Acharya N.G. Ranga Agricultural University, Bapatla-522101, India2ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad-500030, India


In the present scenario of climate change, high temperature has become a major concern challenging the productivity ofrice. Hence identification of tolerant genotypes is pre-requisite to mitigate the adverse effects of climate change andglobal warming. Chlorophyll fluorescence traits gives insights into the ability of a plant to tolerate environmental stresses.Fv/Fm ratio, representing the primary photochemical efficiency of PSII is used as a selection criterion under heat stresscondition. A diverse set of 58 rice genotypes comprising of 2 germplasm lines, 11 green super rice lines, 2 introgressedlines, 8 landraces, 2 tropical japonicas and 33 released varieties were screened based on their maximum quantumefficiency of PSII (or PSII photochemistry), measured by Fv/Fm values. Flag leaves at flowering stage were collected.One set was kept at ambient temperature (control) and another was subjected to high temperature in a growth chambermaintained at a temperature of 420C and light intensity of 300 μmol m-2 s-1 for 2 hours. Florescence measurements weremade using a portable PAM Fluorometer (Walz PAM-210). Exposure to high temperature (420C) resulted in significantreduction in mean Fv/Fm ratio in all the tested genotypes. Significant variation was noticed amongst the genotypes in percent reduction in Fv/Fm. The reduction was less than 5% in Assanchidiya, GSR 330, Khudaridhan, Akshayadhan andRasi, indicating these genotypes are relatively tolerant to high temperature stress. On the contrary, more than 35%reductions were noted in Mahamaya, MTU 1001, RNR 6378 and Pantdhan 18, suggesting that the photosynthetic apparatusof these varieties were sensitive to high temperature stress.

PA120 (IPPC0871)

Identification and characterization of heatstress tolerance in maize (Zea mays L.)

Suprokash Debnath1, Anuradha Gautam2, Pranjal Yadava2, Madan Pal1 and Ishwar Singh2*

1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, Pusa, New Delhi-110012, India2ICAR-Indian Institute of Maize Research, Pusa Campus, New Delhi-110012, India


Among the ever-changing components of the environment, high temperature is one of the most detrimental stresses formaize. Therefore, promising parental lines (inbred) with adaptive traits of tolerance to high temperature needs to beidentified. In the present study, we evaluated 20 promising maize inbred lines of diverse genetic background for their perse performance at high temperature (summer season) as compared to control (kharif season), based on the differentheat-stress adaptive traits like leaf area index, leaf firing, tassel blasting, anthesis-silking-interval and grain yield, Theperformance of DTPYC9F119 was best among all the genotypes under heat-stress, while K64R was found to be highlysusceptible to high temperature. These contrasting genotypes were further analyzed under 6 days heat treatment (38/28°C) at flowering stage under managed stress conditions to understand the mechanisms of heat stress tolerance.Chlorophyll a and b ratio under heat-stress increased significantly in DTPYC9F119 after 6 days high temperature treatment.Net photosynthetic rate reduced under high temperature in K64R but the reduction rate was relatively lower in case ofDTPYC9F119. Stomatal conductance increased under stress treatment in both the genotypes but the rate of increasewas lower in DTPYC9F119. Activity of anti-oxidant enzymes (catalase, peroxidase and superoxide dismutase) and theirgenes showed differential expression in the two contrasting genotypes. Therefore, it seems that the tolerant genotypehas evolved adaptive strategies by modulation of anti-oxidant gene expression to tolerate high temperature.


175

IPPCongress2015


PA121 (IPPC0891)

PGPR- mediated modulation of proline metabolic gene expression in Arabidopsis thaliana under drought

stress

Daipayan Ghosh* and Sridev Mohapatra

Biological Sciences, Birla Institute of Technology and Sciences Pilani Hyderabad Campus, Shamirpet, Hyderabad-500078, India


Plant growth promoting rhizobacteria (PGPR) is a group of soil bacteria that are well known to contribute positivelytowards alleviation of abiotic stress in plants. Although some of the physiological processes involved in plant-PGPRinteraction for stress alleviation are known, the precise molecular mechanisms still remain unclear. One of the mechanismsby which plants abate drought stress is through osmotic adjustment, by accumulating several compatible solutes, suchas the amino acid proline. Here we report novel observations on the modulation of proline metabolic genes under dehydrationconditions in the model plant Arabidopsis thaliana, by a beneficial rhizobacteria, Pseudomonas putida GAP-P45. Arabidopsis

thaliana plants inoculated with Pseudomonas putida GAP-P45 under dehydration conditions exhibited significant delay instress senescence with diminished symptoms of dehydration such as chlorosis, growth stunting and necrosis. Postdrought induction, this particular strain of PGPR was capable of maintaining overall health of plants as opposed to thenon-inoculated plants as concluded through visual and comparative physiological studies (analysis of fresh weight, dryweight, plant water content, total proline content etc.) estimated at different time points. Differences in root structurearchitecture were observed in the inoculated vs. non-inoculated plants under drought conditions. Semi-quantitative andquantitative real-time expression analysis of proline metabolic genes indicated a direct upregulation of proline biosyntheticgenes and steady catabolism rate maintenance in plants by PGPR for long term survival under drought stress. Weconclude that this strain of PGPR helps in drought mitigation of A. thaliana by modulating gene expression in the prolinemetabolic pathway.

PA122 (IPPC0897)

Arsenic induced phytotoxicity in black gram (Vigna mungo L.) during early growth stage

M.Z. Shamim* and A. Pandey

Department of Biotechnology, Motilal Nehru National Institute of Technology Allahabad, Allahabad-211004, U.P., India*Presenting author: [email protected]

Arsenic is known as one of the highly toxic element to biological systems. Present study was designed to investigate thephytotoxic effects of arsenic (As) on different physiological parameter in black gram. Two genotypes of black gram, i.e.,IPU 99-176 and UPU 83-35 were used for the experiment. Plants were irrigated with Hoagland’s hydroponic nutrientsolution. Fifteen days after sowing plants were treated with water (control), 50 and 100 μM sodium arsenite along withHoagland’s solution. The increasing arsenic stress condition significantly decreased root length (8.66-24.41 %), freshroot weight (7.61-89.36 %), shoot length (14.63-25.37 %) and fresh shoot weight (13.73 -26.56 %) with respect to control.At 100 μM As reduction in total chlorophyll was 12.91 and 20.4 5% in IPU 99-176 and UPU 83-35, respectively, withrespect to control. Amount of chlorophyll-a increased (15.49 %) with increasing concentration of As in IPU 99-176,whereas it decreased in UPU 83-35 under increasing concentration of As. The amount of chlorophyll-b drastically decreasedat 50 μM As concentration, but at 100 μM concentration the amount of chlorophyll-b increased with respect to 50 μM Asconcentration in both the genotypes. Carotenoids contentt in both of the genotypes decreased, and reduction was greaterat 50 μM As concentration than 100 μM As concentration. Results indicated that IPU 99-176 black gram genotype mayhave greater tolerance for arsenic toxicity.


176

IPPCongress2015


PA123 (IPPC0911)

Epiphytic orchids of Assam and their adaptation to various abiotic stresses

Ranjan Kumar Bora*, Nabami Basumatary and C.M. Sarma

Department of Botany, Pragjyotish College, Guwahati-781009, Assam, India


Assam, a biodiversity rich state of India, harbours 290-odd species of orchids and of which majority are epiphytic. Innature, epiphytic orchids can survive and grow in minimal environments withstanding various abiotic stresses such asdrought, temperature, light, nutrients etc. A study was conducted on epiphytic orchids of Assam to understand their modeof adaptation to different abiotic stresses during 2005 to 2014. Epiphytic orchids survive in minimal nutrient availability.During rainy season orchid roots absorb dissolved nutrients through stem flow and for which retention of water for longerperiod is necessary. To get better accumulation, epiphytic orchids adopt the following modes such as formation of acavity by the root system, extending long roots to great distance, forming patch, encircling the stems and accumulation ofdebris by upright roots. Though orchids prefer diffused sunlight, the light requirement varies species to species andprefers to establish on the phorophyte according to their requirement. Shade loving orchids usually prefer to grow onbarks near the ground. Tiny orchids usually prefer small twigs under the canopy. High light tolerant species have theability to increase the carotenoids which protect the chlorophyll content from photo oxidation. Many epiphytic orchidshave terete and scale leaves to withstand against extreme drought and high temperature. For better management ofwater, orchids possibly open their stomata during the early morning and in the evening. Otherwise, epiphytic orchids mayact as CAM cycling physiotype during severe drought which helps the uptake of CO

2 and conservation of water efficiently.

PA124 (IPPC0916)

Abiotic stress mediated modulation of chromatin landscape in A. thaliana

Vivek Kumar Raxwal1, Sourav Ghosh2, Somya Sinha1, Surekha Katiyar-Agarwal3, Shailendra Goel1, Arun Jagannath1,Amar Kumar1, Vinod Sacaria4 and Manu Agarwal1*

1Department of Botany, University of Delhi, New Delhi-110007, India2Genomics and Molecular Medicine, CSIR-Institute of Genomics and Integrative Biology, Mathura Road, New Delhi, India3Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India4GN Ramachandran Knowledge Centre for Genome Informatics, CSIR-Institute of Genomics and Integrative, New Delhi, India*Presenting author: [email protected]

Considerable progress has been made in understanding abiotic stress-mediated physiological responses and theircorrelation with changes in candidate gene expression patterns. With significant advancements in genomic technologies,it is now possible to study transcriptional regulation in response to abiotic stresses on a genome-wide scale. One of themajor constraints to transcription is the packaging of DNA by histones, which hinders accessibility of regulatory proteinsto cis-elements. During transcription, chromatin dissociates the bound histones, which is later repackaged on completionof the transcriptional process. Hence, at any given stage, the chromatin is in a dynamic state w.r.t. its association withhistones. We employed two different cross-validating methodologies, DNase-seq and FAIRE-seq (Formaldehyde Isolationof Regulatory Elements) to capture the open chromatin regions in A. thaliana genome upon imposition of four differentabiotic stresses. Open chromatin regions identified in both FAIRE-seq and DNase-seq (OCFD) samples were used toanalyze chromatin states relative to the control samples. Three categories of relative chromatin states were defined viz.,(1) OC-OS in which chromatin was open in both control as well as stress samples (2) OC-CS, which included chromatinregions that were open in control and closed in stress samples and (3) CC-OS having chromatin regions that were closedin control and open in stress samples. Identification of relative chromatin states led to the elucidation of stress specificcategorization of chromatin conformation. In conjunction with mRNA-seq data, this study allowed genome-wide identificationof genes that are significantly up-regulated or down-regulated in response to different abiotic stresses.


177

IPPCongress2015


PA125 (IPPC0931)

A twist in the “plant- rhizobacterial interaction” tale in drought mitigation

Diti Shah1, Sunetra Sen1*, Sridev Mohapatra1, Minakshi Grover2, Shalini Akula3 and Daipayan Ghosh1

1Biological Sciences, Birla Institute of Technology and Sciences Pilani Hyderabad Campus, Shamirpet, Hyderabad-500078, India2Central Research Institute for Dryland Agriculture, Santoshnagar, Saidabad, Hyderabad-500059, India3J.K. Agri- Genetics Ltd., Hyderabad, India*Presenting author: [email protected]

Plant growth promoting rhizobacteria (PGPR) have been known to contribute positively towards abiotic stress mitigationin plants. It is known that several strains of PGPR secrete plant growth promoting chemicals such as phytohormoneswhich may help in modulating plant responses to abiotic stresses. Here, we report some novel observations in context ofthe interaction between the model plant, Arabidopsis thaliana and a Pseudomonas putida strain of PGPR which releasescopious amounts of the auxin, indole acetic acid (IAA) during drought stress. Contrary to previous studies on PGPR, thisparticular strain, on inoculation with A. thaliana under drought conditions, exhibited a strong inhibitory effect on droughttolerance ability of the plant, while promoting plant growth under well-watered conditions. Inoculation of A. thaliana withthis strain of bacteria under dehydration conditions reduced the plant’s inherent ability of transient drought mitigation.Plants exposed to drought with simultaneous P. putida inoculation exhibited enhanced drying, reduced plant water content,root growth inhibition and an overall attrition in drought tolerance as opposed to drought induced plants without PGPRinoculation. We conclude that this strain of P. putida (in the concentration used in the present study), enhances thesusceptibility of A. thaliana to dehydration stress. We hypothesize that excess amounts of secretory/excretory IAA maybe contributing to the negative impact of this strain towards drought mitigation in A. thaliana.

PA126 (IPPC0964)

High temperature tolerance in chickpea genotypes as evaluated by photosynthetic rate, SPAD, PS-II,

canopy temperature depression and yield stability index

Pattipaka Bhasker1, Ajit Singh Nandwal1*, Neeraj Kumar1, Sarita Devi1, Gurdev Chand1, Krishan Kumar2 andSatish Kumar3

1Department of Botany and Plant Physiology, 2Pulses Section, Department of Genetics and Plant Breeding, 3Department of Agronomy,CCS HAU, Hisar-125004, Haryana, India


Chickpea (Cicer arietinum L.) is an important food crop grown in India under rainfed condition. The crop usually encountersterminal high temperature (HT) stress during pod filling stage. The objective of the work was to determine the degree ofHT effect on two contrast genotypes, viz., ICCV 92944 and HC 5 differing in their tolerance on the basis of net photosynthesis(P

n), stomatal conductance (g

s), SPAD, photochemical efficiency of PS-II measures as chlorophyll fluorescence (CF) and

canopy temperature depression (CTD) under field condition. HT stress was given by the changing of two sowing dates,i.e., normal (11th Nov. 2011) and late sown (16th Dec. 2011). Observations were recorded as the temperature rose above35 ºC after 4, 8, 12 days of exposure. Control readings were taken at the temperature below 30 ºC. With increasing daysafter exposure to HT a close relationship in decline in photosynthesis, SPAD and CTD values was observed. ICCV 92944exhibited higher values of Fv/Fm (0.69), SPAD (54.77) but less CTD (-3.56 ºC) as compared to HC-5 having values 0.60,49.20 and -2.95 ºC, respectively. The variance of these parameters was statistically related to yield stability index. Themeasured parameters of ICCV 92944 (identified as HT tolerant), declined relatively more slowly than those of HC-5.Results showed that HT induced stress is detectable from P

n, g

s, SPAD, CF, CTD, and these physiological traits can be

useful tools to help in screening of chickpea germplasm for HT tolerance.


178

IPPCongress2015


PA127 (IPPC0967)

Evaluation of root system architecture for adaptation to water stress in wheat: deeper rooting

Rakesh Pandey1*, Neelu Jain2, Vijay Paul1, G.P. Singh2, R.C. Meena1, Atar Singh1, M.B. Shine2, Vinod Pandey2

and Neha Rai2

1Division of Plant Physiology, 2Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India*Presenting author: [email protected]

Deep rooting is a useful trait for adaptability under water limited environments. It can be studied in terms of maximum rootlength and root growth angle. Genotypic variation in root growth in wheat was observed by trench profile wall method afteranthesis in thirty three genotypes. Root growth at different depths was expressed as % of total root number density. In theupper profile (0-15 cm) root density was higher in genotypes HD2894, PBW343, GW322, Kukri, HD2824, HI1544 etc. Inthe middle profile (16-30 cm) the root density was higher in HD2987, DBW17, C306, HD3043, Wyalkechem, HUW468,HD3016 etc. In the lower profile (31-50 cm) the root density was higher in HI1500, Raj3765, HD2687, HUW468, HD2932,HI1531, WH730 etc. These genotypes having higher root density in the lower profile are more adaptable to water stress.However, measuring the root growth in the field conditions is cumbersome and time consuming. Therefore, visualizationof root system architecture and measurement of the seminal root angle in the agar gel is a much easier alternative. Anarrow root angle between seminal roots is associated with deeper rooting. Variability in the seminal root angle wasobserved from narrow to wider angles (25oC to 87oC). The relatively drought tolerant genotypes viz. N59, HD2985, DBW-14, C-306 etc. showed narrow root angles compared to the genotypes for irrigated conditions e.g. HD2967, HD2643,HD2285, HD2824 etc.

PA128 (IPPC0968)

Heat stress responses in chickpea

Neeraj Kumar1*, Ajit Singh Nandwal1, Krishan Kumar2 and Dharamvir Soni1

1Department of Botany and Plant Physiology, CCS Haryana Agricultural University, Hisar-125004, Haryana, India2Pulses Section, Department of Genetics and Plant Breeding, CCS Haryana Agricultural University, Hisar-125004, Haryana, India

*Presenting author email: [email protected]

High temperature stress (>30ºC) is one of the most evident phenomenon of the global climate change, which limits theplant growth and development in chickpea and leads to drastic reduction in seed yield. With this objective, an experimentwas conducted with 15 chickpea genotypes for screening against high temperature tolerance by studying various morpho-physiological traits and seed yield attributes under late (2nd fortnight of December) sown conditions. A significant variationin the quantum yield of PSII (Fv/Fm) was observed among the tested genotypes. The value for Fv/Fm ranged from 0.644to 0.783 and was highest in ICCV 92944 and lowest in H 09-65. Relative stress injury (RSI %) of leaves varied from 13-20%, and was considerably high in genotype H 10-22 (20%) and lowest in H10-41 (13%). The canopy temperaturedepression (CTD) revealed significant differences among genotypes. CTD values ranged between -0.5 (H10-22) and1.6ºC (H 09-96). Pollen viability was found to be in the range of 65 to 88%, being highest in the genotype ICCV 92944 andlowest in H10-01. Genotypes H09-96, H10-21 and H10-41 gave highest grain yield of 21.7, 19.4 and 18.8 g plant-1,respectively. The genotypes H09-96, H10-21 and H10-41 were identified as heat tolerant lines (>30ºC) based on highvalues of percentage pollen viability, quantum yield (Fv/Fm), yield attributes and yields but low values of RSI % andmaintained cooler canopy temperature.


179

IPPCongress2015


PA129 (IPPC0969)

Brassinosteroids modulate antioxidant defense system under thermal stress in Brassica oleraceae var

broccoli

Spall Kaur Nirmal K* and Geetika Sirhindi

Department of Botany, Punjabi University, Patiala, Punjab-147002, India


Anti-cancerous properties of brocoli are well known, which may be due to the presence of high antioxidants in the form ofenzymes and non-enzymes. Brassinosteroids are known for enhancing antioxidant potential of different crops in pico- tomicro- molar concentrations. Present study was conducted with the aim to explore the ability of 24-epibrassinosteroid(24-EBL) in enhancing the tolerance level of broccoli under extreme temperature stress. Seedlings of broccoli wereprimed with distilled water (control) and different concentrations of 24-EBL (10-6, 10-9, 10-12), and grown in seed germinatorat 25 0C under controlled conditions of plant growth. To 7 days old seedlings temperature stress was given for threeconsecutive days and then harvested on 10th day for investigating the content of proline, ascorbic acid along with activityof antioxidant enzymes SOD, POX and CAT. Brassinosteroids increased accumulation of proline by 15% and ascorbicacid by 5%, while activity level of SOD, POX and CAT were enhanced by 18.7%, 34 % and 31%, respectively. The datasupports to postulate that 24-EBL exhibited a good potential in mitigating the toxic effect of extreme temperatures onplant growth and enhanced the crop thermo-tolerance without degrading their nutritional and medicinal properties andrelatively enhanced the antioxidants of the crop to significant levels thus increasing its anti-cancerous potential.

PA130 (IPPC0983)

Salinity induced variations in growth, photosynthetic performance and ionic balance in desi and kabuli

genotypes of Cicer arietinum L. (chickpea) under salt stress

Ranju Gulati1* and Neera Garg2

1Department of Botany, Dav College, Sector 10, Chandigarh-160036, India2Department of Botany, Panjab University, Chandigarh-160014, India


Salinity is one of the major environmental stresses affecting crop production especially in arid and semi-arid regions ofthe world. Chickpea is the most important legume crop for human nutrition globally but at the same time it is highly saltsensitive. However significant variability in terms of salt tolerance has been reported in chickpea germplasm. The presentstudy was carried out to compare salt induced modulations in growth, photosynthetic characteristics and ion accumulationin salt tolerant kabuli (CSG 9651) and salt sensitive desi (DCP 92-3) genotypes of chickpea. The plants of both genotypeswere raised under control and NaCl salinity (EC = 4, 6 and 8 dSm-1). The saline treatments were applied15 days aftersowing DAS (days after sowing) and continued till harvest. Salinity significantly reduced roots and shoots dry mass anddisturbed the root to shoot ratios in both genotypes with significantly higher reductions recorded in DCP 92-3. CSG 9651possessed higher values for photosynthetic attributes in terms of leaf chlorophyll content with higher Chl a/b ratios andRubisco activity as compared to DCP92-3. Kabuli genotype CSG 9651 recorded lower sodium uptake, higher potassiumand better K/Na ratio which manifested in the form of higher growth and productivity (higher seed yield, HI etc). The studyindicated better tolerance ability of kabuli genotype of chickpea subjected to long term salinity when compared with desi.


180

IPPCongress2015


PA131 (IPPC0986)

Introgression of water use and water use efficiency in rice (Oryza sativa L.) and its relevance to drought

tolerance

N.V. Preethi1*, B.R. Raju, K. Sumanth Kumar, R. Ramachandra and M.S. Sheshshayee

Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India


Rice is the most important cereal of the world population and is an intensively water consuming crop. It is hypothesizedthat maintenance of turgor and positive carbon balance is essential for enhancing crop performance under water limitedconditions. Global opinion strongly emphasise introgression of these complex traits to achieve improved adaptation towater limited conditions. Towards developing such trait introgressed lines, root donor line (IET15963) was crossed withWUE donor line (Thanu) and a large number of recombinant inbred lines were developed. Among a few trait introgressedlines, KMP175 was chosen for further studies. SSR markers associated with WUE and root traits were used to identify theintrogression of the target genomic regions governing the two traits with the integration of water mining and WUE traits.KMP175 displayed superior growth and productivity both under aerobic and water limited conditions. Introgressed linerecorded higher photosynthetic rate and stomatal conductance compared with the parents. The field performance ofKMP175 was significantly higher than the other rice cultivars recommended for aerobic cultivation. The study providedstrong evidence on the importance of introgressing physiological traits to improve water productivity of rice.

PA132 (IPPC1003)

Development of activation-tagged mutant population for enhanced water-use-efficiency (WUE) in Oryza

sativa ssp. indica

Mazahar Moin1*, Achala Bakshi1, A.R. Reddy1, K.V. Rao2, E.A. Siddiqi3, M. Udaya Kumar4 and P.B. Kirti1

1Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India2Centre for Plant Molecular Biology, Osmania University, Hyderabad, India3Honorary Professor, Institute of Agricultural Biotechnology, PJTS, Hyderabad, India4Department of Crop Physiology, GKVK, Bangalore, India


The epitome of any genome research is to identify all the existing genes contained in a genome and investigate theirroles. Rice is the most appropriate model crop for generating mutant resource for functional genomics studies because ofits economic importance, availability of high quality genome sequence and relatively smaller genome size. In the presentstudy to develop an activation-tagged (gain-of-function) mutant resource for high WUE in indica rice Samba Mahsuri(BPT 5204), we have generated 10,000 Ac/Ds and 3,700 stable enhancer based activation-tagged insertions. Theseplants are being screened for high WUE by growing under limited water conditions based on morphological (tillering,plant height, stay green character and seed yield), and physiological characterisation (photosynthetic performance throughMini-PAM and carbon isotope composition through IRMS). Two hundres plants have been selected as for high WUE.Their flanking sequence analysis through TAIL-PCR revealed that the insertions are non-uniform and mostly genic, inparticular inter-genic (60%). Four phenotypically distinct plants were chosen to study the expression of the candidategenes residing within a 20 kb region, the approximate limit of activation of genes by enhancers from the site of insertion.Expression analysis of 16 genes indicated that activation of a gene is independent from the site of insertion of enhancers,although in most of the cases there was activation of only one of the many tagged genes, however, in one plant two geneswere activated. The candidate genes responsible for varied phenotypes are being cloned to further validate their roles inWUE.


181

IPPCongress2015


PA133 (IPPC1011)

Clout of rhizobacterial inoculation on antioxidative enzyme activity in Zea mays under aluminium toxic

conditions

Priyanka Arora1*, Geeta Singh2 and Archana Tiwari1

1School of Sciences, Noida International University, G.B Nagar, Greater Noida-203201, U.P., India2Division of Microbiology Indian Agriculture Research Institute, New Delhi-110012, India*Presenting author: [email protected]

This study explores the role of microorganisms in enhancing antioxidative defense mechanism of plants to combat againstaluminium stress. In the present study, a pot experiment was conducted to evaluate the effect of bacterial inoculation onantioxidative enzyme activity in maize plants under 60 mM aluminium stress condition. Pots were inoculated with twobacterial isolates belonging to genus Burkholderia and Bacillus. Experimental results revealed that although there was asignificant increase in antioxidative enzyme activity under aluminium stressed condition but it was not sufficient enoughfor complete scavenging of ROS. This activity further enhanced on bacterial inoculation to overcome the imposed aluminiumstress. Burkholderial isolate performed better than Bacillus isolate showing an increase of 1.47, 7.1 and 2.17 fold in SOD,CAT and POX activity respectively. Therefore the inoculation of Burkholderial isolate can be suggested for plant growthimprovement in maize plant under aluminium toxic conditions.

PA134 (IPPC1016)

Ecophysiological adaptation of an extreme halophyte Salvadora persica to high salinity: Implications

from ion homeostasis, photosynthesis, PS II photochemistry and antioxidant response

Jaykumar Rangani1,2, Asish Kumar Parida1,2*, Ashok Panda1 and Asha Kumari1,2

1Division of Wasteland Research, 2Academy of Scientific and Innovative Research, CSIR-Central Salt and Marine Chemicals ResearchInstitute, Gijubhai Badheka Marg, Bhavnagar-364002, Gujarat, India*Presenting author: [email protected]

Salinity induced modulations in growth, photosynthetic pigments, relative water content, electrolyte leakage, lipidperoxidation, photosynthesis, photosystem II efficiency, and changes in activity of various antioxidative enzymes werestudied in the halophyte S. persica treated with various levels of salinity (0, 250, 500, 750 and 1000 mM NaCl). Fresh anddry biomass of shoot and root as well as leaf area decreased at all levels of salinity as compared to control. There wasgradual increase in leaf Na+ content with increasing salt concentration up to 750 mM NaCl, but at high salinity(1000 mMNaCl), Na+ content surprisingly decreased and came down to the level observed at 250 mM NaCl. There were non-significant changes in chlorophyll, carotenoid, photosynthetic rate (P

N), stomatal conductance (g

s) and transpiration rate

(E) at low salinity (250 to 500 mM NaCl), however, significant reductions were observed at 750 and 1000 mM NaCl. Therewere non-significant changes in quantum yield of PSII (ÖPSII), photochemical quenching (qP) and electron transport rate(ETR) at low salinity (250-500 mM NaCl), however, decreased significantly at high salinity (750-1000 mM NaCl). Analysisof isoforms of antioxidative enzymes revealed that the leaves of S. persica have three isoforms each of SOD and POX,two isoforms of APX, one isoform of CAT, and five isoforms of glutathione reductase (GR). There were differential changesin different isoforms of antioxidative enzymes. Our results suggests that the absence of pigment degradation, reductionof water loss and maintenance of water use efficiency, and protection of PSII from salinity induced oxidative damage bycoordinated changes in antioxidative enzymes enhanced the high salt tolerance capacity of S. persica.


182

IPPCongress2015


PA135 (IPPC1024)

Expression of the PGPR induced rice OsIAA10 gene improves root growth and confers abiotic tolerancein transgenic Arabidopsis plants

Pallavi Agarwal*, P.S. Chauhan, V.A. Sane and C.S. Nautiyal

Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India*Presenting author: [email protected]

PGPR are plant growth promoting bacteria, which are known to induce plant growth by variety of mechanisms. Microbialassociations within roots and hormonal regulation in the plants have been reported, but the molecular mechanism behindthis increase in growth of plants is still unraveled. PGPRs (NBRI isolated strain) help in inducing growth of potted riceplants. Microarray Data of PGPR treated rice plants suggested up-regulation of 494 genes and down-regulation of 1045genes in PGPR treated rice roots as compared to control rice roots. One of the genes OsIAA10, with 3 folds up-regulationin treated roots, was selected for further studies. OsIAA10 belongs to Aux/IAA family involved in auxin signaling.Phytohormone auxin is known to play a key role in plant development and lateral root formation. Transgenic Arabidopsislines over-expressing OsIAA10 showed longer roots and better growth under various abiotic stresses.

PA135A (IPPC1039)

Elucidation of Glutaredoxin proteins in Dicots.

Nilesh Kumar1*, Minesh Jethva1, Saurabh Yadav2 and Hemant R. Kushwaha1

1Synthetic Biology and Biofuels Group, International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India2Department of Biotechnology, Hemwati Nandan Bahuguna Garhwal University, Srinagar Garhwal, Uttarakhand-246174, India*Presenting author: [email protected]

Glutaredoxins are small proteins (approx. 11 kDa) containing an active site with a redox-active motif. They regeneratethe oxidatively damaged proteins via thiol-disulphide reactions. GRXs acts in anti-oxidant defense by reducingdehydroascorbate, peroxiredoxins, and methionine sulfoxide reductase etc. These proteins are also involved inmaintaining the redox homeostasis in many organisms like bacteria, yeast. Apart from their role in antioxidant defense,GRXs are known to bind iron-sulfur clusters (ISC). Depending on the motif present GRXs are classified in two classes:Monothiol which contain ‘CGFS’ and dithiol which contain ‘CxxC’ motif. In the present study, we have characterizedGRX family members in various dicots. Fact-finding on GRX in many organisms has revealed a surprising variety of vividcorollaries. In view of that, we have analyzed chromosome localization, protein domain architecture, phylogeneticrelationship and subcellular localization of GRX member proteins in various dicots. These analyses have been used toinfer the evolution of GRXs in dicots. This study provides cognizance of the domain structure conservation of GRXs indicots. In analysis provided in the study will assist in developing an understanding of the evolution of redox defensemechanism in plantspecies.

Session B : Signaling and Phototransduction

183

IPPCongress2015


PB136 (IPPC0114)

Exploring signature of natural ageing and CDT by studying AOS and antioxidative enzymes in neem[Azadiracta indica juss] seeds

Alok Kumar Sahu* and Subhas Naithani

Seed Biology Lab., SoS in Life Sciences, Pt. Ravishankar Shukla University, Raipur-492010 (CG), India


Seed longevity is a resultant of the interaction of seed storage conditions with its genetic and physiological potential.Natural ageing [NA] and control deterioration [CDT] tests are widely used to deduce the pathway of ageing. CDT is rapidtest as it accelerates ageing [within days] hence preferred over NA. Natural and CDT protocol revealed conflicting resultsin regard to ageing. Hence, this investigation is an attempt towards resolving the CDT and NA protocols, mechanism ofseed deterioration, and the role of reactive oxygen species (ROS) and their detoxification in the non-orthodox seeds ofneem (Azadirachta indica). During NA the 100% viable neem seeds became non-viable in 150 days, whereas under CDTconditions the seeds survived only 24 h. Initially (100% germination), the ROS producing capacity was minimum butincreased with prolonged storage, showing comparatively higher levels in the CDT conditions. High levels of SOD, APXand CAT protect the neem seeds from ROS damage thus maintaining high viability. ROS levels accumulated in ageingneem seeds due to impaired antioxidative capacity; decline of intensity of isoenzymes or disappearance of isoenzymes.Appearance of new isoenzymes in the ageing seeds [up to 78% germination] in the NA seeds revealed the basis ofintermediate storage behavior of neem seeds. The reduced transcripts of SOD and APX confirmed isoenzymes pattern inNA aged seeds. The pattern of ROS, its antixodative enzymes; quantitative, isoenzymes and gene expression analysiswill be discussed to explain the mechanism of NA and CDT in neem seeds in respect to its intermediate storage behavior.

PB137 (IPPC0137)

The illusive plant osmosensor: OsHK3b as a possible candidate in rice

Ramsong Nongpiur1*, Ashutosh Sharan1, Ratna Karan1, Praveen Soni1, Sneh Singla-Pareek2 and Ashwani Pareek1

1School of Life Science, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

Osmotic stress like drought and salinity greatly reduce crop productivity. Rice, one of the major food crops, is extremelysensitive to osmotic stress and it is imperative that we understand the mechanism of how rice responds to osmotic stress.Till date, there is no comprehensive report which shows how plants perceive osmotic stress and thus no bonafide plantosmosensor has been identified. Our quest for identifying an osmosensor has led to the identification of an osmotic stressinducible rice histidine kinase, OsHK3b as a putative candidate. Our results show that OsHK3b has a high degree ofstructural similarity to EnvZ and SLN1, which are osmosensors in E. coli and yeast, respectively. OsHK3b also shares ahigh degree of similarity with putative osmosensor of Arabidopsis, AtHK1. Heterologous expression of OsHK3b could notonly complement the function of SLN1 in sln1-mutant yeast but also restore osmosensing capabilities to the yeastosmosensing-deficient double mutant, sln1Äsho1Ä, thus showing that OsHK3b can function as an osmosensor in yeast.Overexpression of OsHK3b in rice positively regulates growth as well as tolerance to salinity and drought, while knockingdown of OsHK3b expression through RNAi resulted in hypersensitivity to osmotic stress, indicating its positive regulatoryrole in the osmotic stress response of rice. A probable mechanism of action through the interaction and modulation ofOsPhp3 and, subsequently, OsRr22 and OsRr26 is also observed in yeast-two-hybrid and bifluorescence complementationassays.


184

IPPCongress2015


PB138 (IPPC0165)

Elucidating the two-component mediated signal transduction system in Oryza sativa L.

Ashutosh Sharan1*, Praveen Soni1, Sneh L. Singla-Pareek2 and Ashwani Pareek1

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

Our laboratory has been engaged in working out the intricacies of stress physiology and functional genomics of modelcereal crop, rice. In this endeavor, we have attempted to dissect the complex interactions amongst an important class ofsignaling proteins comprising a group designated as two-component system (TCS). The complex rice genome possesslarge number of TCS genes including 13 histidine kinases (HKs), 5 histidine phosphotransferases (Hpts) and 34 responseregulators (Rrs). TCS signaling include specific protein-protein interactions between its various components for signaling.In present study cytosolic fragments of OsHK4 and OsHK5 were used, as full length HKs are membranous protein whichcan not be used for interaction study using yeast-two hybrid technique (Y2H). We found OsHk4 to interact with OsAhp2,whereas OsHk5 was found to interact with OsAhp1, OsAhp2 and OsPhp1. All Hpts exhibited multiple interactions withdifferent Rrs indicating their function as interaction hubs. We also observed selective interactions between type-A andtype-B Rrs. Different pseudo-response regulators showed interactions among themselves. These results constitute strongarguments in favour of a cross talk between signaling mediated by different sensory histidine kinases. We also re-validated these interactions by bimolecular fluorescence complementation assay (BiFC). Interactions of HKs (cytoplasmicfragments) with Hpts were found to be cytoplasmic as well as nuclear where as those of Hpts with Rrs and those amongRrs were found to be localised to the nucleus only. This study provides starting point for further analysis of the molecularmechanisms for the signal transmitted by the TCS into biological processes.

PB139 (IPPC0218)

A natural resistant associated macrophage protein from Brassica juncea exhibits a novel interactionwith a membrane bound thioredoxin protein

Ananya Marik, Haraprasad Naiya, Gairik Mukherjee, Madhumanti Das and Chinmay Saha and Anindita Seal*

Department of Biotechnology, University of Calcutta, 35,Ballygunj Circular Road, Kolkata-700019, West Bengal, India


NRAMPs are a family of evolutionarily conserved proton coupled transporters, which transport both essential and non-essential metals in plants. Fifty protein interactors of a Brassica juncea NRAMP, BjNRAMP4.1 were identified through asplit ubiquitin yeast two-hybrid screen. Among these interactors, interaction with a membrane bound thioredoxin likeprotein BjHCF164, homologous to Arabidopsis high chlorophyll fluorescence protein 164 (AtHCF164), was studied indetail. AtHCF164 is implicated in the transfer of reducing equivalents from stroma to luminal proteins during photosynthesisand a knockout is seedling lethal in Arabidopsis. BjNRAMP4.1 and BjHCF164 exhibited dual localization in plants.BjHCF164, together with its reported site of function in chloroplasts, was also found to strongly express in Brassica roots.BjNRAMP4.1 localized in chloroplasts and epidermal membranes in leaves and membrane in roots. In keeping with suchdistinct tissue-dependent localization, the proteins interacted in chloroplasts in leaves and membrane in roots. Theinteraction was found to increase in the presence of excess manganese both in yeast as well as in plant roots. Aninteraction network created from Arabidopsis homologs of the BjNRAMP4.1 interactors was highly enriched in proteinscentral to chloroplastic/cellular ROS signaling. Homology modeling and docking studies suggested that BjNRAMP4.1could be potentially redox regulated by BjHCF164, while BjHCF164 could be regulated through hydrophobic interactionthrough residues within the thioredoxin domain. Our results suggest that interaction between BjNRAMP4.1 and BjHCF164may be a potential link between metal homeostasis and chloroplastic/cellular redox function.


185

IPPCongress2015


PB140 (IPPC0296)

Regulation of microRNA biogenesis by mitogen activated protein kinases in plants

Prakash Kumar Bhagat* and Alok Krishna Sinha



MicroRNAs (miRNA) are 21 -24 nucleotide in length transcribed by RNA polymerase II from mir gene locus. The processingof primary miRNA (pri-miRNA) transcript to precursor miRNA (pre-miRNA) followed by generation of mature miRNA/miRNA* duplex is carried out by a microprocessor/biogenesis complex. The microprocessor complex comprises of RNaseIIIendonuclease, DCL1 (DICER –LIKE1 protein), DRB1/ HYL1 (double stranded RNA binding protein /HYPONASTYLEAVES1), SE (zinc finger protein SERRATE) and TGH (G- patch domain protein TOUGH). The mature miRNA/miRNA*duplex is transported to the cytoplasm where it is processed further, finally targeting mRNA trancript to regulate itsexpression. Earlier studies have suggested a role of dephosphorylarion of DRB by a phosphatase, CPL1 is required forthe biogenesis of miRNA. While another report from our lab suggested that DRB1 is phosphorylated by MPK3 and that itnegatively regulates miRNA biogenesis. We here report characterizing of OsDRB1 in rice using surface plasmon resonance(SPR) technology. Deletion construct of DRBs have been prepared to map the MAPK phospho site/s on these proteins.To get an insight of these interactions and to elucidate the biological function of DRBs and OsMPK3, a transient knockdownprotocol using VIGS vector has been standardized. In nut-shell the regulation of DRBs during miRNA biogenesis byMAPK will be discussed.

PB141 (IPPC0300)

Investigation of novel regulators involved in MAP kinase and light signalling pathways during Arabidopsis

seedling development

Alok Krishna Sinha and Deepanjali Verma*

Plant Signaling, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Light modulates the changes in the gene expression that occur at various developmental stages of plants in definedtissues. These changes take place mainly at the transcriptional level and lead to the regulation of transcription of specificgenes that contributes light regulatory developments. MYC2, a basic helix loop helix transcription factor is known as apositive regulator of jasmonic acid mediated inhibition of root growth. In an earlier study it has been reported that a MAPkinase module plays a key role in MYC2 mediated light signalling. The study demonstrates that MPK6 is activated byMKK3 in blue light (BL) and that this process is MYC2 dependent. It is also shown that MPK6 physically interacts with andphosphorylates MYC2 which binds to the promoter of MPK6 and represses its expression. In the present work we haveidentified a phosphatase, MKP1 as an interacting partner of MPK6 and regulator of MPK6-MYC2 module. The interactionwas confirmed by in-vitro phosphorylation assay, which showed that MKP1 dephosphorylates MPK6 in the presence ofblue light thus regulating MPK6-MYC2 module, which is a very crucial step and needs to be tightly controlled. To identifythe upstream MAP3K which activates MKK3 in this pathway, we have selected 6 MAP3Ks showing upregulation in BL onthe basis of in-silico microarray analysis. To further substantiate we performed gene expression analysis which identified3 MAP3Ks that might be involved in the pathway. We intend to identify the specific MAP3K involved in the activation ofMKK3-MPK6-MYC2 module mediated light signalling.


186

IPPCongress2015


PB142 (IPPC0353)

Functional analysis of Arabidopsis FCS-LIKE ZINC FINGER gene family suggests their role in the regulationof plant growth and stress mitigation

Muhammed Jamsheer K., Dhriti Singh*, Chanchal Thomas Mannully, Yadvendra Singh Chattha and Ashverya Laxmi


The FCS-LIKE ZINC FINGER (FLZ) gene family is characterized by the presence of a solitary FCS-LIKE ZINC FINGER(FLZ) domain which is involved in mediating protein: protein interaction. Many members of this gene family are implicatedin sugar signaling, biotic and abiotic stress response. In this study, we identified that the Arabidopsis thaliana FLZ proteinsinteracts with a wide variety of proteins which are involved in vegetative and reproductive development, developmentalphase transition, stress response, etc. In some cases, even the evolutionary distant proteins were found to share commoninteracting proteins suggesting a high degree of functional redundancy in this gene family. Besides, these zinc fingerswere found to be interacting with other members of this protein family and in many cases this interaction was found to beimportant for maintaining proper sub-cellular localization. The FLZ domain was found to be the canonical interactingmodule for interacting with other proteins while the N-terminal region was found to important for interacting with theproteins of the same protein family. The knock-out mutants of these genes were found to have an impact on normalgrowth and adaptive growth under osmotic stress suggesting the role of this gene family in plant growth and developmentand adaptive growth under stressed conditions. Further molecular and physiological analysis need to be done to decipherthe exact molecular functions of these genes.

PB143 (IPPC0367)

Role of stress-induced F-Box proteins in ABA and reactive oxygen species mediated signalling inArabidopsis and rice

Avantika Bhaskar1, Eshan Sharma1* and J.P. Khurana2

1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110021, India2Department of Plant Molecular Biology, Interdisciplinary Centre for Plant Genomics and University of Delhi, South Campus,New Delhi-110021, India*Presenting author: [email protected]

Protein degradation is an important aspect of plant growth and development mediated by the ubiquitin/26S proteasomepathway, allowing cells to rapidly respond to changes in environment. Here, we present functional characterisation of anF-box protein encoding gene OsFBX in rice and its orthologous genes in Arabidopsis. Quantitative RT-PCR expressionanalysis revealed induction of OsFBX under various abiotic stresses with high transcript levels specifically under droughtin Oryza sativa cv IR64. Our analysis also showed high levels of OsFBX during seed development followed by panicledevelopmental stages. Transiently expressed OsFBX (fused with reporter gene encoding GFP) localized to the nucleusindicating it as a part of proteasome complex that is abundant in the nucleus. Ectopic expression of OsFBX in Arabidopsis

thaliana led to a hypersensitive phenotype in response to ABA, glucose and osmotic stress. The corresponding orthologousmutants in Arabidopsis showed hyposensitivity to ABA mediated responses such as inhibition of seed germination, rootand seedling growth in presence of ABA, NaCl and mannitol. Further, short duration ABA treatment increased ROS levelsin Arabidopsis lines overexpressing OsFBX compared to reduced levels in Arabidopsis mutants. The ABA sensitivity ofmutants and transgenic plants also correlated with faster water loss in mutants. Thus, the F-box protein analysed in thisstudy may positively regulate ABA-mediated plant responses under abiotic stress conditions.


187

IPPCongress2015


PB144 (IPPC0378)

Effects of oxidative and photooxidative stress on reactive oxygen species and proline metabolism inleaf discs of Pisum sativum

A.S. Raghavendra and Vetcha Aswani*

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India*Presenting author: [email protected]

Plants are continuously exposed to biotic and abiotic stresses. On exposure to stress, the reactive oxygen species (ROS)accumulate and these ROS should be scavenged. We have examined the patterns of ROS accumulation and theconsequences in leaf discs of pea, Pisum sativum. We employed oxidants, which induce ROS in different cellularcompartments: acifluorfen methyl ester (AFM) and paraquat (PQ) (both in chloroplasts), menadione (MD) (mitochondria)and abscisic acid (ABA) (plasma membrane). The accumulation of H

2O

2 and superoxide were estimated by the oxidation

and reduction of DAB and NBT, respectively. The ROS levels were higher in AFM and PQ treated samples compared tothat of MD and ABA. Among the four antioxidant enzyme activities studied, the increase in the activity of superoxidedismutase (SOD) and glutathione reductase (GR) were higher than that in ascorbate peroxidase (APX) and catalase(CAT). Proline, an osmolyte, increased significantly under stress. Among key enzymes of proline metabolism, slightincrease in the activity of pyrroline 5 carboxylate synthase (P5CS) compared to the marked decrease in prolinedehydrogenase (ProDH) indicated that increased proline levels were due to the restricted proline oxidation. Finally, thechange observed in the antioxidant metabolism was validated by studying physiological response by oxygen electrode.These results indicated that the leaf photosynthesis was decreased in all treatments; maximum was with PQ, whereas,respiration was lowest in MD treatment. The protein levels and gene expression pattern of the antioxidant enzymes (CAT,APX, GR) and of proline metabolism (P5CS and ProDH) are also studied.

PB145 (IPPC0385)

Signaling components and their interaction during stomatal closure by microbial elicitors, in comparisonwith the effects of abscisic acid

Gunja Gayatri1*, Appa Rao Podile1, Mallikarjuna Rao Puli1, Srinivas Agurla1, Kazuyuki Kuchitsu2 and A.S. Raghavendra1

1Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-50046, India2Department of Applied Biological Science, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba


Plant stomata act as the main access points for the entry of the pathogens. Stomatal closure restricts the entry of thepathogens, becoming one of the plant innate immune responses. Such closure is mediated by microbial elicitors, producedfrom either microbes or plant tissue. We therefore studied effect of three elicitos: flg22 (a 22 amino acid peptide fromeubacterial flagellin), harpin (from Pseudomonas syringae pv. syringae), cryptogein (from Phytophthora cryptogea) onstomata of Arabidopsis thaliana. All the three elicitors (flg22, cryptogein and harpin) induced marked stomatal closure.The stomatal closure, for e.g. by ABA, involves several signaling components, including reactive oxygen species (ROS),nitric oxide (NO), G-proteins, calcium and protein kinases/protein phosphatases. We therefore examined the patterns ofROS and NO in guard cells. The elicitors raised markedly the levels of ROS or NO in guard cells. Time course monitoringof ROS or NO, and studies using ROS or NO modulators on stomatal closure revealed that ROS acted upstream of theNO during stomatal signaling by these elicitors, similar to ABA. Participation of signaling components was further confirmedby mutants. Impaired stomatal closure and decreased ROS or NO levels by these elicitors in guard cells of atrbohD/Fconfirmed that ROS was essential during stomatal closure. The partial reversal of stomatal closure in nia1 and nia2,decreased NO production indicated the essentiality of NO production during elicitor triggered stomatal closure. A schemeof signaling events during stomatal closure by microbial elicitors would be proposed.


188

IPPCongress2015


PB146 (IPPC0423)

Abiotic stresses, tissue specificity and diurnal rhythmicity act as multiple regulatory switches to controltranscription of members of two-component system in rice

Praveen Soni1#*, Priyanka Gupta1, Sneh L. Singla-Pareek2 and Ashwani Pareek1

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India#Present Address: Department of Botany, University of Rajasthan, Jaipur-302004, Rajasthan, India*Presenting author: [email protected]

Two-component system is his-to-asp phosphorelay based signaling machinery that mediates perception, signal transductionand response to environmental stresses. Though, TCS members have already been reported in rice but their functionsare yet to be identified. To gain insight into the role of TCS members, their expression responses to environmentalstresses and ABA hormone were analyzed in contrasting genotypes-IR64 and Pokkali at seedling stage. We observedtheir differential regulation in genotype-dependent and stress-specific manner. Transcript profiling distinguished OsHk3,OsPhp3, OsRr21 and pseudo-response regulators as ABA inducible and multiple stress-responsive TCS members. Wealso carried out their transcript analysis in different plant tissues and different time points of a day. This analysis revealedthat TCS machinery is regulated by not only various abiotic stresses, but also by the tissue specificity and diurnal rhythmicity.Some members showed prominent expression in all tissues while some others such as Authentic histidinephosphotransferases, OsRr21 and pseudo-response regulators exhibited high expression in mature panicle and greenparts. Our data suggests that TCS members might play important role in organ development during juvenile stage andpanicle development or grain filling at mature stage. Expression of most of the TCS members showed robust oscillationwith peaks at different time points of a 12h light/12h dark cycle. Thus, we report an intricate and dynamically intertwinedregulatory network of TCS members, as influenced by the abiotic stress, ABA, tissue and diurnal rhythm. This studyprovides important clues for thorough investigationof diverse roles of the various TCS family members in rice.

PB147 (IPPC0438)

Polyamines induce stomatal closure and increase reactive oxygen species and nitric oxide levels inArabidopsis thaliana

Agurla Srinivas*, Gunja Gayatri, A.S. Raghavendra

Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India*Presenting author: [email protected]

Stomata are tiny pores present mostly on leaves and regulate the gas exchange as well as transpirational loss of H2O into

the atmosphere. The opening and closing of stomata are due to changes in turgidity of guard cells. Several abiotic andbiotic factors induce stomatal closure. For example abscisic acid (ABA), a drought inducible phytohormone, inducesstomatal closure in different plant species. Signalling components like reactive oxygen species (ROS), reactive nitrogenspecies (NO), Ca2+, cytosolic pH, sphingolipids, phospholipids and MAP kinases were all involved in ABA induced stomatalclosure. Polyamines (PAs) are ubiquitous polycationic nitrogenous compounds, that are associated with plant adaptationto abiotic factors. We have studied the effect of three polyamines: putrescine (Put), spermidine (Spd) and spermine(Spm), on stomatal movement in Arabidopsis thaliana. Among these, Put was more effective than Spd and Spm ininducing stomatal closure. The levels of ROS and NO in guard cells on exposure to the polyamines were monitored byusing fluorescent probes like CM-H2DCFDA and DAF-2DA respectively. The levels of ROS and NO in guard cells wereincreased upon treatment with the PAs. Thus, the patterns of ROS and NO levels in guard cells, corresponded withstomatal closure. The restriction of stomatal closure by 2-Phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO,NO scavenger) and N-nitro-L-Arg-methyl ester (L-NAME, a nitric oxide synthase inhibitor) confirmed the role of NO duringpolyamines induced stomatal closure. The participation of NADPH oxidase, polyamine oxidase and peroxidases in theROS production and stomatal closure is being examined.


189

IPPCongress2015


PB148 (IPPC0444)

Characterization of a potato class-I KNOX gene- potato homeobox 15

Ameya Mahajan1, Kirtikumar Kondhare1*, Mohit Rajabhoj1, Tejashree Ghate1, Nevedha Ravindran1, Amit Kumar1, FarhatHabib1, Sundaresha Siddappa2 and Anjan Banerjee1

1Indian Institute of Science Education and Research, Pune-411008, Maharashtra, India2Division of Plant Protection (Plant Physiology), Central Potato Research Institute, Shimla-171001, India*Presenting author: [email protected]

Potato Homeobox 15 (POTH15) is orthologous to Shoot Meristemless (STM) in Arabidopsis. POTH15 belongs to potatoKNOX-I family gene. KNOX genes have been characterised in several species, but very few studies are available inpotato. The major objective of this study is to characterise POTH15 and identify its targets involved in potato development.In the first experiment, we investigated whether POTH15 transcript shows differential expression under tuber-inducingshort day v/s tuber non-inducing long day conditions in a photoperiod sensitive potato cultivar. qRT-PCR analysis wasconducted in a tissue-specific manner and it was observed that shoot tips and stolons showed high levels of POTH15

mRNA under short day conditions. In the second experiment, we studied the expression of POTH15 promoter. POTH15

promoter showed a wide expression pattern (e.g. apical and axillary meristems, petiole, stolon tip, tuber eyes, tuber pith)suggesting that POTH15 has a diverse role in potato development. In subsequent experiments, we have undertaken aconstitutive over-expression approach to identify the functions and targets of POTH15. In POTH15 over-expression lines,number of morphological traits, such as leaf, stem, tuber development, were altered. POTH15 over-expression lines alsotuberised earlier than wild-type plants under in vitro conditions. We further performed a comparative RNA-seq analysiswith wild-type and POTH15 over-expression lines, which identified 435 differentially expressed genes involved in keybiological processes. Selective candidate genes were validated by qRT-PCR for their differential expression in wild-typeand POTH15 over-expression lines. To summarise, this study suggests that POTH15 controls diverse developmentalprocesses in potato.

PB149 (IPPC0452)

Functional characterization of OsSnRK2 gene in rice

Sanya Shrivastava*, Santosh Kumar Vinjamuri, Shashank Kumar Yadav, Pooja Dhuppar, Viswanathan Chinnusamy

Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India*Presenting author: [email protected]

Sucrose non-fermenting-1 related protein kinase (SnRK) is a unique family of serine/threonine protein kinases associatedwith energy and abiotic stress signalling in plants. They are divided into three subfamilies SnRK1, SnRK2, and SnRK3. Inrice, 10 members SnRK2 family were identified and shown to be activated by salt stress. However, their role in abioticstress tolerance is poorly understood. Microarray data analysis of OsSnRK2.4 revealed that it is up-regulated mainly bydehydration and salinity treatments. In this study, rice OsSnRK2.4 gene was cloned and characterized. OsSnRK2.4 genewas cloned in pCAMBIA1300 vector under stress inducible AtRD29A promoter and constitutive ZmUBI promoter. Transgenicrice plants were developed using these constructs and confirmed through PCR, qRT-PCR and southern blotting. Post-germination analysis of OsSnRK2.4 overexpressing transgenic plants showed better shoot length and root length ascompared with WT plants. Analysis of drought tolerance of RD29A:OsSnRK2 transgenic rice lines in pot culture undergreenhouse conditions revealed that RD29A:SnRK2 transgenic rice lines are more tolerant than non-transgenic lines interms of maintenance of membrane stability index, RWC, survival and recovery growth. Further studies on detailedcharacterization of SnRK2 will elucidate their role in development and abiotic stress responses of rice.


190

IPPCongress2015


PB150 (IPPC0490)

Transcriptional regulation of BREVIS RADIX (BRX) in progenitors of bread wheat with contrasting roottraits

Pramod Awakale, Sneha Tiwari and Monika Dalal*

ICAR-National Research Centre on Plant Biotechnology, New Delhi-110012, India


Deep root system is an important drought avoidance mechanism in plants. The highly plastic nature and difficulty inphenotyping deters the use of root traits in breeding programs. Identification of genes/alleles and markers associatedwith RSA can expedite the use of this trait to improve yield of crop plants. The diploid progenitor species of bread wheat(T. aestivum L.) namely T. monococcum (A genome), Aegilops tauschii (D genome) and Ae. speltoides (S genome,closest to B genome), are potential resources for various agronomic traits including abiotic stress tolerance. However,these species have not been explored for inductive root traits. Therefore to characterize this untapped genetic resource,seedlings of nine accessions of diploid wheat (three accessions each representing A, B and D genomes) were phenotypedfor inductive root growth under two levels of osmotic stress viz. -0.5 (S1) and -1.48 bars (S2) for 10 days. Significantgenetic variability was observed for root traits among the accessions with different genomes. The BREVIS RADIX (BRX)transcription factor controls cell proliferation and elongation in growth zone of root tip and explained 80% of variance inroot length in Arabidopsis. Hence, BRX expression was analysed under control and stress conditions in the diploidaccessions with contrasting root traits. The analysis of BRX expression and allelic variation will help in understanding theevolutionary role of BRX in root development and growth in wheat.

PB151 (IPPC0493)

Interaction specificity of heterotrimeric G-protein beta (Gâ) and gamma (Gã) subunits in Brassica juncea

Gulab Arya*, Roshan Kumar and Naveen Bisht


Heterotrimeric G-proteins (G-proteins) are guanine nucleotide (GTP and GDP) binding proteins that constitute Gα, Gβand Gγ subunits, and regulate many aspects of physiological development and cell signaling across phylas. Various loss/gain of functional studies in model plants Arabidopsis and rice revealed that G-protein βγ dimers are involved in manybiological processes like hormone signaling, stomatal opening and closing, resistance against necrotrophic fungi, controllingshape and size of leaf and seeds, and root development. However, our understanding about G-protein signaling complexfrom any polyploid crop is limited. In current study we identified multiple homologs of G-protein β and γ subunits fromallotetraploid B. juncea (AABB), an economically important oilseed crop of Brassica genus. Protein-protein interactionbetween different G-protein β and γ subunits using yeast based assays showed a high degree of differential interactionspecificity among the multiple G-protein subunits. Amino acid sequence alignment of multiple G-proteins revealed presenceof divergent key residues across Gγ homologs. Site directed mutagenesis (SDM) followed by yeast based interactionassays could delineate key residues imparting interaction specificity across B. juncea G-protein β and γ subunits.


191

IPPCongress2015


PB152 (IPPC0499)

Arabidopsis G-protein coupled receptor (GCR1) regulates cold, heat and salt stresses

Navjyoti Chakraborty*, Navneet Singh, Nandula Raghuram and Kanwaljeet Kaur

University School of Biotechnology, Guru Gobind Singh Indraprastha University, Sector 16C, Dwarka, New Delhi-110075, India*Presenting author: [email protected]

G-proteins and G-protein coupled receptors (GPCRs) have been implicated in regulating the plant’s response to some orthe other biotic/abiotic stress, but a comprehensive assessment of the role of GPCR in stress response has not beendone so far, partly due to the recent controversy regarding the existence and need for in plant G-protein signalling. Weisolated a GCR1-knock-out mutant (gcr1-5) in Arabidopsis thaliana and performed its transcriptome analysis usingmicroarrays in comparison with the wild type to understand the genome-wide roles of GCR1. Using a stringent cut off oflog

2FC 1.0 and p-value d”0.05, we found 350 differentially expressed genes (DEGs) in the mutant. More than 100 of them

were found to be involved in response to biotic/abiotic stress. Further analyses revealed that out of these, 57 DEGs (49up/8 down) belong to abiotic stress response category. We validated the effects of cold, heat and salt stresses on theGCR1 mutant and WT using standard and biochemical parameters. We also validated some of the stress-responsiveDEGs by qRT-PCR. Overall, we found that gcr1-5 is most tolerant to cold, followed by heat and salt, in terms of all theabove parameters. These results clearly show the important role of GCR1 in mediating the plant’s response to multipleabiotic stresses. Considering that G-proteins are also implicated in many of them, our data also indicate the possibilitythat GCR1 may act as a GPCR in G-protein signalling in stress response.

PB153 (IPPC0558)

Cloning and expression analysis of ABA receptor(s) in rice

Shashank Yadav1, Pooja Dhuppar1*, Santosh Vinjamuri1, Sanya Shrivastava1, Pragya Yadav1, Rakesh Verma1, BhupendraChaudhary2 and Viswanathan Chinnusamy1

1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India,2School of Biotechnology, Gautam Buddha University, Greater Noida, Uttar Pradesh, India*Presenting author: [email protected]

Abscisic acid (ABA) is an essential phytohormone which is involved in a host of biological processes, including plantdevelopment and responses to biotic and abiotic stresses. The hunt for ABA receptors culminated in the identification andcharacterization of PYRABACTIN RESISTANCE1 (PYR1)/PYR1-like (PYL)/ Regulatory Component of ABA Receptor(RCAR) proteins as ABA receptors (ABARs) in Arabidopsis. ABA-bound ABAR inhibits phosphatase type 2C proteins.This results in relieving of SnRK2 inhibition by PP2C, and then SnRN2 mediated activation of effector ion channels andtranscription factors. The specific functions of individual members of ABARs in developmental and stress responses arebeing currently investigated worldwide. This study was conducted to identify and characterize of ABAR family genes fromrice. Phylogenetic analysis grouped these ABARs in to three distinct subfamilies: the monomeric receptors formed subfamilyI and sub-family II, while the dimeric receptors formed subfamily III. Only the subfamily I genes contain introns. Expressionanalysis of ABARs in different tissues and stress conditions revealed that differential expression patterns. This suggestsnon-redundant roles of ABARs in development and stress responses. All the ABA receptors were cloned from Nagina22in pCAMBIA1300 under the control of stress inducible promoter AtRD29A for Agrobacterium mediated genetictransformation of rice. Comparison of abiotic stress tolerance of rice transgenics overexpressing ABAR9/ABAR10/ABAR11with non-transgenic plants suggests that ABARs are potential candidate genes for improving drought tolerance of rice.


192

IPPCongress2015


PB154 (IPPC0576)

Functional characterization of Med14 in Arabidopsis

Sourobh Maji* and Jitendra Thakur


Mediator complex is an important transcriptional co-regulator and consist of 31 subunits in Arabidopsis. AtMed14 is apart of tail modular subunit of plant Mediator complex. It is conserve in eukaryotes and contains N-terminal MED14domain, which is conserve throughout eukaryotic kingdoms. In metazoans it is known to be involved in androgensignaling, intracellular steroid hormone receptor signaling, stem cell maintenance and positive regulation of RNApolymerase II transcriptions. Recent studies in plants revealed its important role in growth, development and survivability.Mutation in Med14 has dramatic alteration in cell proliferation, cold response, salicylic acid induced defense responseand SAR response. We found that a homozygous T-DNA insertion line of Med14 was embryo lethal in which embryo gotarrested at different stages of embryo development. Most of the seed obtained from heterozygous lines were defectivein seed germination. These defects in the seed germination were due to defect in the secretion of mucilage fromcolumella cell of seed coat. This particular defect in seed mucilage secretion phenotype is identical to leunig_homolog(luh) and mucilage modified 2 (mum2) mutants. MED14 physically interacting with LUH and regulate the expression ofMUM2, an enzyme needed for mucilage extrusion. In addition, MED14 also physically interacts with well known auxinregulator LEUNIG (LUG) and AINEGUMENTA (ANT), suggesting its role in transcriptional regulation during auxinsignaling. Thus, our study reveals importance of Med14 in auxin-mediated processes in early stage of plant development.

PB155 (IPPC0675)

Functional characterization of a central effector of light signaling, AtHY5, ortholog in rice

Naini Burman*, Akanksha Bhatnagar and Jitendra P. Khurana

Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular Biology, University of Delhi South Campus,New Delhi-110021, India*Presenting author: [email protected]

Light plays a very important role in plant’s life as they are sessile and photoautotrophic. Plants have developed anintricate network of light signaling components, which interact with almost all the important pathways. This light signalingcascade is highly hierarchical with photoreceptors like phytochromes and cryptochromes being at the top. They arefollowed by early signaling factors like HFR1 and central integrators like COP1. Then lies the downstream effectors likeHY5. Most of the components of light signaling pathway have been characterized in Arabidopsis, a model dicot plant.One of the orthologs of AtHY5 in rice, encoded by OsbZIP48, has been functionally characterized using transgenicapproach. Over-expression transgenics of OsbZIP48 caused dwarf phenotype and accumulation of more chlorophyllcontent. There was a high degree of floret sterility, which severely affected the grain yield. RNAi transgenics of OsbZIP48

segregated into two types of seedlings in T2 generation; one which resembled wild type and vector control in morpologyand the other which showed lethal phenotype. Microarray analysis revealed that a number of hormonal pathways likegibberellin synthesis pathway were affected in these transgenics. These data obtained by analysis of rice transgenics forOsbZIP48 provide evidence that rice ortholog of AtHY5 performs overlapping functions with HY5 as well as it plays somemonocot specific roles in regulating plant development.


193

IPPCongress2015


PB156 (IPPC0689)

OsTLP, an F-box containing TUBBY protein in rice, regulates stress signaling and development

Nitin Jain*, Satyam Vergish and Jitendra P. Khurana

Department of Plant Molecular Biology, Interdisciplinary Centre for Plant Genomics, Delhi University, South Campus, New Delhi-110021, India*Presenting author: [email protected]

The TUBBY genes were identified in mammals as a small group of 4-5 members based on a mutation in mice that causedobesity on maturation. In animals, they also regulate insulin pathway, retinal maintenance, neuronal development, etc.These class of proteins, harboring a C-terminal TUBBY domain, are bi-partite transcription factors that translocate to thenucleus from the plasma membrane on perceiving certain signals and have both DNA binding and transcriptional activationproperties. In plants, these C-terminal TUBBY domain containing proteins are invariably coupled with a conserved N-terminal F-box domain and are thus called as TLPs (TUBBY LIKE PROTEINS). Both Arabidopsis and rice have nearly14 TLPs that regulate diverse functions such as abiotic stress signaling, hormone signaling, plant-pathogen interactionsand so on. Our lab identified a few F-box proteins in rice that are regulated under different light conditions including anOsTLP. This OsTLP has been further characterized and found to be upregulated under blue light and is also undercircadian control as its expression dips at midnight. The gene also showed upregulation under salt and drought stressand in ovary tissues. The protein harbors a canonical F-box domain which interacted with OsSKPs showing that it is acomponent of a functional SCF-type E3 ligase. The protein was found to be localized to the plasma membrane thatshifted to the nucleus under stress conditions. The over-expression lines in Arabidopsis showed better over-all growthand involvement of the gene in various abiotic stresses. The data thus far indicate involvement of the OsTLP in abioticstress signalling and contribution towards overall growth and development.

PB157 (IPPC0690)

Functional characterization of an OsbZIP gene in rice for its possible role in light signaling

Akanksha Bhatnagar*, Naini Burman and Jitendra P. Khurana

Interdisciplinary Centre for Plant Genomics & Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India*Presenting author: [email protected]

Plant growth, development and response to environmental stimuli are determined by programmed expression of itsgenes which are regulated by transcription factors. These transcription factors are usually members of multigene families.One of the largest and most diverse transcription factor families in eukaryotes is the basic leucine zipper (bZIP) transcriptionfactor family. Over 120 bZIP transcription factor genes have been identified in different plants, out of which 89 have beenidentified from rice and 67 from Arabidopsis. Phylogenetic analysis of the bZIP transcription factors across plant specieshas predicted a majority of the rice bZIP transcription factors to be orthologs of Arabidopsis bZIP transcription factors.One of the most well characterized bZIP transcription factor in Arabidopsis, HY5, has been predicted to have threeorthologs in rice, one of which (OsbZIP48) has already been charcaterized in our laboratory for its orthologous function toAtHY5. In this presentation, the function of another member of this sub-group has been presented. This OsbZIP gene toohas been found to functionally complement the hy5 mutant in Arabidopsis. Expression levels of this ortholog of HY5 havebeen checked in different developmental stages as well as in different tissues of rice by real time PCR. Analysis of itsexpression in 2-10 day old light as well as dark grown seedlings has also been done and these results indicate differencein expression patterns of both these orthologs in rice. Transgenic analysis in both rice as well as Arabidopsis, however,points towards their functional similarity.


194

IPPCongress2015


PB158 (IPPC0707)

Promoter analysis of GA2ox1, a gibberellin catabolism gene, reveals its role in blue light-mediatedphotomorphogenesis in Arabidopsis

Sushma Mishra* and Jitendra P. Khurana

Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India*Presenting author: [email protected]

The young light-grown seedlings characteristically display short hypocotyl, open apical hook with expanded cotyledonsand plastid development in dicot systems like Arabidopsis. This photomorphogenic development occurs by the co-ordination of light signals perceived by photoreceptors and endogenous cues like phytohormones. One such exampleis the interaction between cryptochromes (CRYs), the blue light-sensing photoreceptors, and the phytohormone gibberellin(GA). Prompted by the observations made in our laboratory on Brassica CRY1-ox transgenics, which showed reducedplant height accompanied by altered transcript levels of GA metabolism genes, it was decided to work out the intricaciesassociated with this cross-talk in Arabidopsis. The real-time PCR analyses confirmed that blue light stimulates theexpression of AtGA2ox1, a gene encoding GA-catabolism enzyme. Moreover, Arabidopsis transgenics containingGA2ox1 promoter-reporter (GUS) construct showed higher promoter activity in blue light grown seedlings in comparisonto the dark grown ones; it is likely that higher activity of GA catabolism enzyme leads to low availability of bioactive GAlevels and hence causing suppression of elongation, resulting eventually in shorter plant height. The trans-acting factorsinteracting with GA2ox1 promoter were identified by yeast-one-hybrid assay, and the interaction confirmed byelectrophoretic mobility shift assay. Work is in progress to elucidate the precise role of these factors in light regulatedexpression of GA2ox1.

PB159 (IPPC0853)

Nitric oxide in plants: the biosynthesis and cell signalling functions of a fascinating molecule in plants

Luis Mur, R. Ratcliffe2, Werner Kaiser3 and Kapuganti Jagadis4*

1Institute of Biological, Environmental and Rural Sciences, University of Aberswyth, Wales, UK2Department of Plant Sciences, University of Oxford, Oxford, UK3Department of Botany, University of Wuerzburg, Wuerzburg, Germany4National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


The free radical nitric oxide (NO) emerged as an important signal molecule in plants. Various reductive, oxidativepathways operative for NO biosynthesis. The reductive pathway utilizes nitrite as substrate, which is exclusively producedby cytosolic nitrate reductase (NR) and mitochondria. Using plant mitochondria, we show that nitrite reduction to NO isstrongly increased with the decrease in oxygen, which is a general consequence under flooding stress. The reaction islinked to ATP synthesis under hypoxia. NO diffuses from the mitochondria to the cytosol where it is scavenged to nitrateby the non-symbiotic haemoglobins. I will present experimental data supporting these reactions and showing that underlow oxygen, the plant mitochondrion serves as a nitrite: NO reductase and becomes a major component in the anoxicnitrogen cycling where it directly contributes to a decrease of cell reduction level and to a limited ATP synthesis. We alsofound that NO inhibits aconitase and increases in citrate levels which then act as a potent inducer of AOX pathway. TheNO production, inhibition of aconitase, and induction of AOX leads to a shift of plant metabolism towards amino acidbiosynthesis. Under normoxic conditions NO regulates respiration, internal oxygen, carbohydrate utilization, and ROSlevels in roots. We show that a decrease in NO, leads to a drop in internal oxygen, an increase in glucose consumption,further elevation of ROS. Thus, NO is required for maintaining steady-state oxygen concentrations and to keep ROS low.Under pathogen attack nitric oxide is important signal for plant resistance and development of hypersensitive responseand NO also play role in suppression of high concentration of NO induced by root pathogens.


195

IPPCongress2015


PB160 (IPPC0854)

Nitric oxide plays a role in the establishment of aerenchyma formation in wheat (Triticum aestivum L.)

Aakanksha Wany*, Sonal Mishra, Rythm Vanvari and Jagadis Kapuganti


Plant have several adaptive strategies for survival under hypoxic conditions. One of the adaptive strategy is aerenchymaformation in roots. Lysigenous aerenchyma contributes hypoxic tolerance in roots, by providing an internal aerationsystem for the transfer of oxygen from the shoot to root. However, aerenchyma formation requires the death of cells in theroot cortex. Nitric oxide is known to induce cell death in plants during incompatible plant pathogen interactions, thereforethere is a possibility that NO could play role in lysigenous cell death during aerenchyma formation. In order to understandwhether NO play role in aerenchyma formation, we subjected nitrate grown wheat (HD 3086 and HD 2967) plants toanoxia for 24, 48 and 72 hours and then compared aerenchyma formation and nitric oxide formation with normoxicconditions. We found that under anoxic condition NO production has increased which was visualised by DAF-FMfluorescence. Under anoxic treatment roots have increased levels of aerenchyma formation. Application of nitric oxidescavenger cPTIO reduced the levels of aerenchyma formation. Metabolic, gene expression studies revealed that NOplays an important role in establishment of aerenchyma thus contribute for flooding survival.

PB161 (IPPC0862)

Hormone profiling reveals the influence of He-Ne laser on Brinjal (Solanum Melongena L.) var. MattuGulla

P.S. Swathy1*, K.R. Kiran1, K.K. Mahato2, K. Satyamoorthy3 and A. Muthusamy1

1Department of Plant Sciences, 2Department of Biophysics, 3Department of Cellular and Molecular Biology, School of Life Sciences,Manipal University, Planetarium Complex, Manipal-576104, Karnataka, India*Presenting author: [email protected]

Plants are immobile organisms that use various light signals to acclimatize their growth and development to the inconsistentenvironmental conditions. He-Ne is a powerful source of light that has widely been applied in the field of agriculture andit accelerates seed germination rate, crop yield, biomass and resistant to stress conditions. The laser excites the biologicalmolecules, leading to photochemical transformations at molecular level. Therefore, the aim of this study is to understandthe influence of He-Ne laser on endogenous phytohormone level and yield characters of brinjal on in vitro and in vivo

conditions. In the present study, the seeds of brinjal (Solanum melongena L.) var. Mattu Gulla were irradiated with singleexposure of He–Ne laser at different doses of 20, 25 & 30 J/cm2 and germinated aseptically on MS basal medium.Various physiological and growth parameters of the seedlings established in in vitro & in vivo conditions were estimatedand significant enhancement was observed with respect to morphological and yield characteristics. The basis of stimulationprocess by red light could be the synergism between laser beam and the phytochrome receptors. Phytochrome controlsthe seed germination via regulation of two plant hormones, abscisic acid (ABA) and gibberellic acid (GA

3). The hormones

were extracted from the control and laser irradiated seedlings, quantified using HPLC and the endogenous level of thehormones revealed the antagonistic regulation between ABA and GA

3. The results of the present study will be presented

and discussed.


196

IPPCongress2015


PB162 (IPPC0885)

Relevance of WUE in improving growth rates in rice (Oryza sativa L.) lines with comparable root andleaf area

A.B. Priyanka*, D. Pushpa, M. Pratibha, S. Rajesh Kumar, B. Pooja, B.R. Raju, M. Udayakumar and M.S. Sheshsshayee

Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India*Presenting author: [email protected]

Rice is the highest user of irrigation water. With the increasing demand for fresh water and erratic monsoon due to climatechange, rice production is fast becoming unsustainable. Thus rice cultivars with reduced water requirement need to bedeveloped through pyramiding drought adaptive traits. Although water use efficiency is considered as an important trait,its relevance in reducing water requirement and improving yield has remained equivocal. Here we report that WUE isindeed an important determinant of growth rates when other integral traits like root and leaf area are optimised. Thishypothesis was verified using 30 rice genotypes with similar leaf area and root weight but differing in Δ13C, a surrogate forWUE. Trait diversity among these genotypes was revalidated and four contrasting genotypes with significant variation inΔ13C were selected. These lines were raised in pots maintained at 100% FC and 60% FC by gravimetry. A strong positivecorrelation between WUE and TDM indicated the relevance of WUE. Based on the influence of physiological parameterson variability in WUE, genotypes can be classified as conductance and capacity type. The capacity types possess superiorefficiency of photosynthesis and hence can produce more biomass at a given water use. The genotype R-44 emerged assuch a capacity type. This genotype showed superior carboxylation efficiency based on CO

2 response curves and a

better mesophyll conductance as evidenced by chlorophyll fluorescence kinetics. The study provided convincingexperimental evidences to prove the relevance of WUE.

PB163 (IPPC0889)

Proteomic analysis of calcium signallome under dehydration stress in rice

Deepti Gupta1, Yogita Rai1* and Niranjan Chakraborty2

1Department of Biotechnology, TERI University, Vasant Kunj, New Delhi-110070, India2National Institute of Plant Genome Research, Aruna Asif Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

Calcium is a ubiquitous secondary messenger involved in various stress signalling pathways in plants. It has beenreported that exogenously applied calcium leads to alleviation of dehydration stress in plants. The present work was doneto emphasise the role of exogenous calcium by using a calcium channel blocker (Lanthanum chloride) on physiology ofdehydration stressed plants. 2-D coupled with MS/MS was used to generate a calcium signallome map of dehydrationstressed plant. The proteomic investigation led to the identification and characterization of novel protein/s which havebeen grouped into transcription factors, metabolism, signalling and structural proteins. A novel protein having its role inplant development but no reports under abiotic stress response has been identified and characterized. This study presentssignificant step towards understanding of calcium signalling network under water deficit stress in rice.


197

IPPCongress2015


PB164 (IPPC0990)

HOX 10 – A homeodomin transcription factor controlling root length in rice

Rajesh Kumar Singal1*, K. Sumanth Kumara1, H.R. Sowmya1, M.P. Rajanna2, R.P. Sharma3, T. Mohapatra4, N. Sarala5,S. Robin6, A.K. Singh7 and M.S. Sheshshayee1

1Department of Crop Physiology, University of Agricultural Sciences, GKVK, UAS Bengaluru-560065, Karnataka, India2Zonal Agricultural Research Station, College of Agriculture, Mandya3National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India4Central Rice Research Institute, Cuttack-753006, India5Directorate of Rice Research, Rajendranagar-500030, Hyderabad, India6Department of Rice, Tamil Nadu Agricultural Research Institute, Coimbatore, India7National Bureau of Plant Genetic Resources, New Delhi-110012, India*Presenting author: [email protected]

Mutants with single nucleotide variations created by treating seeds with EMS form an excellent resource for functionalgenomics in plants. An upland rice cultivar, Nagina 22 was mutagenized and a set of mutagenized lines were screenedfor variations in root traits and WUE using Δ13C and other standard protocols. One high root volume and one low rootvolume mutant were identified and advanced to stabilize the genotype. These mutants along with the N-22 were used fortargeted re-sequencing of around 120 root and photosynthesis related genes by NGS technology. A non-synonymousnucleotide variation was noticed resulting in a change of guanine to thymine in a homeodomin transcription factor HOX10. These mutants were extensively characterized for physiological alterations under well watered (WW, 100% FC) andwater limited (WL, 70% FC) conditions done by using gravimetric procedure. The high root mutant maintained high rootvolume, low root length and significantly higher water relations as well as canopy gas exchange characteristics under WLconditions compared to the wild type and the shallow rooted mutant. To further investigate the genetic basis of the HOX10 mutant allele, a MUTMAP population was generated by crossing the high root mutant (N22_BADT_392_9_1) with thewild type. Selective genotyping of F

2 MUTMAP progeny differing in root traits was performed by sequencing the HOX 10

gene. The HOX 10 mutant allele was found in all the 10 low root length F2 progeny that had high root volume. The results

clearly demonstrated the role of HOX 10 transcription factor in regulating root architecture by influencing root length andproliferation.


198

IPPCongress2015


PB165 (IPPC0991)

Detoxification of reactive cytotoxic compounds by Aldo-keto reductases (AKR’s) improves seed viability and

seedling vigour in rice

K.N. Nisarga*, S.V. Ramu, N. Ashwini, M. Udaykumar and M.B. Raju

Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India*Presenting author: [email protected]

Seeds deteriorate during storage and in this process, reactive oxygen species (ROS) mediated lipid peroxidation leads togeneration of reactive carbonyl compounds (RCC’S) and non-enzymatic modifications of proteins through maillard andamodari reactions. This leads to early loss of seed viability and vigour. Therefore, managing these compounds hasrelevance in improving the seed viability and seedling vigour. Towards this, an attempt was made to assess the variationin lipid peroxidation induced loss of seed viability through accelerated ageing technique in selected rice genotypes. Theresults showed variation in level of cytotoxic compounds such as melondialdehyde (MDA), Methyl glyaoxal (MG), Miallardand amodari reaction products with longer seed viability genotypes showing less compared to short viable genotypes.NADPH dependent dehydrogensae activity was found to be significantly low in short viable genotypes. Further, a significantinverse relationship was found between the level of cytotoxic compounds and seed viability and seed germination. Thisclearly suggests that, the level cytotoxic compounds in seeds determine the seed longevity. Aldo-keto reductases, areknown to detoxify the cytotoxic compounds and accordingly, AKR1 transgenic was developed in a short viability ricegeneotype (Telahamsa) to check the relevance of AKR in regulating seed viability. AKR1 transgenics when subjected forageing treatment, showed improved seed germination, longer seed viability, vigour with reduced level of cytotoxiccompounds compared to the wild type. This study therefore, provides evidence that, detoxification of reactive cytotoxiccompounds is important in improving the seed viability and AKR’s proved to be effective in regulating the cytotoxiccompounds.

Session C : Genomics, Epigenomics and Regulation Biology

199

IPPCongress2015


PC166 (IPPC0093)

Proteomic, metabolomic and ionomic analysis of a halophyte, Suaeda sp. to understand its molecular

adaptation

Wungrampha Silas1*, Sneh Singla-Pareek2, Jayant Tripathy3 and Ashwani Pareek1

1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India3School of Environmental Sciences, Jawaharlal Nehru University, New Delhi-110067, India*Presenting author: [email protected]

Halophytes, an extremophile plant group, represent about 2% of the plant kingdom. They can grow at salt concentrations

higher than 200 mM. The ability of halophytes to grow in saline soils has enabled them to flourish well in diverse

geographical areas. Research focused on elucidating mechanism of salinity tolerance in halophytes is recently picking

up its pace. Molecular analysis to understand the mechanism of adaptation and further improve their yields is underway.

Proteomics, metabolomics and ionomics of a succulent halophyte, Suaeda sp. grown at the salt extraction site of

Sambhar Lake, Rajasthan were analysed to understand the molecular adaptation for it to grow in that harsh environment.

The plant sample was harvested in the month of June, the driest season of the year, in the lake area when the salinity is

at its highest. And August, which is just after the monsoon and the site is covered with all types of vegetation. The

samples were harvested at various intervals during twenty four hour period to check the circadian molecular expressional

changes occurring in the plant.

PC167 (IPPC0119)

Exploring the roles of cysteine-rich secretory proteins in bacteroid differentiation and nitrogen fixation in

peanut nodules

Senjuti Sinharoy1*, Ivone Torres-Jerez2, Kurtis Cressman3, Josh Clevenger4, Peggy Ozark Atkins4, Brian Scheffler5

and Michael Udvardi2

1Department of Biotechnology, University of Calcutta, Kolkata-700037, India2The Samuel Roberts Noble Foundation, Plant Biology Division, Oklahoma, USA3Oklahoma State University, OK 74074, USA4University of Georgia, College of Agriculture and Environment Science, UGA5USDA-ARS JWDSRC, Stoneville, MS*Presenting author: [email protected]

Legumes develop specialized organs called nodules for NF symbiosis with rhizobia, which are released into nodule

cells via endocytosis, forming a unique organelle called the symbiosome. Inside the symbiosome, rhizobia differentiate

into NF bacteroids. Infected nodule cells each accommodate thousands of bacteroids. Plant cells control bacteroid

differentiation, which in some legume species involves nodule-specific cysteine-rich (NCR) peptides These peptides

promote elongated/Y-shaped symbiosome formation. Peanut (Arachis hypogaea) has a very high nitrogen fixation

capacity by the virtue of forming spherical terminally-differentiated bacteroids. We have performed RNA sequencing

analysis of peanut nodule developmental to understand the molecular mechanism behind peanut spherical symbiosome

formation. The peanut genome encodes around 130 cysteine-rich secretory proteins (CRSPs) that are longer (around

300-350 amino acids) than NCRs and possess a signal peptide and a pathogen-related (PR1) cysteine-rich domain,

unlike the NCRs. We are exploring the possibility that CRSPs play dual roles in the control of symbionts and pathogens

in peanut.


200

IPPCongress2015


PC168 (IPPC0141)

Genome-wide survey and expression analysis of genes encoding seed storage proteins in chickpea (Cicer

arietinum L.)

Subodh Verma*, Chandra Kant and Sabhyata Bhatia



Chickpea seeds are one of the major sources of plant protein for poor and vegetarian population especially in developingcountries. However, considering the importance of chickpea for human nutrition and agricultural commodities, themolecular mechanisms that program chickpea seed development are not well understood. In mature chickpea seeds,storage proteins are the major storage compounds and accumulate during the late phases in the process of seeddevelopment. To understand the regulatory machinery of seed storage proteins (SSPs) synthesis and accumulation, agenome wide analysis of genes encoding SSPs was carried out from three legumes including chikcpea, medicago andsoybean. Publicly available transcriptome data of different chickpea tissues was further utilized for in silico expressionprofiling of SSP encoding genes and genes encoding transcription factors (TFs). Moreover, co-expression analysis ofSSPs and TFs resulted in identification of transcription factors having expression profiles similar to SSPs. These TFsincluded members of B3, bHLH, bZIP and C3H families. This is the first study of genome-wide identification of SSPencoding genes and their expression pattern during seed development in chickpea. This comprehensive study willprovide a useful resource to explore the regulatory networks involved in SSP synthesis and accumulation in chickpeadeveloping seeds.

PC169 (IPPC0245)

Genome-wide comparative profiling of heat shock proteins and heat shock factor during multiple abiotic

stresses reveals superimposed and stress specific adaptational responses in Pennisetum

glaucum and Oryza sativa

Saurabh Pandey* and M.K. Reddy

Plant Molecular Biology Lab, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

Plants possess a flexible and elaborate nexus of heat shock protein (HSPs) and heat shock factors (HSFs) that arewidely involved in cellular response to multiple abiotic stresses besides heat. HSPs are accountable for protein folding,assembly, translocation, protein intracellular localization, secretion and degradation in a broad array of cellular processesin normal and stressed cells. HSPs are transcriptionally activated by HSFs that are transcription factors involved inresponse to different abiotic stresses. To illustrate the functional regulation of Pennisetum glaucum HSP and HSFgenes, we analyzed the changes in their global expression profiles in response to heat, salt, cold and drought stressesin seedlings. Comparative analyses revealed that while high resemblance existed in expression of most HSPs and HSFwith an overlapped response and regulation patterns during different stresses, some genes showed notably stress-specific response. Interpretation of microarray data analysis identified several functionally correlated genes comprisingof stress up-regulated known and new genes that may be implicated in heat and other stresses. Based on the findingson collateral analysis of stress associated variation in transcripts of selected HSPs, genome wide microarray profilingand co-expression analysis, present study is a step ahead in comprehending the mechanism underlying stress responsein Pennisetum glaucum. Moreover, heat-responsive gene profiling differed largely from those under drought, cold andsalt stresses and drought treatment was more effective in up-regulating HSF expression. Results advocate that theconserved expression of HSPs and HSFs in response to multiple stresses may highlight their role as important componentsfacilitating cross-talk between various stress-related signal transduction pathways.


201

IPPCongress2015


PC170 (IPPC0260)

Specificity of interactions between ARFs and ARF-GEFs during vesicle formation at different membranes

in the cell

Manoj Kumar Singh1*, Sandra Richter1, Hauke Beckmann1, Marika Kientz1, Nadine Anders2, York Stierhof1,

Ulrike Mayer1 and Gerd Juergens1

1Center for Plant Molecular Biology, Developmental Genetics, University of Tübingen, Germany2Department of Biochemistry, University of Cambridge, Cambridge CB2 1QW, United Kingdom*Presenting author: [email protected]

Transport of various biomolecules between different subcellular compartments through vesicle trafficking is essential for

optimal growth of plants and for their ability to adapt to different biotic and abiotic stresses factors. Formation of a vesicle

from a donor compartment is regulated by the ARF family of monomeric GTPases. At the site of vesicle budding, GDP-

bound ARF GTPases’ are recruited from the cytosol to the membrane and become activated by replacing the bound

GDP with a GTP. This activation process requires interaction of ARFs with ARF guanine nucleotide exchange factors

(ARF-GEFs). Once GTP-bound, ARFs recruit the coat machinery, which is necessary for formation and scission of the

cargo- containing vesicles. The hydrolysis of bound GTP to GDP inactivates the ARF and results in dissociation of coat

proteins from the vesicle, allowing its fusion with the acceptor compartment. Arabidopsis thaliana genome encodes nine

ARF GTPases and eight ARF-GEFs. Functional studies have revealed the specific roles of different ARF-GEFs in

secretion, recycling of the auxin efflux carrier protein PIN1, trafficking to the cell plate during cytokinesis, and vacuolar

transport. Although ARF-GEFs have been studied extensively, relatively little information is available about the role of

different ARF GTPases and, more importantly, the specificity of ARF-GEF interactions with ARFs at different subcellular

compartments. Our findings from biochemical, genetic and microscopic analysis of transgenic Arabidopsis plants will be

presented and discussed.

PC171 (IPPC0284)

Study on role of Arabidopsis Methyl CpG binding domain protein 4 (AtMBD4) in phosphate starvation

Adwaita Parida, Amrapali Sharma* and Arun Sharma

Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India*Presenting author: [email protected]

Methyl CpG Binding Domain proteins are a special class of protein that recognize and bind to methylated DNA. In

Arabidopsis, mutation in the Methyl CpG Binding Domain protein 4 gene (AtMBD4) shows more lateral root development

as compared to wild type plants. Transcriptome analysis of atmbd4 mutant using microarray reveals that this phenotype

is due to up-regulation of many genes involved in phosphate starvation response. Plants grown in phosphate deficient

condition also show a similar phenotype as that of atmbd4 mutant. Yeast two hybrid screening using AtMBD4 as bait;

found a Ubiquitin conjugating enzyme 36(AtUBC36) that interacts with AtMBD4. AtMBD4:CFP and AtUBC36:YFP fusion

proteins localized in the nucleus. Further in vivo interaction between these two proteins was analyzed in plant cell using

Florescence Resonance Energy transfer (FRET). Both the proteins were co-localized in the nucleus and showed a very

good FRET efficiency that confirmed their association. The mutant atubc36 also showed the similar root phenotype with

more lateral roots like atmbd4. Both the genes were up-regulated during phosphate starvation. Moreover, both atmbd4

and atubc36 mutants were found to accumulate more inorganic phosphate and anthocyanin as compared to wild-type

plants. Our results clearly suggest that these two proteins interact with each other and perform an important role during

phosphate starvation response.


202

IPPCongress2015


PC172 (IPPC0301)

The evolution of pitchers in Nepenthes khasiana: how many and which genes does it take to make a

pitcher?

Jeremy Dkhar*, Yogendra Bhaskar, Andrew Lynn and Ashwani Pareek

School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India*Presenting author: [email protected]

A structural phenomenon seen in certain lineages of angiosperms that has captivated many scholars including CharlesDarwin is the evolution of plant carnivory. We now understand why carnivory evolved but how carnivorous plantsacquired these attributes remain a mystery. In an attempt to understand the evolution of pitchers, a carnivorous traitfound in three flowering plant families, we de novo assembled for the first time the transcriptome sequence of Nepentheskhasiana, an insectivorous plant endemic to the state of Meghalaya, North-east India. We generated more than 180million reads using Illumina sequencing from the highly specialized N. khasiana leaf comprising the leaf base, tendriland the different parts/zones of the pitcher tube viz. digestive zone, waxy zone and lid. We then mapped respective readsinto the de novo assembled reference transcriptome to quantify transcript abundance along the N. khasiana leaf. Wedetected highest number of expressed transcript in the tendril followed by waxy zone, digestive zone, lid and leaf base.Hierarchical clustering of differentially expressed genes indicated functional relationship and similar cellular processesunderlying the leaf base and pitcher tube, thereby implying that the Nepenthes pitcher is indeed a modified leaf. Takingcues from SEM and LM photomicrographs of the five distinct parts/zones of the N. khasiana leaf, we examine theexpression pattern of genes involved in stomatal and trichome development as well as leaf polarity specification genes.Our findings imply that alterations in the expression of key regulatory genes involved in leaf development probably ledto the evolution of Nepenthes pitchers.

PC173 (IPPC0310)

Integrating RNA transcriptome wide and microRNA analyses for the identification of molecular regulators

associated with high salt tolerance in Sesuvium portulacastrum (L.)

Ganesh Nikalje1*, Ashish Kumar Srivastava2, Gaurav Sablok3, Tukaram Nikam1 and Suprasanna Penna2

1Department of Botany, Savitribai Phule Pune University, Pune-413007, India2Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, India3Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, Sydney*Presenting author: [email protected]

Sesuvium portulacastrum, a member of Aizoaceae family, is endowed with its ability to complete its life cycle under highsalinity which is associated with regulation of cellular energetics and redox homeostasis. The present study was conductedto identify transcriptional mediators related to salt adaptation. On the basis of post-germination phenotyping, 100 and250 mM NaCl concentrations were selected at which, growth was either comparable to control or reduced by 50%,respectively. The growth reduction at 250 mM was concomitant with higher Na+ accumulation at 10 d; although, at 24 hafter stress, Na+ accumulation was equally high at both the salt concentrations. The activation of enzymatic cascade wasseen only at 250 mM. At 24 h treatment, total RNA was isolated from root and RNAseq was performed at Illumina HiSeqplatform. A total of 189 million reads were assembled into high quality 104843 unigenes and their functional annotationrevealed key salt responsive genes. To link this with micro regulators of RNA mediation regulation, a set of conservedmiRNAs (miR156, miR159, miR162, miR164, miR167, miR169, miR172, miR390, miR395, miR399 and miR838) andtheir expression was measured using stem-loop RT-PCR. A total of 33 targets were identified using assembledtranscriptome and their expression data confirmed that transcriptional changes were more pronounced at 250 mM NaCl.This suggested that observed effects are more closely related with osmotic components. Thus, present study highlightsthe concentration dependent effects on growth and identifies key genetic elements associated with salt adaptation.


203

IPPCongress2015


PC174 (IPPC0316)

Phylogenomic analysis of AP2/ERF family genes in sorghum

Sushree Sangita1, Supriya Mathur1*, Aleena Francis2, Rita Sharma2 and Manoj Sharma1

1School of Biotechnology, 2School of Computational & Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India*Presenting author: [email protected]

Sorghum bicolor is the fifth most important cereal crop in the world. It not only provides food, feed and fiber but is also

emerging as a leading biofuel crop. It is a low input crop with high water-use efficiency and potential to recover growth

after drought stress. However, changing environmental conditions pose a serious threat to plant growth and productivity.

Therefore, it is imperative to understand the mechanism of stress adaptation and identify potential candidates for

breeding improved varieties in Sorghum. The members of AP2/ERF family transcription factors have particularly emerged

as important candidates for engineering stress tolerance in crop plants. They are characterized by AP2/ERF DNA

binding domain and have been further classified into three subfamilies including AP2, ERF and RAV. To elucidate their

role in plant development and stress response in sorghum, we performed genome-wide identification and phylogenomic

analysis of AP2/ERF family genes in Sorghum. Using HMM and NCBI-CDD analysis, we have identified more than 175

ERF family genes in sorghum. We present here the results of phylogenetic analysis, structural characterization and

expression analysis of candidate AP2/ ERF genes. Expression analysis of the genes was performed at using publicly

available transcriptomic data developmental stages and in response to abiotic stress treatments.

PC175 (IPPC0336)

microRNAs in chickpea (Cicer arietinum L.) root nodulation

Chandra Kant* and Sabhyata Bhatia

National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India*Presenting author: [email protected]

Chickpea (Cicer arietinum L.) is widely grown across the Indian subcontinent. In symbiotic association with Mesorhizobium

ciceri root nodules are formed that play important role in the farming system by fixing atmospheric nitrogen. miRNAs

have emerged as an important regulatory component in a number of developmental processes including legume-

rhizobia symbiosis. In order to understand the role of miRNAs in root nodulation a smallRNA library from root nodules of

chickpea was sequenced using the Illumina GA II X platform. Conserved miRNAs were identified based on their similarity

to miRNA of other plant species present in miRBase, and putative novel miRNAs were identified based on potential

hairpin loop forming precursors from available plant databases. Presence of miRNAs were validated by using northern

blotting and stem loop RT-PCR. In-silico expression profile of miRNAs in nodule in comparison to other tissues and

target identification revealed a set of miRNAs that may have imperative role in nodule development and function, which

were further validated by Quantitative Real Time PCR. The conserved and putative novel miRNA identified in this study

will help reveal intricacies of chickpea-Mesorhizobium symbiosis.


204

IPPCongress2015


PC176 (IPPC0357)

Expression and protein: protein interaction analysis of FCS-LIKE ZINC FINGER 17 and 18 of Arabidopsis

thaliana

Muhammed Jamsheer K, Manvi Sharma*, Chanchal Thomas Mannully and Ashverya Laxmi



The FCS-LIKE ZINC FINGER (FLZ) family is a plant-specific family of zinc finger proteins characterized by the C2-C2

FCS-LIKE ZINC FINGER (FLZ) domain. The FLZ domain acts as a protein-protein interaction module. The FLZ domain

containing proteins are identified from all sequenced spermatophytes out of which the Arabidopsis thaliana genome

possesses 18 FLZ domain genes. Out of theses 18 genes, FLZ17 and FLZ18 are tandem duplicates without any

sequence divergence. Both these proteins are highly diverged from other members of this gene family. FLZ17/18 is

localized in both nuclei as well as cytoplasm and interacts with many other members of the same class of zinc finger.

Moreover, the Y2H analysis identified that they interact with many regulators of processes like flower development,

photosynthesis, photomorphogenesis etc suggesting the possible function of these genes in these processes. Consistent

with this observation, FLZ17/18 found to be profusely expressed in flower and silique stages and during the vegetative-

to-reproductive phase transition. Similarly, the expression of these genes was found to be differentially regulated by

various biotic as well as abiotic stresses. The genes were found to be differentially regulated by hormones involved in

biotic and abiotic stresses. Taken together, the present study gives a comprehensive account of the transcriptional

regulation and protein: protein interaction properties of FLZ17/18 which will be helpful in the molecular and functional

characterization of these genes.

PC177 (IPPC0412)

Variability for root traits in sunflower hybrids and inbreds

Lakshmi Prayaga*, P. Lakshmamma and C. Sarada

Department of Plant Physiology, Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad-500030, India


Fifty four inbreds and 8 hybrids of sunflower were studied for variability in root traits by growing them in polybags (90 cm

length and 40 cm diameter) of 120kg soil capacity @ 2 plants per bag for 65 days during rabi. FYM was mixed with soil

in the ratio of 1: 5 (v/v). Plants were irrigated as and when required. Observations were recorded at the time of harvest.

Wide genetic variability was observed for all the parameters studied. Hybrids recorded higher root weight, volume,

length, total dry matter (TDM), leaf area, leaf number compared to inbreds. Root weight showed strong positive relationship

with TDM (0.77) and leaf area (0.62) compared to root volume (0.74 & 0.61) and root length (0.62 & 0.55). Among the root

traits, root weight and volume showed good correlation (+0.89) compared to their relation with root length. Root traits did

not show any relation with SCMR and SLA (specific leaf area). A selection index was computed using principal component

analysis by taking into consideration root characters along with TDM and leaf area. Based on the index value, hybrids

DRSH-1, NSFH-1001 and inbreds 106A, 335 A, AKSFI-42-I, DS-2, 298-R, CSFI-5075, 856-R, R-7, CMS-335A were

found promising.


205

IPPCongress2015


PC178 (IPPC0417)

Occurance and arrangement of three mutisubunit inorganic carbon transporters in cyanobacteria

Vandana Tomar*, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra and Sandhya Mehrotra

Biological Sciences, Birla Institute of Technology and Science, Qtr.28/1 Pashchim Marg BITS, Pilani-333031, India


The oxygenase reaction catalyzed by Rubisco became an issue only after the evolution of the oxygenic photosynthesisin cyanobacteria. Several strategies were developed by autotrophic organisms to increase oxygen levels in the viscinityof Rubisco to maximize its net carboxylation rate. One of the crucial advancements in this context was the developmentof more efficient inorganic carbon transporters which could help in increasing the influx of inorganic carbon (Ci) at thesite of CO

2 fixation. A survey was done to find out the genes encoding for cyanobacterial Ci transporters in twenty

cyanobacterial phyla with respect to transporters present in Gloeobacter violaceous PCC 7421, an early divergingcyanobacterium. An attempt was also made to correlate the prevalence of the kind of transporter present in the specieswith its habitat. Basically two types of cyanobacterial inorganic carbon transporters exist i.e., bicarbonate transportersand CO

2 uptake systems. The transporters also show variation in context to their structure as some exist as single gene

encoded proteins (BicA and SbtA), while others exist as multisubunit proteins (BCT1, NdhI3 and NdhI4). The phylogenyand distribution of the former have been extensively studied and the present analysis provides an insight into the latterones. The in silico analysis of the genes under study revealed that their distribution was greatly influenced by the habitatand major environmental changes in the course of their evolution viz., great oxidation event. Knowledge of the evolutionof these transporters could be a step towards selection of suitable candidates for plant transformation.

PC179 (IPPC0458)

Transcriptome modulation in Arabidopsis natural variations under arsenic stress

Tapsi Shukla1*, Smita Kumar2, Deepika Sharma1, Deepika Lakhwani1, Ria Khare1, Mehar Asif1 and Prabodh Trivedi1

1Molecular Biology, CSIR- National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India2University of Lucknow, Lucknow, India


Heavy metal contamination primarily due to the industrial processes, agricultural practices, mining and other anthropogenicactivities has resulted in severe threat to environment and human health. Among other heavy metals, arsenic (As) is mosttoxic to biota even at low concentration and is considered to be a potent carcinogen. In view of As toxicity there is anincreasing concern about the molecular mechanisms underlying the uptake, transport and accumulation of As in plants.In recent years, considerable attention has focused on the use of Arabidopsis natural variations to understand adaptationand response to environmental changes. Here, we analysed the response of Arabidopsis accessions towards As stress.Differential response in terms of phenotypic variation (root length) was observed in the accessions exposed to differentconcentrations of As. On the basis of reduction in the primary root length, nine accessions were categorized as tolerant,moderate and sensitive for As(III) stress. The accessions Col-0, Koch-1, Sij-2 were tolerant (< 55% reduction in primaryroot length) whereas Borsk-2, Altneb-2, Sij-1 were moderate (> 55-60% reduction) and Ciste-2, Slavi-1, Ws weresensitive (> 60% reduction) towards As(III) stress. To study the contrasting response of different accessions, genome-wide expression analysis using most tolerant and sensitive accessions was carried out. A group of defence and stress-responsive genes, transporters, and regulatory genes showed differential expression in the contrasting accessions. Themodulation in the expression pattern of the genes that showed the highest contrast in the microarray was furthervalidated by qRT-PCR. Thus, the study provides an insight about the genes and the molecular mechanisms involved inarsenic stress response/ adaptation.


206

IPPCongress2015


PC180 (IPPC0470)

HY5-dependent differential expression of genes involved in flavonol biosynthesis under low temperature

and light

Chitra Bhatia1*, Ashutosh Pandey2 and Prabodh Trivedi

1Plant Genomics Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India2National Agri-Food Biotechnology Institute, Punjab, India


Light and temperature are two most important factors affecting plant growth, development by various biological processes.Flavonoids are a class of secondary metabolites produced by the plants in response to stress conditions throughphenylpropanoid pathway. Studies suggest that biosynthesis of flavonoids is regulated by light through various regulatoryfactors as well as proteosomal machinery. LONG HYPOCOTYL 5 (HY5), a bZIP transcription factor, binds directly to thepromoters of light-inducible genes and plays important regulatory role in flavonoid biosynthesis. During dark conditions,HY5 is degraded in the plant via proteasome-mediated degradation involving CONSTITUTIVEPHOTOMORPHOGENESIS 1 (COP1), RING E3 ubiquitin ligase. Previous reports suggest that during low temperatureand light conditions, enhanced biosynthesis of anthocyanins is regulated through HY5-dependant manner. However,very little information is available on effect of low temperature and light on regulation of flavonol biosynthesis whichshares the same pathway as anthocyanins. In this study, we analyzed expression of regulatory (MYB11, MYB111 andMYB12) and structural genes (CHS, CHI, F3H and FLS) involved in flavonol biosynthesis under different light qualitiesin wild type, hy5-215 and cop1-4 mutants of Arabidopsis thaliana. Similarly, expression analysis of these genes in lowtemperature conditions under white light was also performed. Our results suggest induction of regulatory and structuralgenes involved in flavonol biosynthetic pathway in response to light as well as low temperature conditions in HY5-dependant manner.

PC181 (IPPC0475)

Comparative analysis of methylome under abiotic stress in foxtail millet cultivars [Setaria italica (L.) P.

Beauv.]

Garima Pandey*, Chandrabhan Yadav and Manoj Prasad

Plant Molecular Biology, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Foxtail millet is a C4 Panicoid crop with a relatively small genome, short life-cycle and is closely related to several biofuel

crops. Apart from this, it is a potential abiotic stress tolerant crop which categorizes it as an excellent model system tostudy the stress induced gene regulation mechanisms. DNA methylation is one of the important mechanisms related togene regulation at epigenetic level. As methylation based stress regulatory mechanism is poorly understood in foxtailmillet, which is naturally adapted to abiotic stress, we aimed to study DNA methylation level in two contrasting cultivarsof foxtail millet (i.e., tolerant cv. IC403579 and sensitive cv. IC480117) under salt stress condition using MethylationSensitive Amplification Polymorphism (MSAP) technique. Reduced level of DNA methylation was observed in tolerantcultivar as compared to the sensitive cultivar. Total 69 genes showed differential methylation behaviour in both thecultivars. Majority of which corresponds to transposase related genes which are responsible for activation and transpositionof transposons in the genome. Some of the transposase related genes were subjected to bisulfite sequencing for furthervalidation. Along with this expression analysis of demethylases showed significant upregulation in tolerant cultivarunder salinity stress. These results indicate that on exposure to salinity stress, demethylation of genome by demethylasesmight lead to activation of stress related genes which may be one of the probable reasons for stress tolerance behaviourin foxtail millet cultivar. Further, functional characterization of stress-induced differentially methylated genes will provideprofound understanding of abiotic stress tolerance mechanism in millets.


207

IPPCongress2015


PC182 (IPPC0507)

MicroRNAs: Small nodes of stress management in root apical meristem in chickpea

Hitaishi Khandal* and Debasis Chattopadhyay

Plant Molecular Biology, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Root growth is regulated by cell division at the root apical meristem. Structure of root apical meristem of Fabaceae plants

is notably different from that of Arabidopsis. Histological studies have shown that unlike a closed structure of RAM in

Arabidopsis, RAM of legume plants exhibits a basic-open structure. MicroRNAs have emerged as regulators of many

key biological functions in plants including stress management. To identify key miRNAs involved in abiotic stress

management in root meristem of chickpea, we used deep sequencing to compare miRNA populations among five small

RNA libraries prepared from control and abiotic stress treated chickpea root meristem (~2 mm). A total of 8,03,259 high

quality reads were generated. 458 unique sequences were mapped 100% to the chickpea genome. Statistical analysis

on transcript abundance of predicted annotated and putative miRNAs revealed that almost 90% of them were differentially

regulated by PEG induced dehydration and salinity treatments at early and late stages. We identified a total of 284

putative miRNA sequences out of which 224 were already annotated in miRBASE dataset which belongs to 195 unique

families, while rest of 60 sequences were categorized as predicted putative miRNAs. The chickpea miRNAs targeted a

wide range of mRNAs involved in diverse cellular processes, including transcriptional regulation (transcription factors),

protein modification, signal transduction, and metabolism. This study reveals a comprehensive set of conserved and

novel miRNAs along with their expression patterns and putative targets in chickpea, and provides a framework for

understanding regulation of developmental processes and stress tolerance mechanisms in legumes.

PC183 (IPPC0517)

Proteomic analysis of chickpea (Cicer arietinum L.) reveals dynamic plasma membrane architecture

Praga Barua*, Nilesh Lande, Subhra Chakraborty and Niranjan Chakraborty


Plant cells contain numerous membrane systems that are specialized to perform particular functions. The plasma

membrane (PM), in particular, is an organized system that serves as physical barrier and as a communication interface

with the extracellular environment. Most importantly, the processing of cell signaling reactions that are involved in

responses to biotic and abiotic stressors occurs in the plasma membrane. The multivariate role of plasma membrane

demands its proteome to be highly dynamic, comprising a large number of proteins with highly varied function. Therefore,

characterization of the plant plasma membrane proteome can provide new insights into its multivariate functions. In an

attempt to understand the complexity of plant plasma membrane proteins (PMPs), we developed a proteome reference

map of a food legume, chickpea. Plasma membranes were isolated from 3-week-old seedlings and purity of the fraction

was confirmed by immunoblot screening. The proteome was constructed using plasma membrane enriched fraction

using 1-DE technique. Analysis of 1-D bands as well as in-solution tryptic digests of PMP led to the identification of more

than 2000 proteins including well known intrinsic PMPs such as aquaporins, H+ATPase etc, as well as many predicted

and uncharacterized proteins. This comprehensive overview of the proteome would provide a basis for future comparative

proteomics efforts for other crop species. Our future efforts would focus on identifying differentially regulated proteins

under stress conditions, which may help in the long-term efforts to develop transgenic crops with improved stress

tolerance.


208

IPPCongress2015


PC184 (IPPC0549)

Understanding the epigenetic regulation of molecular memory of infection in Arabidopsis

Zeeshan Zahoor Banday*, Sadan Sharma and Ashis K. Nandi

School of Life Sciences, Jawaharlal Nehru University, New Delhi-110 067, India


Immobility poses a challenge to plants. Plants live in complex milieu where they interact with a wide range of organisms.

These interactions can be beneficial or detrimental. Plants respond to pathogen attack by activating local as well as

systemic defenses that limit the growth and spread of pathogen. The reduced in systemic immunity 1 (rsi1) identified by

the genetic screen of EMS mutagenized wild-type Arabidopsis, fails to activate systemic acquired resistance (SAR).

RSI1 is allelic to Flowering locus D (FLD). The mechanism of FLD mediated SAR activation, however, is not known. FLD

is known to function as a part of transcriptional co-repressor complex. The protein interacting partners of FLD in the co-

repressor complex remain largely unidentified. To elucidate the possible mechanism of FLD mediated SAR activation,

we screened Arabidopsis thaliana cDNA library for FLD interacting proteins (FINs) in yeast. We identified several

putative interacting factors of FLD. Few of these interactors play a relevant role in defense and could be important

candidates for FLD to form a functional complex with, underlining its role in epigenetic modification of SAR. Here we

report the studies on one such FIN, FIN1.We show that FIN1 physically interacts with FLD. We also demonstrate the

nuclear co-localization and in vivo interaction of FIN1 with FLD. The FIN1 T-DNA insertion mutants were subsequently

shown to be compromised in SAR. Understanding the mechanism of SAR regulation will have far-reaching benefits both

for the improvement of crop yields and human health.

PC185 (IPPC0555)

Identification of RSI1 target for systemic acquired resistance in Arabidopsis

Ashis Nandi, Deepjyoti Singh* and Vijayata Singh

School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India*Presenting author: [email protected]

Plants are under attack of large number of pathogens. However due to the presence of well elaborate immune system

they remains mostly healthy. One of such immunity mechanism is Systemic Acquired Resistance (SAR); by which a

partly infected by pathogen becomes highly resistance for secondary infection. Upon infection, infected tissue generates

mobile signals that get transported through vasculature to rest of the plant. These signals are believed to cause epigenetic

modifications to generate the state of alertness in plants. Due to this alertness the experienced plants show the SAR

associated priming, i.e, higher level of defense gene expression than a naive plant. Even though a large number of

mobile signals are known the molecular mechanism by which they turn into epigenetic modifier is not known. Our lab

identified a mutant rsi1 (reduced systemic immunity1), through forward genetic screening of EMS treated wild type seeds

of Arabidopsis (Col-0). This mutant demonstrated normal local resistance but not SAR. RSI1 is allelic to FLD. We

compared the transcriptome profile of WT and rsi1 plants to identify the targets of rsi1 (TORs). By knowing RSI1/FLD is

a negative regulator of transcription, we detected the genes that are specifically upregulated in SAR induced rsi1 plants.

One of such genes TOR1 (Target of RSI1) shows predicted pattern of FLD/RSI1 mediated histone methylation in its

promotor. The mutant phenotype of tor1.


209

IPPCongress2015


PC186 (IPPC0563)

Genome-wide identification and characterization of dehydration-responsive miRNA(s) and targets in Setaria

italica

Amita Yadav* and Manoj Prasad

Plant Molecular Biology, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

miRNA-mediated gene regulatory networks under dehydration stress remain largely unexplored in plant includingfoxtail millet (Setaria italica), which is a natural abiotic stress tolerant crop. To find out the dehydration-responsivemiRNAs at the global level, four small RNA libraries were constructed from control and dehydration (6h) stress-treatedseedlings of two foxtail millet cultivars showing contrasting tolerance behavior towards dehydration stress. Using illuminasequencing technology, 55 known and 136 novel miRNAs were identified. Moreover, 18 known and 33 novel miRNAswere differentially expressed during dehydration stress. After stress treatment, 32 dehydration responsive miRNAs wereup-regulated in tolerant cultivar IC403579 and 22 miRNAs were down-regulated in sensitive cultivar IC480117, suchantagonistic behavior of foxtail millet miRNAs indicated that miRNA-mediated molecular regulation might play importantroles in providing contrasting characteristics to these cultivars. Moreover, northern blot analysis and SL-qRT PCR havebeen performed to validate the expression patterns of some known and novel dehydration-responsive miRNAs. Differentialexpression behavior of few miRNA-target genes were also verified under dehydration stress treatment and miRNA-directed cleavage site was validated by RLM-RACE. One of the candidate miRNA, which is monocot-specific, showedup-regulation in sensitive cultivar and simultaneously, expression of its target was found to be down-regulated afterdehydration treatment. To understand its role, particularly in dehydration stress, rice transgenic lines over-expressing itsfoxtail millet precursor were generated. Expression level and copy number of transgenics were analyzed and evaluationof T2 lines is in progress. This study would help in elucidating one of the dehydration-stress associated post-transcriptionalregulation mechanism.

PC187 (IPPC0570)

Genome wide analysis of ACGT/CRE in Arabidopsis thaliana reveals frequency of ACGT_CRE is higher

than CRE_CRE

Rajesh Mehrotra, Chetna Sangwan*, Sandhya Mehrotra, Vandit Gandotra and Zaiba Hasan Khan

Biological Sciences, Birla Institute of Technology and Science (BITS) Pilani, Jhunjhunu-333031, India*Presenting author: [email protected]

It is inevitable to prevent the outcomes of the abiotic stress in the plants. Drought and salt stress, together with lowtemperature, are the major problems for agriculture because these abiotic environmental factors prevent plants fromrealizing their full genetic potential and affect the plant growth and productivity adversely. To combat these stressesplants have evolved mechanisms where multitude of gene expression profile changes are involved. Synergy betweenthe cis regulatory elements and their cognate transcription factors leads to the exactitude and melange of gene expression.In plant system, 469 cis regulatory elements have been documented as per PLACE database. ACGT cis regulatoryelement forms the core sequence of other cis regulatory elements viz salicylic acid responsive element, abscisic acidresponsive element (ABRE), core sequence of G box, light response element. Earlier study reveals that occurrence ofACGT element is one of the pivotal features of the stress responsive promoter, provided either an additional copy of thesame motif or any other cis element (CREs, DREs, LREs etc.) is present. In our genome and promoter wide analysis ofCREs (Cold responsive elements) focusing on the “CCGAC” and its variants in Arabidopsis thaliana, we found that thefrequency of ACGT_CCGAC & CCGAC_ACGT spacers is higher than two CCGAC spacer_CCGAC in tandem. Wealso found that (ACGT_CCGAC) motifs with spacer lengths of 2, 9, 15, 20, 21, 26, 28 & 29 nucleotides occur at higherfrequency in the promoters regions than other spacer lengths.The data obtained in this study will be presented.


210

IPPCongress2015


PC188 (IPPC0583)

Trancriptional profiling during dormancy release in Apple suggests the role of epigenetic mechanism in

regulating the perception of differential chilling

Gulshan Kumar*, Gagandeep Singh, Mohit Swarnkar, Ram Sharma and Anil Singh

Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, H.P., India*Presenting author: [email protected]

Apple is a temperate fruit crop and low temperature plays a crucial role in floral bud dormancy and release. Therefore,comparative transcriptome profiling of dormant bud, silver tip (mid active) and green tip (active bud) of field grown applegenotype “Royal delicious” under differential chilling availability was performed using RNA-seq approach. The de novoassembly of 113,666,401 paired-end reads resulted in 47,712 contigs. The 32,408 (68%) contigs were successfullyannotated by BLASTP search against non-redundant NCBI database. Higher chilling availability during dormancyperiod was found to be associated with more number of differentially expressed genes in transition from silver tip togreen tip stage. The expression of two DAM (Dormancy Associated MADS) genes was found to be decreased withconcomitant chilling acquisition, however the basic transcript abundance of these DAM genes remained higher underlow chill conditions during the active growth period. Similarly, the expression of FLC-like gene was found to be decreasedwith chilling acquisition under high chill condition, while its expression was induced under low chill condition. In addition,the expression of predicted DRMs and CMTs involved in de novo DNA methylation, and SAM synthase (SAM, a cofactorfor methyltransferases) was found to be higher under low chill conditions. Moreover, the methylation sensitive amplifiedpolymorphism also showed higher methylation events under low chill conditions. The regulation of FLC and DAM geneshas been reported to be regulated through methylation of their promoter regions. Thereby, the role of epigeneticregulation in chilling mediated dormancy release in apple is apparent.

PC189 (IPPC0624)

Role of histone acetyltransferases and deacetylases in epigenetic regulation of cotton fiber development

Sunil Kumar Singh, Babita*, Archana Bhardwaj, Verandra Kumar, Krishan Mohan Rai, Surendra Pratap Singh and

Samir Vishwanath Sawant

Plant Molecular Biology and Genetic Engineering, CSIR-National Botanical Research Institute, Lucknow- 226001, India*Presenting author: [email protected]

The cell differentiation and development largely depends upon the signals generated by dynamic control of differentgene and regulatory networks acting within the cell. Histone modifications play key role in regulating the gene networksand largely contribute to the transitional alterations in expression pattern of genes. The present study aims to decipherthe role of histone acetyltransferases (HATs) and deacetylases (HDACs) in epigenetic regulation of cotton fiberdevelopment. In this study, we used Anacardic acid (inhibitor of p300/PCAF class of HATs) and Trichostatin A (inhibitorof almost all HDACs, except HDA08). We performed ovule culture assays and checked the effects of inhibitors againstvarious histone modifiers at phenotypic, biochemical and transcriptome level. We observed that Anacardic acid promotes,while Trichostatin A inhibits the fiber development in cultured cotton ovules. We identified genome wide targets of thesemodifiers by transcriptome sequencing. Treatment of these inhibitors induces the change in expression of various genesinvolved in auxin biosynthesis and response pathway. Next, we performed ChIP-seq assay to study the histone acetylation(H3K9Ac) landscape at the target genes and expressional changes due to alterations in the histone acetylation patternby inhibitors of HATs and HDACs. Anacardic acid spraying to the developing flowers and bolls in cotton plants improvesfiber yield and quality. The results suggest that histone modifiers play a crucial role in regulation of fiber cell differentiationand development.


211

IPPCongress2015


PC190 (IPPC0634)

Understanding transcriptional and epigenomic regulation of abiotic stress responses in rice

Rama Shankar*, Annapurna Bhattacharjee, Narayana Chevala, Rohini Garg and Mukesh Jain

National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, IndiaPresenting author: [email protected]

Drought and salinity are the major environmental factors affecting the rice productivity. Various rice cultivars have been

identified with evolved mechanism of abiotic stress tolerance. The comparative transcriptome analysis between stress-

tolerant and -sensitive rice cultivars can give us clue about the source of genes and/or genetic variations to improve the

stress tolerance of high yielding rice cultivars. We performed comparative transcriptome analysis between stress-

sensitive (IR64) and -tolerant (N22 and Pokkali) rice cultivars under desiccation and salinity stresses. A large number of

novel transcripts and alternatively spliced transcript isoforms were identified in the rice cultivars. Differential gene

expression analysis resulted in identification of various cultivar-specific transcripts. Functional categorization of these

transcripts revealed enrichment of several biological processes in N22 and Pokkali rice cultivars under stress conditions.

In addition, discovery of SNPs/InDels and polymorphic SSRs were also carried out in IR64/N22, IR64/Pokkali and N22/

Pokkali. Annotation of SNPs/InDels resulted in identification of various genes harboring large-effect SNPs/InDels. These

genes were found to be enriched in defense response and response to abiotic stress terms. In addition, quantitative trait

loci (QTLs) harboring differentially expressed genes and large-effect SNPs/InDels have also been identified. Further,

DNA methylation as a result of SNPs was discovered to find out the allele-specific methylation in the rice cultivars.

Altogether, these analyses provide a comprehensive overview about the transcriptional and epigenetic regulation of

stress responses, and serve as a resource to find out better genes/pathways important for abiotic stress tolerance.

PC191 (IPPC0636)

Understanding the importance of KIX domain proteins

Jitendra Thakur*

National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India*Present author: [email protected]

KIX domain is found in p300/CBP and Mediator subunit Med15 and is important for protein-protein interaction. Sequence,

mutational and structural studies from our and other laboratories suggest that CBP KIX has evolved from Med15 KIX

domain. In fungi and metazoans it has been found that the activation domains of diverse transcription activators target

KIX domain to regulate transcription of their target genes. However, not much is known about KIX domain proteins in

plants. We attempted to characterize all the KIX domain proteins in Arabidopsis and rice. Interestingly, in addition to

p300/CBP- and Med15-like proteins, KIX domain was also found in F-box proteins in rice and DNA helicase in Arabidopsis

suggesting new roles of KIX domain in ubiquitin-mediated proteasomal degradation of protein and genome stability. In

both rice and Arabidopsis, we have found more than fifty proteins interacting with the KIX domain of MED15 suggesting

its involvement in diverse processes. In rice, expression analysis revealed overlapping expression of OsKIX_3, OsKIX_5

and OsKIX_7 in seeds of different stages of development, suggesting their individual or combined role during seed

development. Moreover, the association analysis using data of 136 SNP loci in 23 contrasting rice genotypes and their

grain length-specific phenotypic information identified three non-synonymous SNP loci in these three rice genes showing

strong association with long- and short-grain differentiation. Interestingly, these SNPs are located within KIX domain

encoding genomic regions. It, thus, indicates the additional importance of novel SNP loci/alleles identified in KIX domain

containing genes for determining seed size in rice.


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PC192 (IPPC0637)

Identification of novel and conserved miRNAs from extreme halophyte, Oryza coarctata, a wild relative

of rice

Tapan Kumar Mondal* and Showkat Ahmad Ganie

National Bureau of Plant Genetic Resources, New Delhi-110012, India*Presenting author: [email protected]

Oryza coarctata, a halophyte and wild relative of rice, is grown normally in saline water. MicroRNAs (miRNAs) are non-

coding RNAs that play pivotal roles in every domain of life including stress response. There are very few reports on the

discovery of salt-responsive miRNAs from halophytes. In this study, two small RNA libraries, one each from the control

and salt-treated (450 mM NaCl for 24 h) leaves of O. coarctata were sequenced, which yielded 338 known and 95 novel

miRNAs. Additionally, we used publicly available transcriptomics data of O. coarctata which led to the discovery of

additional 48 conserved miRNAs along with their pre-miRNA sequences through in silico analysis. In total, 36 known

and 7 novel miRNAs were up-regulated whereas 12 known and 7 novel miRNAs were down-regulated under salinity

stress. Further, 233 and 154 target genes were predicted for 48 known and 14 novel differentially regulated miRNAs

respectively, which with the help of gene ontology analysis, showed several important biological processes that could be

involved in salinity tolerance. Relative expression trends of majority of the miRNAs as detected by real time-PCR as well

as predicted by Illumina sequencing were found to be coherent. Additionally, expression of most of the target genes was

negatively correlated with their corresponding miRNAs. Thus, the present study provides an account of miRNA-target

networking that is involved in salinity adaption of O. coarctata plant.

PC193 (IPPC0650)

Genome-wide transcriptome orchestra and involvement of Aux/IAA gene family in chickpea flower

development

Vikash Singh*, Rohini Garg and Mukesh Jain

Functional Genomics, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India*Presenting author: [email protected]

Flower development is one of the key developmental processes that controls seed setting in angiosperms. However,

little is known about the molecular mechanisms regulating flower development in legumes. Employing RNA-seq for 11

tissues/organs/developmental, we identified a large number of differentially expressed genes during flower development

in chickpea. Further, we identified several genes expressed exclusively at a particular flower development stage.

Various transcription factor families and metabolic pathways involved in flower development were also deciphered. The

members of MADS-box family were highly enriched among the transcription factor encoding genes up-regulated during

various stages of flower development. These data provide a resource for exploring the complex molecular mechanisms

underlying flower development and identification of gene targets for functional genomics in legumes. Further, a genome-

wide analysis of Aux/IAA gene family members in legumes identified several members, which expressed specifically at

various stages of flower development. Notably, CaIAA2, CaIAA9 and CaIAA13 revealed distinctively higher transcript

accumulation during stages of flower development, suggesting their possible role in flower development. Overall, these

results would help in investigation of precise functions of Aux/IAA genes in legumes during flower development in

chickpea.


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PC194 (IPPC0662)

Preparation and characterization of resource materials for the isolation of Mediator complex from rice

Subhasis Samanta* and Jitendra K. Thakur

National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India*Presenting author: [email protected]

Mediator complex acts as a molecular bridge transmitting the regulatory signal from the promoter-bound transcription

factor to the RNA polymerase II in the process of transcription. Genetic and molecular analyses have unravelled important

regulatory roles of Mediator subunits at every stage of plant life cycle starting from flowering to embryo and organ

development, to even size determination. The present study is regarding preparation and characterization of resource

materials for the isolation of Mediator complex from rice. We are exploiting both the immuneprecipitation and the ‘Tag’-

mediated purification principles for the isolation of rice Mediator complex. Towards the end, we have tested and validated

quite a few antibodies against rice Mediator subunits, which help us to detect the chromatographic fractions containing

the Mediator complex. Further, the Mediator enriched chromatographic fractions is being used to selectively precipitate

the Mediator complex. On the other hand, we have raised rice transgenic lines over-expressing ‘HaloTagged’ OsMED7

and OsMED17, two important subunits of the Mediator complex. The tagged subunit will act like a bait protein to fish-out

the entire complex from a sea of thousands of other proteins. We have performed pull-down experiment with the

HaloLink resin, which covalently binds with the HaloTagged Mediator subunits like Halo-MED7 and Halo-MED17 of the

transgenic calli. The presence and the purity of the isolated Mediator complex are being analysed. The isolated Mediator

complex would be useful to understand the structural and mechanistic details of the Mediator functioning in the

transcriptional process.

PC195 (IPPC0685)

Early axis growth during germination is gravisensitive and mediated by ROS and calcium

Rup Kumar Kar* and Khangembam Singh

Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva Bharati Univ., Santiniketan, Dist. Birbhum-731235, India*Presenting author: [email protected]

ROS has been explored for their possible role in gravity-induced early axis growth during germination of Vigna radiata

seeds. Axis of seeds incubated at any position tends to aligned itself along gravity this resulted in curvature growth due

to cell elongation at the convex side (upper side) as observed histologically. Such growth was accompanied with ROS

production, visible on the convex side besides accumulation in the apical region of axis, as observed through NBT stain

for O2

•-. The accumulation was inhibited completely by ZnCl2 while DPI inhibited accumulation only in the apical region.

Apart from NADPH oxidase, another O2

•- producing enzyme, NADH oxidase was revealed during spectrophotometric as

well as in gel native PAGE assay. The enzyme was insensitive to DPI and SHAM while inhibited by ZnCl2 as observed

through spectrophotrometric assay. Treatment with IAA inhibited early axis growth (predominantly occurring in the

radicle) as well as ROS accumulation while TIBA, auxin transport inhibitor, inhibited only curvature growth and ROS

accumulation at the convex side without affecting elongation growth and O2

•- accumulation in the elongation zone. Thus

gravitropic growth resulting from unequal cell elongation at the upper and lower sides of the axis was a result of spatial

regulation of ROS production on the opposing sides of the germinating axis. NADH and NADPH oxidases are the main

sources of ROS and the inhibition of NADH oxidase by auxin on the lower side resulted in curvature growth.

Pharmacological studies suggest the possible mediation of cytosolic Ca2+ in curvature growth via ROS.


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PC196 (IPPC0696)

Novel feed-forward loop between two putative plasma membrane enzymes: NADPH oxidase and H+-

ATPase

Arkajo Majumdar* and Rup Kumar Kar

Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva Bharati Univ., Santiniketan, Dist. Birbhum-731235, India*Presenting author: [email protected]

Plants’ growth, development and defence rely largely on the activities of Plasma Membrane (PM) located NADPH

oxidase (NOX) and H+-ATPase enzymes whose potential synchronization has been investigated in the roots of two

days grown Vigna radiata (L.) Wilczek seedlings. Treatments with CCCP (protonophore) and ortho-Vanadate (P-type

ATPase inhibitor) revealed that proton gradient across plasma membrane, generated due to its regulated extrusion by

PM H+-ATPase, is crucial for NOX-dependent ROS metabolism. While intensity of NBT staining decreased in

histolacalization studies, spectrophotometric (XTT) assays of both intact and extract samples showed lower superoxide

production than control and analyses of in-gel Native PAGE assays confirm that NOX-activity of the seedlings grown in

either CCCP or ortho-Vanadate decreases. On the other hand, incubation in different ROS scavengers (KI and CuCl2)

and NOX inhibitor (ZnCl2) lessened PM H+-ATPase activity in spectrophotometric as well as in Native PAGE assays with

the effect of CuCl2 being most pronounced. Interestingly, PM H+-ATPase was more susceptible to CuCl

2 than CCCP.

Activation of NOX and PM H+-ATPase appears to be positively regulated by [Ca+2]cyt from apoplastic source, since

LaCl3 (PM Ca+2-channel blocker) and EGTA (Ca+2 chelator) repressed both the enzymes. Ethylene, probably by stimulating

hyperpolarization-activated Ca+2-channels (HACC), induced the activity of both NOX and PM H+-ATPase whereas,

AgNO3 (ethylene action inhibitor) and CoCl

2 (ethylene synthesis inhibitor) inhibited both the enzymes providing additional

support to the supposition that NOX and PM H+-ATPase are interdependent.

PC197 (IPPC0698)

Characterizing novel miRNA regulating cellulose synthase genes in Arabidopsis thaliana so as to increase

the cell wall biomass and bioenergy production

Chandrashekhar P. Joshi and Yogesh Ahlawat*

Department of Biological Sciences, Michigan Technological University, Houghton, Michigan, USA*Presenting author: [email protected]

Plant cell wall, the formidable, dynamic and complex barrier is governed by more than 1500 genes. Besides providing

define structure plant cell wall plays vital role in transport of solvent, solute, intercellular communication and acts as a first

immune layer against various pathogens. Plant cell wall constitutes majorly of cellulose with small amounts of

hemicellulose, lignin and pectin. Cellulose is synthesized by cellulose synthase complex, rosette terminal domain

embedded in plasma membrane. Different forms of cellulose synthases (CesAs) are responsible for the production of

cellulose in primary and secondary cell wall. So far, 10 CesA genes in Arabidopsis thaliana have been characterized,

which plays an important role in cellulose synthesis. Several attempts have been made to understand the regulation of

CesA genes by tiny non coding RNA based regulators called miRNAs. miRNA regulating the expression of cellulose

synthases is of intensive research because of their prominent applied consequences like bioenergy production. In the

present study using in silico approaches we find out several miRNA which could influence the expression of CesA.

Furthermore, we define a network of in silcio software and manual algorithm which could efficiently pull out biologically

relevant miRNA out of a pool of several miRNAs. These selected miRNAs can be further validated by several routine

molecular biology assays.


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PC198 (IPPC0702)

Investigations on microRNA-mediated regulatory networks during heat stress in tomato

Sombir Rao*, Asosii Paul, Sarita Jha and Saloni Mathur


Heat stress is a major cause of crop loss worldwide, reducing average yields by more than 50%. Plants have evolvedmultidimensional regulatory mechanisms to minimize cell damage and maintain cellular homeostasis during elevatedtemperatures. This hyper-thermal stress is challenged jointly by two major classes of heat stress response (HSR)proteins: the heat shock proteins (HSPs), molecular chaperones and heat stress transcription factors (HSFs) that regulateHSP transcription. Knowledge of the interplay of these HSR components and microRNAs (miRNAs), major class of smallnon-coding regulatory molecules, in regulating heat stress response is very rudimentary. Till date, only a handful of heat-responsive miRNAs have been reported in plants, none of which have been functionally validated to target any HSF orHSP. Moreover, information on role of HSFs/HSPs in miRNA regulation is also limited. We have identified HSF and HSPmembers through rigorous motif-based searches in tomato and performed extensive expression analysis of candidategenes in response to different heat stress regimes as well as under other abiotic stress conditions, phytohormones andvarious plant developmental stages. Conserved as well as novel heat responsive miRNAs have been identified fromNGS data generated from control and heat-stressed tomato leaves. Differentially expressed miRNAs targeting HSFsand HSPs as well as regulation of heat-responsive miRNAs themselves by HSFs has revealed a complex regulatorymechanism involving interplay between miRNAs and HSR genes. Our findings have enhanced the understanding ofvarious miRNA-mediated regulatory networks operating in plants to provide thermo-tolerance, leading to applications incrop improvement.

PC199 (IPPC0704)

Development of genome wide PCR based InDel markers in chickpea

Ankit Jain*, Manish Roorkiwal, Sandip Kale, Mahendar Thudi, Samineni Srinivasan, Pooran M Gaur and

Rajeev K Varshney

International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India


Chickpea (Cicer arietinum) is the second most important food legume globally, which provides nutritional food securityto global population. With the availability of large scale sequencing and re-sequencing data, use of sequence basedInsertion-deletions (InDel) markers are gaining popularity because of their genome wide abundance. In addition, InDelsare also co-dominant in nature, inexpensive, highly polymorphic and high precision sequence specific PCR basedmarkers. With the objective to extend the marker repertoire and utilization in molecular breeding, re-sequencing data onparental lines of chickpea inter- and intra-specific populations were aligned with reference genome and then Dindelprogram was used for identification of InDels. As a result 8,307 InDels with >20 bp size among different parental lineswere selected. Average distribution of InDels was found 1,038 InDels/LG with maximum number of InDels on LG04(1952 InDels) and minimum number of InDels on LG08 (360 InDels). To validate and convert these InDels in to PCRbased markers, primers were designed for 428 InDels across genome using various criteria. These primers were testedfor amplification and polymorphism on the same parental lines. So far we have tested ~200 primers on these parentallines resulting in higher amplification success rate (>90%) with almost 50% polymorphism rate across parents on 3%agarose gel. These gel based markers are expected to contribute and accelerate the genetic studies and support themolecular breeding activities in chickpea.


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PC200 (IPPC0706)

Identification, characterization and expression profiling of the regulators of small RNA biogenesis and

function in response to biotic stresses in select legumes

Vanika Garg*, Gaurav Agarwal, Aamir W Khan, Himabindu Kudapa, Lekha Pazhamala, Dadakhalandar Doddamani

and Rajeev K. Varshney

International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India*Presenting author: [email protected]

Small RNAs have emerged as one of the most versatile performers in regulation of gene expression in plants. Thebiogenesis and functioning of small RNAs is regulated by major proteins like Dicer-like (DCL), RNA dependent RNApolymerase (RDR) and Argonaute (AGO). It is therefore imperative to investigate the role of protein/gene familiesinvolved in regulation of expression of small RNAs in important legumes like chickpea, pigeon pea and peanut. HMM,blastp searches, followed by domain scanning identified 4 DCL, 5 RDR, 13 AGO in chickpea; 4 DCL, 5 RDR, 13 AGOin pigeon pea and 7 DCL, 10 RDR, 22 AGO proteins in peanut. Phylogenetic analysis resulted in clustering of thesemembers into groups based on their orthologs from Arabidopsis and soybean. Subsequently, motif analysis, chromosomaldistribution and gene structure analysis of these families in the above mentioned legumes were carried out. Theseresults are in concurrence with the previous reports in other species indicating the conservation of small RNA functionand structure in plants. Along with conservation, some variations were also observed across the members of the samefamily in the legumes studied. The promoter analysis of the genes revealed the presence of stress responsive andhormone related cis-acting elements. The identified genes are being validated using qRT-PCR for their role in responseto biotic stresses. In summary, this study provides genomic information of the gene families involved in regulation of geneexpression and identification of probable candidate genes imparting resistance against stresses in these legumes.

PC201 (IPPC0723)

Comparative transcriptome analysis of differentially expressed genes in Pearl millet Pennisetum glaucum (L.)

R. Br. during drought stress condition

Rajendra Prasad Meena1, Kishor Gaikwad1, Harinder Vishwakarma1, A.R. Rao2, Sarika Sahu2, C. Tara Satyavathi3,

Madan Pal4 and Jasdeep Chatrath Padaria1

1National Research Centre on Plant Biotechnology, 2IASRI, 3Division of Genetics, 4Division of Plant Physiology, Indian AgriculturalResearch Institute, New Delhi-110012, India


The rapidly changing global climate is leading to enhanced environmental stress conditions which are causing adverseimpacts on agriculture and food security. Drought is one of the main environmental constraints to agricultural productivityworldwide. Drought tolerance is the interactive result of different morphological, physiological and biochemical traits.Pearl (Pennisetum glaucum) is most widely grown millet, which has an efficient adaptive mechanism to cope withdrought and other environmental stresses. In this study P. glaucum cv. 841B drought tolerant and 411B drought susceptiblewere subjected to water withdrawal for 10 days, 15 days at post flowering stage. Total RNA was isolated and RNAseqwas performed using Roche454. A total of 45 GB raw data was obtained (having 1273008 reads), after trimming ofadaptor and unnecessary sequences (showed 1272852). High quality sequences reads were assembled de novo usingCLC genomics (average read length of 315.78bp). Unigenes were identified using RPKM analysis and further categorizedas biological process, cellular component and molecular function. Total 12883 contigs were obtained and 43% transcriptswere annotated using GO database AmiGO2. Around 4361 (78%) of 5583 (~43% of total contigs) had GO category and585 common transcripts were obtained. Genes were assigned to different metabolic pathways using Expath software.For finding the SSR markers MISA software tool was used and a total of 286 and 153 SSRs were found in droughtsusceptible and drought tolerant cv respectively. Outcome of this study would be helpful in gene exploration involved inabiotic stress tolerance.


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PC202 (IPPC0734)

Next-generation sequencing for identification of candidate genes for Fusarium wilt and sterility mosaic

disease in pigeon pea (Cajanus cajan)

Vikas K. Singh1*, Aamir Khan1, Rachit Saxena1, Vinay Kumar1, Sandip Kale1, Pallavi Sinha1, Annapurna Chitikineni1,

Lekha Pazhamala1, Vanika Garg1, Mamta Sharma1, C. Sameer1, Swathi Parupalli1, V. Suryanarayana1, Suyash Patil1, S.

Muniswamy2, Anuradha Ghanta3, K.N. Yamini3, P.S. Dharmaraj2 and Rajeev Varshney1

1International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India,2Agricultural Research Station (ARS)-Gulbarga, University of Agricultural Sciences (UAS), Raichur, Karnataka, India3Institute of Biotechnology, Professor Jayshankar Telangana State Agricultural University (PJTSAU), Telangana, India*Presenting author: [email protected]

To map resistance genes for Fusarium wilt (FW) and sterility mosaic disease (SMD) in pigeon pea, sequencing-basedbulked segregant analysis (Seq-BSA) was used. Resistant (R) and susceptible (S) bulks from the extreme recombinantinbred lines of ICPL 20096 and ICPL 332 were sequenced. Subsequently, SNP index was calculated between R- andS-bulks with the help of draft genome sequence and reference-guided assembly of ICPL 20096 (resistant parent). Seq-BSA has provided seven candidate SNPs for FW and SMD resistance in pigeon pea. In parallel, four additional genotypeswere re-sequenced and their combined analysis with R- and S-bulks provided a total of 8362 non-synonymous (ns)SNPs. Of the 8362 nsSNPs, 60 were found within the 2-Mb flanking regions of seven candidate SNPs identified throughSeq-BSA. Haplotype analysis narrowed down to eight nsSNPs in seven genes. These eight nsSNPs were furthervalidated by re-sequencing 11 genotypes that are resistant and susceptible to FW and SMD. This analysis revealedassociation of four candidate nsSNPs in four genes with FW resistance and four candidate nsSNPs in three genes withSMD resistance. Further, in-silico protein analysis and expression profiling identified two most promising candidategenes namely C.cajan_01839 for SMD resistance and C.cajan_03203 for FW resistance. Identified candidate genomicregions/ SNPs will be useful for genomics-assisted breeding in pigeon pea.

PC203 (IPPC0749)

Transcriptome profiling of developmental stages of rice varieties varying in carotenoid content

Upasna Chettry* and Nikhil Kumar Chrungoo

Department of Botany, North Eastern Hill University, Shillong-793022, Meghalaya, India


Carotenoid level is an important quality trait in rice. We have investigated the relationship between carotenoidaccumulation and expression levels of genes involved in carotenoid biosynthesis during grain filling in selected cultivarsof rice from North East India varying in the bran colour. The grains with coloured pericarp had higher abundance of psy,pds, lcy and βch transcripts. The highest level of expression of these genes was noted in the purple cultivar whichaccumulated 44.3 μg g-1 of β-carotene, 25.07 μg g-1 of zeaxanthine and 18 μg g-1 of lutein. The carotenoid profile showeda change from β-carotene accumulation to β-carotenoid during maturation. The brown cultivars showed no detectablelevel of beta carotene, although they had 21 μg g-1 of zeaxanthine and 36 μg g-1 of lutein. Transcriptome profilesgenerated from two stages of grain filling identified 5812 differentially expressed genes (DEGs) (log2Ratio >1, FDR<0.001) with 1563 transcripts being upregulated while 4249 transcripts were downregulated. While 22% of DEGs couldbe identified with biosynthesis of secondary metabolites, 15% were identified with biotic stress and 14% with pathwaysinvolving signal transduction and starch/sugar metabolism. An insight in carotenoid biosynthetic genes revealed 14transcripts were upregulated while 5 transcripts were downregulated. Of the upregulated transcripts include psy3, zcd,lcy, zep, cyp4f3 which are related to abiotic stress. Compared to the purple variety, the variety of rice with off white branshowed significant downregulation of psy,1, pds, lcy dhrs4 and zep transcripts during transition from milking to maturestage.


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PC204 (IPPC0751)

Structure, expression and evolution of dioscorin (alpha Carbonic Anhydrase) in Dioscoreaceae

Paramananda Barman* and R. Geeta

Department of Botany, University of Delhi, Delhi-110007, India*Presenting author: [email protected]

Dioscorin (an alpha Carbonic Anhydrase; CA) is the major storage protein in the Dioscorea tubers. Dioscorin, a

multifunctional protein, consists of two types, dioscorins A and B, with 70% sequence similarity. Here, we present the

gene structure, expression and evolution of dioscorin. Genomic sequences of dioscorin A and B, with and without

introns, were characterized from Dioscorea and Tacca (Dioscoreaceae). Expression of dioscorin A and B genes in tuber,

shoot and leaf was detected at different developmental stages in three cultivars of D. alata (tuberous) and in medicinal

D. deltoidea (rhizomatous): a first report for aerial tissues and rhizome. Semi-quantitative analysis revealed that levels

of expression of all three paralogs (A1, A2 and B1) were high in tubers in comparison to leaves and shoot tips. Expression

was higher in newly emerging tubers compared to the parent tubers (setts). Phylogenetic analyses of dioscorin and

related sequences across eukaryotes showed that in Dioscorea, dioscorins formed a strongly supported clade that is

distinct from a second group of alpha CAs which are more similar to CAs in grasses. The analyses revealed ancient

duplications and independent rapid diversification of alpha CAs across eukaryotes. In plants, duplications occurred in

the vascular plant ancestor, before the monocot-eudicot splitting event and within major angiosperm lineages, e.g.,

monocots and eudicots. Despite extensive duplication and diversification of this apparently fast evolving gene, active

sites and secondary structure are highly conserved, suggesting conserved function.

PC205 (IPPC0752)

The rice F-box protein, OsFBK, is a component of a novel SCF E3 ligase and is involved in the regulation

of anther wall thickenings

Pratikshya Borah*, Shane W. Rydquist and Jitendra P. Khurana

Interdisciplinary Centre for Plant Genomics, Department of Plant Molecular Biology, UDSC, New Delhi-110021, India


The Ubiquitin-26S proteasome system (UPS) is a large group of proteins involved in the turnover of proteins no longer

necessary in the plant system. One of the E3 ligases involved in this process is known as the SCF complex (Skp1-Cullin-

Rbx1-F-box) that is instrumental in substrate identification and ubiquitination. The substrate adaptor, F-box, in one of the

largest group of proteins found in plants. The present research deals with a rice F-box protein, OsFBK that has a Kelch

domain at its C-terminal, a combination unique only to plants. Y2H analysis showed that OsFBK is a part of a novel SCF

complex. Expression analysis of this gene reveals its up-regulation in the reproductive stages and root tissues of the rice

plant and, also in response to auxin and ABA. Rice transgenics over-expressing OsFBK and its RNAi plants clearly

reveal the role of this gene in anther and root development. Electron and confocal microscopy of the transgenic anthers

demonstrated differences in their morphology and lignification status. The putative interacting partners of OsFBK were

identified from Y2H library of anther tissue and their interactions were validated by Bimolecular Fluorescence

Complementation and Co-immunoprecipitation assays. Thus, it could be hypothesized that OsFBK might be involved in

lignification processes in anthers, and in controlling proliferation of roots.


219

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PC206 (IPPC0758)

Unraveling the genome content of short arm of chromosome 2A of wheat using BAC end sequencing

Chanderkant Chaudhary*, Jitendra P. Khurana and Paramjit Khurana

Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi-110021, India


Wheat (Triticum aestivum) is an allohexaploid by genetic constitution. It has one of the largest genome of 1700 Mbp

which is approximately 40-fold larger than rice. As part of International Consortium on Wheat Genome Sequencing,

India is responsible for sequencing of chromosome 2A BAC ends of wheat. Work on sequencing in India started in 2011

with two BAC libraries obtained from Institute of Experimental Botany (Czech Republic) for chromosome 2A of wheat,

Triticum aestivum cv. Chinese Spring. So far sequencing of 48,504 out of 55,648 BAC clones has been completed from

both the ends generating approximately 97,008 BES sequences. In total, nearly 53.3 Mbp of wheat genomic sequence

having an average read length of ~550bp per BAC end has been generated. These BES were mined for repeat

sequences using REPEATMASKER revealing approximately 81.9% repetitive elements. Thereafter these sequences

were subject to BLAST analyses with different Databases with appropriate E-value (range 1E-10 - 1E-100) and Bit Score

(>150). Blast analysis lead to the conclusion that a sizeable number among the significant hits obtained represent the

genic region but a vast majority of them represent repeat elements. Protein coding genes which are represented in blast

hit are from the diverse biological processes ranging metabolic pathways to signaling pathways. In addition to the above

analysis, the BES generated data will help in physical and genetic mapping of wheat genome.

PC207 (IPPC0759)

Genome-wide survey and expression analysis of genes involved in RFOs synthesis during development

and thermal stress in wheat

Naveen Sharma1*, Paramjit Khurana1

1Department of Plant Molecular Biology, University of Delhi, New Delhi-110021, India*Presenting author: [email protected]

The raffinose family oligosaccharides (RFOs) are synthesized by addition of alpha 1,6 galactosyl to sucrose. Increase

in levels of RFOs has been associated with the thermal stress such as heat and cold stress. The availability of wheat

survey sequence and transcriptome data open avenues to comprehend the genesinvolved in RFOs synthesis during

thermal stress. By searching available survey sequence and flcDNA sequences from IWGSC, we identified genes

involved in synthesis of RFOs eg. Galactinol Synthase (GolS; EC 2.4.1.82), Raffinose Synthase (RS; EC 2.4.1.82),

Stachyose Synthase (STS; EC 2.4.1.67), and Sucrose Synthase (SUS; EC 2.4.1.13). Extensive sequence analysis has

been done of these putative RFOs encoding genes to elucidate their structure, chromosomal location and Phylogenetic

relationships.Their expression profiling by RNA Seq data analysis, revealedRFOs encoding genes showing specific

and co-expression at different stages of wheat plant and in response to thermal stress. The expression analysis of

differentially expressing genes were complimented by Real Time PCR analysis. This study implicates the role of RFOs

during thermal stress in higher plants.


220

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PC208 (IPPC0765)

Identification of long noncoding RNAs (lncRNAs) in different Capsicum species

Vineet Sharma1*, Mukesh Nitin1, Vijaya Bramha2, Sushil Chhapekar1, Toshi Mishra3, Rashmi Gaur1, Sarpras M1, Andrew

W. Lynn2 and Nirala Ramchiary1

1School of Life Sciences, 2School of Computational & Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India3Department of Bioinformatics, Banaras Hindu University, Varanasi-221005, India


Majority of the eukaryotic genome, around 90% is transcribed into noncoding RNAs, only 1-2% is protein coding. Thesenoncoding RNAs constitutes of long noncoding RNAs (lncRNAs) and small noncoding RNAs (sncRNAs) which performsdiverse functions which includes transcriptional interference, chromatin remodelling and histone modifications, alternativesplicing and post-transcriptional gene regulation. It indicates that the noncoding region of the genome which wasconsidered to be “junk”, no longer stands true. Recently, Capsicum whole genome has been sequenced which constitutesapproximately 80% of transposable and repetitive elements. In this study, our aim was to identify lncRNAs whichregulate fruit shape, fruit size, metabolites and develop lncRNA based molecular markers for capsicum breeding program.For this, two Capsicum species with contrasting fruit characteristics and metabolite contents were selected fortranscriptome sequencing using Illumina High Seq 2000 and samples were collected from three different fruit stages(Unripe, breaker and ripe), leaf and flower. The de novo assembly of the transcriptome sequences was done by usingtrinity. The entire dataset from each species after assembly was annotated and protein-coding transcripts were differentiatedfrom noncoding transcripts by using Coding Potential Calculator (CPC). Out of the total transcripts (184975 and 179780),around 85% are noncoding in nature in both species. Some of the differentially expressed lncRNAs are being validatedby RT-PCR. Our data reveals that lncRNAs are differentially expressed in tissue and species specific manner and theirfunctional roles creating phenotypic and metabolite diversity between two capsicum species has to be further studied.

PC209 (IPPC0839)

Partial sequencing and characterization of enzymes involved in galactomannan biosynthesis pathway in

Cyamopsis tetragonoloba L.

Monika Verma*, Promila Sheoran, Anoop Singh, Sweta Soni, Archana Yadav, Chhavi Bhardwaj, Kiran Bala, Priyanka

Khurana, Sonam Yadav, Usha Kantiwal, Mohit Yadav, Divya Tikaniya and Ashok Chaudhury

Dept. of Bio & Nano Technology, Bio & Nano Technology Centre, Guru Jambheshwar University of Science & Tech., Hisar-125001, India


Cluster bean, known as Guar, is a large seeded drought-tolerant annual sub-tropical legume of Leguminosae family.Guar gum or its derivatives have tremendous applications in pharma, textile, paper, petroleum, mining, explosives andfood industries. Galactomannan biosynthesis pathway begins with sucrose. Cleavage of O-glycosidic bond betweenglucose and fructose units of sucrose is catalyzed by invertase and sucrose synthase. Sucrose synthase is a glycosyltransferase, converting sucrose in presence of UDP into UDP-glucose and fructose. Direct precursors for galactomannanbiosynthesis, GDP-D-mannose and UDP-D-galactose, are formed by actions of GDP mannose phosphorylase andUDP-galactose 4-epimerase. Phosphomannose isomerase converts fructose-6-phosphate (Fru-6-P) to mannose-6-phosphate (Man-6-P). In present investigation RNA was isolated from HG365 and HG870 followed by c-DNA synthesis.Specific and degenerate primers were designed for sucrose synthase, UDP-galactose-4-epimerase, β-mannan synthase,phosphomannose isomerase, α-galactosidase and hexokinase enzymes involved in galactomannan biosynthesis.Conditions were optimized for PCR amplification using cDNA as template by employing specific and degenerate primers.Specific amplified products were sequenced using enzyme specific forward and reverse primers. Sequences wereanalyzed using BLAST. High degree of similarity was observed with closely related plant species. Alignment datashowed a high score of >=200. Phylogenetic analysis of derived sequences and pre-existing sequences showedconsiderable relatedness among them. This signifies a high level of similarity between the derived sequence andsequences of the enzymes in closely related plants. Attempts are also underway for identification and characterizationof major enzymes involved in galactomannan biosynthesis through transcriptome and whole genome sequencingapproach in cluster bean.


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IPPCongress2015


PC210 (IPPC0842)

Identification and modulation of chickpea WRKYs in development and stress

Vikas Srivastava*, Kamal Kumar, Savithri Purayannur and Praveen Verma

Plant Immunity Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India


The WRKY proteins are one of the promising transcription factors with significance over biological process, metabolism

and plant defence. The recent investigation of this TF in many genome calls for further functional studies. This family can

be demarcated into group I, II and III WRKYs. The group II have further categorization i.e., IIa, IIb, IIc, IId and IIe. Our

analysis indicated 79 WRKY genes in chickpea genome. Among them, CaWRKY5, CaWRKY38, CaWRKY39, CaWRKY44

and CaWRKY61 represents group IIA. Group IIa is smallest clade and have only three representations in Arabidopsis

thaliana known for negative regulation of plant defence. Moreover, in Glycine max, 8 members signify WRKY-IIA. Their

expression in different developmental tissues suggested mostly ubiquitous expression, with dominance in stem or

leaves. Interestingly, CaWRKY39 was found to express in floral tissues. To assess the potentiality of them in plant

defence, their response under hormonal (Salicylic acid, Jasmonic acid and Abscissic acid) and pathogenic (Ascochyta

rabiei) stresses were investigated. Under such conditions, these were found to up-regulate during SA and ABA treatments;

however mixed response were achieved after JA treatment /Ascochyta rabiei infection. Addition to this, these proteins

were known to interact together and form homo and hetero-dimer. To look into this possibility of CaWRKY-IIA proteins,

Y2H experiments were carried out. The growth on QDO and positive X-gal agarose overlay assay suggested that this

feature is well represented in chickpea. Together our experiment suggests the regulation of CaWRKY-IIa during various

environmental cues and probably their function may mediate through protein-protein interaction.

PC211 (IPPC0866)

Genome-wide transcriptome profiling to unravel mechanism of growth in a sub-tropical bamboo

Abhishek Bhandawat*, Gagandeep Singh and Ram Kumar Sharma

Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, H.P., India


Bamboo is one of the most important forest resources of the world. They are widely distributed in tropical, subtropical and

temperate forests. It comprises of almost 1200 species worldwide, of which, 130 species representing 18 genera are

found in India, making it a major hot spot for bamboo genetic resources. Plant growth is a complex trait determined by

various exogenous and endogenous factors. Earlier, transcriptome studies have been carried out in temperate bamboo

to identify genes involved in growth. However, molecular basis of growth in tropical/sub-tropical species is unknown.

Dendrocalamus hamiltonii (DH) is a commercially important fast growing sub-tropical bamboo. We performed transcriptome

profiling of DH to identify the gene(s) and pathways associated with growth. In total, 56.9 million clean filtered reads were

generated which were assembled into 39,603 contigs. Of these, 29,032 contigs showed significant hits against nr

protein database and 10,256 contigs were differentially expressed during various developmental transitions. We found

genes related to growth, cell size regulation, carbohydrate metabolic process and cell wall were substantially enriched

in growing tissues. The study suggests that environmental cues, cell signaling pathways, transcription factors and

phytohormones were found to regulate expression of genes involved in cell division and elongation leading to overall

growth of the bamboo.


222

IPPCongress2015


PC212 (IPPC0872)

Chromatin remodeler in support for bacterial growth in plants

Shweta Roy*, Priya Gupta and Ashis Nandi



Plants defend themselves from pathogens with their well elaborated immune system. All plants possess basal defense

which is induced upon recognition of evolutionary conserved molecular patterns. In addition, certain plant-pathogen

combinations trigger a strong immune response based on the recognition of effector molecules. Function of most effector

molecules remains unknown. By analysing pathogen induced Arabidopsis transcriptome profile, we identified a PRC2

member, which named here as POLYCOMB REPRESSOR FOR DISEASE 1 (PRD1). Along with other PRC2 complex

subunits, it silences the target genes through repressive histone modification. Pathogenesis-induced expression of

PRD1 is associated with enhanced accumulation of H3K4me3. PRD1 over-expression plants are susceptible to

Pseudomonas syringae carrying AvrRpt2 (Pst-AvrRpt2) effector, and are defective in SA, JA and ABA signaling. Through

tanscriptome analysis we identified RPS2, which provides resistance against AvrRpt2 in gene-for-gene interaction, as a

target of PRD1. PRD1 overexpression plants have reduced basal and pathogen induced expression of RPS2. Chromatin

immuno-precipitation experiment shows that PRD1 physically binds at RPS2 promoter and suppresses its expression

by enhancing accumulation of H3K27me3, a repressive chromatin modification at RPS2 locus. Through screening of

Arabidopsis cDNA library by yeast two-hybrid assay, we identified large numbers of PRD1 interacting partners, and one

of them is DROUGHT INDUCED 19 (DI19). RPS2 promoter has a binding motif for DI19. Our results suggest that

AvrRpt2 enhances expression of PRD1, which along with DI19 and probably with some other unknown partners make

a complex and binds to RPS2 promoter and represses its expression, to support bacterial growth in plants.

PC213 (IPPC0903)

Source-sink dynamics and QTLs based options to improve sucrose content in sugarcane stalks

Amaresh Chandra1*, Indu Verma1, Kriti Roopendra1, Radha Jain1, Ramakant Rai1 and Yong-Bao Pan2

1Indian Institute of Sugarcane Research, Lucknow-226002, U.P., Indian2USDA-ARS, MSA, Sugarcane Research Laboratory, Houma, LA 70360, USA


Improving sucrose content in sugarcane stalk is the prime objective that can be addressed through breeding as well as

source-sink dynamics associated interventions. Since sugarcane cultivars possess >100 chromosomes (2n = 100 -

130), and are genetically complex polyploid and aneuploids, identification of quantitative trait loci (QTLs) associated

with sucrose content is considered the preferable option to improve sucrose content through molecular breeding. In this

direction, a linkage map was constructed using AFLP, target region amplification polymorphic (TRAP) and SSR markers.

This map had a cumulative genome length of 7406.3 cM that included 675 AFLP (69.8%), 90 TRAP (9.3%), and 202

SSR (20.9%) markers. The 202 SSR markers were assigned to 65 CGs and 8 HGs. Based on this map, 24 putative QTLs

affecting sucrose content were identified. Five QTLs were unlinked and the other 19 were located on nine CGs within

four HGs. Of these QTLs, 11 had an effect in both, plant cane (20.33%) and first stubble (25.68%) crops. While addressing

the source-sink dynamics, the signal communication was perturbed by restricting source leaves’ photosynthetic ability

and by improving sink strength with the application of plant hormone like GA3. Results indicated differential accumulation

of sugars, along with expression of some pivotal genes in stalk, source and sink leaf tissues. Results propose a model

and various options to address this important issue in sugarcane genomic research.


223

IPPCongress2015


PC214 (IPPC0926)

Cloning, phylogenetic analysis and expression profiling of Crocus sativus zinc finger transcription factors

Aubid Malik* and Nasheeman Ashraf

Plant Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Sanat Nagar, Srinagar- 190005, India


Crocus sativus is the only plant known to produce appreciable quantities of apocarotenoids, like crocin, picrocrocin and

safranal. These metabolites are produced in stigma tissue in developmental stage specific manner. Although, the

biosynthetic pathway of apocarotenoids in Crocus is pretty well understood, but the mechanisms regulating tissue and

developmental stage specific production of these metabolites is still elusive. Characterization of the transcriptome of

saffron stigmas is sine quo non for elucidating the molecular basis of flavor, color biogenesis, genomic organization and

biology of gynoecium of saffron. In this backdrop, transcriptome-wide survey of Crocus sativus lead to the identification

of 173 zinc finger transcription factors. C3H, C

2H

2 and AN20/AN1 were the most preponderant classes with 42, 24 and 18

members which grouped into 4, 4 and 3 classes respectively. Besides, expression analysis of representative members

of the three classes was carried out in different tissues, at different stages and in response to different stresses and

phytohormones. Member of C3H showed highest expression in anther, C

2H

2 in tapel and AN20/AN1 in Stigma. C

3H

member was induced by salt, C2H

2 by cold and AN20/AN1 by drought. C

2H

2 member was upregulated by auxin and

gibberellic acid, while C2H

2 by Methyl jasmonate and ABA and AN20/AN1 by auxin and ABA. Since AN20/AN1 showed

highest expression in stigma and in anthesis stage commensurate with the apocarotenoid profile of the flower, was

selected for further characterisation. Therefore, we envisage probable role of AN20/AN1 in Crocus sativus specific

metabolite regulation, which is under further evaluation.

PC215 (IPPC0938)

Genome wide identification expression profiling and characterization of Lectin gene super family of

mulberry

Bushra Saeed* and Paramjit Khurana



Lectins are a diverse group of ubiquitously present, highly specific sugar binding proteins. Members of this large gene

family have been assigned broad biological functions from defense to acting as storage proteins. Despite possessing

several interesting characteristics, their functions remain essentially undefined. Mulberry known for its medicinal benefits

is also a rich source of lectins. Using an exhaustive HMM-based search we identified the lectin gene complement in

Morus notabilis with around 197 members. These putative lectin genes were classified into twelve distinct gene families

based on the presence of characteristic sugar-binding domains. Members of this superfamily were assigned varied

gene ontologies to identify putative functions and determine cellular localizations. Interestingly, characteristic expression

patterns were observed across the lectin superfamily in response to a variety of stress conditions. This is suggestive of

specialized functions under diverse conditions possibly by linking the specificity of sugar recognition with mediating

precise stress responses in plants. A Jacalin family member showing a drought specific expression was further

characterized in Arabidopsis. The identification of putative gene family members from the genus Morus developed in this

study can find wide applicability in lectin gene identification and characterization. It can also contribute immensely in the

understanding of lectins from mulberry with potential medicinal uses.


224

IPPCongress2015


PC216 (IPPC0952)

Co-expression of regulatory genes involved in protein synthesis and protection to improve cellular level

tolerance of tobacco plants under abiotic stress

Mahesh Patil*, Chandrashekar Reddy, T.G. Prasad and Udayakumar Makarla

Department of Crop Physiology, University of Agricultural Science, GKVK, Bangalore-560065, Karnataka, India


Plants when experiences stress there will be reduction in metabolic activities of cell leading to reduced growth anddevelopment. In this context, improving cellular level tolerance (CLT) to sustain cell metabolic activities has a phenomenalrelevance under stress. Among several CLT mechanism protein turnover and stability are important, which are getaffected under stress. Among many regulatory checks in protein turnover, translation and protein protection are veryimportant and several studies have showed relevance of these processes under stress. From this context, to maintainprotein homeostasis the best approach will be overexpression of genes/transcription factors involved in these processes.Therefore, an attempt has been made to improve protein turnover and stability by overexpressing three regulatory genesi.e. Pg47 (Pennisetum glaucum 47, a RNA helicase), OseIF4E (Oryza sativa translational initiation factor 4E) andPgHSF4 (Pennisetum glaucum Heat shock factor4) involved in protein synthesis and protection in a model systemtobacco. Since, aim was to address both protein synthesis and protection, further, an attempt has been to co-express allthese genes together in model system tobacco and comparative analysis was also done. Multigene transgenics showedincreased plant growth and tolerance to diverse stresses such as salinity, DTT, PEG, CdCl

2 and cold stress with

improved seedling growth compared to single gene expressing trangenics. Further, multigene transgenics showedsignificantly higher protein content and lesser MDA level and also enhanced expression of few stress responsive genes.These results clearly demonstrate that co-expression of specific genes involved in translation and protein stabilitysubstantially improved stress adaptation of crops.

PC217 (IPPC0966)

Comprehensive transcriptome analysis of Crocus sativus for discovery and expression of genes involved

in apocarotenoid biosynthesis

Shoib Baba, Tabassum Mohiudin, Aubid Hussain, Zahoor Wani, Nazia Abass and Nasheeman Ashraf*

Plant Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Sanat Nagar, Srinagar, India


Crocus sativus stigmas form rich source of apocarotenoids like crocin, picrocrocin and safranal, which besides impartingcolor, flavour and aroma to saffron spice also have tremendous pharmacological properties. Inspite of their importance,the biosynthetic pathway of Crocus apocarotenoids is not fully elucidated. Moreover, the mechanism of their stigmaspecific accumulation remains unknown. Therefore, deep transcriptome sequencing of Crocus stigma was done toidentify the genes and transcriptional regulators involved in the biosynthesis of these compounds. Transcriptome ofstigma was sequenced using Illumina Genome Analyzer IIx platform, which generated 51,350,714 stigma reads.Sequences were assembled de novo using trinity resulting in 64,438 transcripts which were classified into 32,204unigenes comprising of 9,853 clusters and 22,351 singletons. A comprehensive functional annotation and gene ontology(GO) analysis of all the sequences was carried out. Many genes involved in carotenoid/apocarotenoid pathway wereidentified for the first time in this study which includes zeta-carotene isomerase and desaturase, carotenoid isomeraseand lycopene epsilon-cyclase. GO analysis showed that the predominant classes in biological process category includemetabolic processes followed by cellular processes and primary metabolic processes. KEGG mapping analysis indicatedthat pathways involved in ribosome, carbon, starch and sucrose metabolism were highly represented. Differentialexpression analysis indicated that key carotenoid/apocarotenoid pathway genes including phytoene synthase, phytoenedesaturase and carotenoid cleavage dioxygenase 2 are enriched in stigma thereby providing molecular proof for stigmato be the site of apocarotenoid biosynthesis. This data would provide a rich source for understanding the carotenoid/apocarotenoid metabolism and flower development in C. sativus.

Session D : Plant Metabolism and Metabolomics

225

IPPCongress2015


PD218 (IPPC0247)

Increased glyphosate resistance gene (igrA) confers resistance to the herbicide glyphosate in transgenic

indica rice

Vijay Sheri*, Bhabesh Borphukan, Mohan Acharya and M.K. Reddy

Plant Molecular Biology, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


Plants expressing Pseudomonas sp. strain PG2982 increased glyphosate resistance gene (igrA) are known to be resistant

to glyphosate, a potent herbicide that inhibits the activity of the endogenous plant EPSPS. In order to develop herbicide-

resistant indica rice, the disarmed Agrobacterium strain EHA 105 harbouring the binary vector pMDC-99 was used for

transformation. The T-DNA region of the vector carries igrA gene with a DNA segment encoding a chloroplast transit

peptide, which would direct the igrA to the organelle where the shikimate pathway is located, under the control of rice

actinII promoter for constitutive expression, and hygromycin phosphotransferase gene (hpt) as a plant selectable marker

for selection on hygromycin. A large number of transgenic rice plants were obtained with hygromycin and glyphosate,

most of the transformants showed fertile. The integration and expression of igrA gene from regenerated plants was

analyzed by PCR and Southern blot analysis. Transgenic T1 seeds were germinated on hygromycin and transferred

them to glyphosate medium to assess the glyphosate tolerance. The transgenic rice plants had more glyphosate resistance

than the wild-type plants. Field test of transgenic rice plants further confirmed that they survived 100% spray of glyphosate

(Roundup) at a regular dose used for conventional rice weed control.

PD219 (IPPC0261)

Influence of arsenate and phosphate on growth and thiol metabolism in two cultivars of rice

(Oryza sativa L.)

Jayeeta Saha* and Asok Kumar Biswas

Department of Botany, University of Calcutta, Kolkata-700019, West Bengal, India


Arsenic pollution has become a serious problem since the last decade in West Bengal owing to extensive withdrawal of

water for irrigation and portable purposes. Arsenic (As), a metalloid, is toxic to all forms of life including plants. There is a

competitive interaction between arsenate and phosphate for the same uptake system in plants as phosphate is chemically

similar to arsenate, both belonging to group (Va) of the periodic table. The influence of arsenate and phosphate on

growth, total glutathione content and activity of its regulatory enzymes viz. GPX, GR and GST were investigated in 21 day

old rice (cv. Nayanmoni and cv. IR-64) seedlings. Arsenic toxicity triggered formation of reactive oxygen species. Thus,

the levels of oxidative stress markers viz., proline, H2O

2, non-enzymatic antioxidants like glutathione, α-tocopherol,

ascorbate, including activity of enzymatic antioxidants viz. ascorbate peroxidase, catalase were altered in the test cultivars;

the effects being more significant in IR-64 in contrast to cv.Nayanmani. Phytochelatins (PCs), a class of small thiol-rich

peptides, can detoxify the excess of toxic heavy metals by forming a metal-PCs complex in which the heavy metal is

bound to the thiol group of Cys unit and therefore, can be used as a potential biomarker for evaluating metal toxicity.

Different types and levels of PCs were detected in the roots and shoots of the test cultivars grown in arsenate

(Na2HAsO

4.7H

2O) with or without phosphate (KH

2PO

4) by RP-HPLC. Therefore, PCn analysis plays an important role in

the study of metal toxicity and tolerance mechanism.


226

IPPCongress2015


PD220 (IPPC0262)

Physiological, phytochemical and molecular studies on antioxidant accumulation in the medicinal plant

Amaranthus spinosus Linn. as influenced by abiotic stress

G. Garggi* and Roy Stephen

Plant Physiology, Kerala Agricultural University, College of Agriculture, Vellayani, Thiruvananthapuram-695522, Kerala, India


Plants continue to serve mankind as food and medicine since time immemorial. The medicinal effects in plants are due tothe presence of various bio-active principles which are the products of secondary metabolism in plants. Plants haveevolved the secondary metabolic pathway mainly to defend themselves from numerous biotic and abiotic stresses. Thesestresses increases production of reactive oxygen species (ROS), that creates oxidative stress to plants. Antioxidantsmaintain the equilibrium of generation of ROS and its scavenging. It is found that abiotic stress acts as an environmentalcue for the enhanced production of antioxidants, thereby increasing the therapeutic value of medicinal plants. A studywas conducted to investigate the influence of abiotic stress factors viz. low light stress and water stress, on the growthcharacteristics, physiological and biochemical parameters as well as on the various enzymatic and non-enzymaticantioxidants in Amaranthus spinosus Linn. It is an annual herb found throughout India and is potent medicinal plantknown for its antioxidant accumulation. Growth parameters like plant height and leaf area had a positive response withshade. Starch content was high for the plants under open environments. Enzymatic and non-enzymatic antioxidants inthe plant showed variations across different treatment combinations of shade and water stresses. Expression study of thegene chalcone synthase (CHS), the key enzyme in Phenyl Propanoid pathway in the biosynthesis of flavonoids andphenolic compounds, was also done using Reverse Transcriptase-PCR (RT-PCR) and was related with the intensity ofstress.

PD221 (IPPC0335)

Gas-chromatography mass-spectrometry (GC-MS) based metabolite profiling in two contrasting

germplasms of rice in response to salinity stress

Khalid Anwar1*, Nita Lakra2, Sneh Lata Singla-Pareek2 and Ashwani Pareek1


Salinity is a major factor that limits rice production worldwide. Gas chromatography mass spectrometry (GC-MS) isfrequently used to provide the sensitive and reproducible detection of hundreds to thousands of metabolites in a singletissue sample. Untargeted metabolite profiling has the potential to identify numerous novel metabolites. Here, we developeda method for GC-MS based metabolite profiling for rice, in order to gain an overview of the pathway of primary metabolism.In order to reveal the physiological and molecular differences in salt tolerance between wild (Pokkali) and cultivatedvariety of rice (IR64), profiles of more than 100 of key metabolites were studied using GC-MS. In the present study,untargeted and targeted metabolic profiling analyses were carried out in rice tissues. Principal component analysis (PCA)was employed to explore the metabolite profiles from rice shoot samples at eight different time points. Significant changesin metabolite levels were found in both Pokkali and IR64 in response to salinity with tryptophan, proline and lysine beingaccumulated at much higher levels in the Pokkali as compared to IR64. In contrast, glycine, homoserine, threonine andalanine levels were higher in IR64 as compared to Pokkali, as IR64 probably adjusted its cellular homeostasis mainlythrough increasing sugars levels at initial stress exposure. We also found that overall amino acids, sugars were increasedwhereas, organic acid decreases during salinity stress treatment. A possible role of these metabolites in response tosalinity will be discussed.


227

IPPCongress2015


PD222 (IPPC0406)

A comparative evaluation of GC-MS based phytochemicals, in vitro radical scavenging properties,

antioxidant content and yield performance of two promising accessions of Amaranthus hypochondriacus

Aditya Manashi* and Bhattacharjee Soumen

Department of Botany, Centre for Advanced Study, University of Burdwan, Burdwan-713104, West Bengal, India


The present study aims at studying two promising accessions of Amaranthus hypochondriacus (IC107144 and IC94661)based on their antioxidant and radical scavenging properties, types of hydroxyl rich phytochemical availability and yieldpotentials. Quantitative antioxidant profiling of methanolic leaf extracts were done based on in-vitro radical scavengingproperties (ABTS, FRAP and DPPH radical scavenging assay) and availability of individual antioxidants (Total polyphenol,monophenol, diphenol, flavonoids, betacyanine, anthocyanin, carotenoids). A comparative evaluation of antioxidantsfrom leaf extract reveals a significantly higher antioxidant availability in accession IC107144 as compared to IC94661.The same trend of result were observed when the antioxidant potential of the methanolic leaf extracts of both the accessionswere estimated and compared by FRAP, ABTS and DPPH radical scavenging properties. The data of in-vitro radicalscavenging properties of both the accessions exhibited significant correlation between individual antioxidant contents.When GC-MS study was carried out, it exhibited presence of several phytochemicals in both the accessions, some ofwhich were having antioxidant properties. The accession IC107144 exhibited presence of some additional compounds,which contain hydroxyls, responsible for radical scavenging properties. So taken as a whole, the data of quantitativeantioxidant profiling and GC-MS based qualitative evaluation of phytochemicals from leaf extracts of two accessions ofAmaranthus hypochondriacus not only provide evidence of rich sources of antioxidant, radical scavenging properties andavailability of several phytochemicals with hydroxyls but also prove that accession IC107144 as better compared toaccession IC94661.

PD223 (IPPC0419)

Role of phenylalanine for the production of flavonoids in callus cultures of Prosopis cineraria Druce

cultivar K1 and Ziziphus mauritiana cultivars Gola and Seb

Ruchi Nag Shrivastva*, Arjit Chaturvedi and T.N. Nag

Department of Biotechnology, M.N. Institute of Applied Sciences (MGS University) Bikaner-334022, Rajasthan, India


Plant in vitro cultures are able to produce and accumulate many medicinally valuable secondary metabolites. Flavonoidsare an important group of medicinal preventive compounds as well as food additives inhibiting detrimental changes ofeasily oxidizable nutrients. Many different in vitro approaches have been used for increasing biosynthesis and theaccumulation of flavonoid compounds in plant cells. The present study summarized the achievements of in vitro technologyfor the production of flavonoid compounds by focusing on different approaches to improve the production of thesecompounds. Unorganized callus tissues of plant species were established on MS (Murashige and Skoog, 1962) mediumsupplemented with different phytohormones. Callus tissues were transferred to fresh MS medium singly supplementedwith various concentrations [25, 50, 75 and 100 mg (100 ml)-1 of media] of DL-b-phenylalanine and allowed to grow up tomaximum growth indices. Total flavonoid (kaempferol and quercetin) content showed a marked increase in the tissuesfed with phenylalanine and gradual increase in the growth of tissues was observed. Highest amount of flavonoid wasobserved in tissues grown on MS medium supplemented with 100 mg (100 ml)-1 as compared with other concentrationsof phenylalanine in the medium in both the cultivars of Ziziphus mauritiana, viz., Seb (8.45 mg g-1 dry wt.) and Gola (8.29mg g-1 dry wt.) but in the case of P. cineraria highest concentration of 11.23 mg g-1 dry wt. was obtained with 75 mgphenylalanine (100 ml)-1 medium. Enhancement by feeding phenylalanine supports the pervious findings that this aminoacid acts as precursors of flavonoids. The results will be discussed in the light of contemporary data.


228

IPPCongress2015


PD224 (IPPC0420)

Antioxidants in arid zone plants and their tissue cultures

T.N. Nag*

Department of Biotechnology, M.N. Institute of Applied Sciences, MGS University, Bikaner–334022, Rajasthan, India*Presenting author: [email protected]

Antioxidants are an important group of medicinal preventive compounds as well as food additives inhibiting detrimental

changes of easily oxidizable nutrients. Many in vitro approaches have been used to increase biosynthesis and the

accumulation of antioxidant compounds in plant cells. Antioxidants are capable of stabilizing, or deactivating free radicals

before they attack cells, and are critical for maintaining optimal cellular and systemic health and well-being. These are

gaining lot of importance as a panacea for a large number of life-style diseases like aging, cancer, diabetes, cardiovascular

and other degenerative diseases etc. Thus, investigation of various antioxidants present in arid zone plants and their

tissue cultures is the demand of today. Cultures of 21 arid zone plants, established from hypocotyls/seedlings/shoot tips

were maintained on MS medium supplemented with different concentration of hormones for 12 months by frequent sub-

culturing. In this paper various types of antioxidants identified in plant parts and tissue culture will be discussed in the light

of contemporary data. The enhancement of these compounds by addition of different concentration of precursors and

phyto-hormones will also be reviewed.

PD225 (IPPC0441)

LC-MS based approach for metabolic fingerprinting of Capparis moonii, Wight

Nutan Malpathak and Pallavi Yadav*

Department of Botany, Savitribai Phule Pune University, Pune-411007, Maharashtra, India


Capparis moonii, Wight is an endemic plant belonging to biodiversity hotspot region of Western ghats with ethnomedicinal

importance. Therefore, C. moonii emerges as a potential candidate for Bioprospecting. Sequential polarity wise extraction

coupled with sonication was adopted for extraction of secondary metabolites from Capparis moonii. An LC-MS based

metabolomics approach was used to characterize secondary metabolite quantitative and qualitatively in stem and leaves

of C. moonii. PCA, a multivariate technique was used for the analysis of metabolomics data.Current findings are significant

in establishing metabolic fingerprint of an unexplored plant and probably paves way for reverse pharmacognosy approach

in drug discovery.


229

IPPCongress2015


PD226 (IPPC0445)

Modulation of photorespiratory metabolism by oxidative and photooxidative stress in leaf discs of

Pisum sativum

Ramesh B. Bapatla* and A.S. Raghavendra

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India


Photorespiration is known to protect plants from photoinhibition and is considered to be an important component of plant

adaptation to oxidative stress. Reactive oxygen species (ROS), such as H2O

2, are generated under oxidative stress

conditions. Menadione, a redox active quinone, interferes with mitochondrial electron transport and generates free radicals,

causing oxidative stress. We studied the effect of oxidative stress, induced in mitochondria by menadione on enzymes

localized in different cell organelles of pea (Pisum sativum) leaf discs. The enzymes were glycolate oxidae (GO), catalase

(CAT) and hydroxypyruvate reductase (HPR), all in peroxisomes, while aconitase was located in mitochondria and

glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in cytosol. The enzyme activities were examined after incubation

in dark, moderate (300 μE m-2 s-1) and high light (1200 μE m-2 s-1) intensities. The activities of GO and CAT were increased

by menadione upto 40 to 50%, particularly in high light. In contrast, the levels of HPR increased marginally by <20%. The

activity of aconitase was marginally decreased. Our results indicate that oxidative stress condition, induced by menadione,

modulates markedly the redox-sensitive key photorespiratory enzymes, particularly in light. The enzymes located in

mitochondria (e.g. aconitase) or cytosol (GAPDH) did not respond as strongly as those in peroxisomes. We suggest that

oxidative stress, originating in mitochondria can affect photorespiratory components in peroxisomes.

PD227 (IPPC0478)

Waterlogging responsive MaAP2 an AP2/ERF protein from Mentha arvensis regulates carbohydrate

metabolism in Arabidopsis through bidirectional sugar transporter AtSWEET10

Ujjal J. Phukan1*, Sonal Mishra2 and Rakesh K. Shukla1

1Department of Biotechnology, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226015, India2National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Waterlogging is accountable for huge loss of yield as well as quality in commercial and agronomical breeds. MaAP2 an

ERF identified in Mentha arvensis waterlogging subtractive cDNA library was showing induced expression in dehydration,

cold, salt and waterlogging treatment. It was also induced in response to ABA and ACC. We observed that it binds

specifically to different cis-elements DRE, GCC and Jasmonic acid response element which was competed out with

excess of cold probe. We have also mapped a potential transactivation domain in MaAP2 adjacent to the DNA binding

domain. MaAP2 over-expressing Arabidopsis lines displayed increased tolerance to dehydration, salt and waterlogging

treatment in comparision with WT. Microarray analysis of MaAP2 overexpressing transgenic lines revealed 7889 and

4895 up- and down-regulated transcripts, respectively. As carbohydrate metabolism is deeply affected during waterlogging

stress we screened the data for sugar metabolism and transport related genes. AtSWEET10 involved in bi-directional

sugar transport was highly up-regulated along with other SWEET and Sugar transporter family proteins. We showed that

MaAP2 directly binds to the promoter of AtSWEET10 in in vivo and in vitro conditions, allowing MaAP2 to modulate

carbohydrate metabolism which would help the plants to survive under adverse environmental stress condition. The

study provides novel information about the role of AP2/ERF protein and their role in regulation of carbohydrate metabolism

under adverse environmental condition.


230

IPPCongress2015


PD228 (IPPC0528)

Association of galactomannan hydrolyzing enzymes activity with early seed germination in rice

Nakul Gupta1, M.B. Arun Kumar1*, Jacob Sherry Rachel2, S. Gopala Krishnan3, Malavika Dadlani1, C. Viswanthan4

and M. Nagarajan5

1Division of Seed Science and Technology, 2Division of Germplasm Conservation, 3Division of Genetics, 4Division of Plant Physiology,Indian Agricultural Research Institute, New Delhi-110012, India5ICAR-IARI Regional Center, Aduthurai, India


The scarcity of water and labour is forcing the rice farmers to shift from transplanted method of cultivation to direct-seeded cultivation. Faster germination of seed as well as establishment of vigourous crop is of paramount importance indirect-seeded crop. Rice being an albuminous seed, it is hypothesized that galactomannan hydrolysing enzymes playsimportant role in loosening of the embryo-surrounding tissue, which in turn governs the speed of germination. To test thishypothesis and the effect of gibberellins and Abscisic acid on enzyme activity, studies were conducted on four ricegenotypes comprising of two early germinating (<28 hours) and two late germinating (>46 hours) genotypes. Activities ofgalactomannan hydrolysing enzymes (β-mannanase, β-mannosidase and α-galactosidase) increased with germinationin both early and late germinating rice genotypes. The average enzyme activities of the genotypes was higher in earlygerminating group in comparison with the late germinating group for all the three enzymes and at all the three stages ofgermination (dry seed, lemma rupture and radicle emergence). Among the enzymes, activity of β-mannosidase wasfound higher in dry seeds of both the group followed by α-galactosidase and β-mannanase. GA supplement (50 μM) inthe germinating medium has increased the activities of all the three enzymes that manifested in faster lemma rupture andradicle emergence in both early and late germinating rice genotypes by 3-5 hrs and 8-12 hrs, respectively. Supplement ofABA (50 μM) in the germination medium has decreased the activities of all the enzymes that resulted delayed seedgermination.

PD229 (IPPC0569)

Modulation of photorespiratory metabolism by osmotic stress in leaf discs of pea, Pisum sativum

Fareed Khan* and A.S. Raghavendra



Plants need to adapt to stress conditions. It is proposed that photorespiration may be an important component of suchplant adaptation to abiotic stress, but there is no direct evidence. We therefore, studied the effect of osmotic stresscaused by 1.0 M sorbitol on the photorespiratory enzymes in leaf discs of pea, Pisum sativum. To ensure osmotic stress,the pattern and consequences of reactive oxygen species (ROS) in pea leaf discs were monitored on exposure to 1.0 Msorbitol for 3 h under dark, moderate (300 μE m-2 s-1) and high light (1200 μE m-2 s-1) intensities. H

2O

2 and superoxide

levels were estimated by using DAB and NBT, respectively. Upon exposure of pea leaf discs to 1.0 M sorbitol and light thelevels of H

2O

2 as well as superoxide were increased significantly. Levels of H

2O

2 increased by 80 %, while that of superoxide

increased by 60%. We then looked at the activities of three photorespiratory enzymes located in peroxisome, namelyglycolate oxidase, catalase and hydroxypyruvate reductase in pea leaf discs. There was a marked increase in the activityof glycolate oxidase (43%), followed by catalase (20%) and hydroxypyruvate reductase (14%). Experiments to cross-check and compare the activities of aconitase (mitochondrial enzyme) and glyceraldehyde-3-phosphate dehydrogenase(cytosolic) are in progress. Our data suggest that exposure of pea leaf discs to osmotic stress, particularly under highlight, can upregulate key photorespiratory enzymes.


231

IPPCongress2015


PD230 (IPPC0577)

GC-MS evaluation of bioactive compounds of whole plant ethanol extract from Osmunda hugeliana C.

Presl.

Manisha Kale*

Department of Botany, Jaysingpur College, Jaysingpur-416101, India


Osmunda hugeliana C. Presl. belonging to family Osmundaceae is one of the medicinally important pteridophyte used as

tonic, styptic and also for rickets, rheumatism and for intestinal gripping. In this study, the bioactive compounds of O.

hugeliana have been evaluated using Shimadzu make QP-2010 GC-MS. The chemical composition of the whole plant

ethanol extract was investigated. While the mass spectra of the compounds found in the extract were matched with the

NIST library. The compounds were identified by comparing their retention time and peak area with that of literature and

interpretation of mass spectra. GC-MS analysis revealed the presence of sixteen bioactive compounds. The most

compounds were 3, 7, 11, 15-tetramethyl-2-hexadecen-1-ol, tetradecanoic acid (myristic acid), n-hexadecanoic acid,

hexadecanoic acid ethyl ester and octadecanoic acid. Tetradecanoic acid (myristic acid) is having antioxidative, lubricative,

hypercholesterolemic, cancer preventive properties, and also used in cosmetic. N-Hexadecanoic acid (palmitic acid) is

also having antioxidantive, hypochloresterolenic, nematisidic, pesticidal, lubricative, antiandrogenic flavor, hemolytic

properties.

PD231 (IPPC0602)

Stage-specific regulation of triglyceride synthesis during seed development in Jatropha curcas L.

Bharatula Chaitanya*, Sumit Kumar, P.S. Sastry and Attipalli R. Reddy



The present study describes the changes in lipid profiles as well as acyl fluxes occurring in developing seeds of a

potential biofuel tree species, Jatropha curcas. Metabolic labeling with 14C-acetate indicated a relationship between

phosphatidylcholine (PC) and triacylglycerol (TAG) with seed development (34, 37 and 40 days after anthesis, DAA). The

developing endosperm preferred synthesis of phospholipids to neutral lipids at 34 DAA. Study of acyl fluxes showed that

PC, diacylglycerol (DAG) and TAG were rich in palmitic acid (16:0). From 37 to 40 DAA, the developed endosperm was

actively involved in storage TAG synthesis. Fatty acid methyl ester analysis showed presence of oleic acid (C18:1n-9)

rich PC, DAG and TAG molecules. This shift in fatty acid composition of PC molecules (16:0 to 18:1) was concomitant

with synthesis of 18:1n-9-rich TAGs during these stages. Furthermore, distinct pools of DAG were observed with turnover

of acyl-edited C18-rich DAGs showing higher unsaturation at 40 DAA which was confirmed by analysis of molecular

species and positional distribution. The expression of diacylglycerol acyltransferase (DGAT2) was correlated with 16:0-

rich TAGs at 34 DAA, whereas expression of DGAT1 was significant during 37 and 40 DAA, which was related to 18:1n-

9-rich TAGs. Our data demonstrate metabolic reprogramming and existence of alternate pathways during TAG synthesis

involving distinct pools of PC and DAG, presumed to react with each other and contribute to storage TAG synthesis

during seed development in Jatropha.


232

IPPCongress2015


PD232 (IPPC0733)

Non-targeted metabolomics of a medicinal plant cumin (Cuminum cyminum L.) shoots under salinity

Sonika Pandey*, Manish Kumar Patel, Avinash Mishra and Bhavanath Jha

Division of Marine Biotechnology and Ecology, CSIR-CSMCRI, Bhavnagar, Gujarat, India


Spice crops occupy a valuable place in traditional medicinal applications and act as food adjuncts because of distinct

aroma and flavour. Cuminum cyminum is an important aromatic herbaceous spice crop belonging to the Apiaceae family,

cultivated in semi-arid regions of the world. India is the largest producer, consumer and exporter of cumin seeds and

contributes up to 90% of total productivity. The present study is intended to comprehensively analyse the composition of

metabolites under salinity that provides the physiological status of a plant to combat stress conditions. Cumin is an active

reservoir of metabolites (primary and secondary) or phytochemicals that are characterized as amino acids, flavonoids,

polyphenolic and terpenoids etc. Different analytical methods, such as LC MS, GC MS, etc. have been used to elucidate

comprehensive information about metabolites. Cumin shoots contain 53% polyunsaturated fatty acids. Saturated FAs

increased gradually with salinity, whereas unsaturation index and degree of unsaturation changed arbitrarily along with

the percent quantity of unsaturated FAs. A total of 45 differentially expressed metabolites were identified, including

luteolin, salvianolic acid, kaempferol and quercetin, which were phenolic, flavonoid or alkaloids in nature and contain

antioxidant activities. Additionally, metabolites with bioactivity such as anticancerous (docetaxel) and antimicrobial

(megalomicin) were also identified. The study showed that cumin shoots are rich source of metabolites with bioactivities,

essential amino acids, phenolic compounds and fatty acids, which unveil its medicinal characteristics, providing useful

insights about metabolic responses under salinity that creates its utility in nutraceutical industries.

PD233 (IPPC0736)

Metabolomics of medicinal and spice plants: A tool for quality check of herbal products

Agepati Raghavendra and Sarada Tetali*



Metabolomics provide a snap shot of biochemical status of an organism, by elucidating the spectrum of metabolites

within. Plants are estimated to contain more than 200,000 different metabolites. Such wide diversity of phytochemicals

and their occurrence spread across aerial and underground organs makes “plant metabolomics”, a daunting task. However,

recent technological advances in metabolomics made the exploration of such treasure easy and discovery of novel

natural plant products possible. The combination of liquid/gas chromatography and mass spectrometry provides a common

base of metabolomics though there are several other techniques like nuclear magnetic resonance (NMR) in use. Metabolite

libraries of Indian medicinal and spice plants have largely remained unexplored. The metabolite fingerprints of medicinal

and spice plants not only help in the discovery of drug molecules but also can be exploited for quality check of herbal

samples including herbal medicines and spices for commercial use. We have attempted to perform metabolite profiling

using GC-MS and/or LC-ESI-MS/MS of several medicinal plants, which are used for treating human inflammatory diseases,

e.g. Ocimum sanctum (leaves), Terminalia arjuna (stem-bark), Boswellia serrata (gum resin), Hemidesmus indicus (roots)

and of Curcuma longa and C. aromatica (rhizomes). Further, we have tried to compare metabolite variation in rhizomes

of seven cultivars of Curcuma. The possible use of metabolomics data to differentiate the cultivars and to assess the

quality of turmeric samples would be discussed.


233

IPPCongress2015


PD234 (IPPC0901)

CCPM v3.4: towards collaborative metabolomics

Indira Ghosh1, Abhijit Mitra2, Nita Parekh2, VikramPudi2, BrotoChakrabarty2, Prashanthi Dharanipragada2, Ramya

Gurrapu2, Narendra Babu Karuputhula2, Manoj Kumar Sekhwal1, Gaurav Sharma1*, S.R. Kiran Raj2, Manoj Kumar

Maramreddy2, V.P. Srivani2 and V. Dharma Teja2

1School of Computational and Integrative Science, Jawaharlal Nehru University, New JNU Campus, New Delhi-110067, India2Center for Computational Natural Sciences and Bioinformatics, International Institute of Information Technology Hyderabad, Gachibowli,Hyderabad-500032, India*Presenting author: [email protected]

Assisted by advances in high-throughput instrumentation, metabolomics has emerged as a popular research area, whichis witnessing rapid generation of large volumes of high quality data. This demands ready availability of advanced analyticaland computational methods for generating analyzed datasets, as well as advanced tools for data analysis, visualizationand integration; within a collaborative platform which addresses the unique interdisciplinary character of metabolomics.The Computational Core for Plant Metabolomics, CCPM v3.4, is a fully functional, open, state of art web application,addressing the issues of storage, processing, analysis and sharing of data and results of metabolomics experiments.With a python web framework (web2py) backend, CCPM allows rapid prototyping, easy maintenance and role-basedaccess control without compromising on security, scalability and efficiency. Accessible through a user friendly bootstrap-based GUI, CCPM functions as a secure repository with a pipeline for end-to-end analysis of LC/GC-MS data involvingraw data capture, data pre-processing, putative metabolite characterization (linking to METLIN and GOLM), datapretreatment, and a wide range of statistical analysis, with option for customization of parameters from the web interfaceitself. Bypassing the pipeline, the modular architecture of CCPM also allows inputting of externally preprocessed/pretreateddata, directly, to available analysis tools. In contrast with task size restrictions, associated with usual stateless or session-oriented web applications, CCPM supports long-running tasks featuring task completion alerts. Other features includesupport for handling data in all popular file formats, user friendly tools for on the fly data analysis and visualization, andprovision for publication quality image download.

PD235 (IPPC0910)

Comparative physiological basis of variability in germination, seedling vigour and vegetative vigour in

wheat and barley

Madhurima Das*, Rakesh Pandey, Vijay Paul and Attar Singh



Wheat and barley are two important cereal crops having similar phenological stages but different growth and developmentalbehavior. A comparative study on the physiological basis of variability was made in wheat and barley in terms of germinationrate, seedling growth, vegetative growth and its relation to cell wall properties (ferulic acid content), assimilate partitioningto roots, root distribution in soil profile. The study showed that the wheat genotype (HD2824) showed a faster germinationrate, due to its ability to maintain lower critical water potential of seed, higher β amylase activity, shorter duration of theIInd phage in triphasic imbibition curve than the barley genotype (RD2668). However, the seedling and vegetative vigourwas higher in barley than wheat. At 15-21 DAS in field, the seedling vigour of barley was greater as compared to wheatsimilarly, the vegetative vigour was greater in barley than wheat in relation to higher dry matter production, leaf area,specific leaf area, no of nodal root, root length. The higher specific leaf area was negatively associated with cell wallferulic acid, which helps in cell wall lignifications, thus limiting cell expansion. Root distribution study was done by trenchprofile wall method (1 m depth), showed a higher percentage of total root distribution in the upper soil horizon (0-30 cm)in barley as compared to wheat. The present study summarise that wheat genotype showed faster germination rate thanbarley genotype but the seedling and vegetative vigour was higher in barley as compared to wheat.


234

IPPCongress2015


PD236 (IPPC0970)

Interactome analyses in the stress associated signaling cascade: A case for the legume crop chickpea

(Cicer arietinum L.)

Savithri Purayannur*, Kamal Kumar, Vikas Srivastava, Shreenivas K. Singh and Praveen K. Verma



Plants have evolved many signaling cascades that relay multiple environmental and developmental signals thuscompensating their sessile nature, especially in harsh environments. The legume crop chickpea (Cicer arietinum L.) wastaken as a system to identify interactions among the components of MAPK signaling cascade and their targets. Weidentified 7 MAPKKs and 16 MAPKs from the chickpea genome. These gene families were classified into four groupseach, based on the model plant Arabidopsis classification. Many alternatively spliced forms of MAPKs were also identified,which increases the signaling repertoire. The MAPKs had 2, 5, 1 and 8 members in groups A, B, C and D respectively.The MAPKKs group A, B, C and D had 3, 1, 1 and 2 members respectively. The open entry clones of 7 MAPKK, 6MAPKKEE and 21 MAPKs were generated. To identify the interactions by split-ubiquit in based yeast-two hybrid (Y2H)system, we mobilized these ORFs into bait and prey destination vectors. Interaction analysis revealed many uniqueinteractions, some of which were isoform-specific. Our study thus highlighted the importance of alternative splicing inMAPK signaling by direct protein-protein interaction. Selected interactions among these members were confirmed in-planta by bimolecular fluorescence complementation (BiFC) analysis. We also studied the interaction of the MAPKs withthe selected downstream WRKY transcription factors. Therefore, we have generated useful resources in the form of entryclones, gateway compatible split-ubiquit in based Y2H vectors and the legume-specific MAPK cascade interaction data.In near future, the role of individual interactions in relaying the signals will be studied.

PD237 (IPPC0981)

Non-targeted metabolomics approach to decipher biochemical mechanism under drought tolerance in

rice (Oryza sativa. L)

Pushpa Doddaraju1*, M.D. Prathibha1, Sumant Kumar Kambalimath1, A.B. Priyanka1, H.G. Jalendra Kumar2,

H.V. Thulasiram2, M. Udayakumar1 and M.S. Sheshshayee1

1Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India2CSIR-Natioinal Chemical Laboratory, Pune-41008, India


Rice cultivation account for over 50% of water utilized in agriculture and hence increasing water productivity is paramountsignificance. Though aerobic cultivation can save up to 60% of irrigation water, concomitant yield loss of upto 40% isoften noticed is caused primarily due to an increased spikelet sterility. Cellular level tolerance (CLT) associated withproduction of compatible solutes and metabolites that reorganize proteins and cellular structures leading to maintenanceof leaf turgor, cellular redox balance and homeostasis. Comprehensive profiling of solutes and metabolites in rice genotypeswith varied level of drought tolerance and spikelet fertility would leaf to understanding the biochemical mechanisms at thecellular level and to identify genes responsible for high CLT under drought conditions. Fingerprint of biochemicalperturbations could also be a reliable method to phenotype CLT under stress for breeding drought tolerant genotypes. Inthis study, differentially accumulated metabolites among drought tolerant and susceptible rice genotypes under wellwatered and water stress were measured using high resolution LC-MS. Metabolites that accumulated with significantlyhigher abundance in drought tolerant rice genotype compared to susceptible genotype upon stress induction were identifiedas drought tolerance related induced (DTRI) metabolites. Such DTRI metabolites mainly belonged to primary metabolites:sugars, amino acids and secondary metabolites such as phenylproponoids, flavonoids and alkaloids. Primary metabolitesmainly involved in synthesis of stress responsive proteins and osmoprotection and secondary metabolites as antioxidants,reactive oxygen species (ROS) scavengers, coenzymes, UV and excess radiation screen and also as regulatory molecules.


235

IPPCongress2015


PD238 (IPPC1007)

Health benefits and therapeutic properties of dietary rice bran

Irfan A. Ghazi1*, Genevieve M. Forster2, Jan E. Leach3 and Elizabeth P. Ryan4

1Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India2Department of Clinical Sciences, 3Department of Bioagricultural Sciences and Pest Management, 4Department of Environmental andRadiological Health Sciences, Colorado State University, Fort Collins, Colorado-80523, USA


Rice is one of the most important cereal crops that feed more than half of the world population. Rice bran (RB) is adiscarded by-product of rice milling, which was earlier used for animal feed, contain a variety of health promotingcompounds. These compounds have chemotherapeutic, disease protective and several health beneficial properties.These properties include anti-oxidant, anti-cancer, anti-tumor and antidiabetic. RB from diverse varieties containsdifferent bioactive compounds, such as polyphenol, tricin, ferulic acid, carotenoids, vitamin E, lutein and essential aminoacids. RB Oil (RBO) is rich in gamma-oryzanol, phytosterol and sterolins. These compounds help in preventing oxidativedamage of DNA, damage of body tissue, high blood pressure, hyperglycemia, hyperlipidemia and stimulate our immunesystem and protects against UV radiations. It has been proposed by several groups that RB may be used as functionalfood as it has cholesterol lowering properties, anticancer activity and has cardiovascular health benefits. RB derivativesare also used for different types of dermatological and cosmetic applications. Dietary RB intake also induced non-specific gut mucosal immune responses in mice. We have investigated how RB improves intestinal health by increasinginnate resistance against enteric pathogens such as Salmonella that cause diarrhea in mouse model. RB from differentcolored and non-colored varieties can be used in breeding and transgenic to develop enteric disease protective traits inongoing global rice crop improvement programs. Dietary RB is a promising food, which can be further investigated for itsnutraceutical and therapeutic properties.

PD239 (IPPC1017)

Towards understanding the biosynthesis and regulation of withanolides in Withania somnifera

Dinesh A. Nagegowda1*, Anup Kumar Singh1, Varun Dwivedi1, Shaifali Pal2, Seema Meena1, Rajeev Kumar1 and

Ajit Kumar Shasany2*

1Plant Molecular Biology and Biotechnology Lab, CSIR-CIMAP Research Centre, Bangalore-560065, India2Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226015, India


Ashwagandha (Withania somnifera L.) accumulates unique class of triterpenoid steroidal-lactones collectively termedas withanolides, which have diverse biological activities. To enable fast and efficient functional characterization ofgenes in this slow-growing and difficult-to-transform plant, a virus-induced gene silencing (VIGS) was established andits utilization was validated by silencing the gene encoding squalene synthase (SQS), which provides precursors fortriterpenoids. VIGS of SQS resulted in significant reduction in squalene and withanolides, demonstrating its applicationin studying withanolides biosynthesis in W. somnifera leaves. Further, using VIGS approach a WRKY transcriptionfactor (WsWRKY1) involved in regulation of withanolides accumulation in W. somnifera, was characterized. WsWRKY1was induced by phytohormones, predominantly expressed in leaves, and localized to the nucleus. VIGS of WsWRKY1in W. somnifera resulted in stunted plant growth and reduced withanolides, which was brought about by negativeregulation of sterol pathway genes and reduced phytosterols. In contrast, transient over expression of WsWRKY1 in W.somnifera leaves up-regulated sterol pathway genes and enhanced the accumulation of withanolides. Moreover, ectopicexpression of WsWRKY1 in tobacco led to up-regulation of sterol pathway genes with significant increase in stigmasteroland campesterol. Furthermore, while WsWRKY1-silencing led to down-regulation of defense related genes resulting inreduced tolerance to bacterial growth, fungal infection, and insect feeding, its over-expression in tobacco led to improvedtolerance to biotic stresses. Our results demonstrated that WsWRKY1 could be a metabolic engineering tool to improvewithanolides in W. somnifera plants or cultures and to enhance phytosterols and biotic stress tolerance in plants.


236

IPPCongress2015


PD240 (IPPC1037)

Evaluation of flavouring compounds in local and processed potato products

Pinky Raigond*, Brajesh Singh, Alka Joshi and Som Dutt

Central Potato Research Institute, Shimla-171001, India


Market for processed potato products is rising day by day. Because of consumers preferences for better tasting foods,

product flavor plays an important role in decision making by the consumers. In potato and potato products, glutamic acid,

aspartic acid, guanosine 5’-monophosphate (GMP) and adenosine 5’-monophosphate (AMP) are the major umami

compounds which contribute towards its flavour. Therefore, umami 5’ nucleotides (AMP+GMP) were estimated from

local potato products (Samosa Fried, AlooTikki, Lachcha Fried, Lachcha Tokri and Aloo Seekh) as common fried

products in the Indian markets and processed potato products (LAYS- Indian Magic Masala, Classic Salted, Swiss

Grilled, Thai Sweet Chilli, American ASCO, Mccain- Potato Shortz, Masala Fries, Fries, Potato Chilli Bites, Smiles,

AlooBhujia, Dehydrated- Potato Chips, Crax, Lachcha and Samosa Baked) being sold by retailers. The frozen products

were fried before sampling. The analysis was done through HPLC using standardized protocol. Umami 5’ nucleotide

content ranged from 2.63 (Alooseekh) to 8.26 (fried lachcha) μg g-1 fr. wt. in local potato products. Along with aloo seekh,

fried samosa also contained low level of umami 5’ nucleotides, i.e., 4.42 μg g-1 fr. wt. In processed potato products, umami

5’ nucleotides ranged from 2.72 to 14.75 μg g-1 fr. wt. Umami 5’ nucleotides were the highest in aloo bhujia (14.75 μg g-

1 fr. wt.), followed by dehydrated aloo lachcha (11.14 μg g-1 fr. wt.) and dehydrated potato chips (10.13 μg g-1 fr. wt.).

Processed potato products such as McCain Smiles (2.72 μg g-1 fr. wt.), McCain potato shortz (3.40 μg g-1 fr. wt.) and

dehydrated potato crax (3.51 μg g-1 fr. wt.) contained low level of these compounds. The study suggests that the potato

products prepared solely from potato contained higher levels of umami 5’ nucleotides compared to other products

prepared by mixing potato with other cereals and vegetables.

Session E : Biotic Stress Management and Interactions

237

IPPCongress2015


PE241 (IPPC0064)

Exploring a putative plant defensin from Rorippa indica and its integration into Brassica juncea for

enhanced pest management

Poulami Sarkar* and Samir Ranjan Sikdar

Division of Plant Biology, Bose Institute, P-1/12, C.I.T. Scheme VII M, Kolkata- 700054, West Bengal, India


Mustard aphid - Lipaphis erysimi causes severe damage to the crop Brassica juncea not only by sucking the phloem sap

but also by transmitting several disease causing viruses leading to an eventual yield loss from 35.4% to 91.3% annually.

Previously, Rorippa indica - a wild crucifer was identified as aphid tolerant by the present group. To understand the

tolerance mechanism of Rorippa indica - L. erysimi incompatible interaction, cDNA amplified fragment length polymorphism

(cDNA AFLP) analysis was carried out in which a putative EST homologous to PDF1.2c - a defensin from Arabidopsis

was identified. In the present study differential expression of this significantly upregulated EST has been studied using

real time PCR analysis. The full length gene corresponding to the EST has been isolated and cloned from R. indica and

named as Rorippa indica defensin (RiD). L. erysimi mortality in artificial diet based insect-bioassay with the in vitro

expressed RiD indicated that the gene can be deployed for developing aphid tolerance in transgenic crops. The full length

coding sequence of RiD was finally cloned into plant transformation vector pCAMBIA 1301. The pCAMBIA 1301 - RiD

gene construct has been introduced into susceptible cultivar of B. juncea by using Agrobacterium-mediated plant

transformation protocol with an aim to develop aphid tolerance. Molecular analyses of the putative B. juncea transformants

are underway. B. juncea lines with successful integration and expression of RiD might provide enhanced aphid tolerance

resulting in better pest management.

PE242 (IPPC0102)

Potentials of Parthenium to control root knot nematode in cowpea

Nisha Raghava*, Ravindra Pratap Raghava and Jayanti Srivastava

FIST-DST Department of Botany, S.M.M. Town (PG) College, Ballia-277001, U.P., India


Cowpea (Vigna unguiculata (L.) Walp.) is a major pulse crops commonly used by Indians in a different ways as food and

feed to cattle. It is most important crops as fodder and for green vegetable being rich in protein at large amount and many

other nutrients. The roots of cowpea plants are severely galled by Root-Knot Nematode (RKN) at different stages of

growth. Due to RKN formation, the growth of nodules is greatly affected and it causes damage to the plant growth. RKN

are hidden enemies of crop and represent a unique challenge to agricultural research. Plant-parasitic nematode causes

serious economic losses for many important agricultural crops and effective management are essential for sustaining

modern agricultural productivity. Management techniques have relied on chemical control measures since their development

during the middle of the last century. Regulatory restriction on the use of conventional nematicides now known to be

human health and environmental hazards, necessitate the identification and evaluation of alternative non-chemical

management strategies. The allelopathic potentials of Parthenium hysterophorus (L.) family Asteraceae have been utilized

to control root knot nematodes in cowpea. The aqueous extracts of different parts of Parthenium plant were inhibitory to

the number of eggs and active adult nematodes. The data collected on number of galls developed on the root surface

showed remarkable reduction with different concentrations of Parthenium extracts, when sprayed at three stages of

growth.


238

IPPCongress2015


PE243 (IPPC0188)

Induced defense responses in chilli pepper (Capsicum annum L.) against anthracnose pathogen

Colletotrichum gloeosporoides

Rukmini Mishra*, Ellojita Rout, Satyabrata Nanda and Raj Kumar Joshi

Centre of Biotechnology, Siksha ‘O’ Anusandhan University, Bhubaneswar-751003, Odisha, India


Anthracnose, caused by Colletotrichum gloeosporoides is one of the most important diseases of Capsicum worldwide. Inthe present study, expression profiles of nineteen defense responsive genes were compared temporally during compatibleand incompatible interaction with C. gloeosporoides in two capsicum lines. Quantitative real time PCR revealed thatjasmonic acid (JA) and ethylene (ET) biosynthetic pathway related genes such as Plant defensin (PDF1.3), Lypoxygenase1(Lox1), Allene oxide synthase (AOS) and ACC synthase 2 (ACS2) were rapidly induced and to a significantly higher levelin the resistant line Bhoot Jolokia than in the susceptible cultivar Teja Jhal in response to anthracnose infection. Whilesalicylic acid (SA) related gene phenylalanine ammonia-lyase (PAL) showed significant upregulation under incompatibleinteraction, isochorismate synthase (ICS2) showed similar expression for both genotypes. Further, three pathogenesisrelated proteins PR1, Chitinase (ChiIV) and PR5 were moderately responsive and showed similar expression patterns atall time points in both the genotypes. However, β-glucanase was highly expressive during compatible interaction.Additionally, transcription factors associated with disease resistance such as WRKY, MYB, ERF and bZIP were remarkablyinduced in the resistant genotypes as early as 3 dpi and remained significantly high at 9 dpi as compared to susceptiblegenotype. Our results indicated that capsicum adopts multiple strategies by activating several genes from various defenseresponse pathways that play considerable role in resistance to anthracnose. The present study represents a valuableresource for future functional genomics experiments to unravel the molecular mechanisms of capsicum defense againstC. gloeosporoides.

PE244 (IPPC0197)

Parthenium leaf allelochemical preparation as an herbicide for noxious submerged aquatic weed hydrilla

(Hydrilla verticilata L. f. Royle), a weed against weed

Daya K. Pandey*

Principal Scientist (Plant Physiology), ICAR-Directorate of Weed Research, Jabalpur-482004, India


Parthenium (Parthenium hysterophorus L.), one of the world’s worst weeds, continues to spread throughout the countryaffecting human and animal health, agriculture, environment and natural biodiversity with tremendous socio-economicimplications. Phenolics, terpenoids and other constituents in it owe the species a range of biological activities includingtoxic and allelochemic interactions. Hydrilla (Hydrilla verticillata L. f. Royle) is a prominent submerged ubiquitous aquaticweed of waters with wide range of implications on the ecosystems and their services. Present study explores an herbicidalpreparation from parthenium leaf for management of hydrilla. Parthenium leaf allelochemical herbicidal preparation (PLAHP)comprised of acetone water mixture extract of shade dried leaf powder. The PLAHP had about 12% parthenin and 4%phenolics. Majority of these were released instantly into the medium on suspending in water. Pre-weighed hydrilla plantswere placed in suspensions of the PLAHP in 20% (v/v) nutrient medium. Biomass, toxicity, phenolics and terpenoids inthe medium and herbicidal mechanism of action of the preparation were monitored. The experimental results were verifiedthrice by using bulk volume (40 litres) outdoors. The PLAHP was inhibitory at 400–500 ppm and herbicidal at 500-1000ppm, killing the treated hydrilla plants in 5-10 days by involving fragmentation, loss of pigments and massive damage tocellular membranes, with a concurrent rapid dissipation of phenolics and parthenin in the medium. The herbicidal actionof PLAHP on hydrilla opens up avenues for management of the weed as an eco-friendly approach by utilizing a weed likeparthenium.


239

IPPCongress2015


PE245 (IPPC0217)

Concurrent overexpression of OsGS1.1 and OsGS2 for environmental stress resistance and herbicide

tolerance in rice (Oryza sativa cv. japonica)

Donald James*, Dhirendra Fartyal, Mrinalini Manna, Aakrati Agarwal, Baburam Verma, Bhabesh Borphukan, Mohan

Achary and M.K. Reddy

International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-1100167, India


Glutamine synthetase (GS; EC 6.3.1.2) is a key enzyme in the GS-GOGAT (Glutamine synthetase-Glutamate synthase)pathway controlling nitrogen metabolism in plants; it catalyzes the critical incorporation of inorganic ammonium intoglutamine. Glutamine synthetase exists primarily as two isozymes with different subcellular localizations: GS1 in thecytosol and GS2 in chloroplasts/plastids. Several reports have implicated both cytosolic and chloroplastic GS isozymesin conferring resistance to various abiotic stresses in many crop species. Furthermore, glutamine synthetase is the targetenzyme for commonly used non-selective herbicide Glufosinate/BASTA (active ingredient phosphinothricin; PPT). Thisstudy discusses the combined effect of constitutive overexpression of both OsGS1;1 and OsGS2 in rice with respect totolerance to various abiotic stresses and resistance to BASTA. Two full-length cDNAs that encode the rice cytosolicglutamine synthetase1 protein (OsGS1;1) and the chloroplastic glutamine synthetase2 protein (OsGS2) were isolatedfrom rice leaf tissue. The GS genes (OsGS1;1 and OsGS2) were cloned under rice actin constitutive promoters andpyramided invitro (stacked) into pMDC99 plant transformation vector via Gateway® technology. The OsGS1;1 andOsGS2 overexpressing rice (Oryza sativa cv japonica) transgenics were generated using Agrobacterium tumefaciens-EHA105 strain mediated transformation using hygromycin as a selectable marker. The putative transgenic plants werescreened for transgene integration by PCR using hygromycin resistance gene (hptII) primers and further by Southernblotting. The transgenic plants exhibited limited tolerance to BASTA herbicide (Glufosinate/PPT) compared to wild-typeplants as assayed by BASTA leaf paint test. The transgenic plants also showed considerable reduction in ammoniumaccumulation as determined by modified Berthelot assay.

PE246 (IPPC0244)

Triple herbicide tolerant transgenic indica rice for combating weed problems

Dhirendra Fartyal*, Aakrati Agarwal, Donald James, Mrinalini Manna, V. Baburam, Mohan Achary and M.K. Reddy

International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India


Weeds are major biotic constraints in Indian as well as world agriculture systems. In addition to competition for nutrientsand water, sometimes, they are alternate hosts for different kinds of pests and pathogens, which ultimately results inreduced crop production. So, it is necessary to eliminate weeds from the field to obtain higher food production. Sincemanual weed removal is expensive in terms of time, labour and money, it seems logical to use chemicals/herbicideswhich are very effective for weed destruction. But the use of a single herbicide continuously makes the weeds resistant tothat particular herbicide in the long term. Hence, it is always beneficial to use different herbicides alternately. Keeping allthese things in mind, we have developed triple herbicide tolerant indica rice (cv. Swarna). Three genes viz. EPSPS, ALSand BAR, along with a supporting gene igrA, were integrated to impart resistance to herbicides glyphosate, sulphonylurea and glufosinate respectively. The igrA gene also helps to detoxify glyphosate along with EPSPS. All these geneswere stacked in pMDC99 binary vector and transformed into EHA105 Agrobacterium strain. The rice calli were infectedwith these Agrobacterium cells and the transformed calli were selected with hygromycin selection. The selected calliwere transferred to regeneration media and then rooting media to develop the transgenic plants. The putative transgenicplants were screened with PCR and southern blot and the confirmed transgenic plants were further subjected to respectiveherbicide assays.


240

IPPCongress2015


PE247 (IPPC0251)

Targeted mutagenesis of EPSPS gene Using CRISPR/Cas9 platform for the development of Glyphosat-

resistant rice

Bhabesh Borphukan* and V. Mohan Achary, Vijay Reddy, James Donald, Baburam Verma and M.K. Reddy

Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


Development of crops for withstanding marginal lands, resistant/tolerant for different biotic and abiotic stresses and bio-

fortification for nutritional enhancement are the major area of concerned for food security in the coming years. Crop

improvement through transgenic approaches has been arrested due to the continuous criticism on different ethical issues

whereas cost of time and labour has to pay for conventional breeding. Recently CRISPR/Cas9 system has been emerged

as a powerful tool for targeted genome editing. It provides opportunities for unprecedented manipulation of genetic

materials either through gene knock out or knock in via non-homologous end joining (NHEJ) or homologous directed

repair (HDR) pathway, respectively. CRISPR/Cas9 based HDR system provides a better platform for creating or changing

phenotype in a desired way by altering specific DNA sequences, successful application of which will outclass both transgenic

and conventional breeding approaches. Here we have developed Cas9 expressing rice platform under the control of Zm

ubiquitin promoter. This platform was further used for creating T169I and P173S mutation through homologous

recombination in the native OsEPSPS to make glyphosate tolerant indica rice variety Satabdi. Successful demonstration

of Cas9 based HDR mediated genome editing promise efficient and precise engineering of crop plants in near future

through editing native gene or regulatory elements which also assure significant improvement of crop genetic material in

a cisgenic approaches which could be accepted more by the consumers in near future.

PE248 (IPPC0279)

Tolerance assessment of transgenic Brassica overexpresing MPK3 gene against Alternaria blight

Mohd Hassan1, Mohd Tasleem1, Dinesh Pandey1, Anil Kumar1, Vandna A. Kumar2 and Gohar Taj1*

1Depatment of Molecular Biology and Genetic Engineering, 2Department of Biochemistry, G.B. Pant University of Agriculture andTechnology Pantnagar-263145, India*Presenting author: [email protected]

The potential yield of Brassica in India is very high but there is a wide gap between the potential and realized yields,

which is largely because of the biotic and abiotic stresses. Alternaria blight is the most important diseases of rape seed

mustard that leads to major yield losses as well as deterioration in quality of oilseeds. Therefore, in the present study,

transgenic Brassica (BjV2) plant over-expressing MPK3 gene was studied to assess tolerance against Alternaria blight.

Lower disease index and higher antioxidant content found in the transgenic plant (BjV2) as compared to wild Brassica

suggests its tolerant capability to Alternaria blight disease. The similar expression pattern of MPKK9 and MPK3 suggests

that MPKK9 might be an upstream activator of MPK3 and work in a direct cascade for the production of defense related

proteins. MPKK5 was found to support the colonization of the pathogen, as the wild plant showed more expression than

the tolerant transgenic (BjV2) plant. The expression of WRKY transcription factors was also studied, and revealed that

WRKY33 acts downstream of MPK3 in activating defense related genes. With lower disease index, higher ROS scavenging

enzymes content the transgenic Brassica (BjV2) was found to be tolerant to the pathogen Alternaria brassicae.


241

IPPCongress2015


PE249 (IPPC0306)

Identification of Bacillus sp. from algae culture system and its effect on growth of Desmodesmus

ultrasquamatus

Yash Gupte*, Ganesh Iyer, Suhas Pednekar, Vinod Nagle, Sailee Lavekar and Pooja Devadiga

Ramanarain Ruia College, Matunga, Mumbai-400019, Maharashtra, India


Bacillus sp. was isolated from algal culture systems. Presence of bacteria in algal culture system may be a mutually

benefitting relationship. Bacillus being a facultative anaerobe can grow well in the oxygen rich water of algae and in turn

help algae to continue photosynthesis. It often forms a film over the surface of water if the media is not mixed thoroughly.

The gram character was studied to identify the bacterium morphologically as Bacillus coagulans. PCR amplification of

16s rDNA gene was carried out and the sequence was analyzed using BLAST for molecular identification of the bacterium.

Bacillus sp. was found to be resistant to Ampicillin, Kanamycin and Neomycin which may be due to the endospore

forming capacity and change of gram character when passing into stationary phase. The associative growth of algae and

bacteria was found to enhance algal growth by 7-9%, with no difference in the biochemical characters of algae

Desmodesmus ultrasquamatus. Gas Chromatography Mass Spectroscopy (GC-MS) revealed very little change in the

fatty acid profile of the Desmodesmus ultrasquamatus.

PE250 (IPPC0317)

Host-induced gene silencing in tomato and chilli against anthracnose disease caused by Colletotrichum

gloeosporioides

Binod Kumar Mahto* and Swatismita Dhar

The Energy and Resources and Institute (TERI) University, Vasant Kunj, New Delhi-110070, India


Anthracnose disease is caused by ascomycetes fungus, Colletotrichum spp. which is responsible for heavy yield loss for

tomato and chilli production around the worldwide. We are reporting the expression of RNA interference (RNAi) sequences

derived from an essential Colletotrichum gloeosporioides virulence gene, Conidial Morphology 1 (CgCOM1) involved in

fungal conidial and appresorium development, as a method to enhance resistance of tomato (var. Pusa Rohini and Pusa

Early Dwarf) and chilli (var. Pusa Jwala and Pusa Sadabahar) plants against Colletotrichum sp. We have developed

transgenic lines of tomato (Solanum lycopersicum) and chilli (Capsicum annuum) using RNAi construct containing pathogen

gene by Agrobacterium-mediated stable transformation. We have already standardized the Agrobacterium-mediated stable

transformation protocol in tomato and chilli with pCAMBIA2301 vector. We show that HIGS (Host-Induced Gene Silencing)

can be used to silence expression of pathogen genes involved in general function which can confer resistance against

fungal pathogens.


242

IPPCongress2015


PE251 (IPPC0333)

Study on biotic stress interaction on application of a potential herbicidal fungus, Gibbago trianthemae

Pinki Bhandari*, Meenakshi Agarwal and S.K. Raza

Bioscience Division, Institute of Pesticide Formulation Technology, Gurgaon-122016, India


Gibbago trianthemae is a fungus, with potential bioherbicidal activity when applied at high densities onto weed, viz.,

Trianthema, and showed 70-75% weed control. The present study revealed that Gebbago trianthemae working as phyto-

pathogen fungi via selective host specific phytotoxine significantly affect only the plant species Trianthema portulacastrum

of family Aizoaceae. Under biotic stress there is production of singlet oxygen and other photodynamic compounds, such

as hypericin. Plant pathogens produce a variety of phytotoxins that interfere with plant metabolism, ranging from subtle

effects on gene expression to plant mortality. Solutions of Gibbago trianthemae, diluted to different concentrations were

applied in the field with three replications. Soil samples were collected from 7 consecutive depths in (0-15 cm, 15-30 cm,

30-45 cm, 45-60, 60-75 cm, 75-90 cm, 90-120 cm) of soil profile using 5 cm diameter soil core. The soil samples were

enumerated for different group of micro-organisms using different growth media. Collected soil samples diluted and

spread on medium plates, and incubated for 24-48 h, and the growth of micro organism was calculated. It was found that

the Gibbago trianthemae has non-significant effect on soil bacteria and actinomycetes growth.

PE252 (IPPC0344)

Effect of mycoherbicide fungus Gibbago trianthemae for the control of carpet weed & its impact on

certain rabi crops

Meenakshi Agarwal*, P. Bhandari and S.K. Raza

Bioscience Division, Institute of Pesticide Formulation Technology, Gurgaon-122016, India


The recent years there has been a increasing interest in mycoherbicide fungi caused by factors such as increasing

insecticide resistance and environmental concerns over pesticide use. The changing environmental conditions and

increasing demand & consumption of chemical insecticide has created biotic stress in overall agro ecosystem of major

economics crop viz. In present study Gibbago trianthemae simmons a phaeodictyoconidal hyphomycetous fungus reported

on carpet weed, causes leaf spot disease resulting in premature defolation. In present study, a highly virulent fungal

pathogen strain. Gibbago trianthemae was screened out on the basis of its pathogenic potential to develop sporulating

lesion on horse purslane. Seedling/seeds of major rabi crops were planted in the plots. It is liquid suspension formulation

with 1x106 spores ml-1 was found to be most pathogenic on horse purslane. These crops were sprayed with the Gibbago

trianthemae suspensions. Observation on disease incidence and plant vigour were recorded after 5,10,15 and 30 days

after application. There is no disease incidence of Gibbago trianthemae on rabi crops. In host range studied Gibbago was

found pathogenic only to horse purslane and there was no phytotoxicity and any other biochemical change observed on

the main crop viz. cabbage, cauliflower, tomato, chilli, fenugreek, maize, mustard, coriander. It was found that the Gibbago

trianthemae has no significant effect on soil bacterial and actinomycetes growth i.e. (4.44 x 105 CFU/ml) and (3.61x

105 CFU/ml) respectively. Whereas a significant effect on soil fungi at 1.87 x 105 CFU/ml it was also negligible but significant

effect.


243

IPPCongress2015


PE253 (IPPC0431)

Understanding the molecular intricacies of Rice-Rhizoctonia solani interactions

Srayan Ghosh*, Poonam Kanwar and Gopaljee Jha



Rhizoctonia solani is a necrotrophic soil borne pathogen causing sheath blight disease in rice and is responsible for huge

crop loss every year worldwide. In spite of global efforts, till now there have been no reports of even a single R gene

identified against this pathogen. Hence understanding the molecular intricacies of Rice-Rhizoctonia is required to device

better strategies to control this important disease. While mining the available genome sequences of rice and lettuce

isolate of R. solani belonging to AG1-IA and AG1-IB anastomosis groups, respectively we have identified several genes

that are exclusively present in AG1-IA. Many of them encode important biological, molecular functions and exhibit dynamic

expression during in-planta growth of the pathogen in rice. Based upon sequence similarity with genes that are required

for plant and human/zoonotic diseases (PHIbase database), several putative virulence/pathogenicity determinants amongst

AG1-IA specific genes were identified. Further the differentially expressed pathogen and host genes during Rice-Rhizoctonia

interactions have been catalogued while analysing the RNAseq data. We endeavour to understand how R. solani alters

the host’s physiological environment to create a suitable niche to establish itself and cause necrosis.

PE254 (IPPC0442)

The wheat resistance gene Lr34 results in the constitutive induction of multiple defense pathways in

transgenic barley

Harsh Chauhan1*, Rainer Boni2, Liselotte Selter2, Thomas Wicker2, Simon Krattinger2 and Beat Keller2

1Indian Institute of Technology (IIT) Roorkee-247667, Uttarakhand, India2University of Zurich, Switzerland


The wheat gene Lr34 (Yr18/Pm38/Sr57/Ltn1) encodes a putative ABCG-type of transporter and is a unique source of

disease resistance providing durable and partial resistance against multiple fungal pathogens and has been used in

modern wheat breeding for more than 100 years. Lr34 has been found to be functional as a transgene in barley but its

mode of action and biochemical function remained largely unknown both in wheat and barley. Here, we studied gene

expression in uninfected barley lines transgenic for the Lr34 gene. Multiple defense pathways contributing to basal as

well as inducible disease responses such as lignin synthesis, phytoalexin biosynthesis and pathogenesis-related protein

genes were constitutively active both in seedling and mature leaves. In addition, the hormones jasmonic acid and salicylic

acid were induced to high levels in the presence of Lr34. We found that in barley the deletion of a single phenylalanine

residue was sufficient to induce the characteristic Lr34-based responses. Expression of Lr34res alone resulted in a

severe reduction of plant vigor in transgenic barley. These negative effects of Lr34res on barley growth were attenuated

in the presence of the susceptible Lr34sus allele, whereas resistance function was not compromised. These results

demonstrate a strong re-programming of plant metabolism by Lr34 even in the absence of the pathogen. Interestingly,

the negative effects on growth resulting from defense gene induction can be attenuated genetically, suggesting that a

transgenic combination of Lr34res and Lr34sus in heterologous grass species might result in agronomically useful

resistance.


244

IPPCongress2015


PE255 (IPPC0479)

Tomato miR159-SlMyb33 interface modulates R gene activity to control virus infection

Pranav Pankaj Sahu, Manoj Prasad and Namisha Sharma*

National Institute of Plant Genome Research, Aruna Asif Ali Marg, New Delhi-110067, India


Tomato Leaf Curl New Delhi Virus (ToLCNDV), a bipartite geminivirus, causes Tomato leaf curl disease and adverselyaffects the horticultural economy. Natural defense system against geminiviruses includes the phenomenon of RNAinterference. To study pivotal role of miRNAs in plant defense against ToLCNDV, two small RNA libraries were constructedin mock treated and ToLCNDV infected cultivar H-88-78-1. Some of the identified known and novel miRNAs were analyzedby northern blot for their differential expression in two contrasting cultivars (tolerant cv. H-88-78-1 and susceptible cv.Punjab Chhuhara) of tomato, among which 13 miRNAs showed differential expression in both the cultivars. After viralinfection down-regulated expression was observed for sly-miR159 in tolerant cultivar and showed inverse relationshipwith its target SlMyb33, which was also validated by RLM RACE. Silencing of SlMyb33 in cv. H-88-78-1 was performed todetermine the plausible role of SlMyb33 during plant-virus interaction, which revealed that ToLCNDV-infectivity and viraltiter were significantly increased in silenced line in comparison to cv. H-88-78-1 innate state. Promoter binding characteristicof known virus resistant genes was further explored for Myb33 binding sites. In silico study of promoters of eight resistantgenes revealed that three of them (N gene, Sw-5, and Tm2) have Myb33 binding sites. Further, interaction betweenMyb33 and promoter of differentially regulated R gene was validated through EMSA. Till now, miR159 have been shownto be associated with the symptom development upon virus infection, we hypothesize that upon viral infection reducedexpression of sly-miR159 leads to higher accumulation of its target SlMyb33, which in turn activate the transcription of theR gene and provide tolerance against ToLCNDV.

PE256 (IPPC0504)

Functional involvement of a group C mitogen activated protein kinase, OsMPK7 in mediating resistance

against Xanthomonas oryzae in rice

Siddhi Jalmi* and Alok Sinha



Mitogen-activated protein kinases (MAPKs) are highly conserved signaling cascades in eukaryotes, involved in transducingstress related signals from upstream receptor to downstream target by a phosphorelay mechanism. In this study the roleof OsMPK7 is assessed during Xanthomonas oryzae infection, a causal agent of leaf blight disease in rice. X. oryzaetreatment resulted in induction of transcripts of OsMPK7 and OsMKK3, as well as activation of OsMPK7. Further, OsMPK7was found to be target of OsMKK3 and the two proteins interacted in-planta. Overexpression of OsMPK7 and OsMKK3,individually and in combinations resulted in inhibition of disease symptoms caused by X. oryzae, both in leaf and root.Furthermore, the mechanism for disease resistance was exemplified by studying relative expression of defence relatedgenes in rice overexpressing OsMKK3-OsMPK7 module. This revealed that overexpression of OsMKK3-OsMPK7upregulated pathogenesis-related genes and genes involved in cell wall metabolism. These leaves also showed restrictedmovement of the pathogen from the point of infection to other uninfected part. To explore downstream target of OsMPK7involved in mediating X. oryzae related cues, protein-protein interaction studies were performed. OsWRKY30 was identifiedas a putative downstream target of OsMPK7. Taken together, this work suggests a positive involvement of OsMKK3-OsMPK7 module in imparting disease resistance against X. oryzae infection in rice. Further, the role of OsWRKY30 inOsMKK3-OsMPK7 mediated signaling in response to X. oryzae pathogen will be evaluated.


245

IPPCongress2015


PE257 (IPPC0523)

A DNA satellite encoded protein interferes with structure and function of chloroplast leading to

photosynthetic inhibition

Prabu Gnanasekaran*, Dhriti Bhattacharyya, Reddy Kishore Kumar, Nirbhay Kumar Kushwaha, Veerendra Kumar Sharma

and Supriya Chakraborty



Gemini viruses are known for their potency to interfere with plant’s physiology. Recently betasatellite, a group of DNA

satellite has emerged as a major threat to crop production worldwide and acts as a key regulator for symptom induction.

To study host-betasatellite interactions and symptom development, we used Nicotiana benthamiana - Radish leaf curl

betasatellite (RaLCB) as model system. RaLCB initiates leaf curling, vein clearing and severe veinal chlorosis in infected

plants; however, in the absence of βC1 protein, RaLCB failed to induce vein clearing symptoms on host plant. In the

course of infection, βC1 protein localizes into chloroplasts of the host cells and damages chloroplast ultrastructure. In

addition, the betasatellite downregulates expression of genes involved in chlorophyll biosynthesis, chloroplast development

and plastid translocation. Interestingly, the expression of key host genes involved in chlorophyll degradation remains

unaffected indicating selective suppression of host metabolism. Betasatellite infection drastically reduces the numbers of

active reaction centres and plastoquinol pool size in leaves exhibiting vein clearing symptom. Betasatellite mediated

impediments at different levels of chloroplast functionality affects photosynthetic efficiency of N. benthamiana. This study

provides the first evidence of chloroplast localization of a DNA virus encoded protein which in turn affects photosynthesis.

PE258 (IPPC0529)

Identification and characterization of a novel mycophagous bacterium against Rhizoctonia solani, causal

agent of sheath blight disease of rice

Isha Tyagi*, Rajeev Kumar and Gopaljee Jha



Bacterial Mycophagy is a set of phenotypic behaviours that enable bacteria to obtain nutrients from living fungi allowing

conversion of fungal biomass into bacterial biomass. We have isolated a novel yellow-pigmented bacterium from surface

sterilized healthy rice seedlings demonstrating mycophagous behaviour against Rhizoctonia solani (the causal of sheath

blight disease of rice), and broad spectrum antifungal activity against major phytopathogens such as Magnaporthe oryzae

(the rice blast pathogen), Venturia inaequalis (the apple scab pathogen), Fusarium oxysporum (vascular wilt pathogen),

and several other fungal pathogens. 16S rRNA gene sequencing, deep phylogeny, and segregation rooting analyses

identified the bacterium to be Burkholderia gladioli, and henceforth, the strain was named Burkholderia gladioli NGJ1.

The B. gladioli NGJ1’s mycophagous behaviour against R. solani is shown by bacterial growth over fungi, weight loss and

cell death assays. The R. solani on co-inoculation with B. gladioli NGJ1 could not cause disease on rice tillers and tomato

leaves thus suggesting that mycophagous phenomenon could be used as a strong biocontrol agent for sheath blight

disease of rice by R. solani and several other economic important plant diseases. The draft genome sequence of B.

gladioli NGJ1 has been unravelled and the bacterium is enriched in genes encoding many fungal cell wall degrading

enzymes and several novel antibiotics.


246

IPPCongress2015


PE259 (IPPC0532)

Interaction of Allium sativum leaf agglutinin (ASAL) with GroEL related receptor interferes with whitefly

mediated tomato leaf curl New Delhi virus (ToLCNDV) transmission

Ayan Das1*, Amit Roy2, Arunava Mandal1, Pallob Kundu1 and Sampa Das1

1Division of Plant Biology, Bose Institute, P 1/12 C. I. T Scheme VII M, Kolkata- 700054, West Bengal, India2Department of Plant Protection Biology, SLU, PO 102, SE-230 53 Alnarp, Sweden


Allium sativum leaf agglutinin (ASAL), a mannose binding lectin has been established as a promising insecticidal component.Its efficacy was checked on an array of hemipteran insects both in artificial diet based and in planta bioassay on ASALtransgenic plants. A unique receptor of ASAL was previously identified by the present group in brush border membranevesicle (BBMV) of mustard aphid (Lipaphis erysimi) which was found to be GroEL related chaperonin protein- symbionin.GroEL has been reported to play role in persistent, circulative transmission of plant viruses. Both in silico and in plantaexperiments proved the plausible role of ASAL in interference with aphid mediated viral transmission. In the present worksimilar GroEL receptor of ASAL was identified through LC MS/ MS analysis in the BBMV of whitefly, the widely knownvector for several economically important plant viruses including tomato leaf curl New Delhi virus (ToLCNDV). Ligand blotanalyses demonstrates- when whiteflies are pre-fed with ASAL it completely blocks the binding of the ToLCNDV coatprotein to GroEL. Co-localisation of ToLCNDV coat protein and GroEL was detected in whitefly through confocal microscopicanalyses but ToLCNDV coat protein did not co-localise when insects were prior fed with ASAL. Immunoprecipitationfollowed by western analyses also showed that when whiteflies are pre-fed on ASAL supplemented diet it interferes withthe binding of ToLCNDV coat protein to GroEL. These results confirm that ASAL’s interaction with GroEL receptor interfereswith binding of ToLCNDV coat protein and subsequent whitefly mediated viral transmission.

PE260 (IPPC0535)

Drought stress endures Arabidopsis thaliana plants to subsequent Pseudomonas syringae infection

due to priming of basal defence responses

Aarti Gupta*, Sandeep Dixit and Muthappa Senthil-Kumar

National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India


The global climate change is predicted to cause frequent incidences of concurrent abiotic and biotic stresses. Recentstudies indicate that plant responses are adaptively tailored for combating the concurrent stresses, which are otherwisenot elicited under individual stress conditions. Drought potentially influences the occurrence and intensity of a particulardisease by modulating the plant-pathogen interaction. Thus understanding the plant response towards concurrent droughtand pathogen stressors is important. In order to facilitate such studies an accurate protocol for concurrent imposition ofdrought and pathogen is warranted. The prolonged and gradual nature of drought stress imposition protocol poses ahindrance in overlaying with pathogen infection in laboratory conditions to achieve combined stress imposition. Till datean accurate and convenient methodology is lacking, even in model plants like Arabidopsis thaliana. Here wecomprehensively studied impact of drought stress on A. thaliana-Pseudomonas syringae pv. tomato DC3000 interaction.In this study we established a systematic protocol to study the impact of combined drought and bacterial pathogen stress.We also attempted to dissect the contribution of individual stress effect in this stress combination and showed that plantresponses under combined stress are skewed towards the most prominent stressor, namely drought. We also observedthat physiological responses under combined stress are tailored when compared to the plants exposed to individualstresses. Further, our results showed that drought stressed A. thaliana plants better defend oncoming pathogen infection.Both the developed protocol and results from the stress interaction are presented.


247

IPPCongress2015


PE261 (IPPC0537)

Identification of differentially expressed genes under combined drought and Ascochyta rabie infection

in chickpea plants using meta-analysis and their functional validation

Prachi Pandey*, Anjugam Paramanantham, Praveen Verma and Muthappa Senthil-Kumar



Under field conditions, plants are simultaneously exposed to a number of abiotic and biotic stress conditions. Plants’

response to combined stress consists of a combination of shared and unique responses. Whereas shared responses are

the responses common to the individual and combined stress conditions, unique responses are characteristically seen

under combined stress conditions. The transcriptomic information available from the large number of individual abiotic

and biotic stress related microarray datasets can provide valuable insights into the shared responses under the combined

stress conditions. The present study aimed at understanding the shared molecular response of Cicer arietinum under

combined drought and Ascochyta rabiei infection by meta-analysis of microarray data obtained from individual drought

and A. rabiei infection experiments. The study identified a list of differentially expressed genes in chickpea plants which

were common under drought and a fungal pathogen infection. Among all the commonly regulated genes, those encoding

for pathogenesis related factor (PR4), carboxytransferase, linoleate 9 S-lipoxygenase like protein were some of the

commonly upregulated genes. On the other hand, genes encoding for phosphate induced protein, proline oxidase and

photosystem reaction centre proteins were found to be commonly downregulated under drought stress and A. rabiei

infection. Data for the functional validation of few of these genes will be presented. Overall this study presents validation

of the meta-analysis approach and conclude that re-analysis of the transcriptomic data of plants under individual stresses

can provide insights into the shared mechanisms behind tolerance to combined drought and A. rabie infection in Cicer

arietinum.

PE262 (IPPC0543)

Identification and comparative analysis of RNAi suppression activity of different viral proteins

Abhishek Anand*, Sudhanshu Sekhar, Veena Sinha and Neeti Sanan-Mishra

International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India


RNA silencing suppressors have emerged as a relatively new challenge to plant defense system against viruses. The

viruses neutralize the plants defense strategy of RNA silencing by using the suppressor proteins. These proteins are

already present inside the virus, performing their vital functions and have possibly developed the suppression activity

during later stages of evolution.This explains the fact that they do not share any evolutionary homology or pattern. They

are diverse not only in sequence but also in the ways they suppress the silencing machinery. So it has become difficult to

predict any pattern based on their nucleotide or protein sequence or structure to identify and classify the suppressor

proteins. Here we present a simple, quick and reliable strategy to identify a protein with suppressor function. This assay

was used to identify RNAi suppressors from diverse viruses. It is now emerging that a single virus may contain multiple

suppressors, each with a different mechanism of action. This assay can also quantitate the strength of suppression. The

knowledge gained will complement the studies on their mechanism of suppressor action to understand the strategy

adopted by the viruses to evade the host RNAi defense.


248

IPPCongress2015


PE263 (IPPC0546)

Molecular and physiological studies reveal complexities of rice Rhizoctonia solani interactions

Poonam Kanwar*, Srayan Ghosh and Gopaljee Jha



Rice, one of the world’s most important crops faces severe yield loss, due to sheath blight disease caused by a necrotrophic

fungal pathogen Rhizoctonia solani. No source of complete disease resistance is known, till date. We adopted a multi-

pronged approach to study the intricacies of plant pathogen interaction. The interaction transcriptome was explored to

understand the dynamic interplay between host and pathogen genes and processes during infections. The alteration in

photosynthesis, carbohydrate metabolism and induction of cell death were the most prominent host responses. This was

further validated by physiological and biochemical analysis. Collectively, both transcriptome and functional assays revealed

that R. solani causes extensive metabolic re-programming inside the susceptible host. In conclusion, the present study

provides better understanding of this important patho-system and lays foundation for facilitating research towards developing

strategies to combat the disease.

PE264 (IPPC0550)

Artificial microRNA-mediated silencing of ecdysone receptor gene affects growth and development of

Helicoverpa armigera

Sneha Yogindran* and M.V. Rajam

Department of Genetics, University of Delhi, South Campus, New Delhi-110021, India


Food production is seriously affected by various biotic stresses such as insect pests, nematode parasites, pathogens and

weeds. Hence, it is necessary to increase the crop production to feed the ever growing population. Among the different

biotic stresses, insect pests affect the crop productivity to a great extent. Helicoverpa armigera, popularly known as

cotton bollworm is one of the serious agricultural pests which affect more than 360 plant species including 50 crop plants.

This pest is a great threat to the farmers due to its host range, high mobility and fecundity, and the ability to adapt and

develop resistance against all common groups of insecticides. Bt technology paved the way for this problem to a great

extent, but reports came up with insects gaining resistance against Bt toxins. Therefore, there is an urgent need for an

effective and long lasting pest control strategies. The present study focuses on the use of artificial microRNA (amiRNA)

to target ecdysone receptor (EcR) gene of H. armigera. The insect steroid hormone ecdysone and its receptor play

important roles during development and metamorphosis and regulate adult physiology and life span, making it a potential

target for pest control. Insect feeding bioassays with amiRNA-HaEcR caused significant reduction in target gene transcript

level and affected the growth and development of the insect when compared to control. These results suggest that the

use of amiRNA technology may serve as a potential alternative to the current pest management practices.


249

IPPCongress2015


PE265 (IPPC0557)

Identification of ZFD1 interacting factors (ZIFs) which influence immune responses in Arabidopsis thaliana

Anupriya Singh*, Nidhi Singh and Ashis Kumar Nandi



Pathogens put a significant challenge on plant fitness and survival. To combat this challenge plants have evolved well

developed immune system. Plant hormones such as salicylic acid (SA), ethylene (ET) and jasmonic acid (JA) play

incredible role in plant defense. Infection with biotrophic pathogen results in accumulation of SA. SA binds to and activates

non expressor of PR1 (NPR1). NPR1, the master regulator of SA signaling, in turn induces expression of large number of

pathogenesis related (PR) gene. Many PR gene products are antimicrobial, which help in warding off the pathogens. Zinc

Finger protein involved in disease Defense (ZFD1) another positive regulator of disease defense, recently identified in

our lab (unpublished data). ZFD1 mostly influences SA pathway in Arabidopsis. By yeast two hybrid (Y2H) screening

several ZFD1 interacting factors (ZIFs) were identified. Some of these ZIFs contain nuclear localization signal. Here we

report two ZIFs, such as ZIF1 and ZIF2 which influence immune responses. Through bimolecular fluorescence

complementation (BiFC) assay we showed that interaction of ZFD1 with ZIF1 takes place in the nucleus. Disease resistance

studies using Pseudomonas syringae (Psm), we report here that ZIF1 is a negative regulator and ZIF2 is a positive

regulator of immune response. In addition, we also report a third interacting partner ZIF3 which interacts with ZFD1 in ER.

Role of ZIF3 in pathogenesis is under investigation.

PE266 (IPPC0586)

Role of Arabidopsis Polycomb group repressor -2 complex in pathogenesis

Priya Gupta*, Shweta Roy and Ashis K. Nandi



Plants possess highly sophisticated immune response to combat pathogens. They have broader and basal defense

response called pattern triggered immunity (PTI) as well as much more efficient and specific immunity called effector

triggered immunity (ETI). Sometimes plants recruit housekeeping genes or development regulators for participating in

defense. Through transcriptome analysis of WT Arabidopsis plants after HR induction, our lab identified a gene belonging

to PRC2 complex, which supports bacterial growth. We named here as POLYCOMB REPRESSOR FOR DISEASE 1

(PRD1). In PRD1 overexpression plants, disease resistance gene Resistance to Pseudomonas syringae 2 (RPS2)

expression is reduced. The amiRNA lines of PRD1 are resistant against Pseudomonas syringae pv tomato DC3000

carrying AvrRpt2 gene. We show here, by transient expression in Nicotiana benthamiana that PRD1 suppresses expression

of RPS2:GUS. We have also analyzed the promoter of PRD1. Our results identified the region of PRD1 promoter that is

pathogen inducible. Yeast-2-hybrid screening using PRD1 as bait identified a large number of putative interactors of

PRD1. VOLTAGE DEPENDENT ANION CHANNEL 2 (VDAC2) and ABI FIVE BINDING PROTEIN 3 (AFP3) are two such

putative interactors. Through BiFC we have reconfirmed and identified their sub-cellular location of interaction. Mechanism

of PRD1 mediated disease susceptibility is under investigation.


250

IPPCongress2015


PE267 (IPPC0587)

Overexpression of a rice A20/AN1 zinc-finger protein modulates defence response against pathogen

infection in tobacco

Shweta Jha1*, Meenakshi Sharma2, Jitender Giri2 and Akhilesh K. Tyagi2

1Biotechnology Unit, Centre of Advance Studies in Botany, Jai Narain Vyas University, Jodhpur-342001, Rajasthan, India2National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India


Eukaryotic A20/AN1 zinc-finger domain containing proteins play an important role in the regulation of immune responses.

In plants, Stress Associated Proteins (SAPs) are identified as novel A20/AN1 zinc-finger proteins, which are shown to be

regulated by multiple abiotic and biotic stresses and emerging as potential candidates to confer abiotic stress tolerance

in transgenic plants. A total of 18 genes were identified from rice genome encoding SAPs. Overexpression of SAPs from

rice and other plants has been shown to confer abiotic stress tolerance in transgenic plants. In the present study, the

functional role of OsSAP1 in plant defence response has been explored through its constitutive overexpression in transgenic

tobacco plants. OsSAP1 overexpressing tobacco plants showed enhanced basal resistance against virulent bacterial

pathogen, Pseudomonas syringae pv. tabaci, together with the upregulation of known defence-related biotic-stress

responsive genes. Enhanced disease resistance is coupled with increased ROS production in OsSAP1 overexpressing

plants. Although its mechanism of action is poorly understood, emerging evidences suggest that OsSAP1 may act as

ubiquitin ligase, redox sensor and regulator of gene expression during stress. Thus OsSAP1 may be considered as an

ideal candidate for protecting crop plants against multiple stresses and yield loss.

PE268 (IPPC0599)

Responding rapidly to insect herbivores: calcium signalling in plant defense against herbivores

Jyothilakshmi Vadassery1*, Sandra Scholz2, Victoria Kiep3, Edgar Pieter3 and Axel Mithöfer2

1National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi-110067, India2Department Bioorganic Chemistry, Max Planck Institute for Chemical Ecology, Hans-Knöll-Stra&szl3Martin Luther University of Halle-Wittenberg, 06099 Halle (Saale), Germany


In the interaction between Arabidopsis and the generalist herbivorous insect, Spodoptera littoralis, little is known about

early events in defense signaling and link to downstream phytohormone pathways. We demonstrate that cellular calcium

(Ca2+) elevation is involved at multiple stages of interaction between S. littoralis and Arabidopsis in both local and

systemic leaf. Upon insect herbivory S. littoralis oral secretions (OS), and bioactive jasmonic acid-isoleucine conjugate,

induce cytosolic Ca2+ elevation in Arabidopsis. S. littoralis feeding also induces systemic [Ca2+]cyt elevations in Arabidopsis

which are predominantly in leaves with direct vascular connections to the treated leaf and appeared with a delay of 1 to

2 minutes. Simulated herbivory by wounding always induced a local [Ca2+]cyt response, but a systemic one only when the

midrib was wounded. This systemic [Ca2+]cyt response was suppressed by the presence of insect-derived oral secretions

as well as in a mutant of the vacuolar cation channel TPC1. Using microarray, gene expression induced by Spodoptera

oral secretion in Arabidopsis leaf was identified. Calmodulin like proteins (CML), CML42 and CML37, are highly up-

regulated both by oral secretions and insect herbivory in local leaves. Functional analysis of CML42 and CML37 loss-of-

function mutants revealed that they act in antagonistic manner during herbivory defense and modulate jasmonate pathway.

The current study highlights that Ca2+ signaling in insect herbivory is an intricate and complex network with multiple

components involving positive and negative regulators in response to local and systemic Ca2+ elevations and has a direct

impact on jasmonate signaling.


251

IPPCongress2015


PE269 (IPPC0642)

Analysis of the genes involved in terpeneoid pathway during infection in Aquilaria malaccensis

Md Rofiqul Islam* and Sofia Banu

Bioengineering and Technology, Gauhati University, Guwahati-781014, Assam, India


Agarwood is a non-timber dark resinous wood that is found in the stems, branches, and roots of Aquilaria and Gyrinops

trees after they have been wounded or exposed to biotic damage. Resin production is considered as one of the plant

defence mechanisms resulting in accumulation of agarwood oil. Agarwood is widely used in traditional medicines, as a

digestive, sedative, and anti-emetic and also as an incense and perfume, and this huge demand for the agarwood

products has led all Aquilaria spp. being endangered and listed in the Appendix II of the CITES. To promote the production

efficiency and protect the resource of Aquilaria malaccensis, it is critical to understand the regulation mechanisms of

stress-induced agarwood formation. As the major components of agarwood are sesquiterpenes and phenylethylchromones,

the present study aims to understand the molecular basis of wound-induced sesquiterpenes biosynthesis and agarwood

formation. Since, the biosynthesis of sesquiterpenes occurs via the MVA and DXP pathways, involving the following

enzymes in their synthesis, viz., 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), DXP synthase (DXPS),

farnesyl diphosphate synthase (FPS), and sesquiterpenes synthases (SS). The study presents the results of expression

profiles of these four key enzymes as well as other terpenoid genes in different tissues (wood and leaf) of infected and

non- infected A. malaccensis plants, analyzed using semi-quantitative PCR and real time PCR.

PE270 (IPPC0658)

Comparing changes in defence-related gene expression upon root-knot nematode attack in susceptible

and resistant rice cultivars

Chanchal Kumari*, Tushar Kanti Dutta and Uma Rao

Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India


Rice is one of the major staple food crops in the world and an excellent model system for studying monocotyledonous

plants. Diseases caused by nematodes in rice are well documented, and among them root-knot nematode (RKN),

Meloidogyne graminicola, causes extensive damage in yield. It is, therefore, necessary to identify novel sources of

natural resistance to RKN in rice and to investigate the rice-RKN interaction in detail to understand the basal plant

defence mechanisms and nematode manipulation of the host physiology. To this end, six different cultivars of rice were

initially screened for RKN infection and development in pluronic gel medium, which resembles three-dimensional soil

environment. Pusa 1121 was found to be most susceptible and Suraksha and Vandana were most tolerant to RKN

infection. In order to investigate the role of major hormone-regulated plant defence pathways in compatible/incompatible

rice-RKN interaction, some well-identified marker genes involved in each pathway were evaluated for their differential

expression through qRT-PCR. Results suggested that, wound-responsive jasmonate pathway, WRKY transcription factors

and PR-protein machinery are strongly induced in Vandana at early infection, while MAPK cascades, salicylate and

jasmonate pathway-related defence genes are strongly elicited in Suraksha at later stage. However, a consistent down

regulation of defence genes was recorded in Vandana during later stage of infection. As most information on the molecular

interplay between plants and nematodes has been generated on dicotyledonous plants, the current study will strengthen

our basic understanding of plant-nematode interaction in the monocot crops, which will aid in defining future strategies for

best plant health measures.


252

IPPCongress2015


PE271 (IPPC0727)

Understanding the role of CIPK6 in plant defense response

Atish Sardar* and Debasis Chattopadhyay



Calcineurin B-like (CBL) protein family consists of proteins similar to both the regulatory subunit of calcineurin and the

neuronal Ca2

+ sensor in animals. Unlike Calmodulins, which interact with a large variety of target proteins, CBLs specifically

target a family of protein kinases referred as CBL-interacting protein kinases (CIPKs). A database search of the Arabidopsis

thaliana genome sequence revealed 10 CBLs and 26 CIPKs homologues. Expression patterns of these Ca2

+ sensors and

protein kinases suggest their diverse functions in different signaling processes, including light, hormone, sugar, and

abiotic and biotic stress responses. So far, there is only one report in rice on the role of CBL-CIPK pairs in defense

signaling. All the Arabidopsis CIPKs, so far studied, were shown to have roles in abiotic stresses such as high Na+ and

low K+, cold and drought. From our laboratory, it was shown that CIPK6 is involved in growth and development along with

abiotic stress tolerance of plants. Microarray studies showed that atcipk6 knock-down plants had altered expression of

genes related to biotic stresses. Our primary investigation showed that cipk6 knock-out (cipk6) plants were less susceptible

in response to hemibiotrophic pathogen Pseudomonas syringae pv tomato DC3000 (Pst DC3000) in comparison to the

wild-type plants. The expression of several genes involved in the salicylic acid-mediated defense response pathway also

justified and established this observation. Moreover, cipk6 plants were also more sensitive to bacterial elicitors.

PE272 (IPPC0739)

Biochemical characterization of host and non-host herbivore induced proteinase inhibitors in leaves of

black gram (Vigna mungo)

Mariyamma Gujjarlapudi*, Vanka Swaroop, Vadthya Lokya and Kollipara Padmasree

Department of Biotechnology and Bioinformatics, School of Life sciences, University of Hyderabad, Hyderabad-500046, India


Plants have evolved sophisticated defense responses through transcriptional activation of specific factors such as proteinase

inhibitors (PIs) by a network of interconnecting signaling pathways in which oligosaccharides, peptide systemin, salicylic

acid, jasmonic acid, ethylene, and reactive oxygen species (ROS) play an important role during various biotic and abiotic

stresses. PIs are anti-metabolic proteins which interfere with the digestive process of insects. Black gram (Vigna mungo)

contributes to about 10% of the national pulse production. It is a host plant to Helicoverpa armigera, a polyphagous pest

and a non-host plant to Corcyra cephalonica. In the present study, black gram was chosen as a model plant to understand

and characterize the PIs induced in leaves in response to infestation by host and non-host insects. The induction of PIs

was estimated by measuring trypsin and chymotrypsin inhibitory units and SDS-gelatin-PAGE after infestation of leaves

by larvae of H. armigera and C. cephalonica at different time intervals. Treatment of leaves with methyl jasmonate and

wounding to leaves served as positive controls. Electrophoretic profile of induced PIs showed low molecular weight and

possessed both trypsin and chymotrypsin inhibitory activity indicating the possibility of belonging to Bowman-Birk proteinase

inhibitor family. Further, a significant increase in ROS and malondialdehyde content in leaves upon infestation with H.

armigera and C. cephalonica, as well as wounding and methyl jasmonate treatment indicate the role of ROS and its cross

talk with methyl jasmonates in the induction of PIs during both host and non-host insect attack.


253

IPPCongress2015


PE273 (IPPC0750)

Chitinase gene from Xenorhabdus nematophila imparts insect tolerance to plants

Saquib Mahmood1*, Mukesh Kumar1, Punam Kumari1, Nirupama Banerjee2 and Neera Bhalla Sarin1

1Laboratory of Genetic Manipulation for Stress Alleviation and Value Addition in Plants, SLS, JNU, New Delhi-110067, India2School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Plants express a wide variety of genes in response to plant pathogen or infection by pests. The best characterized genes

belonging to PR protein family are those that encode the lytic enzymes such as chitinase, which play a vital role in plant

defense against fungal pathogens due to their ability to hydrolyse the chitin, a major constituent of the lining of gut

membrane of insects and the cell wall of a number of plant pathogenic fungi. Chitinase from different sources has been

used as an important means of biological control for fungal diseases. In the present study, we have isolated a 1.9 kb of

chitinase gene from Xenorhabdus nematophila, a Gram negative, entomopathogenic bacterium. The chitinase gene was

cloned and a 76 kDa protein corresponding to the gene was expressed in E. coli. The purified chitinase protein was

orally insecticidal to the major crop pest, Helicoverpa armigera when it was fed to the larvae of insects. The chitinase

protein was also over expressed in tobacco plants via Agrobacterium mediated transformation. The transgenic lines

were phenotypically normal and healthy. The insect assays showed high mortality of the larvae when fed on transgenic

plant leaves over expressing the chitinase protein and also 60-70% reduction in leaves damage as compared to

untransformed control plants. Our results demonstrate that the chitinase isolated from Xenorhabdus nematophila is

orally toxic to Helicoverpa armigera and can prove to be a potent candidate for pest control in plants in future.

PE274 (IPPC0787)

Exploitation of seed proteinase inhibitors as biopesticides and their biochemical characterization

Kollipara Padmasree*

Department of Biotechnology and Bioinformatics, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India


Proteinase inhibitors (PIs) are proteinaceous molecules which inhibit the catalytic activity of proteolytic enzymes. PIs are

ubiquitously present in plant storage organs like seeds and tubers, since these are the vehicles for the propagation and

survival of the species. They play a major role in regulating endogenous proteolysis, act as reserve storage of essential

amino acids and expressed during biotic as well as abiotic stress tolerance. Seeds usually accumulate PIs during maturation

and their concentration is varied between 1-10% of the total seed proteins. PIs may accumulate constitutively or induced

after pathogen or herbivore attack. They are classified as serine, cysteine, aspartic and metallo PIs according to the type

of proteases which they inhibit. According to the homology, molecular mass and cysteine content, serine PIs are further

classified into Bowman-Birk inhibitor (BBI) type and Kunitz inhibitor families. The BBIs are small cysteine-rich proteins (6-

9 kDa) with two reactive sites typically against trypsin and chymotrypsin, while Kunitz inhibitors are ~20 kDa proteins,

usually made up of two polypeptide chains linked with two disulphide bonds with single reactive site for trypsin or

chymotrypsin. The PIs were known to inhibit digestive gut proteases and thereby affect the growth and development of

insect pests. In the present study, the PIs from the seeds of three leguminous plants were examined for their potential as

biopesticides against three lepidopteran insects Achaea janata, Spodoptera litura and Helicoverpa armigera. Further, the

biochemical properties of the PIs with insecticidal activity would be discussed in detail.


254

IPPCongress2015


PE275 (IPPC0815)

The ZFD1 is a novel positive regulator of salicylic acid (SA) mediated defense in Arabidopsis thaliana

Nidhi Singh* and Ashis Kumar Nandi



Plants are capable of defending pathogen attack for immediate relief from the infection as well as to memorize theinfection for inducing higher level of defense during subsequent infection. Based on the mode of recognition of pathogens,immune systems of plants are broadly classified as pattern triggered immunity (PTI) and effector triggered immunity(ETI). Plant hormone salicylic acid (SA) plays crucial roles in modulating plant defense. Transactivator protein NPR1 isconsidered as master regulator of SA mediated defense. Screening of NPR1 suppressor identified several genes thatfunction as negative regulators of NPR1-independeny pathway. One such mutation identified previously in our lab isconstitutive defense without growth defect 1 (cdd1). Through differential expression profiling of cdd1 and WT plants, weidentified a novel plant defense regulator Zinc Finger protein for Defense (ZFD1). The loss-of-function mutant of ZFD1 issusceptible towards both virulent and avirulent pathogens. Extent of loss of resistance in zfd1 mutants is comparable tothat of npr1 mutants. In agreement with the mutant phenotype, the ZFD1over-expression lines are hyper-resistant tobacterial pathogens and express PR genes at higher level. Expression of ZFD1 gene is induced by pathogen inoculationand exogenous SA application. Pathogen induced accumulation of SA and its glycosylated derivative SAG is largelyunaffected in zfd1 mutant and ZFD1 over-expression lines. The zfd1npr1 double mutant plants were more susceptiblethan single mutants, zfd1 and npr1.The Y2H screening identified two defense regulators NHL3 and VDAC2. The resultssuggest that downstream to SA, ZFD1 positively influences NPR1-independent SA signaling.

PE276 (IPPC0835)

Enhancement of glyphosate tolerance in tobacco by co-expressing CP4 EPSPS and glyhosate detoxifying

enzymes (igrA: Glycine Oxidase)

Amaranatha Vennapusa1*, Ramu S. Vemanna2, Chinta Sudhakar1 and M. Udayakumar3

1Department of Botany, SK University, Anantapur, Andhra Pradesh, India2Plantbiology Division, The Samuel Roberts Noble foundation, Ardmore, OK, USA3Department of Crop physiology, GKVK, UAS, Bangalore, Karnataka, India*Presenting author: [email protected]

Weeds are major biotic constraint for crops yield. The Glyphosate Resistance crops (GR crops) play a vital role in modernagriculture for weed control. The existing commercial GR crops are developed by either mutated or insensitive form ofEPSPS. One of the disadvantages in these crops is glyphosate residues persist in the plant tissue, which is not desirable.Hence, expression of glyphosate degrading enzymes like mutated glycine oxidase (GO) and igrA (Increased glyphosateresistance Aldo keto reductase) are more relevant to achieve glyphosate tolerance besides decreasing residual levels.With this back ground in the present study co- expression of igrA: GO: CP4 EPSPS in tobacco plants has been studiedfor glyphosate bio efficacy and residual glyphosate levels by comparing with single gene (igrA, GO, EPSPS) and doublegene (igrA: GO, igrA: EPSPS) expressing plants. The bio efficacy of transgenics plants assessed based on invitroregeneration of transgenic leaf explants on invitro glyphosate media and also by exposing leaf disc to different concentrationsof glyphosate. The transgenics expressing either single or double and multiple genes showed improved tolerance toglyphosate. These events showed higher chlorophyll content, less membrane and oxidative damage compared to wildtype. However the transgenics co-expressing igrA: Glycine oxidase: CP4EPSPS showed significantly enhanced glyphosatetolerance compared to other transgenic events. The efficacy of detoxifying enzymes was assessed by in vitro studies incrude leaf protein extracts. The efficacy of glyphosate degradation was more in tobacco transgenics co-expressingglyphosate degrading enzymes, compared to other transgenics and wild type.


255

IPPCongress2015


PE277 (IPPC0841)

Identification and expression analysis of a gene associated with defence responses to Sclerotium rolfsii

in groundnut (Arachis hypogaea L.)

Sujit Kumar Bishi1*, Mahesh Kumar Mahatma1 and Dinesh N. Vakharia2

1Directorate of Groundnut Research, Junagadh Agricultural University, Junagadh-362001, Gujarat, India2Department of Biochemistry, Junagadh Agricultural University, Junagadh-362001, Gujarat, India


Groundnut (Arachis hypogaea L.), is an important legume crop and Sclerotium rolfsii is a major soil borne necrotrophicfungal pathogen causing stem rot leading to heavy economic losses worldwide. However the defense responses ingroundnut to this pathogen are poorly understood so far. For implementation of successful molecular strategy to controlstem rot, identification and characterization of important defense related gene(s) are required. Plants produce cell wall-associated polygalacturonase-inhibiting proteins (PGIPs); whose over expression improve the resistance to plant fungaland bacterial necrotrophs by inhibiting the polygalactoronases (PGs), the cell wall degrading enzyme produced by thepathogens. PGIPs are encoded by small gene families, identified in several plants including oil seed crops such assoybean, bean, and mustard; however no reports are available for groundnut. From an unidentified EST sequence(HO115743.1) derived from groundnut seed having sequence similarity with PGIP encoding gene (PGIP) of other crops,specific primers were designed. A partial but putative PGIP was isolated using cDNA as template prepared from stem rotinfected groundnut leaf tissues and the sequence submitted to the NCBI database (Accession No. KP844637). The PGIPexpression showed its higher induction in tolerant as compared to the susceptible groundnut genotypes upon S. rolfsiiinfection suggesting its possible role in stem rot stress tolerance in groundnut. This is identified gene will provide a basisfor further cloning and functional validation of the members of PGIP family and also will aid in an understanding of theregulatory mechanism of groundnut tolerance to S. rolfsii.

PE278 (IPPC0914)

Transient RNAi based gene silencing using CgCOM1 mediated by Agrobacterium confers disease resistance

to chilli against anthracnose

Nanditha Krishnan Vimalakumari*, Binod Kumar Mahto and Swatismita Dhar

The Energy and Resources and Institute (TERI) University, Vasant Kunj, New Delhi-110070, India


Chilli (Capsicum annuum L) is one of the most important crops in the tropics. A major constraint in chilli cultivation is theeconomic losses, often as high as 50%, resulting from anthracnose disease caused by Colletotrichum spp. The genusCapsicum is extremely recalcitrant to stable transformation, which results in poor efficiency and reproducibility. Transienttransformation offers a viable alternative to stable transformation, as it is relatively easy, occur at high frequencies andhelps in the quick analysis of transformants. We have standardized a highly efficient protocol for transient transformationin chilli (var: Pusa Jwala) mediated by Agrobacterium tumefaciens using pCAMBIA 2301 vector which harbours npt IIand GUS genes. Maximum efficiency of transformation was obtained when microinjection, infiltration and application ofvacuum were carried out sequentially to the leaf explants. Transient transformation was then successfully used tointroduce RNAi vector having CgCOM1 gene, from Colletotrichum gloeosporioides, in chilli leaves. The transformedleaves were later challenged with the fungal pathogen. The transformed leaves showed differential resistance toColletotrichum sp. challenge in comparison to wild-type leaves. The analyses of the transformants were carried out byphenotypic characterization, determining the hypersensitive response and by gene expression experiments. The presentstudy provides a reliable tool for functional genomics by enabling rapid assessment of gene function prior to stabletransformation.


256

IPPCongress2015


PE279 (IPPC0915)

Modulation of host immunity and nutrient allocation by a biotrophic pathogen

Divya Chandran*

Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad-121001, Haryana, India


Obligate biotrophic pathogens like the powdery mildew fungi manipulate living plant hosts to acquire nutrients whilelimiting host defense responses. Powdery mildew is one of the most devastating diseases limiting productivity of foodlegumes in India and other developing countries. Chemical methods used to control the disease are neither economicalnor environmentally sustainable. Therefore, there is a need to develop innovative strategies to combat this disease. Todate, few powdery mildew resistant legume varieties have been identified, with the identity of the genes conferring resistanceand the underlying molecular mechanisms not known. Our major goal is to identify and target novel legume host genesassociated with defense that limit growth of the pathogen as well as host compatibility factors required for pathogengrowth and reproduction. We envisage that targeting a combination of such factors would result in dramatically reducedpathogen proliferation and contribute to durable resistance that is less likely to be rapidly overcome by pathogen counter-evolution. To address our goal we are performing infection site-specific analyses via laser microdissection and nextgeneration sequencing in the Medicago truncatula-Erysiphe pisi model pathosystem. We expect to uncover novel hostcomponents, cellular processes, and pathogen effectors impacting pathogen growth, previously hidden in whole-tissueglobal expression analyses. In parallel, we are studying the role of plant sugar transporters in mediating carbon partitioningat the host-pathogen interface, a process critical to both pathogen and host growth. Our long-term goal is to engineerdurable powdery mildew resistance in food legumes of agronomic import.

PE280 (IPPC0918)

Inositol phosphates and their role in regulation and trigger of effector triggered immunity

Jewel Jameeta Noor, Hitika Gulabani* and Saikat bhattacharjee



Plant immunity offers a classic example wherein each invaded cell is sophisticated and sufficiently equipped to possessmechanisms of surveillance, defense regulation and trigger only upon invasion with minimum side-effects. Althoughmanifestation of most responses occur through step wise communication cascades of receivers and transducers, signallingin Effector-Triggered Immunity (ETI) is an exception. Signaling towards transcriptional reprogramming is regulated byintricate and interlinked connections between immune players / regulators at specific cellular locales. The resulting networksoffer a robust and rapid responses characteristic of ETI. Several reports suggest that ETI responses impinge on hormonalsignal transduction pathways. However, even after decades of research molecular processes underlying these signalingevents remain largely unknown. As secondary messengers soluble Inositol-Phosphates (InsPs) influence mRNA export,apoptosis, and control of transcription. Inositol-modified lipids (phosphatidylinositols, PtdIns) determine architecture ofmost eukaryotic membranes. In our assays several tested mutants with unregulated immunity have differential transcriptsfor key inositol biosynthesis genes. In an effort to identify specific InsPs in ETI we are profiling their levels in inducible ETIsystems and in mutants with altered defense capabilities. In parallel, we are trying to identify InsP–interacting proteins inplants. Molecular complexes of central immune regulators with resistance proteins (R) often occur at membranouslocalization likely involving PtdIns. Specific associations in this complex are perturbed by multiple unrelated effectors. Weare further characterizing the this association to gain insights into how ETI is regulated from misfiring in absence of apathogen threat and how effectors target these associations.


257

IPPCongress2015


PE281 (IPPC0921)

Effector-triggered immunity against a phytopathogenic bacterium: regulation, perturbation and signaling

Saikat Bhattacharjee*



Effector-triggered immune responses in plants are induced upon sensing specific effector activities (termed avirulencefactors; Avr) on either a cognate resistance protein (R) directly or on a specific host target which the R protein guards.Although intended for suppressing defenses, in a resistant plant the same effector actions betray a pathogen presence bydisrupting of protein-protein associations in a ‘resistasome’ complexes comprising of a negative modulator that regulateR proteins, and positive modulators that promote defense. Using Arabidopdis thaliana-Pseudomonas syringae pv tomato(DC3000) pathosystem our earlier work has identified one such regulatory association of immune players. Upon disruptionactivated R proteins and positive immune regulators that are resident of this complex signal through intricate cross-linkednetworks that cause massive transcriptional reprogramming that culminate pathogen threat. Whereas many newerresistance specificities are being identified, detailed molecular mechanism of how immune assemblies regulate defensesin an un-invaded plant and how perturbations upon pathogen attack effectively couple to downstream signaling routesremains unknown. To gain further insights into these processes we are characterizing interactions that aid the assemblyof a resistasome complex. Our analysis have identified that extensive post-translational modifications define strategicdeployment of defense regulators and a phytopathogenic effector often impinge on and alter these modifications througha competing/interfering post-translational activity. Interestingly the identification of unrelated effectors targeting a commontarget further support this model. Our studies also propose that production of secondary metabolites that signal immunepathways are also effectively coupled to changes in protein modifications thereby ensuing a robust link between triggerand signaling during plant innate immunity.

PE282 (IPPC0922)

How SWEET is biotrophic life? Role of plant sugar transporters in pathogen nutrition and plant defense

Megha Gupta* and Divya Chandran



Obligate biotrophic pathogens like the powdery mildew fungi modulate plant host metabolism to divert nutrients, especiallysugars, to fuel their own growth at the expense of plant productivity. They form specialized feeding structures (haustoria)within host epidermal cells and create an apoplastic interface through which host sugars, released primarily through theaction of sugar transporters, are absorbed. Recently, a new family of plant sugar efflux transporters, known as SWEET,has been implicated in bacterial and fungal nutrition. For example, the bacterial blight pathogen Xanthomonas oryzaesecretes transcription-activator like (TAL) effectors that directly induce the expression of specific SWEET genes triggeringsugar efflux at the host-pathogen interface. Notably, when SWEET expression is blocked, so is the growth and virulenceof the pathogen. To ascertain the role of SWEET transporters in powdery mildew nutrition we utilized the Medicagotruncatula (legume host)-Erysiphe pisi (pathogen) interaction as a model pathosystem. Expression levels of 24 MtSWEETcandidates (identified through bioinformatics) were quantified via real-time quantitative PCR at various time points post E.pisi inoculation. We found that a number of MtSWEETs exhibited enhanced expression in response to E. pisi in a susceptiblehost with peak expression observed just prior to the onset of fungal conidiation. In contrast, E. pisi-induced expressionwas either absent or reduced in a resistant host with peak expression detected at earlier time points coinciding with fungalgrowth arrest. Our results highlight the possibility of a dual role for plant SWEET transporters in powdery mildew nutritionas well as plant defense.


258

IPPCongress2015


PE283 (IPPC0929)

The ‘Trigger’ in effector-triggered immunity: deregulation via post-translational modification

Kishor Ingole*, Bornali Gohain and Saikat Bhattacharjee



Basal immunity (also called PAMP-triggered immunity, PTI) in plants, induced against conserved microbe determinants,is suppressed by adapted pathogens that secrete effectors and interfere with specific host defense signaling routes(termed virulence). As a counter-defense strategy resistant plants deploy specific resistant (R) proteins that either directlyor indirectly sense the effector-induced modifications and mount effector-triggered immunity (ETI). Pathogens that triggerETI are termed as avirulent. The outcome of ETI is a massive transcriptional reprogramming that culminates pathogencolonization and spread. In many instances unrelated effectors target common virulence target. The specific R proteinthat gets activated depends on the nature of post-translational modifications (PTMs) an effector causes on target. PTM bySUMOylation regulate key processes in normal growth and development such as protein stability, localization, and protein–protein interactions. Transcriptional regulation is also achieved through SUMO-dependent modification of histones. Inplant defenses although SUMO and SUMOylation machinery components are implicated, their precise role remainsunexplored. We are using mutants with altered defense responses to gain insights into specific roles of individual SUMOisoforms in plant defenses. Our preliminary observations suggest induction of a specific Arabidopsis SUMO memberupon effector-triggered immunity. We also observe global upregulation of SUMOylated proteins in plants with uncontrolleddefenses. We are focusing on further characterizing SUMO functions and identifying SUMOylated targets and elucidatingtheir roles in ETI.

PE284 (IPPC0939)

Functional characterization of a key regulator of innate immunity, CaRACK1, in chickpea in response to

necrotrophic fungus Ascochyta rabiei

Sandhya Verma*, Kamal Kumar and Praveen K. Verma

Plant Immunity Laboratory, NIPGR, Aruna Asaf Ali Marg, New Delhi-110067, India


Plants are constantly under threat of infection by myriads of potential pathogens. In order to combat pathogen attack,plants have developed various intricate mechanisms that results in the establishment of effective defense system. A firstlayer of this system is based on perception of PAMPs through pattern recognition receptors (PRRs) at the plant’s cellsurface. Successful pathogens produce effectors to inhibit innate immunity, but plants, in turn, can perceive such effectorsthrough additional receptors to mount induced immunity. Necrotrophic plant pathogenic fungi possess a battery of lyticand degradative enzymes. Moreover, recent studies have shown that they can also subtly manipulate the host defenseduring infection in a manner similar to biotrophic pathogens. In chickpea, the necrotrophic fungus Ascochyta rabiei causesblight disease. Our interest is to study signaling mechanisms required for induced resistance during pathogen interactionin chickpea. Till date, the role of signaling components against fungi has remained elusive. RACK1 (Receptor for ActivatedC-Kinase 1), that has been isolated in our laboratory, functions as a scaffold protein by physically interacting with manyother proteins and facilitating their interactions. During chickpea-Ascochyta infection, CaRACK1 is highly expressedsuggesting its role in plant defense. Moreover, this conserved WD40-repeat-containing protein exists in multi-meric formin vitro. Functional complementation of CaRACK1 in yeast ASC1 mutant rescued the yeast mutant phenotype. It suggestsCaRACK1 is a functional homolog of S. cereviseae ASC1. The importance of CaRACK1 in various key signaling eventsalso suggests that CaRACK1 might be a potential target of Ascochyta rabiei effectors.


259

IPPCongress2015


PE285 (IPPC0974)

Integrated analysis of miRNA and mRNA omics data from diseased tomato plants: possible application

in stress management

Deepti Sarkar, Jayanti Jodder, Sayani Dey, Ranjan Kumar Maji, Rohit Das, Arunava Mandal, Payel Bhattacharjee, Arijita

Sarkar, Zhumur Ghosh, Pallob Kundu*

Division of Plant Biology and Bioinformatics Center, Bose Institute, P1/12 CIT Scheme VII M, Kolkata, West Bengal, 700054, India


Gene expression cascade in a plant is altered in times of stress. Reprogramming of the expression profiles of genes are

required for a robust and specific response. MicroRNA-mediated regulation of gene expression plays a significant role in

this process as a miRNA is capable of regulating expression of multiple genes both at post transcriptional and translational

level. Until now, tomato miRNAs responsive to Alternaria fungal stress were not known. We will present profile of miRNAs

that are differentially expressed during Alternaria stress in local tomato cultivar (Pusa Ruby) and results of integrating the

expression profile with transcriptome analysis of tomato. The knowledge gathered from these omics studies has been

applied to get an insight into the miRNA-mediated regulation of ‘disease-resistance’ genes in tomato. Each miRNA plays

multiple roles, thus deliberate manipulation of their expression profiles for disease management would require thorough

knowledge about the regulation of their biogenesis. We will also present evidence of complex regulation of a miR-gene

expression and suggest how its expression can be manipulated using genome manipulation tools.

PE286 (IPPC0994)

Control of sugarcane mosaic disease using bacterially expressed viral coat protein specific double

strand RNA - A RNA interference approach

Yasir Umar Pattiwala* and Prabu Gajjeraman

Department of Biotechnology, Karpagam Academy of Higher Education, Coimbatore-641021, Tamil Nadu, India


Sugarcane production has declined in recent years due to several adverse factors, including mosaic viral disease.

Sugarcane Streak Mosaic Virus (SCSMV) is a major cause for mosaic disease alaongwith Sugarcane Mosaic Virus

(SMV) and Sorghum Mosaic Virus (SrMV). RNA silencing plays a significant role in plant resistance against viruses,

transposons and other microbes. The phenomenon of RNA interference (RNAi) is involved on sequence-specific gene

regulation driven by the introduction of dsRNA resulting in inhibition of transcriptional or translational suppression. Coat

protein (CP) encoded gene of Sugarcane Streak Mosaic Virus (SCSMV) was amplified in sense and anti-sense orientation

and cloned into RNAi cloning vector to produce double strand RNA (dsRNA). The culture medium and conditions were

optimized for high level bacterial expression of SCSMV_CP dsRNA. The sugarcane sett treatment method was optimized

to induce SCSMV_CP dsRNA mediated RNA interference and resulted 70% reduction of virus titer than the untreated

counterparts. These results provides a valuable means for controlling plant viral disease using bacterially expressed

dsRNA and RNAi approach.


260

IPPCongress2015


PE287 (IPPC1045)

Biochemical changes in different cultivars of Aonla (Emblica officinalis G.) during storage at room

temperature

Jitender Kumar*, Poonam Kumari and Suman Bala

Department of Botany and Plant Physiology, CCS Haryana Agricultural University, Hisar

*Corresponding author: [email protected]

An experiment was conducted to study the biochemical changes in five cultivars of Aonla during storage at room

temperature. Acidity content of fruits decreased during storage from 3rd to 18th day of storage in all the cultivars. More

acidity content was observed in the fruits of cv. Krishna (1.825%) after 18th day of storage. Whereas minimum acidity was

retained in the fruits of cv. Chakaiya (1.574%) after same period of storage. Ascorbic acid content of fruits also decreased

with increasing period of storage in all the cultivars of aonla fruit. Maximum decrease was observed in the fruits of cv.

Krishna (441.2%) after 18 days of storage whereas minimum decrease was observed in Chakaiya fruit (476.7%). TSS

content of fruits also decreased with increasing period of storage in all the cultivars. Minimum TSS was retained in the

fruits of Banarasi (7.70Brix) whereas maximum TSS was retained in Hathijhul fruits (90Brix).

Session F : Plant Nutrition Physiology and Genetics

261

IPPCongress2015


PF288 (IPPC0012)

Physiological characteristics and yield improvement by Arbuscular mycorrhizal fungus in wheat (Triticum

aestivum L.)

Kamal Dutt Sharma1*, Karmal Singh1, Rajesh Gera2 and Jagdev Singh1

1Crop Physiology Lab, Department of Agronomy, CCS Haryana Agricultural University, Hisar-125004, Haryana2Department of Microbiology, CCS Haryana Agricultural University, Hisar-125004, Haryana


Arbuscular mycorrhizal (AM) fungi are ubiquitous root symbionts of more than 90% of vascular plants and are thought to

contribute in plant nutrition, particularly phosphorus. A field experiment was conducted in split plot design, with wheat

genotypes, WH 1021, WH 1105, WH 1123, WH 1124, WH 1158 in main plots and seed treatments practices, i.e., dry

seeding, primed seed (seed soaking overnight in water), primed seed + AM fungi (Glomus mosseae), primed seed +

Azotobacter in the sub plots with three replications under late sown situation. Physiological characters of flag leaves were

measured after anthesis. Wheat genotypes sown with water primed, seed inoculated with AM fungi and Azotobector

increased grain yields by 2.4, 8.1 and 3.8%, respectively over conventional practice (3998 kg ha-1) under late sown

condition. The increase in gain yield was mainly due to early seedling emergence and vigour, more number of spikes per

plant and higher biomass and improved physiological traits like higher membrane stability and photosynthetic efficiency

of flag leaf during anthesis. Among the genotypes, WH 1105, followed by WH 1158 showed significantly higher yield

(4315 and 4278 kg ha-1) over the other tested genotypes due to higher membrane stability, more tillering and test weight.

The increase in seed yield by seed priming and AM fungi was positively associated with days to emergence and heading,

membrane stability and photosynthetic rate.

PF289 (IPPC0142)

Agriculture and nutritional security: Enhancement of lysine contents in rice through genetic manipulation

of aspartate metabolic pathway enzymes

Manjari Mishra1*, Sneh L. Singla Pareek2 and Ashwani Pareek


Malnutrition is a major problem in developing countries of the world. Due to lack of nutritious food, more than one third of

children are dying every year, specially in developing countries. This scenario impose challenges to improve nutritional

status and nutritional security in economically poor regions of the world. Inspite of being a staple food for a large part of

world’s human population, rice is nutritionally poor for human and animal consumption because the proteins are deficient

in certain essential amino acids, especially, lysine. A complex and tightly regulated metabolic pathway is involved in

lysine synthesis in plants where the two enzymes DHDPS (Dihydrodipicolinate Synthase) and LKR/SDH (Lysine

Ketoglutarate Reductase / Saccharopine Dehydrogenase) tightly regulate lysine biosynthesis and degradation, respectively.

In the present work, we have genetically manipulated the lysine pathway enzymes towards metabolic engineering of

lysine content in rice. We have generated overexpression construct, RNAi construct and lysine double construct and

generated transgenic plants of Oryza sativa cv. IR-64. We got maximum increase in lysine content in Lysine Double

Construct (LDC) lines i.e. 58% compared to the wild type. These transgenic lines will be further used for metabolic

profiling to assess the effects of increased lysine content on other metabolites.


262

IPPCongress2015


PF290 (IPPC0152)

Changes in growth, yield, juice quality and biochemical attributes of sugarcane in response to silica

granules

Anshu Singh1*, Smita Singh1, Neeru Jain2, P. Lal1, R.K Singh1, Asha Gaur1, S.K Shukla1, S.P. Singh1, A. Chandra1 and

Radha Jain1

1Indian Institute of Sugarcane Research, Lucknow-226002, India2Privi Life Sciences Pvt. Ltd. Navi Mumbai, India


Silicon (Si), the second most abundant element in the earth crust (27.7%) is usually found in the form of complex silicate

minerals, and less often as silicon dioxide. It plays an important role in inducing resistance to various biotic and abiotic

stresses in plants, helps in controlling Al, Mn and Fe toxicities, increases P availability, reduces lodging and improves

rate of photosynthesis by effective use of sunlight as well as efficient management of plant water economy. Sugarcane,

a typical Si- accumulating plant is known to absorb a large amount of silica from the soil. A field experiment was conducted

to study the effect of Silica granules (obtained from Privi Life Sciences Pvt. Ltd., India) on growth, yield and juice quality

attributes of sugarcane at IISR Experimental Farm, Lucknow during 2013-15 (autumn season). The treatments comprised

of control (T1), silica granules @ 20 (T2), 40 (T3) and 80 kg/ha (T4), and calcium silicate too high (T5). Specific leaf

weight, total dry matter and nitrate reductase activity increased in all treatments relative to control. Silica granules @ 40

kg/ ha (T3) showed highest increase in yield attributes; cane length, girth, NMC and cane yield. oBrix, juice purity, sucrose

% juice, juice extraction, CCS per cent juice, S/R ratio and SPS were comparatively higher in treated plants. Highest

soluble silica content was observed in T2 (leaf) and T5 (root) treatment. Findings obtained suggested that application of

silica granules holds immense potential to improve yield and juice quality attributes of sugar cane.

PF291 (IPPC0158)

Calcium regulates tuberization in potato through enhanced expression of CaM, StCDPK and LOX genes

Devanshi Chandel Upadhyaya*, Abhishek Kumar, Deepak Singh Bagri and Chandrama Prakash Upadhyaya

Department of Biotechnology, Dr Harisingh Gour Central University, Sagar-470003, M.P., India


Calcium plays an important role in plant physiology and various plant cell signaling pathways, and also alters many

biochemical processes by activating specific enzymes. Cytosolic Ca2+ regulates the activity of Ca2+ dependant protein

kinases, which phosphorylate various key metabolic enzymes. In the present study, expression of two Ca2+ dependant

proteins, calmodulin (CaM1) and calcium dependant protein kinase (StCDPK), as well as the lipoxygenase (LOX; EC

1.13.11.12) were studied upon Ca2+ application to the single node segments of potato inoculated for tuberization. Calcium

at higher levels (6 mM) significantly improved the tuber number, tuber growth and tuber yield under in vitro conditions.

The expression levels of the CaM1 and StCDPK were significantly higher in stolons showing positive correlation with

supplemental Ca2+ and tuberization response. Similar trends were observed with LOX enzyme activity, which increased

by 18% and 25% with the addition of Ca2+ at 3 and 6 mM, respectively to the tuber induction medium, when compared to

control. Thus, present study reports that the increase in tuberization, tuber growth and tuber yield with the supplementation

of Ca2+ could be attributed to the increased expression of the Ca2+ dependant proteins and enhanced lipoxygenase

activity.


263

IPPCongress2015


PF292 (IPPC0223)

Influence of silicon solubilizers on yield and biotic stress tolerance in rice (Oryza sativa L.) genotypes

Babita Joshi, Deepti Shankhdhar, Alok Shukla and Shailesh Chandra Shankhdhar*

Department of Plant Physiology, College of Basic Sciences & Humanities, G.B. Pant University of Agriculture & Technology, Pantnagar-263145, Uttarakhand, India


A field experiment was conducted in kharif season of 2014 to investigate the effect of silicon solubilizer on different rice

genotypes namely DRRH-3, PA-6129, PA-6201, PA-6444, PHB-71 and BPT-5204. Soil application of solid silicon solubilizer

(imidazole) was given at the time of maximum tillering and panicle initiation. Various growth, biochemical parameters and

yield attributes were observed. Insect infestations were also recorded at two growth stages (panicle initiation and flowering).

The leaf area index (LAI) was increased 27.55% as compared to control at maximum tillering, 7.05% increased at panicle

initiation and 16.67% increased at flowering in DRRH-3. As compared to control 5.54% increase in economic yield was

obtained in PA-6129 however 5.65% increase in harvest index was found in case of DRRH-3. Silicon solubilizers had a

significant effect on iron content in different rice genotypes. Iron content was increased by 73.36% in grains of BPT-5204

with husk and 78.50% without husk in PA-6201. The zinc content was increased by 52.61% in grains of DRRH-3 with

husk and 29.82% without husk in grains of PA-6129. Pest infestations were also significantly reduced in all genotypes of

rice as compared to control. In BPT-5204, 60.0% decreased in dead heart infestation and 63.64% decreased in white ear

head infestation was observed in PA-6201 as compared to control. Amongst the six genotypes PA-6444 was the most

efficient in utilizing silicon under soil application of imidazole carrier molecule.

PF293 (IPPC0228)

Functional characterization of OsPAP9 under P deficiency

Bipin Pandey*, Poonam Mehra and Jitender Giri



Phosphorus (P) is an essential macronutrient for plant growth and development. Most of the soil P exists in bound form as

insoluble complexes, not readily available to the root system. Thus, P is one of the most limiting nutrients for plant growth.

However, plants have evolved a set of adaptive morphological, physiological, biochemical and molecular responses that

allow them to withstand low P stress in soils. Among these, intracellular purple acid phosphatases (PAPs) play important

role in P recycling from cellular P reserves whereas extracellular PAPs are largely secreted into the rhizosphere to

scavenge P from bound organic sources. In rice, 26 PAPs have been identified, out of which we found ten PAPs being

significantly expressed under P deficiency. However, except OsPAP10a, no detailed study of any of the rice PAPs has

been carried out so far. Therefore, we selected one of the differentially expressed PAPs, OsPAP9 to improve low P

tolerance. Biochemical studies of OsPAP9 revealed that it acts as functional phosphatase at acidic pH and is highly

thermostable. Interestingly, OsPAP9 showed wide spectrum of substrate specificity. Moreover, OsPAP9 requires a

divalent cation Mg2++ as best cofactor for its optimal activity. Constitutive overexpression of OsPAP9 showed improved

growth performance of rice plants as compared to WT under low P conditions. Interestingly, growth of overexpressed

lines was relatively vigorous when low P starved plants were supplemented with ATP as organic P source. Thus,

OsPAP9 transgenic rice will be a useful resource for alleviating P deficiency.


264

IPPCongress2015


PF294 (IPPC0241)

Enhancing flavour and quality of tomatoes by expression of synthetic gene encoding sweet protein

monellin

Tanushri Kaul* and Chinreddy Subramanyam Reddy

Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India


Monellin a sweet-tasting protein exists naturally as a heterodimer of two non-covalently linked subunits chain A and B and

it drops sweetness on denaturation. In this study, we validated the expression of a synthetic monellin gene encoding a

single polypeptide chain covalently linking the two subunits under T7 and fruit-ripening specific promoters in E. coli and

tomato fruits, respectively. Purified recombinant monellin protein retained its sweet flavor at 70 C and pH 2. We developed

15 transgenic T0 tomato plants overexpressing monellin that were devoid of any growth penalty or phenotypic abnormalities

during greenhouse conditions. T-DNA integration and fruit-specific heterologous expression of monellin had occurred in

these transgenic tomato lines. ELISA revealed expression of monellin was 4.5% of total soluble fruit protein. Functional

analyses of transgenic tomatoes of T2-5 and T2-14 lines revealed distinctly strong sweetness as compared to wild type.

Monellin a potential non-carbohydrate sweetener if expressed in high amounts in fruits and vegetables would enhance

their flavour and quality.

PF295 (IPPC0287)

Leaf and grain nitrogen partitioning of a dual-adapted rice variety grown under transplanted and directed

seeded condition

A. Amaregouda*, B.O. Kiran, Supriya Patil, Benoit Clerget and B.K. Desai

Deparment of Crop Physiology, University of Agricultural Sciences, Raichur-584104, Karnataka, India


Fertilizer management practices need to be developed for rice production, a system in which rice is grown under transplanted

and direct seeded rice cultivation. We evaluated the effects of graded nitrogen (N) levels application on the yield of

transplanted and direct seeded rice under rainfed conditions in 2013 wet seasons at Main Agricultural Research Station,

Raichur state, India. Yields increased with increasing N rate from 75-125% RDN (Recommended Dose of Nitrogen).

Better translocation of N was observed with 185 kg N ha-1 as compared to 150 kg N ha-1 in both the methods of crop

establishment. The grain N content was higher (1.52%) in JKPH 3333 under direct seeded condition at 185 kg N ha-1. At

harvest significantly higher leaf nitrogen content (1.01%) was recorded in direct seeded rice compared to that of transplanted

rice (0.66%). Although the yields were higher in transplanted rice, well developed root system enhanced better allocation

of N at harvest in direct seeded rice cultivation. However, among different planting methods, significantly higher nitrogen

content (0.89%) was recorded in transplanted rice as compared to that of direct seeded rice (0.54%). Cultivation of rice

with direct seeded method (42.92 q/ha) resulted in 33.33% yield reduction compared with transplanted method (56.29 q/

ha). Among the yield attributes, number of spikelets/panicle was significantly affected under aerobic method, while test

weight and harvest index were not influenced by the method of water management. The traditional method of rice cultivation

resulted in higher total biomass production as compared to direct seeded rice cultivation. Our result suggested that that

selection for JKPH 3333 N accumulation may have value in variety for nitrogen responsive and stable yields even under

water deficit conditions.


265

IPPCongress2015


PF296 (IPPC0307)

Arbuscular mycorrhiza (Rhizophagus irregularis) influences As accumulation, growth and productivity

in Cajanus cajan (L.) Millsp. (pigeonpea) genotypes exposed to AsIII and AsV stress

Lakita Kashyap* and Neera Garg



As exists naturally in the form of arsenate (AsV) and arsenite (AsIII) in soil which affect crop growth and productivity.Arbuscular mycorrhizal (AM) associations impart abiotic stress tolerance by improving growth and nutrient status (P).However, reports on the relative role of AM in reducing AsIII and AsV toxicities in legumes are scanty. Thus, experimentswere conducted to ascertain the modulatory role of AM in alleviating arsenate (AsV) (25 mg/kg and 50 mg/kg) andarsenite (AsIII) (5 mg/kg and 10 mg/kg) induced responses in terms of growth, water relations and productivity in pigeonpeagenotypes [tolerant Pusa 2002 and sensitive Pusa 991]. Results indicated that at higher concentrations, total As wasdetected in roots, leaves and seeds in AsIII treated plants while in roots and leaves in case of AsV. Moreover, Pusa 991accumulated higher As than Pusa 2002. A significant decline in mycorrhizal colonization (MC), chlorophyll a/b ratio,RWC, root to shoot ratio and ultimate productivity was recorded under both As species, with AsIII more toxic than AsV.However, AM reduced total As accumulation in both AsV and AsIII treated plants which resulted in significant improvementin plant biomass, reduced pigments degeneration and increased ultimate yield. Pusa 2002 was more responsive to AM,established more effective MC and was found to be significantly more tolerant to both AsV and AsIII stresses whencompared with Pusa 991. Study indicated that although R. irregularis alleviated both AsV and AsIII toxicities, percentbenefits of AM were relatively more significant in case of AsIII treated plants of pigeonpea.

PF297 (IPPC0308)

Cumulative effects of Silicon supplementation and mycorrhization in restoring growth, yield and ionic

homeostasis of chickpea genotypes subjected to long term salinity

Purnima Bhandari * and Neera Garg



Silicon (Si) uptake varies among the plant species and has been reported to be low in legumes (<0.5%). Both, Si amendmentto the soil and arbuscular mycorrhizal (AM) symbiosis stimulate plant tolerance to salinity. Few reports indicate that AMfungi increase Si uptake in mycorrhizal plants. However, reports on the comparative role of exogenous application of Siand AM inoculations, as well as their interactive effects through combined treatments, in improving plant resilience to saltstress are lacking. Therefore, pot trials were undertaken to ascertain the influence of AM fungal species - Funneliformismosseae and/or Si supplementation (@ 4mM) on Na+ uptake and translocation, nutrients acquisition (N, P, K+, Ca

2

+) andresultant yield in salt tolerant (HC 3) and sensitive (CSG 9505) Cicer arietinum L. (chickpea) genotypes under salt stress(0-100 mM). Results indicated that although both, Si and mycorrhiza improved plant biomass and enhanced yield understressed conditions, Si-mediated benefits were elicited through significantly reduced Na+ uptake, coupled with enhancedK+, Ca

2

+ contents thereby, restoring K+/Na+, Ca2

+/Na+ ratios, while mycorrhizal effects were directed towards improvedplant growth, enhanced N, P acquisition and the resultant seed yield. Interestingly, under combined treatments, thebenefits achieved were substantially higher than the individual Si as well as AM applications, suggesting that mycorrhiza-mediated Si absorption from the soil contributed towards enhanced salt tolerance and seed yield, more in HC 3 than CSG9505 genotypes. Results strongly highlighted the usage of combined Si-mycorrhization as an effectual strategy in increasingchickpea productivity in salt contaminated areas.


266

IPPCongress2015


PF298 (IPPC0309)

Silicon nutrition and Arbuscular mycorrhizal (AM) inoculation as an effective strategy to improve growth

and nitrogen fixing efficiency of Cajanus cajan (pigeonpea) genotypes under cadmium stress

Sandeep Singh * and Neera Garg



Cadmium (Cd) is amongst the most hazardous elements and its presence at elevated levels in the soils is threatening

agricultural productivity. Role of silicon (Si) nutrition and AM in mitigating Cd stress has gained importance in recent

years. The present study was aimed to evaluate individual and cumulative effects of Si (600 mg/kg) and AM (Rhizophagus

intraradices) on Cd (CdSO4 – 25, 50 mg/kg) induced responses in pigeonpea genotypes (Pusa 2002 and Pusa 991). Cd

stress caused significant reduction in dry matter, with Pusa 991 exhibiting higher decline than Pusa 2002. The impact of

Cd was more pronounced in roots than shoots (decreased R/S ratio) and this decline was proportionate to Cd uptake and

translocation. Decreased root biomass reduced nodulation potential and ultimate N accumulation under metal stress.

Individual applications of both Si and AM fungal inoculations counteracted the negative effects of Cd stress and significantly

improved growth and nitrogen fixing potential, with higher positive effects in Pusa 2002 than Pusa 991. AM was highly

efficient in reducing Cd uptake thereby improving root biomass and N fixing efficiency (ARA) while Si was more effective

in improving shoot biomass. Combined application of Si and AM ameliorated the negative effects of Cd25 with significant

improvement under Cd50. Pusa 2002 was more efficient in establishing mycorrhizal colonization, Si uptake and was

more tolerant to Cd stress than Pusa 991. Thus, the study suggested that combined treatments of Si and AM can be used

as an effective strategy in combating Cd stress in pigeonpea.

PF299 (IPPC0321)

Over expression of MusamiR398a in transgenic banana plants towards improving the iron content

Prashanti Devendra Patel* and Thumballi Ganapathi

Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre,Mumabi-400085, Maharashtra, India


Banana, an important food crop, though macronutrient-rich, is poor in iron. Hence it is imperative to understand the

different mechanisms of Fe homeostasis in the plant so that banana can be enriched in iron to alleviate anaemia. Thus,

the present study was undertaken to understand the roles of micro RNAs in iron homeostasis. The plant miRNAs 397,

398 and 408 are well-known copper-miRNAs that target various Cu- proteins under Cu deficiency. This diverts the precious

metal to the essential Cu protein plastocyanin that can only use copper as a cofactor. However these miRNAs were also

down regulated under Fe deficiency, in common bean and Arabidopsis, suggesting probable roles in the cross talk

between Fe and Cu homeostasis. In this study, one of the two loci of banana miRNA 398- miR398a was retrieved from the

Banana Genome Hub after homology searches. After sequencing, the stem loop sequence along with flanking nucleotides

was cloned into the pCAMBIA 1301 binary vector, under the constitutive ZmUbi promoter. The plant expression vector

was electroporated into Agrobacterium tumefaciens strain EHA 105 and used to transform banana embryogenic cells.

The transgenic plants were regenerated on hygromycin selection medium. Initial screening of plants for transgenic nature,

was done by GUS histochemical staining and subsequently the plants were hardened in the greenhouse. Further analysis

by PCR using ZmUbi and gene specific primers was carried out to confirm their transgenic nature. The analysis of these

transgenic plants would help understand how this miRNA influences iron homeostasis in banana.


267

IPPCongress2015


PF300 (IPPC0326)

Evaluation of organic and inorganic fertilizers on neutral volatile aroma compounds of burley tobacco

Siva Raju Kemisetti*, D. Damodar Reddy and P. Harishukumar

Central Tobacco Research Institute, Rajahmundry-533105, Andhra Pradesh, India


A field experiment was conducted during 2011-13 at Burley Tobacco Research Station, Kalavacherla, East Godavaridistrict, to study the effect of organic and inorganic manures with three formulations of organic manures (Neem cake,Pongamia cake, filter press cake, poultry manure, FYM and vermi compost in different ratios) and two levels of inorganicfertilizers (120 and 160 kg N/ha) on neutral volatile aroma compounds (NVAC) of burley tobacco. Cured leaf samples oftop leaf position from different treatments were analysed for NVAC by GC-MS. The NVAC detected includes phenylalaninemetabolites, carotenoids, thunberganoids, terpenoids and neophytadiene. Among the terpenoids, hydroxysolavetivone,solvitevone, caryophyllene oxide, nerolidiol epoxiacetate, phytol, globulol and epiglobulol are the major constituentswhereas megastigmatrienone and dihydroactinidiolide are major constituents in carotenoid degradation compounds. Asshown by their relative content in NVAC fractions, thunberganoids and cembrenoids content was maximum (310 to 505ppm) followed by terpenoids (255 to347 ppm), neophytadiene (103 to 156 ppm), carotenoids (65 to 90 ppm) andphenylalanine compounds (11.14 to 20.82 ppm). The five isomers of megastigmatrienone were present in all the treatmentsexcept 160 kg N/ha and megastigmatrienone isomer-2 was in maximum content. Megastigmatrienone isomers contentwas significantly higher in manure C and B treatments when compared to inorganic treatments. With increase in nitrogenlevel from 120 to 160 kg N/ha, phenylalanine compounds, terpenoids, carotenoids, neophytadiene and thunberganoidscontents were decreased by 41.81, 10.21, 8.14, 17.6 and 35.8% respectively. The results revealed that increased level ofinorganic nitrogen significantly decreased the NVAC and application of 25% of nitrogen in the form of organic manurealong with the inorganic nitrogen to the burley tobacco significantly increased the NVAC as compared to application ofinorganic nitrogen alone.

PF301 (IPPC0349)

Overexpression of Musa ferritin gene in transgenic banana plants for increasing the iron content

Karuna Yadav* and Thumballi Ganapathi

Plant Cell Culture Tech. Section, Nuclear Agric. & Biotechnology Division Bhabha Atomic Research Centre, Mumbai-400085, India


Banana is one of the most important fruit crop and is a cheap source of nutrients and vitamins. However, the fruit isdeficient in the micronutrient iron and fails to meet the expected daily dietary requirement as per FDA. The iron deficiencymanifests as anaemia in deprived sections of society. In the present study overexpression of native iron storage proteinferritin is undertaken to enhance the iron content in banana plants. Ferritin, an iron storage protein with a capacity tosequester up to 4500 Fe atoms, protects the plant from iron induced oxidative stress. Homology search in banana usingrice sequences predicted two sequences in EST database. These two genes have been amplified using cDNA frombanana leaves. The full length amplified fragments were cloned into pCAMBIA1301 plant expression vector under ZmUbiconstitutive promoter using Pst1 and Kpn1 restriction sites. Agrobacterium tumefaciens mediated transformation of bananaembryogenic cells was carried out. The transformed somatic embryos developed on selection medium containinghygromycin. Putatively transformed regenerated lines were identified by GUS histochemical staining. Genomic DNA PCRanalysis for these lines was carried out to confirm the transgenic nature of the regenerated plants. A total of 22 transgeniclines were regenerated. Copies of each transformed lines are maintained on banana multiplication medium as multipleshoot cultures. One clonal copy was then rooted on MS media supplemented with NAA and hardened in the green housefor further analysis. The results from this study would help in improving the iron content in transgenic banana plants.


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PF302 (IPPC0352)

In silico analysis of EF-hand domain containing protein in five different plant species and analyse of

competitive binding of divalent and monovalent ions for metal binding site

Aditya Nath Jha, Tripathi Kalyani and Ram Singh Purty *

University School of Biotechnology, Guru Gobind Singh Indraprastha University, Sector 16C, Dwarka, New Delhi-110078, India


Metallo-proteins are the proteins that contain a metal ion as a cofactor. Calcium binding proteins are very commonly

found metallo-proteins, which play very significant role in signaling pathways, cell cycle and various metabolic pathways.

There have been many findings that show the role of calcium binding proteins in abiotic stress tolerance like high salinity,

drought and low temperature stress etc. Calcium binding proteins are one of the most important metallo-proteins known

to play role during stress and also inhibits activity of other metallo-enzymes by competing for their binding site. In the

present investigation, calcium binding protein containing EF-hand domain of five different species, i.e., Arabidopsis thaliana,

Oryza sativa, Zea mays, Glycine max and Populus trichocarpa were analyzed. Percentage of calcium binding proteins

was found to be highest in Oryza sativa and least in Populus tricocarpa. Binding sites were higher in Zea mays as

compared to others. An assay was done to analyze the competition of metal for binding sites using alkaline phosphatase.

It was observed that Ca2+ does not competes with the K+ and Na+ for the binding sites and activity of alkaline phosphatase

increases in the presence of calcium only. Percentage decrease in root length of Oryza sativa and Zea mays, was also

analyzed. It was noted that percentage decrease was higher in Zea mays in comparison to Oryza sativa. Also correlation

between the number of binding sites and the level of abiotic stress tolerance was analyzed.

PF303 (IPPC0380)

Morphological, physiological and biochemical responses of maize plants towards nano-micronutrient

fertilization of Fe, Cu and Zn

Rajamanickam Elanchezhian1*, Dameshwar Kumar2, Kulasekaran Ramesh1, Ashish Biswas1 and Ashok Patra1

1Indian Institute of Soil Science, Nabi Bagh, Berasia Road, Bhopal-462038, Madhya Pradesh, India2Indira Gandhi Agricultural University, Raipur-492012, Chhattisgarh, India


Controlled release of fertilizer inputs for enhanced plant growth and improved nutrient utilization can be achieved through

advanced nanotechnological tools. The impact of nano-micronutrient fertilization on growth and metabolism of maize

plants was studied using Fe, Cu and Zn nanoparticles (NPs) based formulations. It was observed that there were differential

plant responses towards the graded concentrations of NPs viz. Fe, Cu and Zn. The responses were evident on

morphological, physiological and biochemical characteristics of maize. The nano-micronutrient fertilization of plants with

lower concentration of Fe NP (27μM) had positively influenced photosynthetic leaf area, leaf area index, leaf area ratio

and proline content of plants. However, plants treated with higher concentration of Fe NP (54μM) had exhibited improved

growth parameters viz. plant height, shoot and root biomass, besides enhancement in relative growth rate, crop growth

rate, net assimilation rate and catalase enzyme activity. In contrast to above, plants treated with Cu NP (0.5 & 0.25μM)

had enhanced root length, membrane stability, total soluble sugar, non-structural carbohydrates, chlorophyll content

while Zn NP (2 & 1μM) had accelerated the biochemical metabolism of plants. The above findings indicated that the effect

of NPs including its dose was crop specific. Moreover it is also observed that NPs at reduced concentration may be useful

for the crop and they may act as catalyst for growth and metabolism which can be utilized for higher yield of plants.


269

IPPCongress2015


PF304 (IPPC0384)

Effect of salt on physiological responses of groundnut to iron deficiency

Anita Mann1*, P.V. Zala2, A.L. Singh2, Vidhya Chaudhari2 and C.B. Patel2

1Division of Crop Improvement, ICAR-Central Soil Salinity Research Institute, Karnal-132001, India2ICAR-Directorate of Groundnut Research, Junagadh-362001, Gujarat, India*Presenting author: [email protected]

Iron chlorosis is very common on alkaline soils such as calcareous ones, since iron availability is limited by high pH.Under these conditions of iron deficiency, graminaceous plant species induce special mechanisms for iron acquisition,involving enhanced release of iron chelators. On the other hand, it is known that most of salt soils have alkaline pH. So,plants growing on this kind of soils are often subjected simultaneously to salinity and iron deficiency. Thus, this workaimed at studying the physiological responses of groundnut (Arachis hypogea L.) to iron deficiency with salinity andevaluating the effect of salt on the iron nutrition and mineral nutrients. For this purpose, seedlings of two groundnutgenotypes, GG-20 and TG-37 were cultivated under controlled conditions, either in a complete nutrient solution with orwithout NaCl, with +Fe and –Fe nutrient solutions. The plant morphological aspects, chlorophyll content of young leaves,iron status, water potential and mineral nutrient contents were assessed. Plants subjected to Fe deficiency exhibited asevere chlorosis, accompanied by a significant biomass reduction. Iron-deficient treatments (-Fe and –Fe/NaCl treatmentsgenerated Fe-deficiency chlorosis on young leaves. Visual Fe deficiency symptoms were observed after one week ofgrowth under Fe starvation. However, the latter was greatly diminished when iron deficiency was associated to salinity. Adepressive effect of salt on iron acquisition in plants subjected only to salt stress was observed which was further confirmedby significant decreased efficiency in iron acquisition. These results suggest that salinity may reduce capacity of plants toacquire iron from alkaline soils.

PF305 (IPPC0396)

Comparative analysis of root proteome in soybean genotypes with contrasting exudation potential under

phosphorus starvation

Krishnapriya V. and Renu Pandey*



Root exudation of carbon compounds is one of the mechanisms evolved by crop plants to adapt to low phosphorus (P)availability. High-molecular weight secretory proteins released by plant roots are involved in hydrolysing P from theorganic pool, while low-molecular weight exudates such as carboxylic acids, phenols, free amino acids and sugarsmobilise P from inorganic sources. This study aims to understand the physiological and molecular basis of root exudationin response to low P stress in soybean. Two soybean genotypes exhibiting contrasting root exudation potential(EC-232019 and EC-113396) were identified from 116 diverse genotypes. Higher P acquisition efficiency observed inEC-232019 compared to EC-113396 was attributed to greater release of root exudates and improved root architecture.EC-232019 exuded more carboxylates, proteins, phenols, sugars and free amino acids at low (4 μM) compared to sufficient(250 μM) P. Root morphological traits viz., root length, surface and volume were higher in EC-2302019 under low P.Further, when grown in soil with low (2 g P kg-1 soil) P, EC-232019 exhibited least reduction in leaf area, photosyntheticrate, total biomass, total P uptake and seed yield compared to sufficient (15 g P kg-1 soil) P. Comparative analyses of rootproteome revealed over 10% of proteins detected on two-dimensional electrophoresis gels with two-fold or more alteredexpression between the genotypes. Out of approximately 225 protein spots detected, 28 differentially expressed proteinsrepresented a large range of functional categories including carbon and energy metabolism, signal transduction and cellcycle regulation.


270

IPPCongress2015


PF306 (IPPC0397)

Regulation of iron deficiency tolerance by sulphur in wheat

Vasundhara Sharma1* and Bhupinder Singh2

1Division of Plant Physiology, 2Nuclear Research Laboratory, Indian Agricultural Research Institute, New Delhi-110012, India


Deficiency of micronutrients in soil particularly, that of iron (Fe) is a major nutritional and production constraint worldwide,

as these play significant role in plant metabolism. Present study elucidates the role and physiological basis of sulphur (S)

mediated regulation of Fe uptake and use efficiency under deficient and sufficient availability of S (0, 1.2 and 2.5 mM) and

Fe (1 and 100 μM) using nutrient solution culture in bread (HD-2967) and durum (HI-8713) wheat, which differ distinctly

in their Fe deficiency tolerance response. Bread wheat, than durum wheat, exhibited a better plant growth under Fe

deficiency which was related to the former’s ability to tolerate Fe deficiency through the production and release of a

higher amount of phytosiderophores (PS) in the rhizosphere. Diurnal variation for the PS release under different treatments

of S and Fe was similar in both wheat types. Further, the transcript expression of S transporter (SULTR1;1) and Fe

transporter (YS1) revealed that SULTR1;1 transient expression requires not only low level of S but also sufficient Fe while

the reverse was true for YS1 expression. Results clearly indicate a complex interplay of physiological, transcriptional and

translational factors that governs the interaction between Fe and S metabolism and subsequently determine the regulatory

role of S nutrition on Fe deficiency tolerance. Further, the requirement of sufficient Fe for the induction of high affinity S

uptake transporter SULTR1;1 is important to gain insight into the regulation of S uptake by Fe.

PF307 (IPPC0414)

Physiological and quality response of bartlett pear to calcium chloride sprays prior to harvest

F.A. Khan*, Hafiza Ahsan, M.A. Dar and S. Narayan

Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Srinagar-190025, (J&K), India


Influence of foliar application of CaCl2 few days before harvest on post-harvest quality of Bartlett pear was investigated.

Plants were sprayed with CaCl2 0.0, 1.0 1.5 and 2.0% at 3, 2 and 1 weeks before harvest. The fruits were harvested at 110

days after full bloom. Bruise and disease free fruits were packed in corrugated fiber board boxes and stored at 1-2°C with

85–90% RH. Fruits showed continuous increase in PLW, TSS, sugars and spoilage with decreased firmness, ascorbic

acid, phenol and acidity during storage and shelf life (20°C). Sprays of CaCl2 close to harvest proved advantageous in

terms of increased fruit firmness, ascorbic acid, phenols and acidity with decrease in PLW, TSS and spoilage. The

beneficial effects of CaCl2 also included reduction in respiration and ethylene production rates and retardation of skin

yellowing. Spray of 1.5% CaCl2 before 14 days of harvest resulted in minimum PLW (9.12%), TSS (15.6%) and total

sugars (12.1%), and respiration (7.3 ml CO2 kg-1 hr-1) and ethylene evolution rates (25.6 μl kg-1 hr-1) with maximum fruit

firmness (11.6 lb inch-2), at 30 days of storage. Calcium treatments (1.5%) also caused an enhanced useful life of Bartlett

pear by about 5-6 days as compared to control.


271

IPPCongress2015


PF308 (IPPC0432)

Effect of putrescine (Put) supplementation and Arbuscular mycorrhizal (AM) fungi on growth, nutrient

status and nodule symbiosis in Cajanus cajan (L.) Millsp (pigeonpea) under salinity stress

Neera Garg and Amrita Sharma*



Salt stress is an important abiotic factor limiting plant growth and productivity in arid and semi-arid regions. Put, a memberof polyamines plays an essential role in imparting salt tolerance in crop plants. In addition AM have the potential to reducenegative effects of salinity and alleviate ionic imbalance. Few reports suggest that Put enhance mycorrhizal root colonization.However, roles of Put in plant-fungal symbiosis are not well understood. Aim of this work was to verify the effects of pre-sowing seed treatment with putrescine and AM (Rhizophagus irregularis) on plant growth, mycorrhizal dependence,nutrient balance and nodulation potential in two genotypes of pigeonpea (Pusa 2001- salt tolerant and Pusa 991- saltsensitive) under NaCl salinity (0-100 mM). Salinity had significant negative effects on roots than shoots which disturbedroot to shoot ratio (RSR). The impact of salinity was directly proportional to increased Na ion uptake and reduced AMpercent root colonization. The decline in root growth affected the nodulation ability of Sinorhizobium fredii AR-4 in bothgenotypes, more in Pusa 991 than Pusa 2001. Individual application of Putrescine and AM improved plant growth,nutrient status and significantly reduced Na+ uptake. AM was found to be more efficient in improving nodulation ability ofPusa 2001 than Put. In addition Put was highly efficient in improving mycorrhizal colonization and imparted significantsalt tolerance to pigeonpea genotypes under combined treatments (Put + AM). Our results demonstrated that the benefitsof AM inoculation under salt stress can be significantly enhanced through Put treatment.

PF309 (IPPC0473)

Effect of biodeg products on growth and biochemical attributes of sugarcane

Smita Singh1*, Anshu Singh1, Abhishek Sharma2, M.K Srivastava2, R.K Singh1, Asha Gaur1, S.K. Shukla1, S.P. Singh1,

A. Chandra1 and Radha Jain1

1Indian Institute of Sugarcane Research, Lucknow-226002, India2Biodeg Chemicals & Allied Industry & Kemtech Polymers, New Delhi, India


In sub-tropical India, cane and sugar productivity are undesirably limited by factors like low germination (35-40%) consuming

more time (about 45 days), low tillering (main cane / tiller ratio1:2) leading to lower NMC, finally culminating in cane and

sugar yield of about 55-60 t/ha and 6-7 t/ha, respectively. In present investigation, a field experiment was conducted at

IISR farm to study the effect of Biodeg products on growth and biochemical attributes of sugarcane. These two products

are enzyme based organic fertilizers obtained by Biodeg Chemical & Allied Industry & Kemtech Polymers, New Delhi.

Treatments comprises of T1 (Biodeg 310, @ 4 kg/acre) and T2 (Bioreg 311, @ 3 kg/acre) with 50% NPK, T3 (Biodeg 310,

@ 4 kg/acre) and T4 ( Bioreg 311, @ 3 kg/acre) with 100% NPK, T5 (Biodeg + Bioreg, @ 4+3 kg/acre) 50% NPK, T6

(Biodeg + Bioreg, @ 4+3 kg/acre), with 100% NPK and T7 and T8 (control) with 50% and 100% NPK respectively. Results

obtained indicated improvement in bud sprouting, shoot population, leaf number, nitrate reductase (NR) activity, chlorophyll,

carotenoids, SPAD reading, amino acids and IAA contents by application of both the products. T1 treatment showed

highest bud sprouting (29.44%) and shoot population (333/plot), however, fresh weight and leaf number were maximum

in T5 at grand growth stage. NR activity ranged from 4.92 to 18.22 ìg nitrite formed per 100 mg fwt, T4 showed highest

activity. IAA and amino acids contents of shoot tip were highest in T4 (75.54 ìg IAA/100 mg fwt) and T6 (168 ìg aa/100 mg

fwt) treatments, respectively.


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IPPCongress2015


PF310 (IPPC0488)

Assessment of genetic variation for nodulation potential in chickpea genotypes under field conditions

Karivaradharajan Swarnalakshmi1*, Subramaniam Gopalakrishnan2, Mohan Singh3, Sushil Kumar Chaturvedi3, Preeti

Chopra1, Vijay Poonia1, Hari Deo Upadhyaya2 and Partha Basu3

1Indian Agricultural Research Institute, New Delhi-110012, India,2International Crops Research Institute for the Semi-Arid-Tropics, Patancheru-5020324, Andhra Pradesh3Indian Institute of Pulses Research, Kanpur-208024, India*Presenting author: [email protected]

Chickpea is an important rainfed crop covering about 9-9.5 m ha area and is largely grown on sandy loam to heavy claysoil with seldom use of irrigation, fertilizers and agrochemicals. The crop has the inherent potential to assimilate atmosphericnitrogen due to symbiotic association with Mesorhizobium ciceri. The contribution of biological nitrogen fixation in chickpeacrop is reported to be around 40-80 kg/ha depending on the plant genotype and soil type. Lower grain yields at fieldconditions have also observed due to poor nodulation and nitrogen fixation. Identification of high nodulating genotypesfor breeding cultivars with greater nitrogen fixing capacity will be useful for sustenance of chickpea productivity underfield conditions. Therefore, evaluating the extent of genotypic variation in nodulation potential and plant growth in availablechickpea minicore collection is required for selection of parental line for high BNF. Efforts were made to phenotypechickpea minicore lines to identify genetic source for high nodulation potential under field conditions. Nodulation, growthand yield at different physiological stages were recorded. Large variation in nodule weight ranged from 2 to 67.5 mg/plantwas observed at early vegetative stage. Chickpea lines ICC13816 (67.5 mg/plant) and ICC4973 (60 mg/plant) showedhighest nodulation compared to the IG7087 and ICC2737 (2 mg/plant). Similarly shoot weight of these lines were 840(ICC13816) and 505 mg/plant (ICC 4973) and 150 mg/plant (IG7087) and 365 mg/plant (ICC2737). Plant genotypedependent variations in nodulation potential of chickpea mini core accessions provide a scope for selecting efficientnodulating lines and using them in varietal improvement programme.

PF311 (IPPC0519)

An efficient element-contamination-free hydroponic system for dissecting phosphate deficiency-mediated

response of the root system and molecular traits in rice

Manisha Negi1*, Raghavendrarao Sanagala1, Vandna Rai1, Shivendra Sahi2 and Ajay Jain1

1National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India2Western Kentucky University, 1906 College Heights Blvd. Bowling Green, KY 42101, USA*Presenting author: [email protected]

Phosphorus (P) is pivotal for plant growth and development, and absorbed by root system in the form of inorganicphosphate (Pi). However, due to slow diffusion rates in rhizospheres it is often a limiting nutrient leading to loss of cropproductivity. Root system often attracted the attention because it exhibits extensive plasticity to various environmentalcues including Pi and other nutrient deficiencies. However, unlike Arabidopsis, rice is largely been grown in pot cultureand /or hydroponics but growth conditions currently being employed are often prone to elemental contamination, and notto facilitate easy documentation of the different traits of roots. Here in this study an element-contamination free aeratedhydroponic system was developed for detailed temporal (2 d, 4 d and 7 d) analysis of the effects of Pi deficiency ondifferent traits of primary, seminal and adventitious roots, of rice seedlings. After 2 d treatment, no significant effects wereobserved on any of these root traits. However 4 d and 7 d treatments, exerted differential effect on these root traits.Further, ICP-MS analysis revealed noticeable influence on ionomic profiles of macro-(P, K, S and Mg) and micro-(Fe, Zn,Mn and Cu) elements during growth under P+ and P- conditions. Further, Pi deficiency trigger significant increase onseveral genes that play a key role in Pi acquisition and or sensing and signaling cascade. Overall, the study provided theempirical evidence towards the fidelity of the system for detail analysis of Pi deficiency on morphophysiological andmolecular responses of rice.


273

IPPCongress2015


PF312 (IPPC0592)

Integrated approach for maximization of seed yield in mustard [Brassica juncea (L.) Czern & Coss]

R.S. Sain*, S.S. Bareth, H.S. Cheema and N.K. Gupta

Seed Technology Research, Rajasthan Agricultural Research Institute (Sri Karan Narendra Agriculture University, Jobner) Durgapura,Jaipur-302018, Rajasthan, India


Quality seed is important for realizing the potential of all other factors without which the improvement on account of

fertilizer, irrigation and pesticides will not pay the desired dividends to the farmers. To study the effect of integrated yield

maximization practices in mustard crop an experiment was conducted at RARI, Durgapura, Jaipur from rabi 2012-13 to

2014-15. The experiment was laid down in a split plot design in four replications with two main treatments based on date

of sowing and sub treatments based on nutrient management through soil application viz., recommended dose (RD) of

NPK; RDNPK + gypsum (RD); RDNPK + gypsum (RD) + Zn (ZnSO4 @ 25 kg/ha); RDNPK + gypsum (RD) Zn (ZnSO

4 @

25 kg/ha + Fe (FeSO4 @ 25 kg/ha. Results showed that seed yield and per cent germination were affected only by

nutrient management and days to maturity by date of sowing significantly. All other characters viz., days to flowering,

plant height, 1000-seed weight, seedling length and seed vigour index were affected by both date of sowing (main effect)

and nutrient management (sub effect) significantly. Seed yield was maximum (23.68 q/ha) in treatment RDNPK + Gypsum

+ Zn (ZnSO4 @ 25 kg/ha) + Fe (FeSO

4 @ 25 kg/ha) being at par with treatment RDNPK + Gypsum + Zn (ZnSO

4 @ 25 kg/

ha)). Thus, timely sowing and optimized nutrient management including critical micronutrients serves to enhances seed

yield and improves quality parameters such as 1000- seed weight, germination per-cent, seedling length and seed vigour

index.

PF313 (IPPC0600)

Effect of abiotic stresses on expression of genes involved in nitrogen uptake and assimilation pathways

in Brassica juncea L.

Parul Goel* and A.K. Singh

Department of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, India*Presenting author: [email protected]

Nitrogen (N) is primary plant nutrient that plays a crucial role in determining plant growth and productivity. Abiotic stresseslike salinity, drought and extreme temperature affects the process of nitrogen uptake and assimilation in plants. However,little is known about the regulation of N pathway genes at transcriptional level under abiotic stress conditions in Brassicajuncea. We have identified and cloned genes encoding nitrate transporters (NRT), ammonium transporters (AMT), nitratereductase (NIA), nitrite reductase (NIR), glutamine synthetase (GLN), glutamate dehydrogenase (GDH), asparaginessynthetase (ASN) and glutamate synthase (GLU) from Brassica juncea L. var. Varuna. Expression of these genes wasstudied under Salinity (150 mM NaCl), Cold (4°C), Heat (42°C), and Osmotic (250 mM Mannitol) stress after 1hr and 24hrof treatments. Most of the genes encoding N transporters and enzymes of assimilation pathway were found to bedownregulated under abiotic stress treatments. The expression of genes encoding BjNRT1.1 and BjNRT2.1, main membersof low and high affinity nitrate transport systems, respectively and genes encoding enzymes of nitrate assimilation areseverely downregulated after 24h of stress treatments. The expression of BjAMT1.1, BjGDH1.1 and BjASN2 wasdownregulated at both, 1hr and 24hr of all the stresses treatments. While, expression of BjGS1.1 and BjGDH1.2 geneswas downregulated after 1hr and expression of BjNRT2.1, BjNR2, and BjNIR1, genes was downregulated after 24h of allthe stress treatments. However, expression of BjNRT1.1, BjNRT1.5 and BjGDH2 was upregulated after 1h of all stresstreatments, while no gene was found to be upregulated after 24h of stress.


274

IPPCongress2015


PF314 (IPPC0645)

Effect of high (CO2) on membrane lipid remodeling and phosphorus uptake efficiency in wheat under

phosphorus stress with different sulphur levels

Milan Kumar Lal*, Krishnapriya Vengavasi, Lakshmi Raj, Sangeeta Khetrapal and Renu Pandey



Phosphorus (P), a non-renewable resource is deficient in most of the agricultural systems while continued rise in atmospheric(CO

2) helps the plants, particularly C

3 in more efficient acquisition and utilization of P. We studied the effect of high (CO

2)

and sulphur (S) levels on alteration of membrane lipid composition under P stress. Different levels of P (5 μM, low-P and500 μM, sufficient-P), (CO

2) (380 ppm, aCO

2 and 700 ppm eCO

2) and S (10 μM, low-S and 2000 μM, sufficient-S) were

used on bread wheat (Triticum aestivum cv. PBW-396) and durum (T. durum cv. PDW-233). The total P uptake increasedin plants grown under e(CO

2) with sufficient-P and S (control) by 32% while in low-S it was 46% higher compared to

a(CO2). In membranes, the amount of phosphalidylglycerol (PG) decreased while there were 2- to 2.5-fold increase in

sulfoquinovosyldiacylglycerol (SQDG) due to interactive effects of low-P and e(CO2). In plants grown at low-S, the level of

SQDG decreased in both varieties but the interactive effects of e(CO2) and low-S resulted in two-fold increase in PG. In

response to nutrient deprived condition, transcript levels of TaNPC4 increased in both root and shoot, increase being 2.6-folds in low-P, 1.5-fold in low-P/S and only 1.3-fold in low-S. The transcripts of TaPLD4 were highly induced in both rootand shoot tissues in response to e(CO

2). Our finding in turn suggests that under high atmospheric (CO

2) plants will

respond to P deficiency by altering its membrane lipid composition provided there is sufficient S fertilization.

PF315 (IPPC0672)

Genome-wide expression of transcriptomes under iron and zinc stresses revealed candidate genes for

kernel iron and zinc accumulation in maize (Zea mays L.)

Mallana Gowdra Mallikarjuna1*, Nepolean Thirunavukkarasu1, Rinku Sharma1, Kaliyugam Shiriga1, Shikha Mittal1,

Firoz Hossain1 and Hari Shanker Gupta2

1Maize Research Lab, Division of Genetics, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India2Borlaug Institute for South Asia, CG Block, NASC Complex, New Delhi-110012, India


Deficiency of iron (Fe) and zinc (Zn) causes micronutrient-malnutrition or hidden-hunger affecting millions of peopleglobally including sub-Saharan Africa, South Asia and Latin America where, maize is one of the major sources of foodand feed. Among several available approaches, genetic-biofortification is a sustainable and cost-effective approach inaddressing hidden-hunger. Identification of key genes involved in Fe and Zn homeostasis is an important prerequisite tobreed micronutrient-rich hybrids. A transcriptome assay was performed, with ~18,000 transcripts distributed across themaize genome, in a SKV-616 inbred after exposing the seedlings to three Fe and Zn stress treatments (+Fe –Zn, –Fe+Zn, –Fe –Zn) along with control (+Fe+Zn). A total of 306 and 715 genes expressed in root and shoot, respectively werefound common across stress treatments, suggesting presence of common transporters controlling both minerals in rootand shoot. Differential expression of genes under stress treatments revealed that Zn association in many pathways wasmore pronounced over Fe pathways and the physiological demand for these minerals in shoot was much higher than inroot. Significant expression of genes involved in mugineic acid pathway, metal-transporters, and phyto-hormonemetabolisms revealed the role of those pathways in understanding the physiological and molecular mechanisms of Feand Zn homeostasis. Co-mapping of DEGs with known QTLs for kernel Fe and Zn concentration identified several candidategenes such as OPT, metal-binding-protein, nramp3 and NAS. The candidate genes identified in the present investigationcould be used for breeding kernel Fe- and Zn-rich maize hybrids.


275

IPPCongress2015


PF316 (IPPC0799)

Effect of Treated distillery spentwash on fodder maize growth and yield

M.B. Doddamani1*, Edna Antony2 and S.K. Gali3

1Department of Environmental Science, College of Agriculture, University of Agricultural Sciences, Dhrwad-580005, Karnataka, India2Indian Grassland and Fodder Institute, Regional Research Institute, Dharwad-580005, Karnataka, India3Dean (Student Welfare), University of Agricultural Sciences, Dhrwad-580005, Karnataka, India


A field experiment on the utility of treated spentwash as source of plant nutrients on fodder maize (African Tall) was

conducted in the year 2014 at MARS, University of Agricultural Sciences, Dharwad. The soil of the experimental field was

Vertisol with clay texture, alkaline in reaction and low electrical conductivity. Graded doses of treated spentwash was

applied alone and incombintaion with fertilizer, based on the nitrogen requirement of the crop. Forty per cent of the

calculated required nitrogen through spentwash was applied 15 days before sowing and remaining 60% was applied after

sowing. Treated spentwash treatment @ 1½ N through spentwash recorded higher plant height, stem diameter, number

of leaves, leaf area, leaf area index, Number of days to 50% tasseling, dry forage yield, seed yield, crude protein and

crude fiber and was on par with 1½ N through fertilizer treatment. The highest plant height (289.6 cm) was recorded with

1½ N through spent-wash followed by 1½ N through fertilizer (286.7 cm). Seed yield (12.5 q/ha), dry fodder (17.0 t/ha),

crude protein and crude fibre content were also higher with 1½ N through spent-wash treatments. Similar trend was also

noticed with respect to stem thickness and leaf to stem ratio and maximum numbers of leaves (13.8) as compared to local

farmers practice. This study is centered on the concept that treated distillery spent wash could be used for one time soil

application as a nutrient source without adversely affecting soil and crop health.

PF317 (IPPC0805)

Cloning and characterization of salt and nutrient stress responsive TaCBL4 gene from wheat

Lekshmy S.*, S.K. Jha2, C. Viswanathan1 and R.K. Sairam1

1Division of Plant Physiology, 2Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India


Worldwide wheat production is impacted by various abiotic stresses, like water deficit, salt, high temperature and nutrient

deficiencies. Genetic improvement of abiotic stress tolerance of wheat can be greatly aided by knowledge of abiotic

stress signaling. Calcium is a ubiquitous signaling molecule mediating signal transduction pathways of diverse

environmental stresses. Calcineurin B-like proteins (CBLs) are related to neuronal calcium sensors of animals. CBL-

interacting protein kinases (CIPKs) act as targets of CBL proteins and mediate stress response pathways in many plants.

However, little is known about CIPKs in wheat and very few CBL-CIPK components are characterized in wheat. CBL4

(Salt Overly Sensitive 3) gene is involved in modulating salinity tolerance of Arabidopsis and Rice. TaCBL4 gene

homologous to salt stress responsive OsCBL4 was cloned from wheat and sequence characterized. Expression of TaCBL4

gene was up regulated by salt stress, nitrogen, phosphorus and potassium deficiency albeit the expression was higher

under salt stress. It is possible that TaCBL4 might mediate crosstalk among different signaling pathways. This work may

facilitate functional studies of TaCBL4 gene and is beneficial for further molecular breeding in wheat.


276

IPPCongress2015


PF318 (IPPC0849)

Potassium induced changes in antioxidant system and growth performance of Trigonella foenum-graecum

L. cultivar RMT

Madhup Shrivastava*, Mohammad A. Ahanger, Megha Tittal, Surendra Argal and R.M. Agarwal

School of Studies in Botany, Jiwaji University, Gwalior-474011, India


Potassium is important for plants because of its active involvement in many functions such as enzyme activation, osmotic

adjustment, maintaining transmembrane gradient and uptake of deleterious ions like Na. Pot experiments were conducted

to find out the effectivity of different potassium salts on certain components of nitrogen metabolism, antioxidant system

and ultimately on growth and yield of Trigonella foenum-graecum L. Varying doses (20, 40, 60 kg ha-1) of three potassium

salts i.e., K2O, KCl and KNO

3 were applied and analysis was done at two different developmental stages i.e., vegetative

(25 DAS) and pre flowering (40 DAS). Obvious increase in growth and chlorophyll content was recorded. Supplementation

of potassium salts enhanced the activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT)

and peroxidase (POD) in Trigonella foenum-graecum L. Added potassium enhanced contents of total phenols and tannins,

thereby probably strengthening both the enzymatic as well as non enzymatic antioxidant system. Potassium application

resulted in considerable increase in synthesis/accumulation of osmolytes like free amino acids, proline and free sugars

which assumes special significance in growth under water stress conditions. Enhancement in the activities of nitrate

reductase and aminotransferases was also noticed indicating better nitrogen assimilation in potassium supplied plants

thereby ensuring better plant growth. Hence, the result indicate that potassium supplementation may contribute towards

qualitative changes in Trigonella foenum-graecum L beneficial from pharmacognostic/ medicinal viewpoint.

PF319 (IPPC0876)

Yield and water production functions of wheat (Triticuma eastivum L.) cultivars and response to exogenous

application of thiourea and ortho-silicic acid

Ratna Kumar Pasala*, P.P. Deokate, G.C. Wakchaure and R.L. Choudhary

National Institute of Abiotic Stress Management (NIAM), ICAR, Malegaon, Baramati-413115, Pune, Maharashtra, India


Supplemental irrigation, drought tolerant cultivars and use of plant bioregulators are now being proposed as the key

strategy to unlock the yield potential and stabilize the yield of wheat grown in rainfed areas. Experiments were therefore

conducted during 2013-2015 using line source sprinklers (LSS) to determine the interactive effects of quantities of

supplemental irrigation and exogenous foliar sprays of plant bioregulators- 10 mM thiourea (TU) and 32 ppm of ortho-

silicic acid (OSA) on wheat (Triticuma estivum L.) varieties viz., HD 2189, LOK 1, NIAW 301, NIAW 34 and PBW 550. The

irrigation quantities were: fully irrigated (I1: 31.8 cm); mild (I

2: 28.9 cm); medium (I

3: 25.9 cm and I

4: 22.7 cm); and severe

(I5: 19.9 cm and I6: 17.2 cm) water stress conditions. The two year field experiments demonstrated that wheat cultivars

varied in their response to water deficits and those e.g. NIAW-301 with higher water productivity under limited irrigation.

Response to TU, OSA also varied across water regimes and was higher under moderate to severe stress. Foliar application

of TU and OSA at root crown index, flag leaf and seed filling stages improved grain yield by 9 and 6 % at fully irrigated; 19

and 18% at medium stress; 17 and 12% at severe water stress conditions and water productivity 0.12- 0.10 mg h-1 with

TU, 0.13-0.09 mg h-1 with OSA at fully irrigated and medium stress; 0.11-0.03 mg h-1 with TU, 0.12-0.03 mg h-1 with OSA

at severe stress conditions. Our major finding was that TU and OSA has various degrees of potential efficacy in alleviation.


277

IPPCongress2015


PF320 (IPPC0877)

Physiological approaches for yield improvement in pigeonpea in Karanataka

Mahalingappa Dhanoji* and Mukesh Kumar Meena

University of Agricultural Sciences, Raichur-584104, Karnataka, India


A field experiment was conducted during kharif-2014-15 to know the effect of hardening and foliar nutrition of essential

elements and PGR’s with a view to mitigate terminal drought and realise yield potential in pigeonpea. Research results

data revealed significant (6.4%) increase in the seed yield in pigeonpea with CaCl2 @ 2.0% seed hardening over control.

Further there was also statistical improvement in the seed yield due to foliar nutrition of (all 19 nutrients including

micronutrients) as compare to control. On the other hand, among the PGRs spray, NAA @ 20 ppm recorded significant

improvement in the seed yield followed by nitrobenzene, BA-6 and CCC over control and tricontinol and water spray. The

Interaction effect due to hardening, foliar spray of essential nutrients and PGRs, revealed significant improvement in the

yield with CaCl2 @ 2.0% seed hardening + foliar spray of nutrients (all 19 nutrients including micronutrients) + foliar spray

NAA @ 20 ppm over control and water spray. The improvement in the seed yield due to hardening, foliar spray of

essential elements and PGRs might be attributed to improvement in the mean value of ancillary growth and physiological

parameters, viz., plant height, number of branches, test weight, pod weight per plant, Dry matter accumulation, light

interception, NDVI and SPAD values.

PF321 (IPPC1008)

Impact of elevated atmospheric carbon dioxide level on uptake of iron and zinc in rice crop

A. Raj1, B Chakrabarti2, H. Pathak3, U. Mina4, R.C. Harit5* and V. Kumar6

Centre for Environment Science and Climate Resilient Agriculture, Indian Agricultural Research Institute, New Delhi-110012, India


Climate change associated with rise in atmospheric greenhouse gases (GHGs) concentration is likely to affect global

food production and agricultural sustainability. Rice crop is also affected by this rise in carbon dioxide (CO2) in terms of

growth, yield and nutrient uptake. An experiment was conducted during the kharif season of year 2013 inside and outside

the Free Air Carbon dioxide Enrichment (FACE) facility at the farm of Indian Agricultural Research Institute, New Delhi to

study the impact of elevated CO2 on uptake of iron (Fe) and zinc (Zn) in rice crop. Two N doses were applied in both

ambient (395 ppm) and elevated CO2

(550 ± 20 ppm) conditions i.e., N0: no N (control), N1: 0.8 g N pot-1(100 %

recommended dose). Results showed that high CO2 significantly enhanced both grain and biomass yields of rice crop.

Uptake of Fe and Zn increased significantly with rise in CO2 level and with N application. With recommended dose of N,

uptake of Fe and Zn in rice grain increased by 64.2% and 19.2% under elevated CO2 condition as compared to ambient

CO2 level. Total uptake of Fe and Zn also followed the same trend with 31.9% and 25.9% increase in elevated CO

2

treatment. Increased yield at higher CO2 level has resulted in higher micronutrient uptake of rice crop.


278

IPPCongress2015


PF322 (IPPC1009)

Ammonia volatilization under different air temperature regimes in wheat crop

Renu Singh1*, Vijay Kumar1, Arti Bhatia1, Renu Pandey2, Vinod Kumar1, Monika Srivastava1 and Himanshu Pathak1

1Centre for Environmental Science and Climate Resilient Agriculture, 2Division of Plant Physiology, Indian Agricultural ResearchInstitute, New Delhi-110012, India


Nitrogen (N) is an essential nutrient for crop production and its efficient use is important for the economic sustainability of

cropping systems. Nitrogen use efficiency (NUE) is one of the major issues in agriculture as it is directly associated with

crop yield, fertilizer costs and N-pollution. In the present scenario of global warming, the increasing air temperature

influences NUE and ultimately crop yield. Ammonia volatilization is the major process leading to loss of N from soil under

increasing temperatures. Wheat being major cereal crop, farmers is bound to apply excessive nitrogenous fertilizer to

increase crop productivity which leads to low NUE as well as lower net returns. An experiment was conducted to evaluate

the effect of high atmospheric temperature on ammonia volatilization and yield under different N levels in wheat crop.

Four different levels of temperature (ambient temperature, temperature gradient tunnel (TGTc) control, TGT+2.5°C and

TGT+3.5°C) and N (0, 90, 120 and 150 kg ha-1) were taken in combinations to grow wheat. The ammonia volatilization

loss ranged from 4.2-20.4 kg ha-1 season-1 while the highest loss was observed in treatment with 150 kg ha-1 N and

TGT+3.5°C. This treatment accounted for 13.6% loss of applied N. The study revealed that the rate of volatilization

significantly increased with rise in air temperature which may lead to decrease in NUE in winter crops like wheat thus

reducing productivity.

Session G : Plant Growth Regulators and Applications

279

IPPCongress2015


PG323 (IPPC0097)

Utilization of fungicide for better seedling growth responses in mung bean [Vigna radiata (L.) Wilczek]

Mukul Kumar* and Prakash Chandra Gupta

Bihar Agricultural University, Sabour, Bhagalpur-813210, Bihar, India


Fungicides are commonly used for seed treatment to prevent seed borne diseases, and thus to establish a good field

stand for high productivity. Non-systemic fungicide thiram (N,N-dimethylcarbamodithioate) and systemic fungicide bavistin

(methyl-N-1H-benzimidazol-2-yl-carbamate) are widely used for seed treatment. However, these fungicides may influence

the plant physiological process and thus may regulate seedling vigour and crop growth. Previous study on wheat showed

that seeds treated with these fungicide delayed germination and decreased percentage germination. In contrast, seed

dressing with thiram has been shown to increase the seedling vigor of sorghum. Hence, we conducted a lab experiment

to study the effect of thiram and bavistin seed treatment on the seedling growth of mung bean, a major rice fallow pulse

in India. A lab experiment in CRD design with three replications was conducted under controlled temperature and relative

humidity in lab conditions during kharif season. The experiment consisted of three treatments, viz., non-treated seeds

(control), thiram and Bavistin treated seeds of mung bean. Morphological characters such as shoot length, root length,

leaf area and dry weight of seedling showed significant differences between treatments. Thiram fungicide treatment-

induced enhancement of the shoot length, root length, leaf area and dry weight of seedlings was higher than that of

Bavistin treatment and control. The results suggest that species specific response of plants to these fungicides. Treatment

with thiram may help enhance yield of rice-fallow mung bean in India.

PG324 (IPPC0111)

Role of ethylene in heavy metal tolerance: Modulation of ascorbate-glutathione pathway and proline

metabolism in mustard plants

M. Iqbal R. Khan* and Nafees A. Khan

Department of Botany, Aligarh Muslim University, Aligarh-202002, U.P., India


Heavy metal contamination of agricultural soil is primarily related to the abrupt anthropogenic activities. Plants frequently

exposed to high concentration of HMs shows adverse effects. In the present study we investigated the role of ethylene

significance in nickel (Ni) and zinc (Zn) stress tolerance in mustard (Brassica juncea) plants. Application of Ni and Zn at

the rate of 200 mg kg-1 soil induced oxidative stress along with increased ethylene production. Under Ni and Zn stress,

photosynthetic attributes and growth characteristics of mustard plants was reduced. Application of ethephon significantly

increased ascorbate-glutathione (AsA-GSH) pathway and proline metabolism and maintained cellular homeostasis and

water relation of mustard plants. Up-regulation of AsA-GSH and proline metabolism resulted in protection of photosynthesis

and growth of the plants under Ni and Zn stress. The result was substantiated by using ethylene action inhibitor,

norbornadiene (NBD), which significantly inhibits both AsA-GSH cycle and proline metabolism resulted in less protection

of photosynthesis and growth of plants. The results of the present study could be exploited in situation where plants are

grown under higher level of Ni and Zn conditions.


280

IPPCongress2015


PG325 (IPPC0129)

Potential uses of homobrassinollide plant growth regulator in increasing berry growth and yield of grape

var. Thompson seedless

Ajeet Singh Tomar*, Burhanuddin Bohra and Girish Dalal

Godrej Agrovet Ltd., R&D Department, Vikhroli-East, Mumbai- 400079, India


Plant growth regulators impart important role towards higher yield of crops. Among different strategies, exogenous

applications of plant growth regulators are involved in promoting plant growth and development under normal and stressful

conditions. Plant growth regulator containing growth hormone homobrassinolide (HBR) enables crop plants to overcome

inefficiency barriers by triggering vital physiological process ensuring higher yields of agricultural and horticultural crops.

In this study, possible role of exogenously application of HBR on berry growth, bunch weight and quality parameters of

table grapes were evaluated. The applications of HBR alone and in combination with Gibberellic acid (GA) in the form of

two dips were given at 2-3 mm and 5-6 mm berry size stages. Clusters were harvested when those of the control vines

reached 16-17 °Brix. Yield and quality parameters were analyzed. It was revealed that first dip with HBR (1 ppm) + GA

( 50 ppm) at 2-3 mm berry size stage followed by second dip with HBR (1 ppm) + GA ( 40 ppm) at 5-6 mm berry size stage

provides significant increase in berry weight, berry size and bunch weight over control and GA treatment. The study

revealed that the use of HBR in combination with GA has a great significance due to its synergistic effects in quality

production berries of grape var. Thompson seedless which has huge potential in export markets.

PG326 (IPPC0144)

Multiple Interactions between glucose and brassinosteroid signalling in regulating plant growth and

development

Aditi Gupta*, Manjul Singh and Ashverya Laxmi



Brassinosteroids (BRs) and sugars/glucose (Glc) are fundamental to plants and regulate similar processes. There are

few evidences for BR and Glc signaling interactions in plants but its molecular basis is not explored. We hypothesize that

there may be an extensive overlap between BR- and Glc- signaling pathways. Whole genome transcriptome analysis was

performed to find out the interdependence/overlap between BR and Glc signal transduction pathways in light grown as

well as etiolated seedlings of Arabidopsis. Glc alone was able to regulate 55% and 71% of BR-regulated genes under

light and dark growth conditions respectively. BR and Glc commonly regulate a large number of genes involved in growth

and development. Physiologically; both Glc and BR could regulate many aspects of early seedling growth and development

such as; hypocotyl and root elongation, root and shoot directional growth, lateral root emergence, root hair development

etc. We utilized both genetic as well as pharmacological approaches to establish hierarchy of multiple signaling components

such as Glc, BR, auxin, cytokinin ethylene etc. in regulating different parameters of seedling development. Briefly, BR

signaling via BRI1 receptor interacts with HXK1-dependent Glc signaling pathway. Intact BR response pathway is essential

for Glc regulation of seedling growth and development. Auxin transport and signaling components mainly act as nodal

points in regulating Glc and BR mediated growth responses. Our results, altogether, suggest that Glc and BR signaling

pathways interact extensively to regulate various growth parameters required for maintaining optimal seedling fitness in

nature.


281

IPPCongress2015


PG327 (IPPC0159)

Protection of growth via antioxidants and endogenous hormones maintenance in response to 24-

epibrassinolide in tomato (Lycopersicon esculentum L.) growing under salt, drought and heavy metal stress

Deepak Singh Bagri* and Chandrama Prakash Upadhyay

Department of Biotechnology, Dr H.S. Gour Central University, Sagar-470003, Madhya Pradesh, India


Brassinosteroids (BRs) are a group of plant growth steroidal hormones present in very low concentrations in seeds,pollen, young plant tissues and are reported to be involved in plant development as well as in protection of plants againstvarious biotic and abiotic stresses. In this study, we report the effect of epibrassinolide (EBR) on in vitro and ex vitrogrown tomato (Lycopersicon esculentum L. cv. Pusa Ruby) seedlings under induced salt (NaCl), drought (mannitol) andheavy metal (ZnCl2) stresses. These stresses hampered the plant development and plant showing decreased shootlength, total leaf area and plant dry matter. However, these parameters changed with application of EBR and enhancedvalues with the same parameters were observed under both in vitro and ex vitro conditions. Significant increases in totalleaf chlorophyll, proline content and nitrate reductase (EC 1.6.6.1) activities were observed in leaves of tomato plantstreated with EBR when compared to their respective stressed controls. The activities of antioxidant enzymes such asascorbate peroxidase (EC 1.11.1.11), catalase (EC 1.11.1.6), dehydroascorbate reductase (EC 1.8.5.1), glutathionereductase (EC 1.6.4.2) and superoxide dismutase (EC 1. 15.1.1) enhanced in tomato plants under these stresses andtheir activities further enhanced with the application of EBR. The endogenous levels of indole-3-acetic acid and abscisicacid elevated significantly in the leaves of tomato plants treated with EBR compared to stressed controls. This is the firstcomprehensive report confirming the protective role of EBR in imparting tomato plant tolerance to abiotic stresses underboth in vitro and ex vitro conditions.

PG328 (IPPC0186)

Silencing of OsCKX2 enhances panicle branching resulting in improved grain yield in rice under both

normal and salinity stress conditions

Rohit Joshi1*, Khirod Kumar Sahoo1, Amit Kumar Tripathi1, Ashwani Pareek2 and Sneh Lata Singla-Pareek1

1Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Plants maintain their cytokinin levels through cytokinin oxidases which catalyze irreversible degradation of cytokinins.Rice cytokinin oxidase 2 (OsCKX2) gene has earlier been identified as a major QTL playing crucial role in determininggrain number. In the present study, we have carried out in-depth studies to unravel the physiological and developmentalfunction of OsCKX2. Via expression analysis using qRT-PCR, we found that OsCKX2 expression is regulated in adevelopmental-stage specific manner. The highest level of OsCKX2 transcript was found between 10-13 days afterpanicle initiation. To further study the effect of reduced OsCKX2 levels, stable RNA interference mechanism was utilizedfor silencing OsCKX2 gene in rice. Ultra-performance liquid chromatography (UPLC)-based estimation of CKs revealed asignificant increase in CKs in OsCKX2-knockdown plants. Enhanced CK levels correlated with enhanced grain yield inthe OsCKX2-knockdown plants. To check if the OsCKX2-knockdown plants maintain their yield even under salinity stressconditions, we assessed their growth, physiology and grain yield under these conditions. OsCKX2-knockdown plantsshowed better relative water content, higher photosynthetic efficiency and reduced electrolyte leakage as compared tothe wild-type plants under 200 mM NaCl stress. Importantly, we found a negative correlation between OsCKX2 expressionand plant productivity as the OsCKX2-knockdown plants showed enhanced panicle branching, more filled grains perplant, higher 1000 grain weight and better harvest index. Together, the results of our study suggest that OsCKX2 may beexplored as a unique regulator of floral primordial activity for enhancing grain production under normal as well as abioticstress conditions.


282

IPPCongress2015


PG329 (IPPC0291)

Auxin plays a critical role in spikelet fertility of rice under drought and high temperature stresses

Laxmi Sharma1*, Rakesh Verma2, Santosh Vinjamuri2, S.R. Kushwaha2, Monika Dalal3 and Viswanathan Chinnusamy2

1ICAR-Vivekanand Parvatiya Krishi Anusandhan Sansthan, Almora-263601, Uttarakhand, India2Division of Plant Physiology, 3NRC on Plant Biotechnology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India*Presenting author: [email protected]

Reproductive processes of plants are highly vulnerable to the environmental stresses, which are increasing due to global

climate change. Drought and high temperature (HT) are the key stress factors limiting the yield potential in rice. Auxin is

considered to be the master regulator of development and patterning. Increasing evidences indicate that auxin plays

critical role in adapting the abiotic stress response. Reports show that HT-induced reduction in the endogenous auxin

content was responsible for loss in pollen viability in barley and Arabidopsis. In this study, we analysed the role of auxin

(indole-3-acetic acid, IAA) in regulating the spikelet fertility of rice under drought and HT stresses. Both these stresses

reduced endogenous IAA levels and the expression of genes coding for auxin biosynthesis and signalling. In addition,

these stresses significantly reduced pollen viability and spikelet fertility in most of the rice genotypes. Application of

exogenous IAA before exposing the plants to drought or HT stress mitigated adverse effect of drought and heat through

restriction of oxidative stress. This was reflected in better pollen viability and spikelet fertility percentage as well as lower

ROS accumulation and membrane injury in the stressed plants. These results suggest that stress induced decline in

endogenous IAA levels is one of the causes of reduced spikelet fertility in rice. Exogenous application of auxin and/or

genetic improvement in the ability of rice to maintain auxin homeostasis under these stresses could help maintain the

spikelet fertility and thus combat the global warming-mediated yield loss in rice.

PG330 (IPPC0345)

Physiological effects of strobilurin compounds in chilli (Capsicum annum L.)

A. Senthil*, S. Punitha and P. Jeyakumar

Department of Crop Physiology, Tamil Nadu Agricultural University, Coimbatore-641003, Tamil Nadu, India


Strobilurins, a new class of fungicides, have shown to be effective in altering the physiological and biochemical traits of

crop plants. Experiment was conducted to study the influence of strobilurin fungicides on morpho-physiological, biochemical

parameters and yield of chilli. The experiment consisted of seven treatments with strobilurin such as trifloxystrobin combined

with tebuconazole as Nativo @ 300, 350 and 400 g/ha, pyraclostrobin combined with metiram as Cabriotop @ 1500 g/ha

and azoxystrobin as Amistar @ 500 ml/ha, along with carbendazim @ 500 g/ha and untreated control. The treatments

were given as foliar spray at 45 and 60 DAT of chilli crop. The results revealed that application of Nativo @ 400 g/ha

recorded the highest root length, root shoot ratio and more number of branches per plant among all the treatments.

Maximum leaf area, LAI and SLW were recorded in Cabriotop @ 1500 g/ha followed by Nativo @ 400 g/ha. The CGR and

TDMA were significantly influenced by Nativo @ 400 g/ha. Among the treatments, Nativo @ 400 g/ha recorded the

highest chlorophyll index, photosynthetic rate and recorded the maximum green chilli yield of 20.44 t/ha which was

followed by Cabriotop @ 1500 g/ha (19.40 t/ha). Hence, it is concluded that foliar application of trifloxystrobin combined

with tebuconazole as Nativo @ 400 g/ha at 45 and 60 DAT was found optimum for increased yield with high economic

returns in chilli crop.


283

IPPCongress2015


PG331 (IPPC0372)

Effect of foliar application of plant growth regulators on photosynthetic rate, stomatal density and grain

yield in relation to nitrous oxide (N2O) emission from field grown wheat

Nirmali Bordoloi* and K.K. Baruah

Department of Environmental Science, Tezpur University, Tezpur-784028, Assam, India


Agricultural fields are the major anthropogenic source of greenhouse gases nitrous oxide (N2O) and methane (CH

4)

emission to the atmosphere. The N2O emission from wheat grown field was studied for two years to investigate the impact

of different plant growth regulators (PGR) on photosynthesis, grain yield and N2O emission reduction. The growth regulators

namely Abscisic acid (ABA) and Cytozyme (20 ppm), Kinetin (10 ppm and 20 ppm) and wet tea extract (1:20 w/w) along

with distilled water as control were sprayed on wheat canopy at tillering and panicle initiation stages and N2O emission

was estimated from the treated plants. Cytozyme and tea extract markedly enhanced the plant dry weight in term of

biomass over control, while abscisic acid and kinetin increased the plant photosynthetic rate at panicle development and

flowering stages. The increased rate of photosynthesis in response to ABA and kinetin indicate efficient photosynthate

partitioning towards the developing grain. ABA (20 ppm) and Kinetin (10 ppm) also decreased the N2O emission over

control along with better grain production. Leaf area index, stomatal conductance, stomatal frequency and xylem vessel

size are found to be associated with N2O transport and emission to the atmosphere. Use of selective PGR like ABA and

Kinetin can enhance the grain productivity of wheat and also reduce the N2O emission from wheat cropping system.

Approaches to increase wheat productivity and reduce N2O emission may be particularly beneficial in a future climate

with rising temperature.

PG332 (IPPC0387)

Seasonal effects of explant collection on bud break and culture establishment from mature nodes of

Tecomella undulata (Sm.) Seem.

Sidhika Chhajer, R. Sharma, R.K. Bhatt and Rajwant K. Kalia*

Central Arid Zone Research Institute, Jodhpur-342003, Rajasthan, India


Tecomella undulata (Family Bignoniaceae), a medicinally and economically important timber tree of the dry regions of

India, Pakistan and Arabia, has been considered as endangered due to over exploitation and negligible conservation

efforts. Micropropagation can produce large number of disease free clones, therefore, to study the seasonal effect of

explant collection on bud break and shoot length, nodal segments were collected from 31 year old trees every month and

cultured on MS medium supplemented with BAP (10 uM), ascorbic acid (50 mg/l), arginine (25 mg/l) and citric acid

(25 mg/l). Highest percentage of explants (73%) showing response was recorded in explants collected during the months

of August-September. Fresh flush of shoots sprouted after rains in July are best for in vitro establishment of cultures.

Decline in response from November to February could be due to transition of actively growing spring buds to winter buds

during which cytokinin supply to shoot system falls down. Decreased response in the months of March-May could be due

to sinking of nutrients in pods for filling. Explants collected during rainy season were more prone to contamination while

it was not a major problem in other months. Shoots collected during dormant months showed increased callusing at the

explant base with lesser bud induction. A progressive reduction in callusing at explant base and increase in number and

length of sprouted buds was observed in explants collected during months of active growth. Therefore, the study concluded

that explants collected from actively growing shoots were best for culture establishment.


284

IPPCongress2015


PG333 (IPPC0390)

Influence of different 1 methyl cyclopropene doses and storage conditions on quality and organoleptic

evaluation of alphonso mangoes

Nitin Gokhale1*, Arun Mane1, M.B. Chetti2, Sandip Mahadik1, Murad Burondkar1 and Sanjay Bhave1

1Dr. B.S. Konkan Krishi Vidyapeeth, Dapoli-415712, Maharashtra, India2Indian Council of Agriculture Research, Krishi Anusandhan Bhawan-II, New Delhi-110012, India


Investigations were carried out, for two consecutive years (2011 and 2012), with an objective to test the effect of 1-methylcyclopropene (1-MCP) under different storage conditions on physico-chemical properties and organoleptic evaluation ofAlphonso mango fruit observed after prolonged shelf life during transportation. After harvesting at physiological maturity,followed by pre-cooling at 130C, fruit were exposed to vapors of 1-MCP @ 70 mg/m3 (1000 ppb) for 18 hrs in air tight(plastic) treatment chamber. The fruit quality assessed at different intervals viz. after 7, 14, 21 and 26 days of storage andshelf life behavior. Treatment combination of 1-MCP at 2000 ppb concentration and cold storage at 13 ±10C foundeffective in maintaining the quality of fruits even after 26 days storage (17.90 0B, 0.38%. 12.50%, 4.63% respectively) ascompared to control in 21 DAT (12.62 0B, 0.22%, 12.95, 3.875% respectively). The fruits treated with higher concentrationsof 1-MCP and kept under cold storage yielded slow rise in vitamin A content with further decline after reaching the peakvalues till the end of storage whereas the ascorbic acid content untreated ambient control fruits was significantly decreasingas compared to other treatment combinations. The organoleptic evaluation obtained at optimum ripening shown nosignificant difference in taste, aroma and flavor amongst all the treatment combinations, whereas 1-MCP treated fruitswith cold storage shown less occurrence of spongy tissue (physiological disorder) with better colour visibility when fruitswere treated with 2000 ppb 1-MCP.

PG334 (IPPC0391)

Ripening and storage behavior of alphonso mangoes as influenced by 1 methyl cyclopropene doses

under different storage conditions

Arun Mane1*, M.B. Chetti2, Sanjay Bhave1, Murad Burondkar1 and Nitin Gokhale1

1Dr. B.S. Konkan Krishi Vidyapeeth, Dapoli-415712, Maharashtra, India2Indian Council of Agriculture Research, Krishi Anusandhan Bhawan-II, New Delhi-110012, India


Investigations were carried out for two consecutive years (2011 and 2012), with an objective to test the efficacy ofdifferent doses of 1-methyl cyclopropene (1-MCP) under different storage conditions, in reducing the ripening rate andprolonging the storage and shelf life of Alphonso mango fruit to suit export by sea transportation from India. After harvestingat physiological maturity, followed by pre-cooling at 130C, fruit were exposed to vapors of 1-MCP @ 70 mg m-3 (1000 ppb)for 18 hrs in air tight (plastic) treatment chamber. The treated and untreated fruits were then transferred to differentstorage conditions for different intervals (7, 14, 21, and 26 days). Effect of 1-MCP on ripening pattern of fruits examinedat six difference stages (viz. green, turning yellow, ½ ripe, ripe, diseased and shrivelled) indicated that 1-MCP treatmentsignificantly arrested the ripening till 8 days and prolonged the subsequent stages from 12, 16, 20 and 24 days, overuntreated control fruit. The physical changes evaluated at different intervals shown the reduction in loss of colour, fruitfirmness and physiological weight under cold storage with an effective dose of 1-MCP at 2000 ppb, ultimately increasingthe shelf life of Alphonso mangoes. Whereas, physiological changes such as chlorophyll content, total carotenoids, rateof respiration and release of ethylene recorded at different intervals viz., after 7, 14, 21 and 26 days of storage found tosustain the minimum values till the end of storage period in case of cold storage treatments with 1-MCP at 2000 ppbconcentration.


285

IPPCongress2015


PG335 (IPPC0401)

Juglone promotes shooting and inhibits rooting in leaf explants of in vitro raised tomato (Lycopersicon

esculentum Miller var. Pusa Ruby) seedlings

Kiran Bamel1*, Rajendra Gupta2 and Shrish C. Gupta2

1Department of Botany, Shivaji College (University of Delhi), Raja Garden, Delhi-110007, India2Department of Botany, University of Delhi, Delhi-110007, India


The effect of juglone was evaluated on morphogenesis of leaf explants excised from in vitro raised 30-d-old seedlings of

tomato, Lycopersicon esculentum var. Pusa Ruby. To test the dose response different levels (10-9-10-3 M) of juglone

was added to the controls. Juglone caused significant inhibition of rhizogenesis as reflected by the reduced rooting

potential of the leaf explants and transition from rooting to shoot formation at 10-7-10-6 M. Juglone increased the

caulogenesis and callusing when added to a shoot regeneration medium. The optimum response was observed at 10-5

M. The results of the present investigation indicated that juglone acts as a stimulator of callusing and shoot formation.

PG336 (IPPC0409)

Effect of JA and Me-JA on chlorophyll fluorescence and photosynthetic pigments of three cultivars of

Brassica oleracea L.

Geetika Sirhindi* and Ruqia Mushtaq

Department of Botany, Punjabi University, Patiala-147002, India


The present study focuses on the effect of Me-JA and JA on chlorophyll fluorescence and photosynthetic pigments in

three cultivars of Brassica oleracea L. cv. Italica, Capitata and Botrytis. In cv. Botrytis maximum yield of PSII (Fv/Fm) was

enhanced by 17%, 13% and 12% in 1μM Me-JA, 1 nM Me-JA and JA treated seedlings, respectively. Effective quantum

yield [Φ PSII (Y)] enhanced in all treatments of Me-JA and JA in Botrytis. No effective enhancement of Me-JA and JA

treatment was experiential in Italica and Capitata on Fv/Fm and Φ PSII (Y). JA and Me-JA augmented photochemical

quenching (qP) in Italica to 281% in 1 nM JA. In Capitata 1μM JA and 1 nM Me-JA showed 14% and 21% enhancement

in qP. Application of JA and MeJA decreased non photochemical quenching (NPQ) in Capitata and Botrytis, but in Italica

1 pM JA and MeJA increased NPQ by 111% and 67% respectively. Total chlorophyll, chl a, chl b and carotenoids showed

significant enhancement in Italica in presence of JA and MeJA whereas in Capitata and Botrytis this enhancement was in

1 nM JA. From the above results it can be suggested that MeJA and JA have a potential to protect the plant from

photoinhibition but in dose dependent manner and also depends on cultivar.


286

IPPCongress2015


PG337 (IPPC0428)

Isolation and characterization of phosphate solubilising bacteria from sugarcane rhizospheric soil for

screening of plant growth promoting activities

Deepmala Katiyar1*, A. Hemantaranjan1 and Bharti Singh2

1Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi- 221005, Uttar Pradesh, India2Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221005, Uttar Pradesh, India


Now a days new practices using beneficial microbes are being developed and employed for sustainable agriculture. The

present study is limited to plant growth promoting rhizobacteria (PGPR). PGPRs are a group of beneficial soil bacteria

that can be found in the rhizosphere, in association with roots which can enhance the growth by a wide variety of

mechanisms. The use of PGPR is gradually increasing in agriculture and offers an attractive way to replace chemical

fertilizers. A large number of bacteria including species of Pseudomonas, Azospirillum, Azotobacter, Klebsiella,

Enterobacter, Bacillus and Rhizobium have reported to enhance plant growth. The study was conducted to isolate phosphate

solubilising bacteria (PSB) from sugarcane rhizosphere and investigate their potential for plant growth promotion. Twenty

nine PSB were isolated by serial dilution in selective media from three places of Uttar Pradesh. Isolated rhizobacteria

were characterized by morphological, physiological and biochemical method. The study was further preceded for molecular

identification of bacteria by 16S rRNA sequencing, and in vitro screening of plant growth promoting traits viz., production

of Indole acetic acid (IAA), Hydrogen cyanide (HCN) Ammonia (NH3) production and antifungal activity against

Colletotrichum falcatum. Seven isolates (PSC1, PSC2, PSC3, KSC1, KSC2, P13, P35) containing Enterobacter sp.,

Pseudomonas sp. and Bacillus sp. have showed high ability in phosphate solubilising and plant growth promoting activities

out of 19 isolates. Enterobacter sp. (PSC3) showed highest plant growth promoting activities among three rhizobacteria.

This study, therefore, suggest that the use of PGPR isolates as inoculants might be beneficial for sugarcane cultivation.

PG338 (IPPC0434)

Intrinsic salicylic acid mediated delay in postharvest life of tomato: ROS metabolism

Tushita Shukla*, Chunoti Changwal, Ajay Arora and V.P. Singh



Postharvest issues are common around the world. The extent of the loss of horticultural products after harvest can vary

in different countries. A remarkable association between exogenous salicylic acid (SA) and fruit ripening has been reported

in various fruits including tomato. SA is known to affect various physiological and biochemical activities of plants and may

play a key role in regulating their growth and productivity. It behaves in a concentration and time dependant manner.

Tomato is a good model system for studying the ripening process. The endogenous SA is directly correlated with postharvest

life of tomato, this hypothesis, has to be tested in 55 tomato germplasm at different stages of fruit ripening vis-á-vis shelf

life. We screened 15 contrasting tomato germplasm for further validation of the hypothesis in association with ROS

metabolism which has been reported previously that external application of SA can alleviate the oxidative stress and

enhance the antioxidative capacity. We observed that oxidative stress increased from green to red ripen stage of tomato

and antioxidative defence rise up to certain level then it decline in later stages of tomato. We will discuss the potent ROS

species, antioxidants (enzymatic or nonenzymatic) and osmoprotectants that play key role in process of ripening in

comparison to SA and shelf life.


287

IPPCongress2015


PG339 (IPPC0435)

Effect of salicylic acid on physiological traits, antioxidant enzymes and grain yield of wheat genotypes

under water deficit stress conditions

D.C. Saxena1*, S.V. Sai Prasad2, Iti Rathi2 and Renu Parashar2

1Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India2IARI-Regional Station, Indore-452001, M.P., India


Drought or water deficit stress is the main constraint limiting crop production globally. An experiment was conducted to

study the effect of exogenous foliar application of salicylic acid (SA) (0.5 mM concentration) applied at anthesis stage of

five contrasting wheat genotypes viz. HI 1581, C 306, Raj 4083, HI 8731 and NI 5439 under water deficit stress conditions.

Physiological traits viz. relative water content, membrane stability index and chlorophyll content were decreased under

water deficit conditions, but the effect was mitigated in salicylic acid treated plants and there was increase in these

characteristics. The activities of antioxidant enzymes viz. superoxide dismutase, catalase, peroxidase and ascorbate

peroxidase was upregulated by water deficit stress and further enhanced by SA treatment which in turn protected plants

against ROS generation and membrane injury. The yield attributes also showed significant reduction under water deficit

stress whereas application of SA enhanced the yield and its components. Hence, salicylic acid can ameliorate the deleterious

effect of water deficit stress by increasing the physiological characteristics and antioxidant enzymes.

PG340 (IPPC0448)

Endogenous salicylic acid associated regulation of fruit ripening in tomato: ethylene and softening

enzymes

Chunoti Changwal*, Tushita Shukla and Ajay Arora



Salicylic acid (SA) is involved in regulation of many processes in plant growth and development such as ripening, abscission,

senescence etc. There are many reports in favour of exogenous application of SA acting as a plant growth regulator for

enhancing the shelf life of perishables. But, so far no literature is available on endogenous SA in relation to postharvest

life. Fruit ripening is a complex process that lead to changes in fruit, increased activity of cell wall-degrading enzymes,

increased respiration and ethylene production. Climacteric fruits, harvested at full maturity, can be ripened even after

detachment from parent plant. The respiration rate and ethylene formation though minimal at maturity, rise at the onset of

ripening, after which it declines. The most studied model of climacteric fruit ripening is tomato. Present study has been

carried out on 55 tomato germplasm at different developmental stages. These were further narrowed down to 15 germplasm

on the basis of physiological traits. Cell wall degrading enzymes, ethylene evolution and ethylene metabolism associated

enzymes were assayed and compared with endogenous SA at various stages of tomato fruit development. Results from

this and earlier studies from our lab. suggest that SA play an important role in fruit ripening. Further, molecular mechanism

of SA regulating postharvest life in model plant will be elucidated.


288

IPPCongress2015


PG341 (IPPC0486)

Salicylic acid induced salt tolerance in chickpea (Cicer arietinum L.)

R. Dhandapani*, H.P. Vijayakumar, G. Somasundaram, S. Natarajan and S.R. Prasad

Plant Physiology, Indian Council of Agricultural Research-Directorate of Seed Research, Mau-275101, India


Salinity is the major abiotic stress affecting germination and early seedling establishment of chickpea. We demonstrated

the differential performance of saline sensitive (PG186) and saline tolerant (Karnal chenna) chickpea cultivar under 150

mM Nacl salt stress. Salt stress adversely affected the germination of PG 186 cultivar. Sand matrix priming along with

Salicylic Acid (SA) proved to enhance the germination up to 40%. To identify the cellular mechanism of SA-induced salt

tolerance, various antioxidant enzymes (AOE) were estimated from cotyledon during germination. Results indicated that,

unprimed PG 186 seeds accumulated more AOE than SA-primed seeds. Like SA-primed seeds, unprimed seeds of

cultivar Karnal chenna accumulated less AOE in cotyledon and in term balanced the accumulates of more H2O

2 signal for

better germination. The enhanced H2O

2 signal activated several processes including reserve protein mobilisation during

germination. Cotyledons of both unprimed and SA-primed seeds were found to contain same level of protein 8 days after

were sowing. The initial protein content of SA-primed seeds was more than the unprimed seeds before sowing. These

results showed that, SA negatively influenced AOE and in term improved H2O

2 signal under high salt stress. SA also

induced stress tolerant protein synthesis during first phase of germination and mobilised them during the final phase of

germination.

PG342 (IPPC0509)

In vitro production of microtuber of potato (Solanum tuberosum L.): Effect of cultivar, sucrose and

inorganic nitrogen nutrition

Sunil J. Macwan1*, Yogesh M. Shukla2, Natvarlal V. Upadhyay3 and Piyush R. Vaishnav4

1Department of Genetics & Plant Breeding; 2Department of Biochemistry; 3Plant Physiology; 4Department of Agricultural Statistics;B.A. College of Agriculture, Anand Agricultural University, Anand-388110, Gujarat, India


The potato (Solanum tuberosum L.) belongs to the family solanaceae, is a crop of worldwide importance. Two-stage in

vitro tuberization process comprising first micro propagation via nodal explants and then tuber induction in the resultant

in vitro plantlets was studied using three cultivars of potato, Kufri Chipsona-1, Kufri Badshah and Kufri Pukhrajs. In vitro

cultures were established with MS (1962) in absence of growth regulators, prior to surface sterilization with 0.1% HgCl2

for 8 minitues. Axenic cultures were evaluated for maintaining and proliferation of in vitro shoots and concluded that plant

growth regulators and nutrients in the medium are key factors for in vitro multiplication of potato micro propagation. Effect

of sucrose concentration for induction and in vitro development of micro tuber were studied and concluded that optimum

level of sucrose (8%) found to be best for free growth regulators. Further it was noted that stimulation and induction of

microtuber initially with full light was essential. Effect of inorganic nitrogen nutrition on the induction and development of

microtubers was studied with an objective to investigate whether a reduction in total nitrogen level in the Murashige &

Skoog medium (MS, 1962) would improve tuberization rate. It was found that lower level of inorganic nitrogen by nitrate

concentration in media gave better response for the microtubrisation. Interaction between cultivar and other factors were

found to be significant indicating the importance of developing variety specific protocol to optimize microtubrisation.


289

IPPCongress2015


PG343 (IPPC0531)

Regeneration of shoot and induction of flowers in Saffron plant (Crocus sativus L.) under in vitro and ex

vitro conditions

Khirod Sahoo*, Chhavi Sharma and Madhu Sharma

Department of Botany, Ravenshaw University, College Sqaure, Cuttack-753003, India


The trifid stigma of Crocus sativus L. flower is valued for the costliest spice of the world-Saffron. Principally, it is used in

medicine, colouring and flavouring confectionary items. It is a summer dormant crop and conventionally propagated

through corms. The total yield of saffron reduces up to 70% due to various biotic stresses. To develop the genetic

transformation method for improvement of saffron quality and quantity, an efficient shoot regeneration protocol is needed.

To overcome these issues, we have successfully standardized protocols for breaking the dormancy of corm, multiple

bud-sprouting, regeneration of multiple shoots, production of sterile corm and cormlets under in-vitro condition. Importantly,

we have standardized a universal protocol to develop multiple shoots from different explants through somatic

embryogenesis. This will help to produce pathogen-free germplasms for field level cultivation as well as to develop a

genetic transformation method in saffron in future. We have also established methods to induce flower from corms under

in-vitro and ex-vitro conditions to understand the hormonal regulation in saffron plants. We found that hormonal regulation

for flowering in saffron plant under in-vitro and ex-vitro conditions is partially correlated with each other. The reason for

minimal uncorrelation could be due to the difference in environmental conditions. Hence, in-depth molecular study will be

needed to comment on the exact hormonal regulation in flower development of saffron plants.

PG344 (IPPC0542)

RNAi-mediated knockdown of tobacco ODC gene results in altered plant regeneration, development and

stress responses

Ami Choubey* and M.V. Rajam

Polyamine, Transgenic and RNAi Research Lab, Department of Genetics University of Delhi, South Campus Benito Juarez Marg,New Delhi-110021, India


Polyamines (PAs) are ubiquitously present polycationic compounds that play a critical role in various biological processes.

In addition to participating in various growth and developmental processes, PAs are known to elicit stress responses in

plants. Yet their specific functions and mode of action remain largely unknown. In plants, complexities intensify due to the

existence of two parallel biosynthetic pathways for putrescine (Put) production; one mediated by the universally present

ornithine decarboxylase (ODC) and another by arginine decarboxylase (ADC). In the present study, we have targeted

ODC by RNA interference to modulate PA levels and study the effects at different developmental time points. Significant

physiological and morphological alterations seen in response to ODC knockdown include poor regeneration response

from leaf explants, reduced leaf size, reduced chlorophyll and carotene content, increased vulnerability to abiotic stress

conditions, delayed flowering, pollen sterility, reduced seed setting, reduced seed viability and delayed seed germination.

Also, for the first time, microarray analysis has been attempted in an ODC knockdown line to study genome-wide gene

expression changes in response to ODC downregulation and changes in PA titres. On the whole, it presents a detailed

study of trying to decipher functions of PAs in tobacco growth and development.


290

IPPCongress2015


PG345 (IPPC0573)

Tiller dynamics of rice in relation to physiological control of panicle grain yield

Kariali Ekamber*

School of Life Sciences, Sambalpur University, Jyotivihar, Sambalpur-768019, Odisha, India


Tiller development in rice is asynchronous. During the early growth phase, rice plant produces a large number of tillers in

an acropetal fashion form the base upwards. The tillers which originate later are highly unproductive and do not contribute

much to the overall grain yield. Leaf longevity, growth duration and pattern of senescence of the flag leaf in a tiller are

indicative of competence of the organ for survival and provision of assimilates for grain filling in rice. However, the

mechanism by which tiller senescence is induced in the plant for termination of the life cycle is not known hitherto.

Therefore, the objective of the present work was to analyze the pattern of senescence, photosynthetic efficiency and lipid

peroxidation of the flag leaf in different tiller groups and their impact on grain yield. The results revealed that origin of

tillers in the stem node occurred in a temporal fashion. The late-formed tillers had short vegetative period, reduced growth

duration, low photosynthetic pigment concentration and higher membrane lipid peroxidation compared to early-formed

tillers; the former had limited source activity compared to the latter. Hence, it can be concluded that asynchronous tiller

origin might be responsible for production of smaller source area for short growth duration in the late–tillers leading to

maintenance of reduced photosynthetic pigments level. High disintegration of membrane lipids leads to the restriction in

the primary production of the source to the detriment of grain filling and gain yield in rice.

PG346 (IPPC0595)

Improving photosynthetic responses during recovery from heat treatments with brassinosteroid and

calcium chloride in Indian bread wheat cultivars

Suboot Hairat* and Paramjit Khurana



Climate change is expected to unleash severe and frequent heat waves in future, adversely affecting crop productivity.

The aim of this study was to examine the effect of two separate episodes of heat stress, mimicking heat wave conditions

on the physiology of two contrasting Indian bread wheat cultivars based on their level of thermotolerance and to study the

ameliorating effects of epibrassinolide (BR) and calcium chloride on the recovery of these cultivars. The thermo-tolerant

cultivars C306 suffered less inhibition of photosystem II efficiency as compared to the thermo-susceptible cultivars HD2329.

Application of BR and calcium chloride resulted in faster recovery in both the cultivars. Measurement of the minimum

fluorescence (Fo) versus temperature curves revealed a higher inflection temperature of Fo (Ti) for the tolerant cultivar as

compared to the susceptible cultivar, emphasizing greater thermo-stability of the photosynthetic apparatus. The thermo-

tolerant cultivar showed higher photochemistry (PSII) relative to the susceptible cultivar. An increase in the steady state

fluorescence was observed in the susceptible cultivar HD2329 as compared to the tolerant cultivar C306. Expression

analysis revealed faster recovery of the transcripts involved in photosynthesis in tolerant cultivar as compared to the

susceptible cultivar. Exogenous application of the ameliorating compounds resulted in faster recovery of transcripts in

both the cultivars. The result suggest that under severe stress conditions tolerant cultivar C306 showed faster recovery

and a better thermo-stability of its photosynthetic apparatus as compared to susceptible cultivar HD2329 and application

of epibrassinolide and calcium chloride can ameliorate the damaging effect of severe temperature.


291

IPPCongress2015


PG347 (IPPC0596)

Radiation-processed chitosan enhances the crop productivity, essential oil and citral production in

lemongrass [Cymbopogon flexuosus (Steud.) Wats.]

M. Masroor A. Khan1*, Hassan Jaleel1, Moin Uddin1 and Lalit Varshney2

1Department of Botany, Aligarh Muslim University, Aligarh-202002, India2 Radiation Technology Development Division, BARC, Mumbai- 400 085, India


Lemongrass is an important medicinal and aromatic plant. Its essential oil (EO) is an excellent source of natural citral,which is used as a raw material for β-ionone synthesis. β-ionone is used for the synthesis of vitamin-A and other compounds.Hence, increased production of lemongrass EO is highly desirable. Recently, a new agro-technique has been evolved inwhich it has been proved that if natural polysaccharides like alginate, carrageenan and chitosan are exposed to gammaradiation, they are depolymerized into various oligomers. When aqueous solution of these oligomers is applied to theplants, it proved as potent plant growth promoter. Keeping this in mind, a field trial was conducted on lemongrass[Cymbopogon flexuosus (Steud.) Wats.] to study the effect of different concentrations (0, 40, 60, 80 and 100 mg L-1) ofirradiated chitosan (ICH) on the growth, biochemical, quality and yield attributes. The foliar sprays were made five timesduring the crop tenure. The sampling was done at 150 days after planting. The EO was extracted using Clevenger’sapparatus and analyzed using gas chromatography. The effect of ICH was significant on most of the parameters studied,with 80 mg L-1 (ICH-80) proving the best concentration for growth parameters, chlorophyll content, leaf-N, -P, -Kcocentrations, carbonic anhydrase activity, nitrate reductase activity as well as EO content and EO yield as compared tocontrol. However, effect of ICH on citral content was insignificant. Conclusively, application of ICH-80 could improve theperformance of the crop including EO production of the lemongrass.

PG348 (IPPC0680)

High throughput somatic embryogenesis system and stage specific expression of SERK1 gene during

somatic embryogenesis in cultures of greengram, Vigna radiata (L.) Wilczek

Kumariah Manoharan1*, Vajravel Sindhuja1, Muniraj Gnanaraj1, Nagan Udhayakumar1, Maluventhen Viji2,

Sellakumar Maheswari1 and Thirupathi Karuppanapandian3

1Department of Plant Morphology and Algology, School of Biological Sciences, Madurai Kamaraj Univ., Madurai-625021, TN, India2Department of Botany, Thiagarajar College, Madurai-625009, Tamil Nadu, India3Department of Experimental Biology, Faculty of Science, Masaryk University, Brno-625 00, Czech Repub


A protocol for attaining high frequency somatic embryogenesis (SE) in green gram (Vigna radiata (L.) Wilczek) wasworked out. Type of explants and age of source seedlings for obtaining explants were found to influence the formation ofembryogenic callus. Various combinations and concentrations of 2,4-dichlorophenoxyacetic acid and indole-3-aceticacid with kinetin were optimized for developing embryogenic callus. Embryogenic callus when exposed to osmotic stresscreated by D-mannitol and sorbitol and dehydration stress imposed by polyethylene glycol were found to produce somaticembryos. Callus incubated for 6 h in specified hormone free nutrient medium supplemented with 4% polyethylene glycolwas found optimal for the induction of high frequency SE. Subsequent to stress incubation, the cultures formed only earlystage somatic embryos with the absence of cotyledonary stage somatic embryos. Supplementation of proline (150 mM)was found essential for the maturation of somatic embryos. The cotyledonary stage somatic embryos were converted intoplantlets and subsequently established in garden soil. Semi-quantitative Reverse Transcription-PCR based transcriptlevel analysis of SERK1 gene expression was carried out employing embryogenic callus, callus containing differentstages of somatic embryo. Expression of SERK1 was specifically associated with the embryogenic callus and callusenriched with globular stage somatic embryos.


292

IPPCongress2015


PG349 (IPPC0708)

The role of ethylene biosynthesis inhibitors in modulating in vitro growth of Cicer arietinum seedlings

induced by thidiazuron

Rachana Dinesh*

Department of Botany, Jai Narain Vyas University, Jodhpur-342001, Rajasthan, India


The study aims at critically analyzing the role of ethylene biosynthesis inhibitors in modulating in vitro growth of chickpea

seedlings induced by thidiazuron. Since, TDZ has also been implicated in the formation of ethylene hence present

investigation was undertaken to evaluate the role of ethylene biosynthesis inhibitors (aminooxy acetate-AOA;

aminoethoxyvinyl glycine- AVG; cobalt-CO++) in modulation of the effect of TDZ on growth and hyperhydricity in chickpea

seedlings. TDZ inhibited root and shoot elongation and the expansion of leaflets in in-vitro grown Cicer arietinum seedlings.

The roots became thick and green, and developed extensive nodular callus. TDZ also promoted prolific formation of

shoots from the cotyledonary node, and induced hyperhydricity. Simultaneous addition of inhibitors of ethylene synthesis

(AOA, AVG, Co++) together with TDZ resulted in a significant modulation of the effects of the latter chemical on the

formation of shoots from the cotyledonary nodes, development of callus on the roots and the induction of hyperhydricity

in the seedlings. These results indicate that, to some extent, ethylene may be involved in the morphogenetic responses

ilicited by chickpea seedlings in response to TDZ.

PG350 (IPPC0719)

Effect of plant growth regulators and nutrients on alleviation of salinity stress effect in tomato physiology

and yield

R. Sivakumar*, K. Nandhitha and P. Jeyakumar

Crop Physiology, Tamil Nadu Agricultural University, Maruthamalai Road, Coimbatore- 641003, Tamil Nadu, India


Agricultural production and soil health is agitated by the accumulation of diverse substances to soil mainly high levels of

salts. The main factors that contribute to this problem are the salt load in the water used for irrigation, lack of sufficient

rainfall and the use of vast and versatile fertilizers. The soil salinity may cause several deleterious effects on physiological

and biochemical parameters, growth and development of crop ultimately yield. Pot culture experiment was conducted to

study the physiological changes in tomato genotypes sprayed with PGRs and nutrients under salinity condition (100 mM)

in tomato genotypes (PKM 1 and TNAU THCO 3). Foliar application of PGRs like brassinolide (0.5 ppm), salicylic acid

(100 ppm), benzyl amino purine (50 ppm), ascorbic acid (100 ppm), gultathione (50 ppm), KNO3 (0.5%) + FeSO

4 (0.3%)

+ Borax (0.2%) and nutrient PGR concoction (K2SO

4 (0.5%) + CaSO

4 (0.5%) + Borax (0.2%) + NAA (20 ppm) were used

at 20 and 40 DAT. Brassinolide recorded highest leaf water potential (-1.51 MPa), maximum relative water content

(76.50%), highest soluble protein content (13.53 mg g-1), maximum catalase activity (5.48 μg H2O

2 g-1 min-1), minimum Na/

K ratio (0.43), increasing the leaf magnesium content (0.69) at 60 DAT and superior fruit yield (574.55 g plant-1) followed

by salicylic acid and ascorbic acid compared to control. However, brassinolide performed better in yield, salicylic acid

recorded highest BC ratio of 3.2 and 2.5 in TNAU THCO 3 and PKM 1 respectively followed by brassinolide (3.1 and 2.3)

under salinity.


293

IPPCongress2015


PG351 (IPPC0724)

De-hulling induced ethylene breaks seed dormancy in wild rice

Jayanta Barik*, Ekamber Kariali and Pravat Mohapatra

School of Life Sciences, Sambalpur University, Jyoti Vihar, Sambalpur-768019, Odisha, India


Pre-harvest sprouting or viviparous seed germination in rice is one of the major factors causing massive yield losses to

rice farmers in many rice growing nations of the world, when climate change-induced erratic rains coincide with grain

maturation period. However, many species of wild rice such as Oryza rufipogon and Oryza nivara including some traditional

varieties exhibit immunity against viviparous germination in spite of the availability of most suitable environmental conditions.

These recalcitrant rice genotypes maintain a fixed dormancy period and germinate only afterwards with the commencement

of suitable environmental conditions. This versatile trait is of prime importance for introgression into the non-dormant high

yielding rice cultivars. The objective of the present investigation was to induce germination in three different (fully-dormant,

semi-dormant and non-dormant) classes of rice seeds by de-hulling their endosperm including the latent embryo to

assess the responsible physiological mechanisms. The treatment imposed promoted germination, seed vigour index,

ethylene production and mobilisation of soluble assimilates to the culminating sprout. Soluble sugars and starch

concentrations were positively correlated with ethylene production. However, ethylene production was negatively correlated

with the lag period for seed germination. The results obtained provided evidence for the existence of lemma and palea as

a strong physical barrier for germination. Their removal from the seed caryopsis promoted high ethylene production for

the access of germination process. De-hulling effects on the genesis of ethylene and its possible role in breaking seed

dormancy and related activities has been discussed.

PG352 (IPPC0743)

Induction of parthenocarpy in jackfruit (Artocarpus heterophyllus Lam)

T.D. Nikam*

Department of Botany, Savitribai Phule Pune University, Pune-411007, Maharashtra, India*Presenting author: [email protected]

The jackfruit (Artocarpus heterophyllus Lam) is the largest tree-borne fruit in the world and are excellent source of

carbohydrates, minerals, vitamins, fibres, antioxidant and low fat. It is a widely cultivated and popular food item throughout

the tropical regions of the world. It is one of the principal fruit trees that offers potential for fruit marketing and income

generation in the sector of agroforestry and agriculture.The Jackfruit tree can produce about 100 to 200 fruits in a year. In

the present investigation, the female inflorescence at different stages of developement were treated with various

concentrations (00, 10, 100, 1000, 10000 ppm) of gibberellic acid, auxins (IAA, NAA, IBA and 2,4-D) and cytokinins

(kinetin and BAP). Iduction of parthenocrpic fruit development was observed in 57 treated samples. The morphological,

anatomical and biochemical investgations were carried out.Morphological differences were not distinct in parthenocarpic

and control fruits but parthenocarpic fruits were lighter in weight. Maturation and ripening period for both remain in the

range of 90 to 140 days. Dissection of parthenocarpic fruit revealed that, compared to control, the wall of the fruit was not

thick and firm. The pedicel was normal but in parthenocarpic fruits the parianth remains thin. Both types of fruits were

characteized for carbohydrate, protein, lipid, minerals, vitamins, and fibre content.


294

IPPCongress2015


PG353 (IPPC018)

Elevated CO2 ameliorates drought and high temperature stress effects in rice

Madan Pal*, Ashish K. Chaturvedi and Divya Shah



Apart from rising CO2, high temperature and drought stress are important climate change factors, which are predicted to

increase in frequency and magnitude under changing climate. These factors often interact with each other under fieldconditions and their resulting impacts may be different compared to individual exposure. This work is focused onphysiological impact of combinations of these factors compared to independent exposure. The experiments were conductedto analyse the role of elevated CO

2 (eCO

2, 550±34 μmol mol–1) in combination with drought and high temperature stress

on growth, gaseous exchange, carbon metabolism, grain yield and quality attributes in Nerica L-44 and Pusa 1121 ricegenotypes, raised inside FACE and open top chambers. Rate of photosynthesis, non-structural carbohydrates andADPGase and starch synthase enzyme activity was stimulated under eCO

2, decreased under drought and high temperature

stress and reduction was less under combined exposure of eCO2 with drought or high temperature. Similar response was

recorded for dry matter production and grain yield. Chalkiness in grains was more under eCO2 individually and further

enhanced under combined exposure of drought and high temperature. Grain protein concentration decreased undereCO

2 individually

but reduction was less under combined exposure of drought and high temperature with eCO

2. Amylose

concentration in grains was less under combined exposure of drought and high temperature with eCO2

compared toexposure to eCO

2 individually. These findings suggest that negative impacts of drought and high temperature stress on

grain yield and quality in rice can be lowered if combined with eCO2 exposure.

PG354 (IPPC0777)

Employing some PGRs and elicitors for improved artemisinin production in Artemisia annua L. under

abiotic stress

M. Masroor Khan, M. Naeem*, Tariq Aftab and Moin Uddin

Aligarh Muslim University, Aligarh-202002, Uttar Pradesh, India


Artemisia annua L. (family Asteraceae), is the only source of a potent anti-malarial drug, artemisinin. Artemisinin, asesquiterpene lactone containing an endoperoxide bridge, has been recognized as an effective and safe remedy againstmalaria parasite Plasmodium falciparum. However, the concentration of artemisinin in the plant is very low, ranging from0.01 to 0.8%. Exposure of A. annua plants to abiotic stresses, such as of light, temperature, salinity, heavy metals and UVlight, have been reported to enhance ROS generation, which augment artemisinin yield by facilitating rapid conversion ofartemisinic acid to artemisinin. In fact, generation rate of H

2O

2 increased consistently under abiotic stress conditions,

suggesting a direct relationship between generation of H2O

2 and artemisinin biosynthesis in A. annua. Various scientific

approaches have been attempted to increase artemisinin production including chemical synthesis and genetic engineeringof the genes involved in artemisinin biosynthesis pathway in A. annua; but not much success has been accomplished asyet because of the high cost of procedure or complex nature of regulation and expression of the genes responsible forartemisinin biosynthesis. The PGRs exhibit a broad spectrum of physiological effects in plants, may influence the inter-relationships between primary and secondary metabolism leading to increased biosynthesis of secondary metabolites. Apot and field study employing PGRs (gibberellic acid, salicylic acid and methyl jasmonate) and radiation-processedpolysaccharides (sodium alginate and carrageenan) proved as non-conventional elicitors that enhanced the overall yieldof artemisinin under normal and abiotic stress conditions. Artemisinin content was analyzed though HPLC. In order toincrease the desired production of artemisinin and utilize the land unsuitable for food crops, cultivation of A. annua mightbe employed with new scientific approaches and convenient strategies.


295

IPPCongress2015


PG355 (IPPC0824)

Elucidating the role of strobilurin in combination with tebuconazole on physiology of crops

Jeyakumar Prabhakaran* and Jidhu Vaishnavi Sivaprakasam

Crop Physiology, Tamil Nadu Agricultural University, Maruthamalai Road, Coimbatore- 641003, Tamil Nadu, India


The new generation fungicides have been found to have phytotonic effects in many crops. Strobilurin fungicides can

supplement nitrogen demand of plants by strongly activating nitrate reductase and in turn nitrate assimilation. In the

present study, strobilurin (trifloxystrobin) in combination with triazole is reported to exhibit significant physiological and

biochemical changes in groundnut, cotton and chillies. Foliar spray of Nativo (trifloxystrobin + tebuconazole) at different

concentrations was taken up at different growth stages of crops. Chlorophyll index, photosynthetic rate, nitrate reductase

activity, nitrogen uptake by the plants and their content in soil were recorded at selected stages up to harvest. Nativo @

300-400 g ha-1 was found effective in enhancing the yield of crops due to changes in physiological characters studied.

The combined application of trifloxystrobin and tebuconazole enhanced the ‘greenness’ of the crop by showing higher

chlorophyll index with an increase of more than nine per cent compared to untreated control. The improvement in ‘greenness’

can be ascribed to higher uptake of nitrogen by the crops and efficient nitrate reductase activity. The nitrogen uptake and

nitrate reductase activity were found higher by more than 10 and 14% compared to untreated control. The better nitrogen

assimilation favoured for enhanced photosynthetic efficiency. The photosynthetic rate was significantly higher by recording

39.79 and 35.92 μmol CO2 m-2 s-1 in groundnut and chillies. In cotton, higher soluble protein content (10.57 mg g-1)

favoured for enhanced photosynthesis. The combined effect of strobilurin and triazole thus enhanced physiological efficiency

and in turn.

PG356 (IPPC0827)

Efficacy of plantozyme on physiological parameters and yield components in thompson seedless grapes

under pune condition

S.D. Ramteke*

ICAR-National Research Centre for Grapes, Pune-412307, Maharashtra, India


An investigation was carried out to study the efficacy of Plantozyme on physiological parameters, quality and yield

components in Thompson Seedless grape at NRC Grapes, Pune during 2012-13, 2013-14 and 2014-15. Eleven treatments

were applied in replicated RBD experiment. Morphological and biochemical parameters were recorded at 90 days after

pruning and yield parameters were recorded at harvest. The results revealed greater values of morphological characters,

i.e., shoot length (138.8 cm) and leaf area (243.8 cm2) and quality parameters like berry diameter (17.62 mm), berry skin

thickness (30.94 μ), and berry length (21.62 mm) in treatment T8, while cane thickness (8 mm), leaf thickness (0.22 mm),

biochemical parameters such as reducing sugar (3.75 mg/g), total protein (2.25 mg/g) and total phenol (2.25mg/g) were

higher in T9. The highest yield per vine (16.01 kg) and BC ratio (3.24) were obtained in T10. This study concluded that

plantozyme increased berry characters, quality parameters and biochemical parameters as compared to control.

Photosynthetic parameters were also influenced with the application of Plantozyme. Application of Plantozyme through

soil @ 50 g/ vine, drip @ 2.5 ml/ vine and spray @ 2 ml/ l improved overall quality and yield of Thompson Seedless grapes

under Pune conditions.


296

IPPCongress2015


PG357 (IPPC0859)

Effect of micronutrient and growth regulators through foliar application on chickpea and wheat under

late sown conditions

S.R. Kushwaha*, C. Viswanathan, Chandrapal, Rakesh Verma, Suchitra Puskar and D.V. Singh



Foliar nutrition is known to correct the nutrient deficiencies usually observed at seed development in crops. A field trials

was conducted at Indian Agricultural Research Institute, New Delhi to find out effectiveness and feasibility of different

nutrient combination and growth regulators applied through foliar spary at 50% flowering in chickpea and wheat. Chickpea

cultivar Pusa 362 and wheat HD 2967 were grown in field condition under late condition during 2013-14 and 2014-15.

Observation was recorded on physiological traits like chlorophyll content, membrane stability index (MSI), yield and yield

attributes. Results showed that physiological parameters i.e. RWC, MSI, chlorophyll content were significantly enhanced

(12-22%) by the foliar application of micronutrient combination. The combination of micronutrient increased yield over

control by 39.34% while application of growth regulators the yield was improved by 18 to 22% under late planting conditions.

The yield attributes of chickpea and wheat were also increased by micronutrient combination and growth regulators over

the control under late sown conditions.

PG358 (IPPC0870)

Role of glucose in regulating directional growth responses in Arabidopsis

Manjul Singh*, Aditi Gupta and Ashverya Laxmi



Phytohormones and sugars/glucose (Glc) affect a number of developmental, physiological and metabolic processes in

plants. Here, we try to understand the role of glucose and its interaction with phytohormones in controlling root directional

responses. Presence of Glc in the medium could extensively modulate seedling root growth direction. Whole genome

transcription profiling revealed that in Arabidopsis roots growing in sugar free medium, gravity reorientation could

differentially regulate a significant number of genes. Whereas, a completely different set of genes were found to be

affected upon gravity reorientation in presence of Glc. Interestingly, Glc alone could also affect a large number of these

gravity reorientation regulated genes under unstimulated condition. The effects of Glc upon transcript levels of gravity-

reorientation specific genes were largely antagonistic. These results suggested the existence of a tightly regulated genetic

network of Glc modulated gravitropic response in roots. Glc could also regulate several phytohormone related genes

previously known to be involved in gravitropic responses even in unstimulated condition indicating Glc and hormone

interplay during gravitropic growth. BR signaling works downstream to Glc in regulating root directional growth. Glc could

also induce BR receptor endocytosis. Mutant phenotypes as well as pharmacological assays suggested an important

role for protein dephosphorylation during Glc-BR induced root-deviation response. Auxin signaling and polar transport

components are also involved in Glc induced changes in root directional growth. Altogether, Glc mediated change in root

direction is an adaptive feature which is a result of a collaborative effort integrating phytohormonal signaling cues.


297

IPPCongress2015


PG359 (IPPC0882)

Hormonal impacts on physiological processes of rice in relation to methane emission

Leena Borah* and K.K. Baruah

Department of Environmental Science, Tezpur University, Tezpur-784028, Assam, India


Rice paddies are major sources of the non-CO2 greenhouse gas methane (CH

4), which contributes significantly to global

warming and climate change. The rice plant mediates the transport of CH4 from soil to the atmosphere. Therefore,

physiological and anatomical characteristics of the rice plant play a regulatory role in CH4 transport. Impact of exogenous

application of growth hormones viz. gibberellic acid (GA), indole acetic acid (IAA), indole butyric acid (IBA) and kinetin

(KIN) (@ 20 mg L-1) on physiological and anatomical characteristics in relation to CH4 emission from rice paddy (variety

Luit) was worked out in a two-year field study. IAA and KIN application significantly enhanced the rate of flag leaf

photosynthesis in the rice plants over untreated (control) plants. The rate of flag leaf transpiration was lower in IAA and

KIN treated plants in comparison to untreated plants which might be attributed to the lower stomatal frequency in the flag

leaf and smaller xylem vessel diameter of the topmost node observed in hormone treated plants. IAA and KIN applications

were found to bring about a reduction in cumulative CH4 emission by ~10% over control. Remarkable improvement of

grain yield and yield attributing characteristics were observed due to application of auxin (IAA) and kinetin. Physiological

manipulations in rice plant by growth hormone application can serve as an effective strategy for improvement of rice

productivity and simultaneous mitigation of CH4 emissions.

PG360 (IPPC0933)

Physiological and biochemical effects of 24-epibrassinolide on heat stress adaptation in maize

(Zea mays L.)

Jyoti Kaushal1, Anuradha Gautam2, Hemangini Parmar2, Ishwar Singh2 and Pranjal Yadava2*

1Sam Higginbottom Institute of Agriculture, Technology & Sciences, Allahabad-211007, Uttar Pradesh, India2ICAR-Indian Institute of Maize Research, Pusa Campus, New Delhi-110012, India*Presenting author: [email protected]

Climate change and associated extreme weather events have made high temperature a major threat to crop production

worldwide. Brassinosteroids (BRs) are a new type of polyhydroxy steroidal phytohormones that regulate a number of

physiological processes in plants. Among these, Brassinolide (BL), 28-homobrassinolide (28-homoBL) and 24-

epibrassinolide (24-epiBL) are commonly used. The present study aims at studying the usefulness of 24-epiBL in

ameliorating the negative impacts of heat-stress in maize along with its role in regulating cellular antioxidant defence

system. PMH 3 maize hybrids were grown in pots in green house maintained at 14 hours day (25oC) /10 hours night

(17oC). 24-epiBL solution (1 μM) was applied externally at V4 stage. Leaf tissues were sampled from both treated and

control plants. Subsequently, both the groups of pots were placed in plant growth chamber maintained at high temperature

(48oC; RH 50%). Plants were sampled for biochemical analysis after 3, 6, 9, 24 and 48 hours of high temperature exposure.

Exogenous application of 24-epiBL arrested protein degradation and enhanced membrane stability, as compared to the

control. The activity of antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT) was also found to

be increased post 24-epiBL treatment. Thus, the study supports the role of BRs as anti-stress agents. However,

investigations are required to gain further insights into the molecular-genetic mechanisms of BRs action in modulating the

plant defence system under different stresses.


298

IPPCongress2015


PG361 (IPPC0971)

Depolymerised sodium alginate and methyl jasmonate improve the growth, physiological attributes and

alkaloids production in Catharanthus roseus (L.) G. Don.

Moin Uddin*, M. Masroor A. Khan and Lalit Varshney

Aligarh Muslim University, Aligarh-202002, Uttar Pradesh, India


Irradiated sodium alginate (ISA) as well as methyl jasmonate (MeJ) acts as endogenous growth elicitors, triggering the

synthesis of different enzymes and modulating various plant responses by exploiting the gene expression. Two pot

experiments were conducted to test whether the foliar application of ISA and that of ISA+MeJ could augment the active

constituents as well as growth, physiological and yield attributes of Catharanthus roseus, which carries anticancer alkaloids

(vincristine and vinblastine) in its leaves in addition to various other useful alkaloids. Totally, 5 spray treatments, comprising

various aqueous solutions of ISA [20, 40, 80 and 160 mg L-1 (Experiment 1)], and those of ISA+MeJ [40+20, 40+30,

80+20, 80+30, 160+20 and 160+30 mg L-1 (Experiment 2)], were applied at seven days interval. Leaf-alkaloids content

and growth, physiological and yield parameters, evaluated at 120 days after sowing, were significantly improved by ISA

application. ISA application could not increase the leaf-content of vincristine and vinblastine; however, it significantly

augmented the yield of these alkaloids because of enhancing leaf dry matter. Compared to the control (water-spray

treatment), collective application of ISA (80 mg L-1) and MeJ (20 mg L-1) resulted in the highest values of most parameters

studied. However, 80 mg L-1 of ISA applied with 30 mg L-1 of MeJ maximally improved the content and yield of total as well

as anticancer leaf-alkaloids. Comparing the control, it increased the content and yield of total leaf-alkaloids (30 and 73%,

respectively) and those of vincristine (55 and 134%, respectively) and vinblastine (19 and 79%, respectively).

PG362 (IPPC1013)

Artificial seed production of Gentiana kurroo Royle

Payal Kotvi*, EneshVashist, Shivam Sharma and Hemant Sood

Department of Biotechnology and Bioinformatics, Jaypee University of Information and Tech. Waknaghat, Solan-173234, HP, India


Gentiana kurroo royle is a critically endangered plant, which is known to be a bitter drug plant in Indian medicine. It has

been widely distributed in places like Jammu and Kashmir, Himachal Pradesh, in northern western Himalayas between

the altitude ranges of 1500 and 3500 m, Nepal and Pakistan. It has been confirmed to be the major producer of secondary

metabolites belonging to the family Gentianaceae. In this study somatic embryogenesis was conducted as a source of

artificial seed production of Gentiana kurroo royle. Artificial seed production has led to increase in the viability rates and

germination capabilities of Gentiana kurroo royle seeds in soil, thereby providing efficient means of germplasm conservation,

easy transportation and large scale propagation of endangered species. Morphogenetic capabilities of leaf explants have

been optimised on MS media containing different concentrations and combination of KN+2, 4-D and out of these KN 1 mg

l-1 and 2,4-D 0.5 mg l-1 gave best results for callus initiation and proliferation. Once the callus was induced torpedo shaped

somatic embryos were selected for artificial seed production. The artificial seed were produced using the sodium alginate

(2%) and calcium chloride solution (5%), which led to calcium alginate bead formation, which were tested for germination

under in vitro conditions for direct shoot formation. Hence this technique would contribute in protecting the endangered

plant species of Gentiana kurroo royle in short duration of time and provide platform for production of important medicinal

compounds.


299

IPPCongress2015


PG363 (IPPC1025)

Transcriptomic analysis of two differentially ripening mango varieties reveals differences in gene

expression of major pathways

Smriti Srivastava*, Garima Pathak, Ridhi Goel, M.H. Asif and V.A. Sane

Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India


Ripening of fruit, known to be governed by plant hormone ethylene (in case of climacteric fruits), involves a complex

interplay of several ripening related genes known to be involved in ethylene biosynthesis and its signaling along with

genes related to cell wall hydrolysis. Mango, a typical climacteric fruit ripens with a burst of ethylene. There are more than

1000 varieties of mango differing in their ripening pattern. Dashehari (and many other varieties) shows a characteristic

pattern, wherein ripening and softening is initiated from the stone and moves towards the periphery. In contrast, ripening

in Banganpalli (south Indian variety) is initiated at the periphery and moves towards the stone. In an attempt to decipher

the molecular basis of these differences in ripening pattern between the two varieties along with complex spatio-temporal

control of ripening, transcriptomic studies were carried out using hybrid assembly of 454 pyrosequencing and Illumina.

From this we obtained 86,083 transcripts. Out of these approx 200 were ripening related. Detailed analysis revealed large

scale changes in gene expression during the course of ripening between the two varieties. Which includes differential

expression patterns of several ACSs and ACOs along with cell wall hydrolases and some aroma and flavour related

genes. Detailed studies are in progress to know regulators which cause these changes.

PG364 (IPPC1026)

Understanding the role of gluconokinase in regulating developmental flower senescence

Sadaf Khan*, Abdul Azeez, Pravendra Nath and A.P. Sane



Gladiolus is a popular garden ornamental, however, the individual flowers of a gladiolus spike suffer from a short vase

life, which is undesirable in floriculture industry. We had previously identified a gluconokinase gene that was up-regulated

during floral senescence in Gladiolus. The enzyme gluconokinase is a component of the pentose phosphate pathway,

where it catalyzes the phosphorylation of gluconate. But there is very little information regarding the role of gluconokinase

in plants. This study was carried out to investigate the role of gluconokinase in floral senescence. Gluconokinase activity

was found to progressively increase during the course of floral senescence starting from stage 0 (open flower) onwards,

concomitantly with increase in transcript level. Maximum activity was observed during late senescence at stage 2. The

gluconate level exhibited a peak at stage 1, whereas the 6-phosphogluconate level showed no such stage dependent

variation. Feeding gluconate to excised leaves and flowers induced petal and leaf wilting and loss of structure within 36

h. A decrease in Fv/Fm values was observed which could be attributed to the decrease in chlorophyll content upon

gluconate treatment. No such change in Fv/Fm and chlorophyll content was observed in flowers and leaves treated with

water or with an equivalent concentration of glucose. These studies provide evidence for a hitherto unknown role for

gluconokinase-gluconate in senescence in plants.


300

IPPCongress2015


PG365 (IPPC1027)

Hormonal and genetic control of root development in tomato

Vinod Kumar*, Deepika Singh, A.P. Sane, V.A. Sane and C.S. Nautiyal



Root development is a complex process governed by a fine balance of different hormones that regulate primary and

lateral roots differently. To obtain an insight into the factors that govern root development in tomato, a differential

transcriptome from tissues such as root, stem, leaves, fruit and flowers was first developed to identify genes that are

primarily expressed in root tissues. Through bioinformatic analysis 279 genes that were predominantly expressed in root

samples at levels 3-10 fold higher than the rest of the tissues were identified. These included about 92 genes that

encoded transcription factors and regulatory proteins. Some of the genes encoding members of the ERF family, Myb

domain family, START domain family and WRKY family were chosen for further studies. Expression analysis at different

stages of root development confirmed the high expression of these genes primarily in the root tissue. Full length cDNAs

and promoters of these have been isolated for further studies. Simultaneously, a hydroponic system for root analysis was

developed. The effect of different hormones such as auxin, ABA, ethylene, cytokinin and some of their inhibitors was

studied on primary and lateral root development.

PG366 (IPPC1028)

Petal abscission in fragrant and non-fragrant roses is associated with differential expression of ethylene

biosynthesis and signalling genes

Priya Singh*, Amar Pal Singh and A.P. Sane

Plant Gene Expression Lab, CSIR-National Botanical Research Institute (CSIR), Lucknow-226001, India


Organ abscission is an important developmental process governed by ethylene. In rose, abscission in the fragrant variety

R. bourboniana is highly sensitive to ethylene, while the hybrid variety, R. hybrida shows greatly reduced sensitivity. To

gain an insight into ethylene responses during petal abscission, the expression of several ethylene biosynthesis/receptor/

signalling genes was studied during the course of abscission in fragrant and non-fragrant roses. Transcript accumulation

of most ethylene biosynthesis genes increased rapidly in petal abscission zones of R. bourboniana within 4-8 h of ethylene

treatment, followed by a decrease. A similar kinetics of transcript accumulation was also observed for most receptor and

signalling genes. Under field conditions where abscission is delayed, a delay in peak transcript accumulation could be

observed in most ethylene biosynthesis/signal transduction genes. Surprisingly, transcript accumulation of most ethylene

biosynthesis/signalling genes decreased considerably in ethylene treated petal abscission zones of R. hybrid, with the

exception of ACS1 and EIN3. These results suggest that the activation of the ethylene biosynthesis /signal pathway is

most likely responsible for the rapid abscission of petals seen in Rosa bourboniana. In contrast its suppression in R.

hybrida (except for ETR2, ACS1 and EIN3) through a hitherto unknown mechanism might be responsible for the delayed

abscission in this species.


301

IPPCongress2015


PG367 (IPPC1032)

A negative regulator of ABA responses governs fruit ripening in tomato

Asmita Gupta*, Rakesh Upadhyay, Rashmi Garg, Pravendra Nath and A.P. Sane



Abscisic acid (ABA) is a plant hormone, which regulates seed dormancy, plant growth, and responses to environmental

stresses. In recent years, a role for ABA in regulating fruit ripening has also been indicated. We have previously identified

SlERF6 as a ripening related AP2/ERF domain gene in tomato. In order to get an insight into its function lines over-

expressing and under expressing SlERF6 were generated. SlERF6 over-expressors showed early seed germination,

reduced ABA inhibition of root growth and reduced stomatal closure in response to water stress, suggesting reduced ABA

sensitivity conferred by SlERF6. Expression of SlERF6 is suppressed by ABA. Manipulation of its expression also affected

fruit ripening and fruit size, with delayed ripening and reduced fruit size being observed in over-expressors and early

ripening being seen in antisense lines. The shift in ripening by about 5 days was most prominent in the breaker stages of

sense and antisense lines. The change in expression of SlERF6 was associated with changes in expression of some

signaling related genes as well as SlCYP707A3. Analysis of expression of SlERF6 in ABA biosynthesis mutants like

notabilis and flacca revealed much higher transcript levels at the immature green, mature green and breaker stages of

their fruit compared to Ailsa Craig fruits. The results suggest a crosstalk between ABA and ethylene during fruit ripening

that might be mediated by SlERF6.

PG368 (IPPC1036)

Early wound induction responses in chickpea are partly controlled by JA dependent and JA independent

manner

S.P. Pandey, Shruti Srivastava* and A.P. Sane



Chickpea (Cicer arietinum) is an important pulse crop in India and other Asian countries that suffers from severe yield

losses due to insect attack from pod borers such as H. armigera. To prevent losses from insects, a detailed understanding

about early wound responses is required. Towards this aim, a comparative analysis of transcriptomes of unwounded and

insect saliva treated and mechanically wounded chickpea leaves was performed. About 10% of the transcriptome was

found to be differentially expressed upon wounding. In silico analysis showed the differential expression of several kinases,

phosphatases and transcription factors belonging to the WRKY, MYB, ERFs, bZIP families etc. Genes encoding components

of jasmonic acid and ethylene pathways were up-regulated, while growth associated hormone pathways like auxin were

suppressed. Validation of expression of several genes was performed and revealed a similar trend of transcript abundance

over a detailed time course study as in transcriptome sequencing analyses, indicating that changes determined by

transcriptome sequencing were true to nature. The expression of these genes was also tested in response to JA and SA

treatments and revealed that not all genes were activated by JA and SA. Moreover, even those that were activated

showed a time delay as well as reduced intensity at peak activation compared to mechanical wounding. These results

suggest that early wound responses are partly dependent but largely independent of jasmonic acid.


302

IPPCongress2015


PG369 (IPPC1041)

Biochemical basis of resistance to Alternaria leaf spot in groundnut breeding lines

Narendra Kumar*, Nidhi Radadiya, M.K. Mahatma, K.S. Jadon, A.L. Rathanakumar M.C. Dagla, S.K. Bishi,

Lokesh Kumar and B.M. Chikani

ICAR-Directorate of Groundnut Research, P.B.No.5, Ivnagar road, Junagadh-362 001, Gujarat, India


Groundnut (Arachis hypogaea L.) is an important food legume crop. Several biotic stresses limit groundnut productivity,

among which the Alternaria leaf spot (ALS) is becoming a major disease of groundnut in rabi-summer. To understand the

biochemical basis of resistance, few advanced groundnut breeding lines showing resistance and susceptible to ALS

were screened during summer-2015. Disease severity was assessed using a scale of 1 to 9, where 1 (no symptoms)

represents resistant and 9 is 81% to 100% defoliation represents susceptible. The identified lines were characterized by

assaying the induction of polyphenol oxidase (PPO), peroxidase (POX) and phenol profiles. Leaf samples were collected

at 75 days after sowing from field. Breeding lines showing higher disease incidence (PBS-18057) had minimum activities

of PPO and POX enzymes and levels of coumaric and salicylic acid. On the contrary, breeding lines with less disease

incidence had higher activities of PPO and POX enzymes along with higher contents of coumaric and salicylic acid (about

2-3 fold). As higher induction of PPO, POX, coumaric and salicylic acid were negatively correlated with disease severity.

Hence these parameters can be used as biochemical markers for screening groundnut genotypes resistant to Alternaria

leaf spot in future breeding programmes.

PG370 (IPPC1043)

Application of paclobutrazol to ameliorate water deficit stress effects in chickpea

Soumya P.R.*, Pramod Kumar and Madan Pal Singh



Chickpea is an important winter season pulse crop of arid and semi arid regions and is sensitive to water deficit stress. A

study was conducted to analyse role of Paclobutrzol to ameliorate water deficit stress effects in chickpea. Experiment

was conducted using two contrasting chickpea varieties viz. desi (Pusa 362) and kabuli (Pusa1108). There were two

main treatments (i.e. well watered and water deficit condition) and four sub-treatments (PBZ foliar spray (60 ppm), PBZ

drenching (120 ppm), water spray and water drenching). Optimum dose of PBZ for foliar and drenching was identified

based on preliminary study. The results showed decreased RWC, MSI, rate of photosynthesis, Total chlorophylls and

carotenoids, grain yield and disrupted chloroplast grana under imposed water deficit stress. However, application of PBZ

in general, maintained higher values of all above parameters and maintained the integrity of chloroplast structure under

imposed water stress and also stimulated faster recovery after stress termination in both the varieties. PBZ reduced the

lipid peroxidation and membrane leakage. It enhanced the activity of antioxidant enzymes and improved stress tolerance

ability in both chickpea varieties. In general PBZ drenching was found better over the foliar application. The findings

depict that PBZ enhanced the yield by maintaining RWC, MSI, photosynthetic activity, photosynthetic pigments and

protected the photosynthetic machinery by restricting lipid peroxidation, increasing antioxidant activity and ABA level,

thereby enhanced water deficit stress tolerance.

Session H : Photosynthesis

303

IPPCongress2015


PH371 (IPPC0132)

Chlorophyll a fluorescence changes in response to short and long term high light stress in rice seedlings

P. Faseela and Jos Puthur*

Department of Botany, University of Calicut, Calicut-673635, Kerala, India


An investigations was carried out to study the high light stress effects on photosystem (PS) II efficiency in intact leaves offour rice (Oryza sativa L.) varieties viz. Jyothi, Aiswarya, Swarnaprabha and Kanchana. Rice seedlings were exposed tohigh light stress [2,000 μmol (photon) m-2 s-1] and the photochemistry of the four rice varieties was studied by the applicationof qualitative and quantitative analysis of chlorophyll (Chl) a fluorescence at 0 (control), 2 and 8 h. The results showedthat some Chl a fluorescence parameters were highly affected under high light stress. It was found that Fv/Fo, performanceindex, area above the ûourescence curve, φo/(1- φo), φDo, φEo and Vj are indicators found to be more reliable forexploring the effect of changes in PSII activity after high light stress application for short term (2 h), which can aid indistinguishing the tolerant and sensitive rice varieties. Some Chl a fluorescence parameters were modified only afterexposing the plants to high light stress for 8 h. The decline in photochemistry was more pronounced in Kanchana andlower in Jyothi as compared to other varieties and the order of high light stress tolerance of these varieties on the basisof Chl a fluorescence parameters was Jyothi > Aiswarya > Swarnaprabha > Kanchana.

PH372 (IPPC0347)

Enhanced photosynthesis is associated with efficient source-sink interaction and nitrogen use in Jatropha

curcas L. under elevated CO2

Sumit Kumar* and Attipalli Reddy

Department of Plant Sciences, University of Hyderabad, Hyderabad- 500046, Andhra Pradesh, India


The present work demonstrates leaf gas exchange characteristics and related morphological and biochemical parametersin Jatropha curcas on a potential biofuel plant exposed to elevated [CO

2] (~550 ppm). Light saturated net photosynthetic

rates of upper-canopy leaves were 45% higher under elevated [CO2] compared with ambient [CO

2] over the 3-yr period in

spite of reduced stomatal conductance (gs) and decreased transpiration rate (E). There were no significant CO2 treatment

effects on maximum photosynthetic (Amax) or biochemical capacity (in vivo rate of RuBP carboxylation, Vcmax or in vivo

rate of electron transport driving RuBP regeneration, Jmax) of J. curcas leaves as evidenced from A/Ci curve analysis.Chlorophyll a fluorescence measurements, indicated by Fv/Fm and ÄF/Fm ratios and photosystem-II associated electrontransport rate (ETRmax), positively correlated with enhanced photosynthesis and growth of the plants in elevated [CO

2]

conditions. Despite increased area-based leaf sugar (13%) and starch content (25%), and reduced mass and area-basedleaf nitrogen concentration (NM, 10%; NA, 6%), the plants showed better photosynthetic nitrogen use efficiency (PNUE)in elevated CO

2 environment. The study also revealed that CO

2 enrichment significantly influenced female to male flower

ratio, fruit and seed yield, above ground biomass and carbon sequestration potential (24 kg carbon per tree per year) inJatropha. Our data demonstrate that J. curcas was able to sustain enhanced rate of photosynthesis in elevated CO

2

conditions as it had sufficient sink strength to balance the increased biomass yields.


304

IPPCongress2015


PH373 (IPPC0373)

The comparative analysis of PSII protein profiles of some plants, grown under enhanced solar UV-B

(280-320 nm) radiation

N. Shanthi1* and S. Jannita Nithia

1Department of Botany, Pachaiyappa’s College, Chennai, Tamil Nadu, India2Department of Botany, Sri Meenakshi Government College for Women, Madurai, Tamil Nadu, India


The ozone layer depletion and UV-B radiation fluctuation in the earth’s surface is make great attention today world. Theplants like Black gram, Green gram .Radish, Carrot was growing in field condition under enhanced UV-B radiation. Liketropical plants the temperate plants also successfully grown in tropical region. The UVB radiation affects growth,Development, photosynthetic activity of plants. In photosynthetic protein the PSII is the main target for UV-B radiation.Enhanced UV-B radiation leads to a change in the organization of PSII complex. It specifically damaged the D1 polypeptide.The UVB radiation damage and affect PSII protein expression in sensitive plant. In Black gram and green gram the UVBradiation did not produce any significant difference in the PSII polypeptides as compared to other plant. Where as inRadish the UV-B radiation increase accumulation of 33-28kDa protein and affect express the 68kDa protein. In the caseof Carrot the large degradation of 32 and 33 kDa, respectively D1 and D2 protein were observed. This severe sensitivityof the D1 protein is undoubtedly a major contributor to the loss of PSII function on exposure to UV-B radiation. From thisstudy, we confirmed the tropical plant naturally resistant to UVB radiation and restore its activity from damage. WhereasCarrot plant more sensitive to UVB radiation the fail to restore it activity from damage. In Radish moderately resistant toUVB radiation they try to stabilize its activity from damage.

PH374 (IPPC0375)

Drought reduces photochemical efficiency and gene expression of photosynthesis

JoãoAraújo Jr, Thayssa Schley* and Luiz Fernando Almeida

State University of São Paulo - Campus Botucatu, Brazil


Current climate changes have resulted in water scarcity in agriculture. Drought interferes in crops productivity. It affectsfood demand for human population. Therefore, our objective was to evaluate drought effects on physiological and molecularresponses of sorghum (Sorghum bicolor L.). For this, one group of plants were subjected to daily watering (control), whilein other group watering was withheld. We determined last day of experiment when photochemical process and stomatalconductance (g

s) variables showed severe stress. During water deficit stress (29 days), plants showed decline in g

s and

leaf water potential as compared to control. It indicates sorghum has stomatal closure and osmotic adjustment to preventwater loss. Stress also reduced photochemical dissipation by 50.4% and heat dissipation in reaction centers by 37.1 %,and increased heat dissipation in antenna by 40.2%. It indicates prejudice in photochemical phase and activation inphoto-protection process. We also compared mRNA differentially expressed between dry and irrigated conditions. Instressed group, preliminary results of transcriptome analysis indicate increased expression of genes ERD10, HSP70 andTpt_PEP_trans_dom related to cell protection under dehydration. Stressed group also had greatly reduced expression ingenes involved in photosynthesis, viz., LHB1B2 and LHCB4 genes involved in antenna structure and RBC51A (rubiscoenzyme). Detected physiological changes are validated by differential expression of genes related mainly to photosyntheticprocess and drought tolerance. Drought decreased plant productivity not only by stomatal restrictions but also by inducingchanges in gene associated with photosynthetic activity. These results help to understand how sorghum tolerates droughtand damages that stress can cause in species.


305

IPPCongress2015


PH375 (IPPC0379)

Differential sensitivity to nitric oxide (NO) and hydrogen peroxide (H2O

2) of photosynthesis and respiration

in mesophyll cell protoplasts of pea

Sunil Bobba* and A.S. Raghavendra

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, Andhra Pradesh, India


The direct effects of nitric oxide (NO) and reactive oxygen species (H2O

2) on photosynthesis and respiration in leaf

tissues are debatable. We studied the consequences of exogenous application of NO (in the form of SNP) and H2O

2 on

the patterns of photosynthesis and respiration in mesophyll protoplasts of pea plants. Increased levels of NO severelyinhibited the photosynthesis of mesophyll protoplasts, while exhibiting marginal or no effect on dark respiration. Theinhibition of photosynthesis by SNP was completely reversed by cPTIO, a NO specific scavenger. In contrast, externallyadded H

2O

2 resulted in drastic decrease in the rates of respiration, with only a marginal effect on photosynthesis. At the

concentration used in our study, H2O

2 didn’t affect the electron transport activities of PS I or PS II. However, both these

photochemical reactions showed marked sensitivity to NO in cells. Our observations suggest that chloroplasts could beprimary targets of NO and mitochondria for ROS in plant cells. We also attempted to assess the role of crosstalk betweenchloroplast and mitochondria, during the effects of NO and H

2O

2. Modulation of NO did not alter much the sensitivity of

photosynthesis in mesophyll protoplasts, to antimycin A or salicylhydroxamic acid (SHAM). On the other hand, the presenceof H

2O

2 decreased the sensitivity of photosynthesis to mitochondrial inhibitors, particularly SHAM, suggesting the role of

alternative oxidase pathway (AOX) during H2O

2 exposure. It appears that H

2O

2 could mediate the crosstalk between

chloroplasts and mitochondria.

PH376 (IPPC0410)

Photosynthesis, dry matter partitioning and grain yield in relation to methane emission in a rice (Oryza

sativa) ecosystem

Ashmita Bharali*, Kushal Kumar Baruah and Sunitee Gohain Baruah

Department of Environmental Science, Tezpur Central University, Napaam-784028, Assam, India


Methane (CH4) is second most potent green house gas in the atmosphere after carbon dioxide (CO

2). Rice paddies are

the largest anthropogenic methane source and produce 7–17% of atmospheric CH4. A field experiment was conducted at

Tezpur University during monsoon rice ecosystem (from June-November, 2014), to assess the integrated effects oforganic amendments on methane emission and its relationship with photosynthesis, dry matter partitioning and grainproductivity. Soil organic amendments resulted in differential methane emission from the rice field and had significantimpact on leaf photosynthesis in rice. Flag leaf photosynthesis revealed a strong association with yield and had aninverse relationship with CH

4 flux. There was a good correlation between the photosynthesis and sucrose-phosphate

synthase (SPS) activity of the flag leaves. Leaf SPS activity increased with the proportion of dry matter in leaves and wasinversely related with the dry matter of the roots and cumulative CH4 emission. Application of Azolla compost resulted insuperior yield attributing parameters along with higher flag leaf photosynthesis over other treatments. SPS activity in theleaves plays a pivotal role in carbon partitioning and therefore high SPS levels boosted photosynthetic rates underfavorable conditions and may indirectly contribute to reduction in CH

4 emission.


306

IPPCongress2015


PH377 (IPPC0421)

Screening of mango (Mangifera indica L.) varieties for photosynthetic light response curve in Konkan

region of India

Mahesh Kulkarni*, Murad Burondkar, Parag Haldankar, Kuruva Mallikarjuna and Kaustubh Kelkar

Department of Horticulture, Dr. Balasaheb Sawant Konkan Agriculture University, Dapoli-415712, Maharashtra, India


Konkan region on west coast of India, is one of the biggest (0.182 million ha.) mango growing belts in India. Heavy rainfall(3500-4000 mm), low sun light during monsoon period (June to September) are considered as the major environmentalfactors, limiting the carbon assimilation and yield. An experiment was conducted during 2014 to screen the commerciallyimportant nine mango cultivars of the Konkan for their photosynthetic response at different levels of light intensity i.e. 100,250, 400, 500, 600, 700, 800, 1000, 1250, 1500, 1750, 2000 μmol mol-1 PAR. Study revealed that irrespective of variety,mean value of light saturation point (LSP) was 711 μmol mol-1, with mean net photosynthetic rate (Pn) of 8.8 μmol CO

2 m2

s-1. Among the nine varieties, Amrapali and Tomy Atkins showed maximum LSP and highest Pn. of 13.28 and 11.53 μmolCO

2 m2 s-1, respectively, while cv. Kesar and Hybrid Ratna had lower LSP of 400 and 500 μmol mol-1, Pn. of 5.15 and 5.11,

μmol CO2 m2 s-1, respectively. Under low light intensity of 100 μmol mol-1, cv. Amrapali and Tomy Atkins showed maximum

Pn. (3.51 and 2.82 μmol CO2

m2 s-1) hence found more suitable for sustaining diffuse sunlight conditions during rainyseason of Konkan.

PH378 (IPPC0451)

Modulation of phosphoenolpyruvate carboxylase by abscisic acid and fusicoccin in the leaf discs of

Amaranthus hypochondriacus

Bindu Prasuna Aloor*, Uday Kiran Avasthi and A.S. Raghavendra

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, Andhra Pradesh, India


The kinetic and regulatory properties of phosphoenolpyruvate carboxylase (PEPC) are modulated by dark/light transitionsand variation in temperature. Since the reports on hormonal modulation are limited, we examined the activity/regulationof PEPC by abscisic acid (ABA, a plant hormone) and fusicoccin (FC, an antagonist of ABA action) in the leaf discs ofAmaranthus hypochondriacus. Incubation with 20 μM ABA for 1 h increased the activity of PEPC in leaf discs by > 40%in light and by 30% in dark. Butyric acid decreased PEPC activity and restricted the stimulation by ABA, while methylamineincreased the PEPC activity and stimulated the effect of ABA. ABA enhanced the levels of PEPC-protein as well as itsmRNA, both in dark and in light. Butyric acid/methylamine modulated the changes induced by ABA of PEPC-protein andmRNA levels, indicating that acidification/alkalization of leaf discs was very important. Since FC is known to antagonise/reverse the effects of ABA, we have tested the effect of FC with or without ABA on PEPC properties. FC (10 μM) inhibitedPEPC activity by 50% in dark as well as under illumination. The responses of PEPC to FC were subdued in the presenceof 20 μM ABA. Significant increase in protein levels were observed in ABA treated leaf discs, while there was a decreasein PEPC protein levels in FC and butyric acid treated ones. We conclude that the enhancement by ABA of PEPC activitywas due to alkalization and an increased protein levels.


307

IPPCongress2015


PH379 (IPPC0468)

Variations in photosynthesis and light induced respiration in rice under different nitrogen treatments

and in temperature gradient tunnel

Rajesh Kondamudi*, Narasimha Swamy Konduri, Sailaja Bhogireddy, Veerendra Jaldhani, Suchandranath B Munnam,Satendra K Mangrauthia, Raghuveer Rao Puskur, Subrahmanyam Desiraju, Sarla Neelamraju and Sirapati Rao Voleti

Indian Institute of Rice Research, Rajendranagar, Hyderabad-500030, Telangana, India


Rice crop was observed to be vulnerable to the climate change. Breeding for climate resilience in rice should includemorpho-physiological characterization of existing germplasm. Five heat tolerant rice varieties were identified earlier (Somaly-2-023-3-5-1-2-1, IR 55178, GQ-25, SG-26-120, IR-82310-B-B-67-2) along with a tolerant check Nagina 22 and a susceptiblecheck Vandana were studied for seven consecutive seasons in four nitrogen treatments with elevated temperature (5±1°Cabove ambient) to understand the impact of elevated temperature and nitrogen fertilization. Leaf characteristics,morphological features, yield and yield attributes were recorded. Photosynthesis (A) and light induced respiration (LIR)were also studied using oxygraph. The A decreased with time in leaf samples, 14.57 μmoles m-2 s-1 (1d), 14.37 μmolesm-2 s-1 (2d) and 10.03 μmoles m-2 s-1 (3d) and A and total carbon were found to be in positive association. Conversely, theLIR varied greatly with 10.44 (1d), 13.11 (2d) and 9.60 μmoles min-1 s-1 (3d). The C/N ratio was decreased in leaf with timein almost all the treatments and varieties and increased with increased nitrogen doses but not under elevated temperature.Under heat stress, gene expression was also studied. Heat shock transcription factor (OsHsfA2a) cell wall integrityprotein (CWIP) and sucrose phosphate synthase (SPS) genes showed increased expression. The expression of Osfdgene (iron-sulphur cluster binding-protein) showed a definite pattern between heat tolerant and susceptible germplasms.GQ-25 and Vandana showed down-regulation but other germplasm showed increased expression under elevatedtemperature. The interaction among N, P, K and Si elements and their role in making the rice crop resilient to hightemperature was focussed.

PH380 (IPPC0494)

Nitrogen assimilation is well coordinated with photosynthetic efficiency in pigeonpea grown under

elevated CO2

Rachapudi Venkata Sreeharsha* and Attipalli Ramachandra Reddy

Department of Plant Sciences, University of Hyderabad, Hyderabad- 500046, Andhra Pradesh, India


Pigeonpea (Cajanus cajan (L.) Millspaugh; Fabaceae) is an important food legume crop of equatorial and semi-arid partsof the world. A short duration pigeonpea cultivar (ICPL 15011) was grown under elevated CO

2 (550 ppm) in open top

chambers to understand the relation between photosynthetic physiology and N assimilation. Elevated CO2, pigeonpea

grown showed significantly high net photosynthetic rates (A’) and foliar carbohydrate content (36% and 43%, respectively),which further manifested in dry biomass at harvest, with an increment of 29% over the ambient plants. Mass and numberof nodules exhibited prodigious accretion in elevated CO

2 grown plants, showing 58% increase in nodule mass ratio

thereby overcoming the N limitation and resulting lack of photosynthetic acclimation. The high levels of N and amino acidcontents in the leaf and root tissues of plants grown under elevated CO

2 can be attributed to the increased levels of

glutamine synthetase (GS) as evidenced by western blotting and qRT PCR studies. Further, GS and glutamate synthase(GOGAT) showed significantly high activities in leaf and root tissues in elevated CO

2 grown plants. To understand the

transcriptional regulation of GS, the ORF sequence was characterized by cloning into E.coli BL21 cells, the expressedprotein was purified and the sequence was confirmed by MALDI-TOF. In conclusion, this study revealed that lack ofphotosynthetic acclimation in pigeonpea under elevated CO

2, can be attributed to the effective functioning of the N

assimilatory enzymes including GS and GOGAT.


308

IPPCongress2015


PH381 (IPPC0510)

Carbon sequestration and biomass yield in short rotation coppice mulberry (Morus spp.) grown under

CO2-enriched atmosphere

K. Madhana Sekhar* and Attipalli R. Reddy



Present study was conducted to elucidate the responses of mulberry under predicted 2050 levels of elevated CO2

atmosphere (550 μmol mol-1). Elevated CO2 grown mulberry plants showed significantincrease in light saturated

photosynthetic rates (A’) throughout the experimental period by increasing intercellular CO2 concentration (C

i) despite

reduced stomatal conductance (gs). Reduced g

s was linked to decrease in transpiration (E) resulting in improved water

use efficiency (WUEi). Increased in vivo maximum rate of RuBP carboxylation (V

cmax), in vivo maximum rate of electron

transport driving RuBP regeneration (Jmax

), light and CO2 saturated photosynthetic rates (A

max), apparent quantum efficiency

(AQE), photosynthetic nitrogen use efficiency (PNUE) and photosystem-II associated electron transport rate (ETRmax

)clearly showed lack ofphotosynthetic acclimation in elevated CO

2 grown mulberry. Changes in V

cmax and J

max were positively

correlated with foliar nitrogen (NA) levels. Significant rise in transcript levels associated with key regulatory enzymes of

Calvin cycle, which are involved in both RuBP carboxylation and its regeneration,were believed to facilitate efficientcarbon sequestration resulting in superior biomass yields (45%) in elevated CO

2 grown plants compared to their controls.

Our data strongly suggest that short rotation (<1 year) with mulberry plantations should be effective to mitigate rising CO2

levels as well as for the production of carbon neutral renewable bio-energy.

PH382 (IPPC0514)

Photothermal indices in relation to physiology, phenology and yield traits of rice genotypes under different

sowing conditions

V. Ravichandran1*, S. Jeyapriya2 and S. Robin1

1Department of Rice, 2Department of Crop Physiology, Tamil Nadu Agricultural University, Coimbatore-641003, India


Rice productivity is highly dependent upon prevailing weather parameters like temperature, day length and solar radiationare important natural resources which affect the rice productivity to a greater extent. Sowing time and selection of varietymay play the deciding role in performance of rice production under changing climatic scenario. The proposed study wasconducted to reveal the photothermal indices on physiology, phenology and yield components of ten rice genotypesunder three sowing windows (June 9th, June 23rd and July 7th) for identifying the climate resilient rice genotypes suitablefor wider climatic change. Among the sowing dates, the rice genotypes sown on June 9th reflected significantly higherplant height and total dry matter production. Physiological characteristics like gas exchange parameter and light transmissionratio was desirable in rice genotypes sown under June 9th and the long duration genotypes surpassed other genotypes forphysiological characteristics, especially the genotypes CB-05-022 and IET 20924. Regarding phenology June 9th consumedmore number of days to reach different growth stages in the rice genotype CB-05-022. The photothermal indices i.e.growing degree days, photothermal units, heliothermal units, heat use efficiency and radiation use efficiency were recordedmaximum under June 9th sowing where, they took longer vegetative growth duration to produce higher grain yield byutilizing heat units and natural resources efficiently under favorable weather conditions. To conclude sowing date June 9th

favoured better growth and yield in rice genotypes CB-05-022 which utilize heat units efficiently under favourableenvironment to provide better yield than other genotypes used.


309

IPPCongress2015


PH383 (IPPC0521)

High light induced changes in organization, protein profile and function of photosynthetic machinery in

Chlamydomonas reinhardtii

Sai Kiran Madireddi, Elsin Raju Devadasu, Rajagopal Subramanyam and Srilatha Nama*

Department of Plant Science, University of Hyderabad, Hyderabad-500046, Andhra Pradesh, India


The green alga Chlamydomonas (C.) reinhardtii is used as a model organism to understand the efficiency of photosynthesisalong with the organization and protein profile of photosynthetic apparatus under one how exposure to various intensitiesof high light. Chlorophyll (Chl) a fluorescence induction, OJIPSMT transient decreased with increase in light intensityindicating the reduction in photochemical efficiency. Further, circular dichroism studies of isolated thylakoids from highlight exposed cells showed considerable change in the pigment-pigment interactions and pigment-proteins interactions.Furthermore, the organization of supercomplexes from thylakoids was studied, in which, one of the hetero-trimer of lightharvesting complex (LHC) II was affected significantly in comparison to other complexes of LHC’s monomers. Othersupercomplexes, PSII reaction center dimer and PSI complexes was reduced. Additionally, immunoblot analysis of thylakoidproteins revealed that PSII core proteins D1 and D2 were significantly decreased during high light treatment. Similarly,the PSI core proteins PsaC, PsaD and PsaG were drastically changed. Further, the LHC antenna proteins of PSI and PSIIwere differentially affected. From our results it is clear that LHC’s are damaged significantly, consequently the excitationenergy is not efficiently transferred to the reaction center. Thus, the photochemical energy transfer from PSII to PSI isreduced. The inference of the study deciphers the structural and functional changes driven by light influence; it maytherefore provide plants/alga to regulate the light harvesting capacity in excess light conditions.

PH384 (IPPC0575)

Photosynthesis and growth responses of mustard (Brassica juncea L. cv Pusa Bold) plants under free

air carbon dioxide enrichment (FACE) condition

Kamal Ruhil*, Sheeba, Altaf Ahmad, Muhammad Iqbal and B.C. Tripathy



Increased atmospheric (CO2) is likely to affect photosynthesis, plant growth, and yield potential of plants. Mustard (Brassica

juncea L.) is an important oil seed crop that is widely grown in India. Therefore, the impact of elevated (CO2) (585 μmol

mol-1) on pigment and protein content, chlorophyll a fluorescence, photosynthetic electron transport reactions, CO2

assimilation, biomass production, and seed yield potential was measured in B. juncea cv. Pusa Bold grown inside free aircarbon dioxide enrichment (FACE) rings installed on the campus of Jawaharlal Nehru University, New Delhi, India. Plantswere grown for three consecutive winter seasons (2010-11, 2011-12 and 2012-2013) in ambient (385 μmol mol-1) orelevated (585 μmol mol-1) (CO

2), in field conditions. Elevated (CO

2) had no significant effect on the minimal chlorophyll

fluorescence (F0), while the quantum efficiency of Photosystem II, measured as the ratio of variable fluorescence (Fv=Fm–F0) to maximum fluoresence (Fm) (Fv/Fm), increased by 3%. Electron transport rate of photosystem I and photosystemII, and the whole chain electron transport rates increased by 8% under elevated (CO

2). However, the net photosynthesis

rate increased by ≈50% in three growing seasons under elevated (CO2) condition. The stomatal conductance and

transpiration rate decreased resulting in higher photosynthetic water use efficiency. The photosynthesizing surface, i.e.,leaf area index substantially increased leading to higher biomass and seed yield under elevated (CO

2) condition. Acclimatory

down regulation of photosynthesis and plant productivity was not observed in three consecutive growing years, suggestingthat in the absence of nutrient limitation, B. juncea is highly responsive to elevated CO

2, whose yield potential shall

increase in changing climatic conditions.


310

IPPCongress2015


PH385 (IPPC0610)

Size effect of zinc oxide nanoparticles on photosynthesis and oxidant response in Oryza sativa

Prabhat Sharma and Maria Vera Da Costa*

Department of Botany, Goa University, Taleigao Plateau, Panaji-403206, Goa, India


Inevitable release of nanoparticles (NPs) in the environment due to waste generated by nanodevices or during nanomaterialmanufacturing process is a major concern. The physiological behaviour of Oryza sativa var. Jyoti treated with zinc oxidenanoparticles (ZnO NPs) of 50 and 100 nm size were hydroponically studied. Both sizes of ZnO NPs showed negligibleeffect on germination. Uptake of NPs was accompanied by reduction in shoot, root length, and shoot biomass, also,number and size of stomata decreased and number of trichomes increased. In contrast to non-metallo nanoparticles, nodeposition of ZnO NPs in the intercellular spaces of root parenchyma was seen, however, a decrease in the thylakoidnumber per granum of chloroplast at 1000 mg L-1 ZnO (100 nm) was observed. The maximum quantum efficiency of PSII(Fv/Fm ratio), photochemical quenching (qP), photosynthetic rate (PN), stomatal conductance (gs), transpiration rate (E)and photosynthetic pigment content were more significantly affected by 50 nm compared to 100 nm ZnO NPs. Prolineand ascorbate content increased while lipid peroxidation showed no significant increase on exposure with both sizes ofZnO NPs. Dissolution of ZnO NPs into metal ions was found to be far below the toxicity threshold attributing the phytotoxiceffect observed to NPs per se. Our results demonstrate that 50 nm sized NPs were more toxic compared to 100 nm sizedZnO NPs on the photosynthetic processes with significant increase in osmotic stress without causing lipid peroxidation.

PH386 (IPPC0626)

Inhibitory effects of metallo nanoparticle on photosynthesis are due to reduction in thylakoid membrane

in the grana region of chloroplasts rather than oxidative damage to proteins and lipids

Prabhat Kumar Sharma*

Department of Botany, University of Goa, Goa, India


The physiological and biochemical behaviour of rice (Oryza sativa var. Jyoti) treated with CuO and ZnO nanoparticles(NPs) in the range of 0-1000 mg L-1 was studied hydroponically. The treatment resulted in accumulation of the NPs, manyfold higher in roots than shoots indicating exclusion of metallo nanoparticle at root level. The accumulation was furtherobserved inside cells, especially in the chloroplasts and caused decreased stacking and structural deformation of thylakoidsper granum. Photosynthetic rate, transpiration rate, stomatal conductance, maximal quantum yield of PSII, biomass andphotosynthetic pigment contents decreased, with a complete loss of PSII photochemical quenching at 1000 mg L-1 of NPstreatment. Significant level of osmotic stress was evident by increased proline content as a result of the treatment,however, little or no oxidation to protein and membrane lipids, studied as protein carbonyl and MDA content respectively,was observed. Elevated expression levels of ascorbate peroxidase and superoxide dismutase but no oxidative damageto protein and membrane lipids suggest that production of ROS due to the treatment was within the inherent capacity ofits metabolism. Results showed that dissolution level of NPs into ions was much below the threshold of ion toxicity,indicating toxic effect due to the NPs rather than their ions. Results demonstrated that NPs caused inhibitory effect mainlydue to the decreased stacking of thylakoid membrane in the grana region of chloroplasts leading to smaller LHC II thuslower production of ROS, and not due to the oxidative damage as seen with bulk particles.


311

IPPCongress2015


PH387 (IPPC1005)

Antioxidant system-mediated regulation of chlorophyll biosynthesis pathway in vascular plant Cucumis

sativus upon exposure to methyl viologen and high light stress

Aarti P. Dhepe* and Komal A. Joshi

Institute of Bioinformatics and Biotechnology, S.P. Pune University, Pune-411007, India


Reactive Oxygen Species (ROS), being highly reactive in nature can cause oxidative damage to various biologicalmolecules. However, they can also act as a signalling molecule in diverse cellular processes. So, their generation andaccumulation has been found to be very crucial for plant growth, development as well as defense. This outstandingpotential of ROS is due to the development of extremely competent antioxidant systems, which either by over expressionor depletion maintain their appropriate concentration in plants. In this study, a regulatory relation between antioxidantsystem and chlorophyll biosynthesis pathway has been studied. Cucumber cotyledons, etiolated (E) and green (G, 14hilluminated), were treated with methyl viologen (MV) or high light (HL, 400-500 uE m-2 s-1) and the effects were studied onchlorophyll biosynthesis pathway as well as on antioxidant system. ROS generation was confirmed by proline and H

2O

2

concentration measurement. It was observed that chlorophyll biosynthesis was affected at the level of ALA synthesis andprotoporphyrinogen IX. Measurement of MDA contents showed that inhibition observed was not due to loss of membraneintegrity. When the activities of several antioxidant enzymes were assayed, it showed that ascorbate peroxidase activitywas reduced in E and G cotyledons upon MV treatment and in HL-treated E cotyledons. Catalse activity was also foundto be severely impaired in HL-exposed E cotyledons. Ascorabate and lutein contents were found to be decreasedsignificantly in MV treated E and G cotyledons and E-HL cotyledons. This suggested that significant decrease in theactivities of H

2O

2–scavenging antioxidants might have resulted in increased concentration of H

2O

2 which in turn might

have caused inhibition of chlorophyll biosynthesis pathway.

PH388 (IPPC1038)

Identification and characterization of sugarcane genotypes for photosynthetic efficiency, moisture stress

tolerance and high yield

T. Sujatha*, V.S.P. Lalitha and K. Jhansi

Crop Physiology, Sugarcane Research Station, Vuyyuru-521165, India


Six early pre-release sugarcane clones were tested against standard Co 6907 for their suitability under moisture stressconditions at Sugarcane Research Station, Vuyyuru during 2013-14 and 2014-15 seasons. Only two irrigations weregiven at 10th and 40th day after planting in addition to at the time of planting. Among the clones tested 2007 V 127 (97.77t/ha) and 2007 V 131 (96.13 t/ha) recorded significantly higher cane yield when compared to standard Co 6907 (80.84 t/ha) where as the clone 2007 V 129 (86.60 t/ha) was on par with the standard. Johari etal 1998 also reported that growthand yield affected due to soil moisture tension but the magnitude varied with the clones. Length of the millable cane washigher in 2007 V 131 and 2007 V 129. Higher diameter of the cane was recorded in 2007 V 131. The clones 2007 V 127(79.55%) and 2007 V 131 (79.53%) maintained more leaf sheath moisture percent, the same clones 2007 V 127 (79.20)and 2007 V 131 (79.07) were also recorded more relative water content percent during formative stage which coincidedwith summer. Dead canes recorded in those clones were also less compared to other cones. Higher percent juice sucrosewas recorded in 2007 V 127(21.39 %) followed by 2007 V 131 (21.35 %) and 2007 V 129 (21.04 %). The enzymesuperoxide dismutase was significantly higher in 2007 V 131 (1.17) followed by 2007 V 121 (1.14) and 2007 V 127 (1.08)which shows the tolerance of these clones to moisture stress. The clones 2007 V 127 and 2007 V 131 with higher RWC% leaf sheath moisture %, juice sucrose % length of the millable cane and cane yield were found tolerant to moisturestress conditions.

Session I : Genomics-assisted Plant Improvement

312

IPPCongress2015


PI389 (IPPC0023)

Quantitative proteomic analysis for elucidation of mechanism governing salt stress tolerance in

germinated seedlings of pigeonpea

Vandna Rai*

National Research Centre on Plant Biotechnology, New Delhi-110012, India


Pigeonpea (Cajanus cajan), a major source of dietary protein, is often subjected to salinity stress under different agroclimatic

conditions thereby severely limiting its productivity. Proteomic analysis is a viable approach for identifying set of proteins

that may have potential roles in conferring tolerance to salinity and/or other abiotic stresses. In this study, the effect of

varying concentrations of NaCl treatment for 7 d were evaluated on the developmental responses of root system architecture

(RSA) and shoots of pigeonpea. Developmental responses of these traits were significantly affected during growth at 150

mM NaCl. A label-free comparative Q TOF, LC-MS/MS technique was employed for qualitative and quantitative proteomics

analysis of roots and shoot grown under saline-free (0 mM NaCl) and saline-stress (150 mM) conditions. A total of 119

differentially expressed (DE) proteins were identified of which 78% of them were represented in both roots and shoots.

Interestingly, some of them such as Ras related and RNA-dependent RNA polymerase were detected only in the roots of

the seedling that were subjected to salinity stress. In addition, a few proteins i.e., Ribulose-1,5-bisphosphate carboxylase/

oxygenase large subunit and Dof 34 were detected in saline-stress shoot. The analysis suggested that these proteins in

roots and shoots may have a role in salinity stress response. Further, protein interaction network (PIN) was developed

using the String software with Confidence Scores >0.7 for identifying key regulators of salt stress tolerance in pigeonpea.

PI390 (IPPC0187)

Dynamics of long non-coding RNAs responsive to Sclerotinia sclerotiorum infection in canola

(Brassica napus)

Raj Kumar Joshi1*, Swati Megha2, Urmila Basu2 and Nataraj Kav2

1Centre of Biotechnology, Siksha O Anusandhan University, Bhubaneswar-751003, Odisha, India2Department of Agricultural Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada


The biggest constraint for canola (Brassica napus) production worldwide is the Sclerotinia stem rot caused by necrotrophic,

non host specific phytopathogen Sclerotinia sclerotiorum. Short non coding RNAs have been proven to play important

roles in the regulation of stress responses in canola but the functions of long non coding RNAs (lncRNAs) largely remains

unknown. In the present study, we used a strand specific RNA sequencing approach to identify lncRNAs responsive to S.

sclerotiorum infection in B. napus. A large set of B. napus RNA-seq data were examined with high sequencing depth

under control and inoculated conditions and 3181 lncRNA candidates were identified. A total of 931 lncRNAs were found

to be pathogen responsive, 12 of which were confirmed by RT-qPCR. Additionally, RT-qPCR verification of a set of

sense/antisense transcript pairs revealed contrasting expression patterns suggesting steric clashes of transcriptional

machinery. Forty one lncRNAs served as precursors for several stress responsive microRNAs having significant role in

the regulation of pathogen defense. B. napus lncRNA, TCONS_00000966 antisense to plant defensin gene PDF1.2 was

significantly induced suggesting its involvement in the transcriptional regulation of defense responsive genes during S.

sclerotiorum infection. Our results suggested that, RNA-Seq provide a comprehensive view of long non coding transcripts

and lncRNAs are important constituent of the antifungal network in B. napus.


313

IPPCongress2015


PI391 (IPPC0220)

LeasyScan - a novel semi-field platform to phenotype traits controlling plant water budget

Grégoire Hummel1*, Vincent Vadez2 and Jana Kholova2

1Phenospex B.V., Jan Campertstraat 11, 6416 SG Heerlen, Germany2International Crops Research Institute for Semi-Aird Tropics (ICRISAT), Patancheru-502324, Telangana, India


We describe the concept and realisation of a high-throughput phenotyping platform (LeasyScan) combined with lysimetric

capacity, to assess canopy traits affecting water use. The platform is based on a novel 3D scanning technique to capture

canopy development, a scanner-to-plant concept to increase throughput (2500 plants/h), and analytical scales to combine

gravimetric transpiration measurements. We present how the technology functions, how data are visualised via a web-

based interface, and how data extraction and analysis is interfaced through ‘R’ libraries. Close agreement between

scanned and observed leaf area data of individual plants in different crops was found (R2 between 0.86 and 0.94). Similar

agreement was found when comparing scanned and observed area of plants cultivated at densities reflecting the field

conditions. Example of the monitoring, the plant transpiration by the analytical scales is presented. Moreover, we present

some on-going applications of the platform to target key phenotypes: (i) the comparison of the leaf area development

pattern of fine mapping recombinants of pearl millet; (ii) the leaf area development pattern of pearl millet breeding material

targeted to different agro-ecological zones; (iii) the assessment of the transpiration response to high VPD in sorghum and

pearl millet.

PI392 (IPPC0227)

Genome-wide DNA polymorphisms in rice genotypes with contrasting behaviour under low Phosphate

Poonam Mehra*, Bipin Pandey and Jitender Giri



Soil phosphorus (P) deficiency is one of the major challenges to the rice cultivation world-wide. Modern rice genotypes

are highly P-responsive and rely on high input of P fertilizers. However, low P tolerant traditional cultivars and landraces

have genetic potential to sustain well under low P conditions. Identification of high resolution DNA polymorphisms (SNPs

and InDels) in such contrasting genotypes can serve as good resource for marker assisted breeding for improvement of

low P sensitive high yielding cultivars. Therefore, high quality DNA markers in low P sensitive modern genotype, PB1 and

tolerant traditional genotype, Dular were identified by whole genome resequencing. Approximately, 2.3 million and 2.9

million high quality polymorphisms with an average read depth of >24X were identified in PB1 and Dular, respectively with

reference to Nipponbare genome. Several non-synonymous and regulatory DNA polymorphisms having potential functional

significance were mapped to key Phosphate Starvation Responsive (PSR) and root architecture genes/QTLs in rice. To

deduce more promising SNPs/InDels potentially associated with low P tolerance, all Dular and PB1-specific variants

underlying PSR genes were compared with variants in another low P tolerant rice genotype, Kasalath. Further, all

polymorphisms underlying PSR genes were classified on the basis of their presence in low P sensitive genotypes (PB1

and Nipponbare) or low P tolerant genotypes (Dular and Kasalath). These identified variants can serve as a good source

of genetic variability for improving low P tolerance and root architecture of high yielding modern genotypes.


314

IPPCongress2015


PI393 (IPPC0237)

Ectopic expression of Pennisetum glaucum AsA-GSH pathway in ripened tomato fruits for enhancing

postharvest shelf life by reduction in oxidative damage

Krishnamurthy Sathelly*, Tanushri Kaul and Murtaza Malik

Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-67, India


Globalization has led to an increasing demand for quality improvement and the wider adoption of quality standards forfruits and vegetables. Plant transformation and regulated gene expression offers an integrated approach to tackle complextraits such as post-ripening oxidative deterioration of fruits. Genes that code for enzymes involved in key oxidative stressdetoxification via the ascorbate-glutathione pathway reveal a decline in their activities and also non-enzymatic antioxidantsascorbate and glutathione content decreases during post ripening of tomato fruits. Our strategy to deal with postharvestoxidative damage in tomatoes involves the overexpression of ascorbate-glutathione pathway genes from a stress adaptedC

4 plant – Pennisetum glaucum driven by different fruit ripening-specific promoters for scavenging the excess ROS

generated in tomato fruits during postharvest storage through agro-mediated transformation. Stable integration oftransgenes was confirmed by PCR and Southern analyses in T0 transgenic plants. Northern analysis in red-ripe stage ofT1 fruits revealed elevated transcript levels of transgenes in comparison to transgenic mature green and WT fruits.Activities of AsA-GSH pathway enzymes significantly increased (80-100%) in red-ripe tomato fruits in comparison to WTfruits. Postharvest decay symptoms appeared within 7 ± 0.8 days in WT fruits, whereas these developed after 31.8 ± 0.5days in transgenic red-ripe fruits that indicated delayed postharvest senescence. Results suggested an upregulation ofAsA-GSH cycle in red-ripe fruits during postharvest stress conditions. Fruit-specific enhanced expression of antioxidantenzymes from AsA-GSH pathway in tomato fruits effectively scavenged terminal ROS and minimized postharvestdeterioration, in turn enhancing their shelf life.

PI394 (IPPC0243)

Protein-Protein Interaction and gene co-expression network of rice TCPs

Vasudha Bhardwaj1*, Aleena Francis2, Ananda Rao Saketi3, Abhishek Kumar Anuj1, Suchismita Roy3, Manoj K. Sharma1,

Ashwani Pareek3 and Rita Sharma2

1School of Biotechnology, 2School of Computational and Integrative Sciences, 3School of Life Sciences, Jawaharlal Nehru University,New Delhi-110067, India


TCP is a plant-specific family of transcription factors with its name derived from the founding members, Teosinte branched1 (TB1) of maize, Cycloidea (CYC) of Antirrhinum and Proliferating cell factors (PCFs) of rice. Initially, identified as keyplayers in regulating plant growth and development, members of TCP family have recently emerged as multifacetedplayers engaged in abiotic stress tolerance as well as plant immunity. They can modulate wide range of biological processesby regulating variety of hormonal pathways and signaling cascades. Rice genome encodes 23 TCP proteins divided intotwo classes. Class I factors activate plant growth; whereas, class II factors have been shown to negatively regulategrowth-associated genes. The ability of class I and II factors to antagonize each other reflect a complex mechanism ofgene regulation. However, genetic redundancy is a major hindrance in elucidating the precise mechanism of their action.Also, TCP proteins are known to form homo and heterodimers for their function but how the choice of interacting partnersdictate the transcriptional outcome remains unresolved. In this study, we have identified rice interologs derived fromArabidopsis TCP interactome and validated the pairwise-interactions using yeast two-hybrid assay. Further, to dissectthe functionally redundant gene pairs, we have performed a comprehensive expression and coexpression analysis ofTCP genes in rice. The experimentally verified interactome coupled with sequence analysis, interologs, transcriptionalprofiling and coexpression analysis would be highly useful to elucidate redundancy and gene functions.


315

IPPCongress2015


PI395 (IPPC0255)

Genomic assisted breeding for introgression of gynoecious trait in Indian monoecious cucumber

(Cucumis sativus) cv. Pusa Uday

Tusarkanti Behera, Harshawardhan Choudhary*, Anilabh Dasmunshi, Rajni Karn and Brihama Dev

Indian Agricultural Research Institute New Delhi-110012, India


Indian cucumber cultivars exhibit monoecious sex expression coupled with crown fruit inhibition which is considered asone of the reasons for lower productivity. Gynoecism has been extensively exploited in heterosis breeding for enhancingproductivity of cucumber worldwide but exotic gynoecious lines are not stable at higher temperature and they possessvery few lateral branches which are not suitable for open field conditions. The introgression of gynoecious trait withmarker assisted breeding can improve the productivity of Indian cucumber. The F

2 population from cross G 421 (exotic

gynoecious) x Pusa Uday (Indian monoecious) was used for mapping, with 6 markers from the genomic region of F locus.The markers SSR-13251 and SSR- 15516 were found to be linked to F locus at 1.5 and 4.5 cM respectively. One genebased SCAR marker clearly distinguished gynoecious and monoecious plants in all F

2 population. These 3 markers were

used in foreground selection from 192 BC1F1 individuals and 18 heterozygous plants for F locus phenotypically similar toPusa Uday were selected. These plants were selfed to produce the BC1F2 and backcrossed with Pusa Uday to developBC2F1 population. The genotyping of these two populations was performed with these 3 markers and phenotypic selectionwas made for the traits similar to Pusa Uday with gynoecism. Genotypes with homozygous for F locus, stable phenotypicexpression of gynoecious trait and growth and fruit characters matching to Pusa Uday were selected. These plants will befurther evaluated for horticultural traits in different environment for varietal development.

PI396 (IPPC0264)

Engineering rice with PgOGG1, a bifunctional DNA glycosylase/AP lyase enhances abiotic stress tolerance

and seed longevity

V. Mohan Achary*, James Donald, Dhirendra Fartyal, Mrinalini Manna, Vijay Reddy, Ravi Kumar, Bhabesh Borphukan

and M.K. Reddy

Plant Molecular Biology Group, International Centre for Genetic Engineering & Biotech., Aruna Asaf Ali Marg, New Delhi-110067, India


Reactive oxygen species (ROS) are the by-products of normal cell metabolism in plants. However, under stress conditionsincluding seed desiccation, storage and germination, ROS homeostasis is disturbed resulting in seed deterioration andtherefore, decreased seed longevity. Notably damage to the genome induced by ROS has been recognized as an importantcause of seed deterioration. The prominent DNA lesion induced by ROS is 7,8-dihydro-8-oxoguanine (8-oxo-G), that formbase pair with adenine instead of cytosine during DNA replication and leads to GC/TA transversion. This altered base isremoved by 8-oxoguanine DNA glycosylase and repaired by the subsequent BER enzymes. In this report, we identifiedand characterized OGG1 from the C4 plant Pennisetum glaucum (PgOGG1) that encodes 45.3 kDa protein showingsignificant sequence identity with plant orthologues as well as yeast and human OGG1 protein. The transcript of PgOGG1was strongly up-regulated during seed desiccation and imbibition. Purified PgOGG1 specifically cleaves 8-OxoG:C mispairDNA duplex which is a characteristic of all bifunctional DNA-glycosylases/lyases. PgOgg1 was found to localize both innucleus and mitochondria. Expression of PgOGG1 in rice showed reduced level of 8-oxo-dG and enhanced seed resistanceto controlled deterioration treatment indicating a DNA damage-repair function of PgOGG1 in vivo. Furthermore, transgenicseeds exhibited increased germination ability under abiotic stresses induced by MV, NaCl and high-temperatures. Takentogether, our results demonstrate that ectopic expression of PgOGG1 enhances seed longevity and abiotic stress tolerancein rice mediated by repair of the damaged DNA through elimination of 8-oxoG from the genome.


316

IPPCongress2015


PI397 (IPPC0285)

Genome wide study of two component system and identification of key members of cytokinin signaling

during nodule development in chickpea

Manish Tiwari* and Sabhyata Bhatia



Chickpea is the world’s second most grown legume, with Indian subcontinent being the principal chickpea producing andconsuming region. Nitrogen fixation by symbiotic association increases its importance. Symbiotic association is in form ofroot nodule, which harbors bacteria and facilitates nitrogen fixation. Several studies have been carried out on the rootnodule formation; and it has been clearly observed that cytokinin has very important role in nodulation. The expression oftrans – zeatin gene in nodulation-deficient phenotype of Rhizobium mutant initiates nodulation. Cytokinin application alsoinduced the early nodulin gene and formation of false nodules. Cytokinin signaling is achieved by two component system(TCS) in plants which is composed of sensor histidine kinases (HKs), histidine phosphotransferases (HPs), and responseregulators (RRs) and it would be interesting to investigate their roles in cytokinin mediated nodulation in chickpea. Towardsthis, an exhaustive genome wide search of the chickpea genome for TCS members was conducted. Analysis of theirconserved domains, and phylogenetic relationships of these protein-coding genes was carried out. The transcriptionlevels of TCS genes in various organs and different nodule developmental stages along with environmental stresseswere investigated. Our systematic analyses provide insights into the characterization of the TCS genes in chickpea, theirinvolvement in root nodule formation and basis for further functional studies of such genes.

PI398 (IPPC0415)

Phenotypic/genotypic evaluation of, and conservation strategy for, natural populations of endangered

Boswellia Serrata Roxb.

Shashank Mahesh1, Pramod Kumar1, Vivek Vaishnaw1, Naseer Mohammad1, Ashwani Kumar2 and Shamim Ansari3*

1Tropical Forest Research Institute, Jabalpur-482021, Madhya Pradesh, India2Indian Council of Forestry Research and Education, Dehra Dun-248006, India3Institute of Forest Productivity, Ranchi-835303, Jharkhand, India


Boswellia serrata Roxb. (salai) is an important species of dry deciduous forest and has been over exploited for its oleogum-resin and excellent paper pulp in the central India. Consequently, there exists fragmentation of populations and poornatural regeneration. We investigated variability and status of populations in terms of wood fiber length and tree girth bysampling 20 trees each of 12 natural Madhya Pradesh populations of B. serrata existing in different agro-climatic conditions.The wood fiber length and tree girth exhibited 59.7% and 505.2% variations, respectively, across populations. There wassmall positive and negative kurtosis respectively for both parameters, which indicates uneven loss of genotypes or geneticdrift. Khandwa population with the longest wood fiber length was 18.2% superior over Jabalpur population with theshortest wood fiber length. Tree girth of Chhindwara population was maximum and 109.2% more than that of Damohpopulation. The broad sense heritability values for wood fiber length (0.201) and tree girth (0.334), indicate that both traitsare associated with fitness in natural populations. Further, QST estimates for both parameters exceeding FST estimatesobtained with neutral molecular markers (RAPD and ISSR), indicate towards operation of divergent natural selection.Nevertheless, the findings encourage for transfer of diverse genotypes from one population to the other for facilitating outcrossing within populations as well as production of viable seeds, circumventing inbreeding depression and genetic drift.The proposed strategy would promote natural regeneration and in situ germplasm conservation/enrichment, which wouldhelp rescue the species from endangered status.


317

IPPCongress2015


PI499 (IPPC0457)

Transcription factor gene families from Papaver somniferum: Identification of regulatory factors involved

in benzylisoquinoline alkaloid biosynthesis

Parul Agarwal1*, Sumya Pathak2, Deepika Lakhwani1, Parul Gupta1, Mehar Asif1 and Prabodh Trivedi1

1Molecular Biology and Genetic Engineering, CSIR-National Botanical Research Institute, Lucknow-226001, India2University of Lucknow, Lucknow-226007, India


Opium poppy (Papaver somniferum L.) is known for the biosynthesis of structurally diverse and pharmaceutically valuablebenzylisoquinoline alkaloids (BIAs) which includes narcotic analgesic morphine, the cough suppressant codeine, themuscle relaxant papaverine and the anti-microbial agent sanguinarine and berberine. Coordinated transcriptional regulationof biosynthetic genes has emerged as a major mechanism controlling the production of secondary metabolites in plantcells. Transcription factors, one of the prime regulators of secondary plant product biosynthesis, might be involved incontrolled biosynthesis of BIAs. In this study, identification of members of different transcription factor gene families usingtranscriptome datasets of 10 cultivars of P. somniferum with distinct chemoprofile has been carried out. WRKY wasidentified as the most represented transcription factor gene family in all the poppy cultivars. Comparative transcriptomeanalysis revealed differential expression pattern of the members of a set of transcription factor gene families among 10cultivars. Utilizing the integrated transcript and metabolite profiling strategy, two members of WRKY and one member ofC3H gene family were identified as potential regulators of the papaverine and thebaine biosynthesis, respectively. Thephylogenetic analysis suggest their close relationship with members of these gene families from other plants species.The regulatory role of these transcription factors and their involvement in specific alkaloids biosynthesis is being investigatedusing virus induced gene silencing.

PI400 (IPPC0459)

Identification and expression analysis of genes involved in biosynthesis of Azadirachtin and related

triterpenoids in Azadirachta indica (Neem)

Sweta Bhambhani1*, Deepika Lakhwani1, Ashutosh Pandey2, Mehar Asif1 and Prabodh Trivedi1

1Molecular Biology and Genetic Engineering, CSIR-National Botanical Research Institute, Lucknow-226001, India2National Agri-Food Biotechnology Institute, Mohali, Punjab, India


Azadirachta indica, commonly known as ‘Neem’ contains tetranoterpenoids synthesized by isoprenoid pathway, whichplay various functional roles in plant defense. The major bioactive components include Azadirachtin, Nimbin and Salannin,which serve as ecofriendly biopesticides and insecticides due to their various important roles as insect feeding deterrants,toxicants, disruptants of growth and development against a variety of insects, pests and nematodes. Numerous studieshave been carried out in relation to phytochemistry and biomedical activity of the plant but limited information is availableon genes involved in the biosynthetic pathway of azadirachtin and related triterpenoids. In the present study, we haveidentified genes involved in triterpenoid backbone and analyzed their expression in various plant parts of A. indica.Structural analysis of genes suggests significant conserved structure and high sequence homology with homologuesgenes from other plants. Expression analysis of genes in various plant parts suggests a weak correlation with phytochemicalaccumulation suggesting involvement of enzymes responsible for secondary transformation. As putative biosyntheticpathway of Azadirachtin suggests involvement of cytochrome P450 (CYP450) enzymes in secondary transformation, weidentified putative CYP450s having significant correlation between Azadirachtin content and its expression. This is thefirst study to identify and correlate expression of putative genes involved in biosynthetic pathway of Azadirachtin biosynthesisfrom neem.


318

IPPCongress2015


PI401 (IPPC0467)

Characterisation of Oxidosqualene Cyclase encoding genes involved in withanolide biosynthesis from

Withania somnifera

Aditya Vikram Agarwal1*, Parul Gupta1, Rahul Michael1, Deepak Chandra2 and Prabodh Kumar Trivedi1

1CSIR-National Botanical Research Institute, Lucknow-226001; 2Department of Biochemistry, Lucknow University, Lucknow-226007


Tri-terpenes, a diverse group of plant secondary metabolites, possess variety of biological activities related to thepharmaceutical, cosmetic and food sectors. Withania somnifera (L.) Dunal, a medicinal shrub has various medicinalproperties as anti-arthritic, anti-aging, anticancer and physiological and metabolic restoration. These pharmacologicalactivities have been correlated with the withanolides, C28-steroidal lactones derived from tri-terpenoids. Biosynthesis ofwithanolides involves one or both of committed upstream routes of isoprenogenesis-the cytosolic mevalonate (MVA)pathway and plastid localized 2-C-methyl-D-erythritol- 4-phosphate (MEP) pathway. These pathways direct the flux of theisoprene (C5) units into the tri-terpenoid pathway via a common C30-intermediate 2,3-oxidosqualene. In Withania, furtherbiosynthesis from this C30 intermediate is partitioned by cycloartenol synthase (CAS) and an array of other oxidosqualenecyclases (OSCs), which together form a myriad of diverse triterpenoids. CAS leads to the formation of cycloartenol, atetracyclic tri-terpene, which acts as a precursor for phytosterols and apparently withanolides. OSCs are positioned at akey metabolic sub-dividing junction executing an indispensable step in varied tri-terpenoid biosynthesis thus presentingfavourable gene targets for redirecting metabolic flux towards specific secondary metabolites. However, involvement ofspecific OSCs towards biosynthesis of withanolides has not been elucidated yet in planta. In this study, involvement ofgenes encoding OSCs in withanolide biosynthesis has been demonstrated through virus-induced gene silencing (VIGS)approach. Expression analysis and phytochemical analysis suggest decreased level of transcripts for specific OSCs inVIGS silenced lines as well as their involvement in directing flux towards withanolide biosynthesis.

PI402 (IPPC0497)

Mobilizing landrace variations for next generation wheat improvement

Prashant Vikram1*, Juan Burgueno Ferrera1, Jorge Franco2, Carolina Saint-pierre1, Carolina Paola Sansaloni1, Amita

Mohan3, Mathew Reynolds1, Carlos Guzman1, Pawan Kumar Singh1, Peter Wenzl1, Thomas Payne1,

Kulvinder Singh Gill3, José Crossa1 and Sukhwinder Singh1

1International Maize and Wheat Improvement Center (CIMMYT), Mexico; 2Departamento de Biometría, Estadística y Computación,Facultad de Agronom&iacut; 3Washington State University, Washington, USA


Slow yield potential gain, climate change effects, expected increase in future food demand and narrowed breedinggermplasm pool has emphasized an increased use of gene bank resources. Food and agriculture organization (FAO) hasaccentuated on enhanced use of on-farm and gene bank diversity. Semi-dwarfing wheat varieties revolutionized grainproduction; however, these varieties could not deliver significant impact in rainfed areas. Preliminary evaluation of pre-breeding populations derived from the most popular wheat variety, PBW343 revealed the importance of semi-dwarfinggenes for rainfed environments and suggested for enhanced infusion of useful landrace variations to the breedinggermplasm pool. Unique efforts have been made in ‘Seeds of discovery’ project at International center for maize andwheat improvement (CIMMYT) to perform physiological, genetic and molecular characterization of gene bank resourcesfocusing mainly on landraces. Genotyping-by-sequencing of 11000 wheat landraces (from Mexico and Iran) was carriedout along with trait characterization for heat, drought and grain quality. Landrace core set was formulated following aunique strategy which involved a data matrix representing phenotypic and genotypic variables simultaneously. Genomicassociations and novel alleles for agronomically important traits have been identified. Trait donors were identified forheat; drought, diseases, quality and yield. These resources are being strategically deployed through pre-breeding toachieve short term, medium term and long term goals. Landrace and other resources are being leveraged with NARS inSouth Asia, Africa and USA for multiple location evaluation. We welcome global wheat community to join hands anddeliver high impact in respective environments.


319

IPPCongress2015


PI403 (IPPC0513)

Unravelling the role of miR159-MYB65 module for regulating reproductive organ development in Brassica

Saurabh Anand*, Neer Singh and Sandip Das

Department of Botany, University of Delhi, Delhi-110007, India


Reproductive development in plants is an outcome of complex interplay of genes and genetic networks. Unravelling the

components of this network and understanding these interactions has helped in gaining knowledge about the genes and

small RNAs involved in androecium and gynoecium development. MicroRNA159 that targets two GA-MYB transcription

factors MYB33 and MYB65, were shown to affect reproductive development in Arabidopsis thaliana. To understand their

precise role, miR159 and MYB65 have been identified from Brassica rapa and B. juncea utilizing comparative genomics

approaches, and cloned. Transgenic lines over-expressing miR159 in A. thaliana Col-0 genetic background exhibit defects

in flowering time, anther dehiscence, anther and pistil morphology, pollen-pistil interaction, pollen viability and fertility,

and seed set. We have performed transcriptome analysis including RNA-sequencing and semi-qPCR of the transgenic

and the WT lines to gain an insight into the global changes and perturbation accompanying developmental defects.

Additionally, the role of hormones in reproductive organ development, especially GA, is being studied through experiments

involving external applications. Experiments are also being performed using Brassica juncea owing to its importance as

oil-yielding crop. Towards this goal, transgenic plants have been generated and are currently being analysed.

PI404 (IPPC0516)

Analysis of root system architecture in chickpea

Vikas Dwivedi*, Swarup Kumar Parida and Debasis Chattopadhyay

National Institute of Plant Genome Research, Aruna Asif Ali Marg, New Delhi-110067, India


Chickpea (Cicer arietinum) is the third most consumed legume crop in the world. It is a major source of protein and,

therefore, a very important food crop in developing countries. Due to several biotic and abiotic factors, annual crop yield

is severely affected all over the world. To develop agricultural sustainability, chickpea is an important legume crop having

excellent ability to fix atmospheric nitrogen biologically. A correlation between root system architecture and resistance to

water stress has been found in several crop plants and breeding attempts have focused on obtaining cultivars with larger

root systems. Development of root biomass and its architecture helps the plant to access water more efficiently from soil.

The manipulation of root architecture could minimize the negative impact of a number of abiotic factors and can improve

yield. Root System Architecture (RSA) is different between two species and also within the species. RSA is a highly

plastic trait. Many methods are used for study of the root system. The development and architecture of roots hold potential

for the use and manipulation of root characteristics to both increase yield and optimize agricultural land use. To understand

the variation in chickpea root system architecture, we have studied about 41 different varieties of chickpea. Root system

architecture (RSA) was analyzed for these varieties using different tools. We have also studied the root architecture of

one mapping population (JGK3 X Himchana 1). Our main objective was concentrating on identification of genetic loci for

root development in two chickpea genotypes with contrasting root traits.


320

IPPCongress2015


PI405 (IPPC0554)

Functional characterization of MYC2 in disease defense

Janesh Kumar Gautam1*, Mrunmay Giri2, Sudip Chattopadhyay3 and Ashis Kumar Nandi1

1 School of Life Sciences, Jawaharlal Nehru University, New Delhi-110 067, India2University of North Texas, USA3National Institute of Technology, Durgapur-713209, West Bengal, India


Plants have evolved a complex immune system that is tightly regulated at transcriptional level by myriad of transcription

factors. MYC2, a basic helix loop helix (bHLH) transcription factor plays important roles in growth, development and

disease defense. MYC2 negatively regulates blue light mediated hypocotyl elongation. Reports suggest that MYC2 also

acts as a negative regulator of basal defense against bacterial pathogens. Our results show that MYC2 OEX plants have

very strong level of resistance than myc2 mutants against Pseudomonas syringae. MYC2 binds to G-box motif (CACGTG).

By the presence of canonical G-box motif, we identified a set of genes that influence defense response and are likely to

be targets of MYC2. Out of the identified genes, we selected ICS1, MKK4, PEPR1 and RIN4 for further studies.The

transcript level of these genes were highly induced after Pseudomonas syringae in MYC2 dependent manner. By gel shift

assay, we show that MYC2 protein physically associates with the target gene promoters. Experiments are underway to

confirm the role of MYC2 binding in regulating transcription of target genes.

PI406 (IPPC0566)

Marker assisted backcross breeding for transfer of a major QTL, qDTY1.1 determining grain yield under

lowland drought into an elite Basmati rice variety “Pusa Basmati 1”

Gaurav Dhawan1*, Priyanka Dwivedi1, S. Gopala Krishnan1, M. Nagarajan2, P.K. Bhowmick1, Madan Pal3 and A.K. Singh1

1Division of Genetics, 3Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India2Indian Agricultural Research Institute, Rice Breeding & Genetics Research Centre, Aduthurai-612101, Tamil Nadu, India


Drought is a major abiotic stress limiting the production of rice. It is gaining importance in India even in traditionally

irrigated ecosystems, owing to decrease in availability of irrigation water primarily due to decline in ground water level as

well as fluctuation in monsoons. Pusa Basmati 1 (PB1), the first semi-dwarf high yielding Basmati rice variety is popular

among the farmers even today, due to its advantages of semi-dwarf growth habit, photoperiod insensitiveness and its

exceptional grain and cooking quality characters. Since, it has been bred for irrigated ecosystem with assured irrigations;

it is susceptible drought, which limits both yield as well as grain quality under drought stress. Marker assisted backcross

breeding (MABB) was adopted to incorporate a major QTL for grain yield under drought, qDTY1.1 from a non-basmati

rice variety “Nagina 22” into Basmati rice variety PB1 for enhancing grain yield under the drought stress at reproductive

stage. Foreground selection was carried out at every generation using the SSR marker RM431, linked to qDTY1.1.

Additionally, at each generation stringent selection for the recurrent parent phenome, grain and cooking quality characters

was carried out to identify the best plant for further backcrossing. Advanced backcross progenies in BC3F

2 harboring

qDTY1.1 in the genetic background of PB1 has been identified. Further testing of the developed backcross derivatives

under drought will help in identifying potential lines with drought tolerance as well as other desirable traits, which will be

invaluable source of drought tolerance in Basmati rice improvement.


321

IPPCongress2015


PI407 (IPPC0584)

Identification of genomic regions and genes for nitrogen use efficiency in rice

P. Vijayalakshmi*, T. Vishnukiran, B. Ramana Kumari, K.N. Swamy, V. Jaldhani, M. Suchendranathbabu, P. Raghuveer

Rao, C.N. Neeraja and S.R. Voleti

ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad, India

*Participanting author: [email protected]

Association mapping is a notable strategy for identifying genes underlying quantitative traits in plants. The main aim of

the present study is to find out genomic regions/genes for NUE at different N levels viz., application of N fertilizer (N100)

and without application of N fertilizer (N0) during wet (2011) and dry (2012) seasons. Total 96 rice genotypes were

characterized for physiological, morphological, yield and NUE related traits. Population structure or hierarchical cluster

analysis was carried with 52 SSRs markers, which resulted in two major clusters. Marker traits associations were identified

with general linear model (Q+K) using Tassel 2.1 software. A total 49 markers were shown significant association with

multiple traits at two N levels during two seasons. Out of which, 37 markers are common for both the seasons resulting

eight and four specific markers for wet and dry season. In wet season, 30 markers were significantly associated in both

N levels with 9 and 6specific markers in N100 and N0, where as in dry season, 21 markers were significantly associated

in both N levels with six and fourteen specific markers in N100 and N0 respectively. Finally, total ten markers were shown

significant association in both N levels and seasons. Many marker associated genomic regions were coincided with

already known genes and also predicted some of candidate genes for NUE with in the window of 1Mb marker region. The

findings from the present study will help to identified and also provide a basis for breeding genotypes that are tolerant to

low-N conditions.

PI408 (IPPC0604)

RNA-Seq analysis for the study of global transcriptome in rice under induced osmotic stress

Narottam Dey*

Centre for Biotechnology, Visva-Bharati, Santiniketan-731235, West Bengal, India*Presenting author: [email protected]

High-throughput mRNA sequencing (RNA-Seq) offers a number of significant information of which discovery of new

genes and measurements of transcript expression in a single assay is the most important one. The most crucial steps

involving RNA-Seq analysis are the appropriate treatment of biological samples, sufficient biological replica, quality

control of reads, proper alignments, and setting up of downstream statistical analysis. NCBI-SRA database is a valuable

global resource for raw transcripts, generated through recent sequencers. The SRA data have been used to validate

experimental results, determine variance and open up new avenues of Bioinformatics research. Rice, a model crop with

known genome sequence has been used in a number of RNA-Seq experiments has an enriched SRA data base linked

with induced osmotic stress experiments. In the present work, a comparative Bioinformatics analysis was carried out

among a set of trascriptome data available in NCBI-SRA database, conducted under varied induced osmotic stresses on

different rice tissues. In analysis pipeline, the raw sequence data collected in from SRA were transformed into reads

(using fastqdump), quality checked (using fastqc), aligned with rice reference genome (using bowtie2 and Tophat2),

assembly of transcripts (with Cufflinks and Cuffmerge), differential expression study (with Cuffdiff) and final visualization

through CummeRbund. In a parallel pipeline, SRA database generated sraRunInfo.csv files were used for samples.

RData which was used in a downstream analysis in R using DESeq and edgeR packages. Finally, the information generated

from two parallel pipelines was compared to generate a final expression profiles with associated information.


322

IPPCongress2015


PI409 (IPPC0679)

A first generation HapMap and diversity analysis in a collection of 129 cultivars of chickpea

Annapurna Chitikineni1*, Manish Roorkiwal1, Weiming He2, Pooran Gaur1, Dadakhalandar Doddamani1, Jianbo Jian2,

Abhishek Rathore1, Rachit K. Saxena1, Narendra Singh3, S.K. Chaturvedi4, Swapan K. Datta5, Gengyun Zhang2,

Jun Wang2 and Rajeev K. Varsheny1

1International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India2Beijing Genomics Institute (BGI) - Shenzhen, China3All India Coordinated Research Project on Chickpea (AICRP), Indian Council of Agricultural Research, Kanpur, India4Indian Institute of Pulses Research (IIPR), Indian Council of Agricultural Research (ICAR), Kanpur, India5Visva-Bharati, Santiniketan, India*Presenting author: [email protected]

Chickpea (Cicer arietinum L., 2n=16) is the second most important food legume crop in the world. It contributes to a larger

part of human food and animal feed in developing countries. Due to exposure of the crop to a range of biotic and abiotic

stresses, the crop productivity, however, is <1 ton per hectare. Furthermore, because of a very narrow genetic diversity

in the elite genepool, molecular breeding approaches have also not been in routine for crop improvement in chickpea.

With an objective to estimate variations and linkage disequilibrium (LD) in elite germplasm, a non-redundant set of 129

varieties released in 12 countries (Australia, Bangladesh, Spain, Canada, USA, Ethiopia, India, Nepal, Kenya, Tanzania,

Myanmar and Sudan) were subjected to whole genome re-sequencing (WGRS). While 5X-12X (with an average 7.3X)

coverage WGRS data were generated for 129 lines. Alignment of sequence data for these cultivars with the draft genome

sequence of chickpea has provided >1.3 million SNPs in genome, >108 K in mRNA and >42 K in CDS regions. By using

these structural variations (SNPs, Indels, PAVs, CNVs), a comprehensive analysis has been undertaken on LD decay

and temporal diversity in terms of released varieties in different decades, as well as market type (desi and kabuli). In

addition, a first generation HapMap for chickpea has been developed that provides a resource of large-scale variations

for facilitating chickpea breeding.

PI410 (IPPC0701)

Characterization and development of marker for discriminating waxy and non-waxy rice germplasm

Gayle Kharshiing* and N.K. Chrungoo

Centre of advanced studies, North Eastern hill University, Shillong-793022, Meghalaya, India


Rice starch consists of amylopectin and amylose, the latter component being controlled by the Waxy (Wx) gene. Apparent

Amylose Content (AAC), regulated by the Waxy gene, represents the key determinant of rice cooking properties. Studies

pertaining to the waxy locus are not only of interest in understanding gene regulation but are also important for agricultural

applications. Waxy varieties of the diploid cereal rice (Oryzae sativa L.) have endosperm starch granules lacking detectable

amylose. In this study, the effectiveness of available molecular markers and comparative studies was conducted in

predicting the AAC values in which 70 rice accessions were characterized into waxy and non-waxy using iodine staining

of starch granules. Two alleles Wxa and Wxb were found to regulate the quantitative level of the Wx protein as well as the

amylose content. A comparative sequence analysis of the locus revealed the unique 23 bp duplication event which is

associated with the base substitution found in the 5’intron1 splice site found in the waxy locus which discriminates the

waxy from the non-waxy. The relationship of the 5’intron1 splice site with the 23 bp duplication event can further validate

their association with the waxy phenotype. Further, a marker was developed to distinguish between waxy and non-waxy

in rice germplasm based on the 23 bp duplication.


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IPPCongress2015


PI411 (IPPC0729)

Prediction of breeding values using genome wide markers for yield related traits in chickpea

Manish Roorkiwal1*, Abhishek Rathore1, Roma Das1, Muneendra Singh1, Samineni Srinivasan1, Pooran M. Gaur1,

Chellapilla Bharadwaj2, Shailesh Tripathi2, Yongle Li3, John M. Hickey4, Aaron Lorenz5, Tim Sutton6, Jean-Luc Jannink7

and Rajeev K. Varshney1

1International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India; 2Indian Agricultural Research Institute(IARI), Delhi, India; 3Australian Centre for Plant Functional Genomics (ACPFG), University of Adelaide, SA, Australia; 4The RoslinInstitute, The University of Edinburgh, Scotland, UK; 5University of Nebraska, Lincoln, USA; 6South Australian Research andDevelopment Institute (SARDI), SA, Australia; 7Cornell University, Ithaca, USA*Presenting author: [email protected]

Genomic selection (GS) unlike MABC (marker assisted backcross) predicts breeding values of lines and makes selectionprior to phenotyping using genome-wide marker profiling. To address the low productivity in chickpea owning to severalbiotic and abiotic stresses, a collection of 320 elite breeding lines was selected as the “training population”. Trainingpopulation was phenotyped extensively for yield and yield related traits at ICRISAT, Patancheru and IARI, Delhi duringcrop season 2011-12 and 2012-13 under rain-fed and irrigated conditions. In parallel, the training population was genotypedusing DArTseq arrays (15,360 features). Collected phenotypic data and generated genome-wide marker profiling datawere used with six statistical models including RR-BLUP, kinship based ridge regression, BayesCπ, BayesB, BayesianLASSO and random forest to predict genomic estimated breeding values (GEBVs). GS models were tested for four yieldrelated traits namely seed yield, 100 seed weight, days to 50% flowering and days to maturity. Prediction accuracy for themodels tested varied from 0.912 (SDW) to 0.138 (seed yield). Heat map analysis using genotyping data to understand therelationship within these lines suggested possibility of two different groups similar to cluster analysis. In order to understandthe effect of population structure on accuracy, analysis was re-performed by implementing population structure forcalculation of GEBV. Population structure did not have significantly effect on prediction accuracy. Results obtained areencouraging for deploying GS in chickpea breeding.

PI412 (IPPC0753)

In silico analysis of an endosperm-specific legumin-type seed storage protein gene and 5’UTR of common

buckwheat (Fagopyrum esculentum Moench)

Lashaihun Dohtdong1*, Kamlesh Kant Nutan2, Nikhil Kumar Chrungoo1 and Ashwani Pareek2

1Department of Botany, North Eastern Hill University, Shillong-793022, India2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India*Presenting author: [email protected]

The nutritional quality of food and its availability are the key factors to food security. We report the cloning of a full length1.7 kb gene (acc. no. KM488332) coding for a legumin type seed storage protein of common buckwheat (Fagopyrumesculentum Moench). BLAST analysis of the sequence revealed >90% homology with legumin gene nucleotide sequencesbearing accession numbers D87980, GQ358524 and AY245536.The deduced amino acid sequence from the open readingframe code for a 64 kDa preprotein. BLASTP analysis of the sequence revealed 98% homology with 13S globulin seedstorage protein from common buckwheat (acc.no.O23878). Analysis of the deduced amino acid sequence revealed thepresence of an α and a β subunit linked together ASN-GLU linkage with the β subunit showing 5.57% lysine, which isamongst the highest recorded for legumin type proteins. Using the gene walking approach, we have isolated the 1kb5’UTR (acc. no.EU595873) of the gene as identified by GENSCAN 1.0. NNPP predicted three probable transcription startsites out of which the TSS at P’733 is located closest to the predicted ATG start codon (P’774) and follows the YR rule.Other cis-elements identified in the sequence included the -30 TATA box, an endosperm specific motif identified asprolamin box in cereals ‘TGTAAAG’, three RY elements ‘CATGCA’, core of ‘AACA’ motif for regulating endosperm specificand quantitative expression, CAAT’ promoter consensus sequence and ‘CACA’ box.We suggest a strong potential for thegene and its promoter in enhancing the nutritional levels of lysine in conventional crops such as rice through transgenicapproaches.


324

IPPCongress2015


PI413 (IPPC0801)

Inheritance of red kernel colour in rice (Oryza sativa L.) and genotypic screening of F2 population through

molecular markers

Ashish Jondhale, Sanjay Bhave*, Jalindar Devmore, Murad Burondkar and Vijay Dalvi

Dr. B.S. Konkan Krishi Vidyapeeth Dapoli-415712, Maharashtra, India


Inheritance of red kernel colour in rice (Oryza sativa L.) and genotypic screening of F2 population through molecular

markers was studied in two crosses involving one white kernel and another red kernel parents. Result indicated that the

red kernel colour of rice was governed by a single dominant gene. Interactions confirmed by making testcrosses of the

F1s with the white kernel parents which were recessive for the kernel colour. In molecular screening, 13 SSR markers

linked to the red kernel colour trait were utilized and among these, two SSR markers (RM 206 and RM 251) showed

parental polymorphism. A representative 100 individuals derived from the crosses Karjat-2 x Munga and their reciprocal

was estimated for white and red kernel colour. Estimation of nutritional aspects especially micronutrients viz; iron, zinc,

manganese and copper exhibited wide variation in both, white and red kernel parents and their hybrids. The iron, zinc,

manganese and copper contents in the red rice were found to be higher than that of white rice. It clearly indicated that red

rice was highly nutritive than that of white rice. Beside this, generation mean analysis, revealed the significant Mather’s

individual scaling tests and joint scaling test for all the characters in both the crosses. Among non allelic gene interactions,

additive x additive and dominant x dominant were more prominent and played significant role in inheritance of the several

traits studied. The additive x dominant interaction were significant but had lesser preponderance for governing the traits.

PI414 (IPPC0829)

Isolation and characterization of UDP-glucose pyrophosphorylase (UGPase) promoter from potato

(Solanum tuberosum L.)

Shashank Yadav1*, Lekshmy S.1, Rakesh Verma1, Santosh Vinjamuri1, Monika Dalal2 and Viswanathan Chinnusamy1

1Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India2National Research Centre on Plant Biotechnology, New Delhi-110012, India


Low temperature storage of potato is inevitable to prevent sprouting and diseases, retention of dry matter and extended

marketability. However, Cold Induced Sweetening (CIS) is a problem associated with cold storage. Towards unravelling

the genotypic differences in CIS, we analyzed allelic variation in genes involved in CIS, and characterized UGPase

promoter. UGPase catalyzes reversible production of UDPG and pyrophosphate (PPi) from Glc-1-P and UTP. UGPase

expression was analyzed in potato genotypes with contrasting CIS. Previously reported allelic variations in UGPase,

apoplastic invertase and invertase inhibitor were analyzed in Indian potato genotypes. Absence of UGPase “B”, apoplastic

invertase inhibitor StINHap-a and apoplastic invertase Stm1051-255 alleles, and presence of apoplastic invertase Stm1051-

221 were associated with CIS tolerance of potato. Expression of UGPase gene was highly induced by cold storage in CIS

tolerant genotypes Kufri Chipsona 1 and Kufri Frysona, whereas the expression was low in CIS sensitive genotype Kufri

Pukhraj. To unravel the differential expression of UGPase genes between CIS tolerant Kufri Chipsona 3 and CIS sensitive

Kufri Pukhraj, the promoter of UGPase was cloned and sequenced. These two genotypes differ by 49 and 6 base InDels

in UGPase promoter. Based on this InDel, a PCR marker was developed and validated in 38 potato genotypes. CIS

tolerant genotypes showed 55 bp deletion in UGPase promoter. Identification of trans-acting factors and cis-elements

involved in cold-induction of CIS genes will further illuminate the mechanisms of CIS tolerance.


325

IPPCongress2015


PI415 (IPPC0865)

Next generation transcriptome sequencing to elucidate molecular mechanism of Self-incompatibility

in tea

Romit Seth*, Aparna Shree Raina, Abhishek Bhandawat, Rajni Parmar, Gagandeep Singh and Ram Kumar Sharma

CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, (H.P.), India


Self-incompatibility is a genetically controlled mechanism existing in higher plants, especially in angiosperms and plays

critical role to inhibit inbreeding depression, henceforth, enhancing heterozygosity, resulting in diversification and speciation

of plant species. Tea (Camellia sinensis (L) O. Kuntze), being an obligate outcross is highly heterogeneous thus hindering

the maintenance and propagation of quality lines for commercial cultivation. To unravel the complexity of this trait, whole

transcriptome sequencing of self-incompatible and cross-compatible samples were performed using Illumina platform. In

total, 98.5 million raw reads were generated followed by de novo assembly of 73.3 million clean reads resulted 26Mb non-

redundant data (48,893 contigs). Functional annotation identified 32,563 unique contigs using sequence search against

KEGG, GO, SWISSPROT, TAIR, KOG, Transcription Factor Database. This study revealed that contigs encoding S-

Locus and apoptosis related genes were found to be differentially expressed, which might be playing role in regulating the

pathways of self-incompatibility. This trait specific study will help in predicting the possible molecular pathways responsible

in providing the self-incompatibility in tea. It may be used for future genomic studies, which in the downstream, will help

in maintenance and propagation of quality lines in tea.

PI416 (IPPC0867)

Global transcriptome analysis for dissecting drought tolerance in tea

Rajni Parmar*, Abhishek Bhandawat, Gagandeep Singh, Kuldip Jayaswall and Ram Sharma

Biotechnology Division, CSIR, Institute of Himalayan Bioresource Technology, Palampur-176061, Himachal Pradesh, India

*Presenting author: [email protected], [email protected]

Abiotic stresses are environmental conditions that reduce growth and yield of crop below optimum level and especially,

drought stress is primary cause of crop loss worldwide. Tea [Camellia sinensis (L.) O. Kuntze] is perennial evergreen

plant of genus Camellia belongs to Theaceae family. Drought stress is one of severe abiotic stress that limits growth, yield

and geographical distribution of tea. Field evaluation studies over the years identified potential source of drought tolerance

in tea. Interestingly, these drought tolerant genotypes capture moderate to high level of diversity. Although, few molecular

studies on drought have been conducted in the recent past but genome wide transcriptomic studies have not been

undertaken so far. Moreover, multiple genotypes having recorded different response to drought (tolerant/susceptible)

have not been studied in tea. During this study, sequencing of five diverse genotypes having background knowledge of

drought response have been done with induced drought at the transcriptome level. A total of 116,666,747 filtered read

were generated. After de novo assembly 36,216 contigs were obtained with an average length of 847 bp. Analysis of

these contigs using non-redundant protein database annotated 28,951 unigenes showed number of genes have significant

role in drought stress. Additionally 2,42,846 high quality SNPs were ascertained from this dataset. Present study will help

to elucidate the drought responsive key molecular players and also in gaining deeper understanding of behavior of

diverse genotypes of tea under drought stress.


326

IPPCongress2015


PI417 (IPPC0898)

Transcriptome sequencing, genome-wide miRNA prediction and their target identification from horsegram

accessions with contrasting drought tolerance

Jeshima Khan Yasin1*, Vivek Thiruvettai, M. Arumugam Pillai2, K.V. Bhat1, Viswanathan Chinnusamy3 and K.C. Bansal1

1National Bureau of Plant Genetic Resources, New Delhi-110012, India2Department of Plant Breeding, Killikulam, Tamil Nadu Agricultural University, Tamil Nadu, India3Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India


Horsegram (Macrotyloma uniflorum) is an unexploited legume with high adaptability to abiotic stresses. A total of 3300accessions of horsegram from national gene bank were phenotyped and 50 accessions with contrasting drought tolerancewere identified. Two accessions with contrasting reactive oxygen scavenging potential were selected and subjected todrought stress. Transcriptome data were generated for control and stressed plants. De-novo transcriptome annotationidentified genes for functional categories such as stress–related (2.87%), signalling (4.31%), hormone metabolism (2.1%)protein metabolism (13.64%) and RNA metabolism (8.33%). Confirming our earlier finding that horsegram employs energyconservation mechanisms, which were explained by transcript abundance for glycolysis (0.47%), gluconeogenesis/glyoxylate cycle (0.06%), TCA (0.38%), and mitochondrial electron transport/ATP synthesis (0.55%). Annotation ofhorsegram transcriptome was helpful in determining the interplay of multiple metabolic activities involved in stress responsesas compared with soybean. Non-coding and weak-coding sequences were analysed for miRNAs based on their secondarystructure and drought stress responsive miRNAs were identified. The targets for the miRNAs were predicted. miRNAsfrom other seven related legumes were collected from miRBase to find their targets in horsegram transcriptome. Predictedstress responsive miRNAs were confirmed to regulate drought tolerance and validated in other legume crops. The novelmiRNAs predicted in this study needs further validation. Reference based genome mapping was performed using Glycinemax as reference genome to identify likely QTLs, candidate gene based markers, exon junctions, etc. The genomicresources generated in this study will be highly useful in genetic improvement of horsegram and related pulses.

PI418 (IPPC0917)

Prospecting candidate genes associated with abiotic stress tolerance from proteins of unknown function

(PUFs) identified from Indian mulberry (Morus sp.)

K.H. Dhanyalakshmi*, R.S. Sajeevan and Karaba N. Nataraja

Department of Crop Physiology, University of Agricultural Science, GKVK, Bangalore-560065, Karnataka, India


Abiotic and biotic stresses have become serious challenges to crop production. Plant growth, development and responsesunder these conditions are tightly regulated by the coordinated expression of diverse molecular, biochemical, cellular andphysiological events that help them to grow and survive. At molecular level, all stress adaptive processes are mainlymediated by the expression of several stress responsive genes, their products and pathways and the roles of some ofsuch genes have been well characterized. But, a vast majority of proteins in public databases still remain uncharacterizedor unknown or hypothetical, collectively called as proteins of unknown functions (PUFs) which might be novel and unique.Understanding their relevance might help to address complex mechanisms of stress adaptation in a better way. In thiscontext, an attempt was made to validate the role of selected PUFs identified from drought transcriptome (SRA accessionSRP047446) of Indian mulberry. The PUFs were uncharacterized according to Uniprot blast analysis. We used diversesequence as well as structure level computational analysis tools to assign probable function to over 100 selected PUFs.The response of a few selected PUFs to different abiotic stresses was examined under laboratory conditions in mulberry.The selected PUFs were up-regulated under these stresses and further validation might uncover novel pathways associatedwith stress tolerance in mulberry.


327

IPPCongress2015


PI419 (IPPC0920)

Expression analysis of genes involved in stem reserve accumulation and mobilization in rice under soil

moisture deficit

Puja Rai1*, A.S. Sakhare1, Chandarpal Vishwakarma1, P.C. Nautyal2 and Viswanathan Chinnusamy1

1Division of Plant Physiology, 2Division of Seed Science and Technology, Indian Agricultural Res. Institute, New Delhi-110012, India


Starch is the major storage carbohydrate in rice and is stored in the stem tissues when photosynthates production ishigher than sink demand. The stored non-structural carbohydrates (starch and sucrose) provide an alternative sourceduring later stage of grain filling or under abiotic stresses when current photosynthesis is reduced. Genes coding forstarch biosynthesis and degradation, sucrose synthesis and sucrose transport have been studied in mainly in cerealgrains. However, their regulation and involvement in stem reserve accumulation and mobilization under soil moisturedeficit stress is poorly understood. This study was aimed to understand the regulation of genes involved in stem reserveaccumulation and mobilization in rice and their regulation under soil moisture deficit stress. Real time RT-PCR expressionanalysis was performed to study the gene expression in different tissues namely leaf, leaf sheath and stem of rice cv.Pusa Sugandh 2 under control and drought stress conditions. The results identified specific members of AGPase, starchsynthase and sucrose transporters (SUT) gene families that predominately express in stem and their regulation underdrought stress. Expression of genes involved in stem reserve accumulation and mobilization under control and droughtconditions in two contrasting rice cultivars is being studied. Understanding of the regulation of genes involved in stemreserve mobilization under drought will open up new possibilities to select high yielding drought tolerant rice varieties infuture.

PI420 (IPPC0944)

Introgression of diverse physiological traits to improve drought adaption in rice by marker assisted

multi-parent backcross breeding strategy

M.D. Prathibha1*, M.V. Mohan Kumar1, N.M. Mallikarjun2, B.R. Raju1, D. Pushpa1, H.G. Jalendra Kumar1, K. Sumanth

Kumar1, J. Smitharani1, M.P. Rajanna3, M. Udayakumar1 and M.S. Sheshshayee1

1Department of Crop Physiology, 2Department of Genetics and Plant Breeding, University of Agricultural Science, GKVK, Bangalore-560065, Karnataka, India; 3ZARS, VC Farm, Mandya-571401


With the rapidly decreasing water resources, devising water saving agronomic practices for rice cultivation is the mostcrucial need of the hour. Global opinion strongly favours a trait based strategy over selection for absolute yield understress in crop improvement. Traits associated with maintenance of water relationships and superior metabolism underwater limited conditions deserve emphasis. Therefore water mining traits (root), efficient use of water and Cellular LevelTolerance (CLT) have greatest relevance. In an earlier investigation, a panel of germplasm was characterised for molecularand phenotypic diversity leading to the discovery of DNA markers by Association Mapping and trait donor genotypes (AC-39020 for roots and CLT, IET-16348 for WUE). We initiated a focused marker assisted backcross breeding to pyramidthese complex physiological traits into the elite genetic background of IR-64. Of the markers discovered earlier by associationmapping, 20 polymorphic markers were used for foreground selection and a genome wide set of 120 polymorphic markerswere used to reconstitute the recurrent parent genome. Of the 10000 DCBC3F2 lines generated, a set of 1500 lines werescreened under aerobic field conditions and the best lines (250) were advanced to DCBC3F3. These lines were extensivelyphenotyped for root traits and Δ13C to verify trait introgression. This study significantly contributed for the development oftrait introgressed lines that can be used for commercial cultivation and QTL-NILS that have great potential in academicunderstanding of QTL interactions.


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IPPCongress2015


PI421 (IPPC0945)

Study of molecular basis of heterosis revealed dominant and additive gene expression patterns in

Capsicum hybrids

Sushil Chhapekar*, Rashmi Gaur, Mukesh Nitin, M. Sarpras, Vineet Sharma and Nirala Ramchiary



Capsicum species commonly known as chilli peppers are economically important vegetable and spice crop of the world.

Chilli fruits are rich source of vitamins, antioxidants and nutrients along with anti-cancer and several medicinal properties.

India is the world’s largest producer, consumer and exporter of chilli peppers; however, the yield potential is very low due

to poor yielding varieties and high incidence of pests and diseases. One of the methods to achieve quantum jump in yield

and quality is through heterosis breeding. Although heterosis has been widely exploited in other crops, the study of

molecular and genetic mechanisms underlying the superiority of F1s over parental lines are poorly understood in Capsicum

plants. An attempt has been made to dissect the molecular basis of intra (Capsicum chinense X C. chinense) and inter-

specific heterotic F1 hybrids (C. chinense X C. frutescens and C. annuum X C. chinense) in capsicum species. The

interspecifc F1 hybrids were observed more heterotic compared to intra-specific hybrids in overall plant phenotype except

for fruit traits including plant growth (height and area), vigor, chlorophyll content, and other metabolites. Transcriptome

sequencing of early heterotic F1 plants showed more dominant and additive gene expression differences compared to

parental lines which might be the possible reason(s) for superiority of performance of heterotic F1 hybrids. Detailed

functional analysis of genes showed that most of the differentially expressed genes belong to photosynthetic pathways,

carbohydrate metabolism, hormonal signalling and code for transcription factors. Some of the differentially expressed

genes are being validated which might be helpful for designing effective breeding strategies to enhance capsicum

productivity.

PI422 (IPPC0951)

Discovery of QTLs and validation of markers for drought adaptive traits through characterization of trait

specific mapping population in rice (Oryza sativa L.)

Pooja Bharti1*, B.R. Raju1, R. Ramachandra, M.P. Rajanna2 and M.S. Sheshshayee

1Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore-560065, Karnataka, India2AICRIP (Rice), ZARS, V.C. Farm, Mandya-571405, Karnataka, India


Introgression of physiological traits like water mining and WUE onto a single elite background is one of the most appropriate

strategies to improve productivity under water limited conditions. Accelerating such trait introgression can only be achieved

by the adoption of focused molecular breeding practices. Identification of DNA markers associated and/or linked with

such complex physiological traits is therefore the primary pre-requisite. Towards identifying markers and QTL, we adopted

association mapping as well as bi-parental mapping strategies. A set of 230 F8 RIL population developed by crossing IET

15963 (high root) and Thanu (high WUE) was extensively phenotyped for root, WUE traits and spikelet fertility under

aerobic conditions. A linkage map consisting of 120 linked markers was constructed and robust QTL regions governing

root, WUE, spikelet fertility and other drought adaptive traits. The bi-parental mapping led to the validation of markers for

WUE and root traits previously discovered by association mapping. Several trait introgressed transgressive segregants

were identified and were characterized for field level performance under aerobic conditions.


329

IPPCongress2015


PI423 (IPPC0953)

Strategy for Meta QTL analysis by phenotyping diverse trait specific mapping populations for drought

adaptive traits in rice (Oryza sativa L.)

J.A. Smitharani1*, M.S. Sheshshayee1, B.R. Raju1, M. Raveendran2, S. Robin2, N. Sarala3, M.D. Prathiba1 and

M. Udayakumar1

1Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore-560065, India; 2Tamil Nadu AgriculturalUniversity, Coimbatore-641003, India; 3Indian Institute of Rice Research, Hyderabad, India*Presenting author: [email protected]

Emperical selection for drought adaptive traits is extremely complex which slows down crop improvement. This led to aspurt of research attempting to discover QTL. Though marker assisted breeding has the potential in accelerating cropimprovement, the diverse traits used by investigators and the diverse genetic background of populations used hassignificantly complicated the process of breeding. In recent years increased efforts are being made to validate QTL byidentifying consensus regions by “Meta QTL” analysis. In these analyses published data on similar traits across variousgenetic backgrounds are considered to identify common genomic regions, governing variability in a specific trait. In thisstudy, we report a strategy to identify consensus genomic regions conferring variability in physiological traits associatedwith drought adaptation in rice. The parental genotypes that have been used in developing mapping populations wereinitially phenotyped to assess variability in drought adaptive traits like roots and WUE. Specific mapping populations fromcontrasting parents were identified. RILs developed from IR 64 x Norungan, IR 64 x Kallurandaikar, Swarna x Madhukarand Thanu x IET 15963 were extensively phenotyped for drought adaptive traits. Root structure experiments were conductedto assess variability in root traits and the Delta 13 C was used to measure WUE. More than 1500 QTL reported from ourCentre and from elsewhere were used to initially determine the parental polymorphism. Selective genotyping of thecontrasting groups of RILs differing in specific traits would lead to a more precise Meta QTL analysis and validation ofmarkers for marker assisted breeding.

PI424 (IPPC0985)

Physiological characterization of an upland rice Nagina-22 derived EMS mutants–contrasting for leaf size

H.R. Sowmya1,2*, K. Sumanth Kumar1, Rajesh Kumar Singal1, P. Niranjana2, M.P. Rajanna3, R.P. Sharma4, T. Mohapatra5,

N. Sarala6, S. Robin7, A.K. Singh8 and M.S. Sheshshayee1

1Department of Crop Physiology, University of Agricultural Sciences, GKVK, UAS Bengaluru-560065, Karnataka, India; 2Departmentof Biochemistry, Kuvempu University, Shivgoga; 3Zonal Agricultural Research Station, College of Agriculture, Mandya; 4NationalResearch Centre on Plant Biotechnology, IARI, New Delhi-110012, India; 5Central Rice Research Institute, Cuttack-753006, India;6Directorate of Rice Research, Rajendranagar-500030, Hyderabad, India; 7Department of Rice, Tamil Nadu Agricultural ResearchInstitute, Coimbatore, India; 8National Bureau of Plant Genetic Resources, New Delhi-110012, India


A greater understanding of the molecular basis of regulation of canopy architecture has great relevance in determininggrowth rates. In this investigation, we report the identification of EMS induced mutants of rice in the background ofNagina-22 with significant differences in leaf size and their physiological characterization. The narrow leaf mutant(N22_BADT_380_4) showed lower stomatal conductance (g

s) than broad leaf mutant (N22_BADT_437_1) while

photosynthetic rate was comparable. This influence was verified by comparing whole plant cumulative water transpired(CWT). The narrow leaf mutant recorded lower CWT than broad leaf mutant and Nagina 22 (wild type). On the other hand,the narrow leaf mutant would maintain relatively higher g

s and A under stress, while a significant reduction was noticed in

broad leaf mutant and the wild type when the plants experienced moisture stress. The leaf thickness as indicated by SLWwas significantly higher in narrow leaf mutant which also recorded higher epicuticular wax load (EWL). With an increasedmesophyll cell number combined with reduced heat load because of EWL, the narrow leaf mutant was capable of maintaininghigher photosynthetic rate per unit area. However, because of a significant reduction in tiller number, the total canopy leafarea was less and hence TDM of narrow leaf mutant was also less. These leaf size mutants represent an excellentresource for generating greater clarity on the factors contributing to canopy architecture and canopy carbon exchangerate in rice.


330

IPPCongress2015


PI425 (IPPC0996)

RNASeq study to understand molecular basis of rice- Rhizoctonia interaction

Archana Kumari1*, Priyanka Jain2, T.R. Sharma2

1Division of Plant Physiology, IARI, New Delhi-110012, India2National Research Centre on Plant Biotechnology, Pusa Campus, New Delhi- 110012, India


Sheath blight, caused by Rhizoctonia solani, is one of the most serious diseases of rice next to rice blast. For understanding

plant pathogen interaction, advanced genetic and statistical methods of analysis have been used in the present study to

identify new resistance genes through transcriptional profiling. RNA Seq analysis of resistant and susceptible rice lines

after infection with R. solani was performed at 0, 12, & 24 hours post infection. RNA Seq data revealed many differentially

expressed genes at 12 hrs and 24 hrs post infection. We obtained 84 (78 upregulated & 6 downregulated), 298 (200

upregulated & 98 downregulated) and 196 (175 upregulated & 21 downregulated) significant (FDR adjusted p value

<0.05) differentially expressed locus (SDEL) in resistant rice line at 0, 12 & 24 hpi respectively. In comparison we could

get 342 (335 upregulated & 7 downregulated), 231 (146 upregulated & 185 downregulted) and 873 (632 upregulated &

241 downregulated) SDEL in susceptible rice lines at 0, 12 & 24 hpi, respectively. Gene ontology (GO) annotation and

pathway analysis of SDEL showed that genes involved in signalling, transcription factors, response to stress and defense

related genes are enhanced in resistant line compared to susceptible one at 12 & 24 hpi. We report most significant

changes in gene expression in rice sheath for the first time and reveal the genes associated with various pathways and

processes in plants during infection. Present study of rice- Rhizoctonia interactions have been undertaken to uncover the

mechanisms by which disease resistance is achieved.

Session J : Flowering and Post Harvest Physiology

331

IPPCongress2015


PJ426 (IPPC0239)

Deciphering the biochemical basis of flower colour variation in palas (Butea monosperma Lam) taubert

Vaibhav D. Lohot1*, A. Roy Chowdhury2, Nandkishore Thombare2, Jyotirmoy Ghosh1, Thamilarasi K1, A Mohanasundaram1,

Gunjan1 and Kewal K. Sharma1

1Lac Production Division, IINRG, Ranchi-834010, Jharkhand, India2Process and Product Development Division, IINRG, Ranchi-834010, Jharkhand, India*Presenting author: [email protected]

Angiosperms exhibits a tremendous diversity of flower colours, with sister species often differing in the intensity, hue orpatterning of the corolla. This flower colour trait is of paramount importance in the ecology and evolution of plants due toits ability to attract pollinators. The colouration of flowers is due to accumulation of flavonoids (including anthocyanins).Palas (B. monosperma) belongs to the family Papilionaceae, enduring beautiful scarlet (orange-red) coloured flowersduring early summer. In Jharkhand Palas is extensively utilized for lac cultivation and is the single largest producer ofrangeeni strain of Indian lac insect (Kerria lacca Kerr). Occasionally in Palas different shades of flower colours rangingfrom brick-red to pure white with several shades of intermediate orange-yellow and yellow colours are seen in wild. Anattempt was made to decipher the basis of colour variation in Palas flowers by quantitative estimation of flavonoids andanthocyanins. Variants were classified into three categories based on colour similarity viz. orange-yellow, yellow andscarlet. The study revealed that negative values were obtained for anthocyanin production in yellow coloured flowersregardless of flavonoid production which indicates that plant is unable to convert flavonoids to anthocyanins. In orange-yellow variants, the conversion of flavonoids to anthocyanins was ranged from 35-60 percent. Though the scarlet producesmaximum flavonoid content among all the variants, conversion percentage recorded very low. Thus the study revealedthat intensity of the flower colour variation in Palas could be due to interplay of flavonoid content and its extent of conversionto anthocyanins.

PJ427 (IPPC0254)

Functional characterization of flowering inducer FT-like gene family in banana (Musa acuminata L.)

Akhilesh Kumar Chaurasia1*, Hemant Patil1, Bal Krishna1, V.R. Subramaniam1, A. Sane2 and P.V. Sane1

1Plant Molecular Biology Lab, Agri Park, Jain Hills, Shirsoli Road, Jalgaon–425001, India2Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India


The phase transition from vegetative to reproductive stage is an important developmental step in the life of floweringplants. It is controlled by exogenous factors such as photoperiod, temperature and abiotic stresses and endogenousfactors such as hormone levels and C/N ratio. The key regulator of flowering is FLOWERING LOCUS T (FT), a memberof phosphatidylethanolamine-binding protein (PEBP) super family. The FT protein, believed to be the elusive ‘florigen’, isa mobile protein which moves from the leaf to the shoot apex through phloem where it interacts with another protein FDto activate downstream targets LEAFY and APETALA1 which are responsible for converting the apical meristem to aninflorescence meristem. Phase transition/flower induction in ever green herbaceous perennials like banana is not known.In this study, we have identified and characterized 12 genes, designated as MaFT1-12, from the FT-like group of thePEBP family in banana (Musa acuminata L. AAA group cultivar Grand Nain). Time-course dependent expression duringthe ten months of plant growth suggests an increase in expression of MaFT2-5 and MaFT12 just prior to initiation offlowering. Functional analysis in Arabidopsis through over expression shows early flowering by MaFT1-5, MaFT8 andMaFT12 compared to wild type. These genes are also able to complement the late flowering ft-10 mutant with MaFT2 andMaFT5 exhibiting a stronger phenotype. Our study, the first of its kind in banana, shows that MaFT2 and MaFT5 (alongwith other genes) are key flowering inducers in banana.


332

IPPCongress2015


PJ428 (IPPC0278)

Studies on modified atmosphere storage of spikes Gladiolus spp.

Harsimran Kaur*, Kushal Singh and Seema Bedi

Department of Botany, Punjab Agricultural University Ludhiana-141013, Punjab, India


The present study was undertaken to find out the storage response of spikes of different cultivars of gladiolus undermodified atmosphere storage. The spikes of four cultivars of gladiolus viz. Jacksonville Gold, White Prosperity, Sancerreand Nova Lux were harvested when coloured tepals were visible in 6 florets per spike. The spikes were grouped intobundles of three each and packed in polypropylene (PP) and low density polyethylene (LDPE) film sleeves (25 μ thick).The sleeves were hermetically sealed at both ends. The packages were subjected to modified atmosphere (MA) storagein a cool chamber (4±0.50C; 90-95 % R.H.) for 3, 6, 9, 12, 15 and 18 days, respectively, thereafter vase life was studiedat room temperature. Vase life and percent opening of florets showed decline with the progress of storage duration in allthe four cultivars. Cultivar Jacksonville Gold took minimum number of days (2.40) for opening of the basal floret while thecv. White Prosperity took maximum number of days (3.14) for opening of the basal floret. Vase life and percent openingof florets were also maximum in cv. Jacksonville Gold. Spikes stored in PP packages exhibited slightly higher vase lifeand floret opening than those stored in LDPE packages. The spikes exhibited considerable increase in peroxidase andcatalase activities in the tepals during storage, the increase being more prominent in cv. Jacksonville Gold. The increasein activity of these enzymes indicates the activation of antioxidant enzyme system to alleviate oxidative stress during therefrigerated storage.

PJ429 (IPPC0377)

Regulatory evolution in homeologs of suppressor of overexpression of constans1 and its role in floral

transition

Tanu Sri*, Pratiksha Mayee, Shikha Tyagi, S.M. Shivaraj, Anupama Atri and Anandita Singh

Department of Biotechnology, TERI University, New Delhi-110070, India


Successful floral transition of an angiosperm is achieved by a complex interplay of signalling cascades involving multipleflowering time genes. This study is focussed on a floral pathway integrator SOC1, a MADS-box transcription factor. Wehave examined its evolution and characterised its role in flowering in Indian Brassicas. Microsyntenic status of Brassicarapa regions containing three homeologs of SOC1 conformed to differentially fractionated homeologous sub-genomes.Screening 18 cultivars of 6 Brassica species led to identification of 8 genomic and 27 transcript variants of SOC1, includingsplice-forms and intron-less SOC1 genes. Phylogenetic analyses revealed the diploid base genome and sub-genome oforigin of each Brassica SOC1 homologs, indicating sub-genomic organization in un-characterized Brassica genomes.Expression analyses of two B. juncea SOC1 homeologs (MF2 and LF) revealed near identical expression pattern. However,MF2-specific homeolog exhibited significantly higher expression implying regulatory diversification. To further investigate,promoters of Brassica SOC1 homologs were isolated and analysed for divergence in cis-regulatory motifs. Cumulativeexpression analysis of B. juncea SOC1 revealed increased levels of SOC1 transcript in various organs during flowering,highest in apical bud, establishing its role in floral transition. Furthermore, B. juncea SOC1 was functionally characterizedby gain-of-function Arabidopsis mutants showing earliness in flowering. Knock-down lines of A. thaliana and Brassicajuncea expressing artificial miRNA against SOC1 displayed delayed flowering along with concomitant presence of matureartificial miRNA and cleaved SOC1 transcripts. Overall, this study highlights the importance of SOC1 in floral transition inBrassica and provides insights into evolution of homeologs of developmental genes.


333

IPPCongress2015


PJ430 (IPPC0676)

The bZIP transcription factor, OsbZIP, in rice is a functional ortholog of Flowering Locus D (FD) in

Arabidopsis

Amarjot Kaur*, Aashima Nijhawan, Mahesh Yadav and Jitendra Khurana

Dept. of Plant Molecular Biology & Interdisciplinary Centre for Plant Genomics, Univ. of Delhi South Campus, New Delhi-110021, India


Flowering is an intricate pathway in plant life cycle and a plethora of genes are involved in bringing about transition from

vegetative to reproductive stage. This pathway has been well characterised in Arabidopsis wherein Flowering Locus D

(FD) interacts with Flowering Locus T (FT) and activate the transcription of floral meristem identity genes. However, this

pathway is less understood in rice and a number of other factors are involved in flowering. Our lab identified a few bZIPs

that may be involved in regulating flowering in rice employing whole genome transcriptome analysis. Among these, one

of the OsbZIPs showed some promising results as it was able to complement fd mutant of Arabidopsis. In contrast to

earlier reports that FD and FT interact directly, it has now been shown that their interaction is mediated by an intracellular

receptor GF14-3-3 protein, which serves as a chaperone. Thus in our study too, yeast one-to-one interaction of OsbZIP

(identified in this study) with OsFTLs and GF14-3-3 was performed and a few of them did show interaction. Also, yeast-

three-hybrid assay involving 14-3-3 proteins showed similar results thereby suggesting the possible involvement of GF14-

3-3 protein in flowering in rice. Rice RNAi transgenics for OsbZIP showed a slight delay in flowering time in comparison

to WT. These results thus provide evidence that OsbZIP might be acting as a putative ortholog of Arabidopsis FD and

may be involved in regulating transition to flowering.

PJ431 (IPPC0858)

Soil moisture deficit stress effect on flowering and fruit development in banana cv. Grand Naine

Iyyakutty Ravi* and M. Mayil Vaganan

Crop Production Section, ICAR-National Research Centre for Banana, Trichy-620102, Tamil Nadu, India


India is the largest producer of bananas. The banana crop cycle extends a minimum of one year and it requires a

continuous source of soil moisture for optimum growth. Due to long crop cycle, the banana may face soil moisture deficits

any time as a consequence of the seasonal decline in soil water availability. A field study was conducted to identify the

effects of soil moisture deficit stress in banana cv. Grand Naine at NRCB farm. The stress was imposed by withholding

irrigation at 3rd, 5th month after planting and at flowering to reach the soil moisture stress level of -0.6 to -0.7 MPa,

thereafter stress was relieved by irrigation till harvest. At harvest, in irrigated control, the average bunch weight was 21.5

kg. The stress imposed at 3rd month after planting did not affect the bunch weight. However, stress imposed at 5th month

after planting had a severe impact on bunch morphology, size and weight. Around 35-40% of bunches expressed choking

and were malformed, and mostly weighed around 9.1 kg, with small fingers. Stress imposed at flowering resulted in poor

development of fingers and became unmarketable bunches. The results revealed that irrigation at 5-6th month and at

flowering was critical for normal bunch and fruit development in banana cv. Grand Naine. Foliar priming of the banana

plant with acetylsalicylic acid (0.1 mM) and butylated hydroxy toluene (100 ppm) alleviated the negative effect of soil

moisture deficit stress.


334

IPPCongress2015


PJ432 (IPPC0982)

Identification of less photo-sensitive activation tagged mutants of rice and their flanking sequence analysis

to identify the possible genes

Anita Pitagi1*, N.B. Shashibhushan1, Appanna V. Koppad1, Rohini Sreevathsa2, Mahantesha Naika1, B.M. Raju1 and

M. Udayakumar

1Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bengaluru-560065, Karnataka, India2National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India


Many rice genotypes with improved adaptation to abiotic stresses and more specifically, for aerobic cultivation are

photosensitive. JBT36/14, an upland rice genotype adapted to aerobic cultivation has superior water relations (high yield,

high roots and better WUE). However, this genotype is highly photosensitive. With an objective of identifying mutants with

less photosensitivity and genes associated with it, a gain-in-function activation tagged mutants were developed in the

background of JBT36/14 using activation tagging vector, pMN20 having 4x35s enhancers and glyphosate selection

marker, mEPSPS. The putative mutants were identified based on the response to glyphosate and the positive mutants

were screened for several phenotypic characteristics including flowering duration, tillering, seed color, biomass, plant

architecture, yield and 234 activation tagged mutants were selected based on the distinct variations in phenotypic characters.

Six mutants that showed distinct variation in photoperiodic response showed almost similar flowering time both under

optimal (kharif) and non-optimal (long day) photoperiodic conditions with wild type remained as non-flowered genotype

under non-optimal conditions. Further, the flanking sequence analysis of one of the mutants showed integration of the

enhancer elements on chromosome 11 flanked by several genes such as Os11g0219000, Os11g0219400, Os11g0218000,

Os11g0218100, and Os11g0218500. The bioinformatic analysis of these genes showed that, one of the genes is a bZIP

transcription factor which seems to have some role in regulating photoperiodic response. The expression analysis of the

flanking genes and their functional validation is under progress.

Session K : Computational and Systems Biology

335

IPPCongress2015


PK433 (IPPC0110)

Characterization of glutaredoxin proteins in monocots

Minesh Jethva1*, Saurabh Yadav2 and Hemant R. Kushwaha1

1Synthetic Biology & Biofuels Group, International Centre for Genetic Engineering & Biotech., Aruna Asaf Ali Marg,New Delhi-110067, India2Department of Biotechnology, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India


Glutaredoxins (GRXs) are small heat-stable disulfide oxido-reductases and members of the thioredoxin protein family

and are involved in maintaining the redox homeostasis in many organisms like bacteria, yeast, and mammalian cells.

GRXs exist in entire green lineage and they are classified into dithiol and monothiol by used on the type of active site

present. Monothiol and dithiol GRXs contain ‘CGFS’ and ‘CXXC’ as a redox active motif, respectively. The availability of

completely sequenced plant genomes allows the comparative study of GRXs distribution across species. In the present

analysis, we have studied 19 monocot plant genomes to reveal sequence and domain level diversity within GRX proteins.

We have characterized GRX family including chromosome locations of coding genes, protein domain structures,

phylogenetic relation and their sub-cellular localization. We have analyzed the phylogenetic distribution of GRXs proteins

to reveal evolutionary events like HGT and duplication. This study provides perception of the domain structure conservation

of GRXs in monocots. The number of GRXs varies from 39 to 96 in these species but domain structures are relatively

conserved.

PK434 (IPPC0199)

Hyperspectral reflectance based non-invasive assessment of tomato fruit firmness

Vijay Paul1*, Rajeev Kumar1, Rakesh Pandey1, Rabi Narayan Sahoo2, S.K. Jha3, Ram Asrey3, V.K. Gupta2, Atar Singh1

and Tauqueer Ahmad4

1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Food Science & PHT, Indian Agricultural Research Institute,New Delhi-110012, India4Division of Sample Survey, IASR, New Delhi-110012, India


Firmness of tomato fruit is a key parameter to ascertain its quality, transportability, marketing, storage period/shelf-life

and preference by consumers. To identify hyperspectral reflectance based indices and to develop models for non-invasive

estimation of tomato fruit firmness, freshly harvested and stored tomato fruits of 2 varieties and 2 hybrids representing

eight ripening stages were sampled. Reflectance of fruits was taken with spectroradiometer. Reflectance at wavelengths

ranging from 350 to 2500 nm was recorded. Fruit firmness (Newton, N) was then determined by texture analyser. Data

obtained is analysed for dimension reduction, index identification and model development. Firmness ranged from 11.74

N (immature fruits) to 1.13 N (over ripe fruits). Out of many indices, two best indices were: R501

(1st derivative) and R521

.

Model based on R501

(1st derivative) (= x) is y (firmness, N) = 1260.8 x + 3.3087 with RMSEP = 11.05 N and bias = -0.01.

The second model based on R521 (= x) is y = - 5.3768 x2 + 18.209x + 2.9509 having RMSEP = 1.11 N with bias = -0.002.

Models were found to be valid irrespective of varieties/hybrids, ripening stages, fresh and stored fruits. Model based on

the index R521

is simple (no derivation step) and therefore can also be considered as rapid one. This study will prove

useful in developing cost-effective tools. Such tools can then be applied for rapid/automated phenotyping, screening and

sorting of tomato fruits based on the firmness at small (lab) or large (industries) scales.


336

IPPCongress2015


PK435 (IPPC0226)

Gravitational biology: significance of the characteristics of statenchyma in dicotyledonous plants

A.D. Barnabas1* and P. Dayanandan2

1Post Graduate Department and Research Centre of Botany, The American College, Madurai-625002, Tamil Nadu, India2Rtd. from the Department of Botany, Madras Christian College, Tambaram, Chennai-600059, India


A specialized group of cells, statocytes, containing starch statoliths (= amyloplasts) appear to be involved in the initial

stages of plant gravitropism. The direction of transport of auxin, lateral or radial, can be resolved if we understand the

significance of the relative position of the statenchyma (RPS) cells. 141 species of dicotyledonous plants were investigated

to characterise the number, dimensions, and relative position of the statoliths and statocytes. Seedlings and young

shoots were gravistimulated and the regions of maximal response were sectioned and observed after staining with I2KI.

All 141 species possessed statenchyma with sedimentable amyloplasts. The average number of statoliths per statocyte

is 10. The statenchyma may be one or more layered and continuous or interrupted. RPS is expressed as an r/R value

where r is the radius of the circle defined by statenchyma as the outer limit and R is the radius of the transverse section

of the shoot with the epidermis as the outer limit. Very thin shoots have small RPS values, whereas very thick ones have

a value of 0.5. In general, the RPS value varies from 0.18 to 0.93. In Peperomia which has scattered vascular bundles,

each bundle is surrounded by a layer of statenchyma. In Gomphrena globosa statoliths sediment normally only in young

shoots that respond to gravity. The significance of RPS will be discussed in relation to auxin transport and elongation

response. Basic research in gravitropism has implications of gravitational biology and space life sciences.

PK436 (IPPC0229)

Differential nucleosome occupancy in plant genomes; Are promoter regions significantly depleted of

nucleosomes?

Amish Kumar* and Gitanjali Yadav



Wrapping of DNA around the nucleosomes core particle is the key to the chromatin packaging as well as for the regulation

of gene expression. A comparative study of nucleosome occupancy patterns, at the genome wide level may help to

understand chromatin structure and its functional control over the cell. Whole genome nucleosome occupancy maps are

available for several organisms and provide sufficient knowledge base to accurately predict the site of occurrence of

nucleosomes over a new genome sequence. For example, yeast genome shows differential organization of nucleosomes

in the upstream regulatory region of genes, resulting in the existence of distinct nucleosome free regions (NFRs) just

before the transcription start site (TSS). This study was designed to test whether such NFRs are present in plant genomes

as well, and whether the DNA itself guides differential nucleosome occupancy. Thus in order to understand the role of

DNA sequence in nucleosome occupancy, we hypothesized that “There are fewer nucleosomes in promoters compared

to other regions of the plant genome”. For this, we analyzed two plant genomes namely, Zea mays (a monocot) and

Solanum tuberosum L. (a dicot) to construct genome wide nucleosome prediction maps. Per base nucleosome positioning

was compared over the promoter and non-promoter regions of both the genomes. Statistical tests reveal that nucleosomes

are indeed depleted in promoters as compared to random, or other genomic regions of plants.


337

IPPCongress2015


PK437 (IPPC0268)

Bioinformatics of KaiC, a cyanobacterial circadian clock protein

Rakesh Yadav* and Namita Singh

Department of Bio & Nano Technology, Guru Jambheshwar University of Science & Technology, Hisar-125001, Haryana, India


Circadian clocks are endogenous biological timers that rhythmically control several processes with an around 24-h period.

They exist in diverse organisms, but cyanobacteria are the simplest organisms known to possess a clock. The circadian

clock protein KaiC, is encoded in the kaiABC operon that regulates circadian rhythms. KaiC have been extensively

studied and are widely distributed among Cyanobacteria. In this study, bioinformatics analyses of KaiC proteins were

performed in 53 Genera of Cyanobacteria. According to the conserved motifs obtained by MEME and MAST tools, two

motifs were common in all proteins. The structural and functional analyses of KaiC proteins were investigated by ProtParam,

SOPMA, SignalP 4.1, TMHMM 2.0 and ProDom tools in ExPASy database. The protein sequences were aligned with

ClustalW algorithm by MEGA 6.06 software and phylogenetic tree was constructed using the Neighbor-joining (NJ) method.

In protein–protein interactions analysis by STRING 9.1 tool, 10 significant protein interaction groups were identified in

KaiC. According to the results, there is a high identity among KaiCs of different species in Cyanobacteria, so that they

have to be derived from a common ancestor. The approaches and results reported here would be useful to understand

biochemical and structural dissection of the molecular basis of a biological clockwork.

PK438 (IPPC0329)

Mining Picrorhiza kurroa transcriptomes for transcription factors contributing to picrosides biosynthesis

Ira Vashisht*, Tarun Pal and Rajinder Singh Chauhan

Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, HP, India


Picrorhiza kurroa is a valuable medicinal herb, recognized for its hepatoprotective, anti-inflammatory, anti-oxidant,

immunomodulatory and several other curative properties. Iridoid glycosides such as picroside-I and picroside-II are major

ingredients of various pharmaceutical formulations. No information exists on transcriptional regulation of picrosides

biosynthesis in P. kurroa. Deciphering complete biosynthetic pathway coupled with expression analysis of genes has

revealed that multiple genes are contributing to the biosynthesis of picrosides, thereby, suggesting that common regulatory

control affect the biosynthesis of picrosides. Field-grown plants showed significantly higher picroside-I (shoots) and

picroside-II (roots) contents compared to in vitro -grown plants. The NGS transcriptomes of P. kurroa tissues with varying

amounts of picrosides, were mined for WRKY, MYB, bhlh, bzip, zinc finger, NAC and PHD classes of transcription factors,

which have been implicated in regulating secondary metabolites biosynthesis in plants. Among all classes, WRKY exhibited

maximum fold variation in field grown shoots (42 folds) and roots (5 folds) as compared to their in vitro counterparts. AP2/

ERF and PHD family of TFs showed negligible expression under in vitro conditions, thereby suggesting their possible

involvement in the regulation of picrosides biosynthesis. Functional validation of TFs can provide suitable candidates for

enhancing picrosides contents in P. kurroa.


338

IPPCongress2015


PK439 (IPPC0351)

KIXBASE: A KIX domain database and webserver for prediction and analysis

Archana Yadav*, Jitendra K. Thakur and Gitanjali Yadav



In today’s era of ever-increasing biological data, it is essential to maintain databases that not only integrate and clusterthe data as a warehouse but as well help analyze it. Such resources are invaluable tools for researchers as they providea useful platform to retrieve biological information like sequences, structures, structure classes, pathways etc., thusaiding in the research to a great extent. KIXBASE is a global repository as well as a prediction tool for KIX domains. Thesedomains are present in co-activators and play a central role in the transcription process. KIXBASE has two main parts, aweb server and a database. To date, there is no other web resource that provides information on KIX domains. The KIXprediction program detects KIX domains in any organism on the basis of profile hidden markov models developed throughalignment of known KIX sequences along with additional filtering criteria for improving accuracy. KIXBASE incorporatesthe most widely used programs like PSIPRED-3.5 and CLUSTALW2 in the web server for further annotation and qualityassessment of the predictions. Users can upload batch entries for protein, genomic or EST sequences in FASTA formatand carry out the detection of potential KIX domains, generate the secondary structure for domain of interest and examineconservation with other KIX domains. The backend prediction algorithm is also used for development of the KIX database,which contains 1891 KIX proteins representing 427 organisms spanning metazoans, fungi and plants, comprising thelargest online collection of KIX sequences. The prediction algorithm is highly accurate, robust and very efficient in handlingmultiple input types for KIX prediction.

PK440 (IPPC0360)

Non-destructive assessment of tomato colour and ripeness by using reflectance at single wavelength

Rajeev Kumar1*, Vijay Paul1, Rakesh Pandey1, Rabi Narayan Sahoo2, Vinod Kumar Gupta2, Anil Dahuja3, Atar Singh1 and

Tauqueer Ahmad4

1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Biochemistry, IARI, New Delhi-110012, India4Division of Sample Survey, Indian Agricultural Statistics Research Institute, New Delhi-110012, India


Quality and consumers preference of tomato fruits are primarily determined by its colour and appearance. Rapid andnon-destructive methods of assessing tomato colour and ripeness are therefore of immense significance for basic researchand applied aspects (processing and value addition). To identify hyperspectral reflectance based indices and developingmodels for non-destructive estimation of colour and ripeness/maturity of tomato fruits, tomato fruits of different varieties/hybrids coming from direct harvest or storage were used. Reflectance of individual fruit was taken in a spectrum of 350 to2500 nm by using a hand-held spectroradiometer. These fruits (immature to over-ripe stages) were then ranked from 0 to8 based on standard colour chart. Obtained data were subjected to chemometric analysis. All the basic and statisticalsteps as required for model development were followed. Two models were developed. First model was based on theindex R521 (x) i.e., y (colour/ripeness) = -2.456 ln (x) -1.0933. This can accurately predict the colour/ripeness for adifference of > 0.86 (RMSEP) with bias of -0.09. The second model y = 2.582 ln (x) - 0.805 was based on the index R546(x). This model had RMSEP = 0.89 and bias of 0.12. Models could distinguish different ripening stages of tomato acrossvarieties/hybrids, plant harvested (fresh) and stored (aged) fruits. These findings will prove useful in developing cost-effective tools for rapid sorting and screening of huge collection of harvested tomato fruits as desired for monitoring andby the processing and pharmaceutical industries.


339

IPPCongress2015


PK441 (IPPC0400)

Evolutionary significance of the CBM49-modulated Type C GH9-cellulase activity in plants

Siddhartha Kundu* and Rita Sharma

School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India


Members of the GH9 (endo-1, 4-glucanase, EC 3.2.1.4) clan have been ascribed roles in the generation of biofuels, as

initiators and facilitators of plant development (elongation, abscission), and as mediators of abiotic stress response

pathways. Catalytically, these enzymes cleave the β (1-4) glycosidic linkage between pairs of β-D-glucopyranose residues

from the non-reducing ends of cellulose in regions best described as amorphous. The Type C subfamily possesses a

unique carbohydrate binding domain (CBM49), which is cleaved during processing of the primary transcript. Evidence

suggests that the delineation from the classical Type B domain is fuzzy, thereby, raising the possibility of a truncated

contribution to the active site of Type C enzymes. To comprehend the significance of the CBM49-modifying Type C GH9-

cellulase activity, an evolutionary profile spanning several millennia was constructed. Previous work has suggested that

the division into sub-families (Types –A, -B, -C) is entirely anagenetic (intra-lineage). Here, we generate HMM-based

datasets of putative sequences spread over several taxa. Our sequence-centric approach is comparatively coarse-grained,

and focuses on patterns of bases/ amino acids. The significance of these intra-sequence clusters is a relatively novel

annotation schema for GH9-orthologs in existing genomic data. These, when subjected to evolutionary analysis, could

offer insights into speciation-level events as well as posit functional roles for the phyletic subgrouping of A-, B- and C-

members. The upgradation of Type C endo-glucanases to include micro-crystalline cellulose as a substrate, could, possibly

simplify the search for the perfect biofuel.

PK442 (IPPC0471)

A smart soft tool to integrate climate model and grid based weather data

Gayathri Chamanthula*, B. Sailaja, S.R. Voleti and D. Subrahmanyam

ICAR-Directorate of Rice Research, Rajendranagar, Hyderabad-500030, AP, India


Rice crop is climatically the most adaptable cereal. Management of rice crop is difficult as it is grown over a large spatial

domain and wide range of landscape types. The projected changes in temperature will further complicate the management

of rice crop. Forecasting grid based future temperatures and identifying vulnerable temperature grids to raise rice yields

at regional level in varied rice growing environments. With this background, ClimGen climate model has been chosen to

integrate with the grid based temperature Map. Grid wise temperature data was collected from IMD for 15 years and total

346 grid points were there for temperature. A software program was developed to convert individual text files supplied by

IMD into one single database and computing day and night temperatures. These files were converted to geo database

files using Arcatalog and thessien polygons were generated for each point using spatial interpolation method of ArcGIS

package. An interface has been created to integrate this grid based weather map with climate model. P-mapper an open

source software is used for this program. ClimGen model was executed for each grid and generated long term series up

to 2050. Weekly maps were generated for mean temperature and difference in day and night temperature. Vulnerable

temperature zones were identified at the time of flowering (February and March) during rabi season for the years 2030

and 2050. This software is user friendly and further integration with any kind of maps (point/polygon wise) and models is

possible with this interface.


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IPPCongress2015


PK443 (IPPC0607)

Integrative co-expression network analysis to infer the immune response in rice during the invasion of

rice blast fungus

Preeti Arya* and Vishal Acharya

CSIR-Institute of Himalayan Bioresoruce Technology, Palampur-1760061 (H.P.), India


Rice is an important staple cereal crop grown world-wide and is among the highly consumed food commodity. Thevarious diseases in rice cost for an average of 37% reduction in the yield globally. Rice blast fungus (Magnaporthe) is themost common phyto-pathogenic fungi responsible for substantial loss in production. The mechanism by which pathogensinfect its host is the most important and still unresolved aspect for many host-pathogen systems. Various studies pertainto decipher the mechanism of host-pathogen interaction; however, in vivo/in vitro methods are laborious, time consumingand are not high-throughput. In silico approach may be the best possible alternative to mine the useful informationregarding pathogenesis using data generated through microarray and highly advanced RNA sequencing. In the presentstudy, the publicly available transcriptome data for rice and Magnaporthe was retrieved from NCBI GEO and used toidentify the expression profiles of differentially expressed gene (DEGs) sets. The co-expression network was constructedby means of the Pearson correlation coefficient (PCC) between expression profiles of DEG(s) using weighted correlationnetwork analysis (WGCNA) that were further employed for finding the highly correlated gene modules, the biologicaltargets of interest. Moreover, the protein-protein interaction and interlog mapping was used to validate interacted genepairs obtained from the co-expression networks. The resulted interactions will be further evaluated statistically usingcomputational strategies and also on basis of experimentally known pathogen-host protein interactions. This rice-Magnaporthe interactome hence will provide valuable insights to understand the mechanism of pathogenesis by targetinghubs responsible for the disease development.

PK444 (IPPC0677)

Tissue specificity gene expression database

Ambarish Kumar*, Vijya Brahma and Andrew Lymm

Jawaharlal Nehru University, New Delhi-110067, India

*Present author: [email protected]

The RNA-Seq is the next generation sequencing technology to get a snapshot of RNA present and quantify (transcriptomeexpression profile) from a genome at a given moment of time. The expression data from the whole transcriptome canprovide important information about mRNA levels revealing stage specific and localized gene expression. Although manytissue specific expression databases are available (TIGER, TiSGeD, Very Gene etc.) but till date no work has beenreported with respect to the plant transcriptome expression, ensembled, based on the tissues (leaves, flower, fruit, rootsetc.), stages of development and localization. The database specific to plant tissue specificity can shed light upon thedisease biology, applied medical research, pharmaceutical drug development , biomarker discovery in the field oftranslational medicine, proteome classification on the basis of tissue specificity, nutritional development as well as increasein the productivity of certain crops. These information from the known repository will be collected, classified and its liaisonwith the assembled and annotated transcriptome will be surmised. The curated dataset for clade of Viridiplantaeencompassing all the green plants was retrieved from UNIPROTKB (http://www.uniprot.org/). A total of 3353453 fastasequences and their corresponding tissue specificity, stages of development, localization and pathways data was usedfor creating the database. We propose to integrate onto Galaxy based platform, wherein the RNA-seq data will be assembled(de novo/reference), analyzed for differential expression, followed by the annotations using BLAST algorithm and thePTSDb will be linked to the annotations.


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IPPCongress2015


PK445 (IPPC0821)

Molecular interaction pattern of sugarcane streak mosaic virus encoded micro RNA, SCSMV_miR16 in

sugarcane - A In silico approach

Prabu Gajjeraman* and Yasir Umar Pattiwala

Plant Functional Genomics Unit, Department of Biotechnology, Karpagam Academy of Higher Education, Coimbatore-641021, India


Sugarcane Streak Mosaic Virus (SCSMV) is a distinct member of Poacevirus in Potyviridae family which has a single

stranded positive-sense RNA genome causing mosaic disease in sugarcane. Among the various regulatory mechanisms,

gene silencing was found to play a major regulatory role during disease progression. Micro RNAs (miRNAs) are recently

identified small RNA molecules (~24 nt bp in length) which are involved in regulation of host plant gene expression and

vice versa in association with argonaute (AGO) proteins via RNA silencing. To study the gene regulatory pattern, we

analyzed the most preferential SCSMV_ miR16 – target mRNA – argonate protein interaction at molecular level by in

silico approach. The hydrogen bonding and other interactions were analyzed and discussed. These finding considerably

broaden future investigation of the AGO-assisted miRNA based gene regulation during SCSMV pathogenesis in sugarcane.

PK446 (IPPC0873)

Morpho-physiological characterization of cashew accessions in west coast region of Karnataka

Babli Mog1*, P.L. Saroj1, T.G. Prasad2 and Vishwanathan Chinnusamy3

1Plant Physiology, Directorate of Cashew Research, Puttur-574202, Karnataka, India2University of Agricultural Sciences, Bangalore, Karnataka, India3Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India


Cashew (Anacardium occidentale L.) is an important nut crop of social and economic importance for India. The low

productivity of raw cashew nut is the major constraint. As detailed physiological characterization of cashew accessions is

the key step for yield improvement, this study was carried out to analyze genotypic variation in yield and yield components

of 13 cashew accessions in twelve year old plantation. Significant differences in leaf traits were observed among cashew

accessions. Maximum leaf ratio was observed in VRI-3 while Bhaskara showed maximum leaf length and width. Maximum

of 28 main veins were recorded in Ullal-3. Highest leaf area and specific leaf weight (SLW) were recorded in VTH-30/

4(112.67 cm2) and Vengurla-3 (17.43 mg/cm2), respectively. The number of leaves per lateral shoot was more in VTH-

174, Bhaskara, VRI-3 and VTH-30/4. Ullal-3 exhibited higher RWC and lower rate of excised leaf water loss (10.8%)

followed by Bhaskara. The contents of Chl a, Chl b, total chlorophyll and carotenoids were highest in Bhaskara. VTH-174

showed highest auxillary bud differentiation into reproductive structures followed by Bhaskara. Significant variation for

nut development observed among accessions. VTH-174 took least time for apple development (56 days) as well as final

harvest (60 days). VTH-30/4 recorded highest apple with nut weight, apple and nut fresh weight. Brix value of apple was

recorded maximum in VTH-174 indicating it’s sweetness with less tannin content. The relationship between component

physiological traits and yield will be presented.


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IPPCongress2015


PK447 (IPPC0900)

Elucidating the alkaline tolerance mechanism of Arthrospira platensis: An In-silico study

Monikam Jangir* and Jyoti kashyap

BITS Pilani, Hyderabad Campus, Hyderabad, India


Alkalophiles, a class of extremophiles are capable of sustaining a high alkaline environment (~pH 11) but their exact

mechanism is unknown. The alkalotolerant mechanism could be extrapolated to improve productivity of higher plants

facing stressful environment. Out of various suggested hypothesis, Na+/H+ antiporters mediated regulation is the widely

accepted one which corroborates their role in pH homeostasis. Sequence and structural analysis was performed to

understand precise role of antiporters. We analyze antiporter sequences of alkalophilic Arthrospira platensis to understand

the underlying mechanism while the antiporters of alkalotolerant Synechocystis spp are considered as a control. Difference

in copy number and the presence of various functional domain(s) indicates the differential regulation of pH by these

antiporters. Greater than 50% identity was obtained between 3 of the antiporters sequences of the two organisms. NhaP,

KefB were the domains present in both organisms along with a unique UspA domain (Universal stress Protein A) only in

Arthrospira platensis. Transmembrane regions were also deduced using online servers in which Arthrospira platensis

showed presence of 11-12 helices while Synechocystis spp showed 9-12 helices. This study pointed out possible role of

antiporters on alkali tolerance.

PK448 (IPPC0947)

The sorghum gene expression database

Muhammad Ali Imam1, Vinay Singh2*, Manoj K. Sharma2 and Rita Sharma1

1School of Computational & Integrative Sciences, 2School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India


Sorghum bicolor, recognized as an alternate feedstock for biofuel production in many parts of the world, is a multipurpose

crop used for food, feed, fodder as well as fibre. Due to relatively small genome, availability of whole genome sequence

and low level of gene duplication, it is also being used as a model for understanding the genetic mechanism underlying

C4 photosynthesis in grasses.Till date, both microarrays and RNA sequencing technologies have been used to elucidate

transcriptional dynamics during different developmental stages of sorghum as well as in response to stress. To facilitate

functional genomics studies and development of improved varieties, we are developing a freely available user-friendly

community resource using publicly-available sorghum expression data. Both microarrays and RNA sequencing-based

transcriptomic data has been downloaded from NCBI-Gene Expression Omnibus and normalized using a standard pipeline.

The data can be retrieved using microarray elements IDs or gene accession numbers. Our web interface will allow

visualization and retrieval of gene expression profiling data via heat map and bar graphs in selected experiments/samples.

Additional tools for coexpression, differential expression and pathway analysis with the genes of interest will also be

provided. Therefore, this database will serve as a useful reference for elucidating gene expression and functions of

sorghum genes.


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PK449 (IPPC0949)

An approach for high throughput phenotyping of tomato genotypes using Plant Phenomics Platform

R.H. Laxman*, P. Hemamalini, R.M. Bhatt and A.T. Sadashiva

ICAR-Indian Institute of Horticultural Research, Bengaluru, Karnataka


Physiologists constantly strive to phenotype plants for identification of desirable traits for their utility in crop improvement.

The plants’ performance is quantified through measurement of various parameters under challenging conditions both in

field and controlled conditions. Both, destructive and non-destructive approaches are adopted in this endeavour. Since,

the detailed traditional methods of plant phenotyping are time consuming, laborious and involve human errors; the focus

is being shifted towards non-invasive imaging and image analysis which are high throughput in nature. The high throughput

phenotyping approaches being non-destructive enable imaging of the same plant at different developmental phases.

Considering these inherent advantages, studies were conducted in the Plant Phenomics facility with the objective to

evaluate the tomato cultivars, hybrids and germplasm lines at various developmental stages. The container grown plants

were imaged and growth performance was quantified. The imaged plants were sampled for measurement of the actual

biomass. The study showed significant relationship between actual biomass and digital biomass and through imaging

technique it was possible to quantify biomass across the genotypes and also at different growth stages. The utility of Plant

phenomics facility for high throughput evaluation and phenotyping of tomato genotypes are discussed in this paper.

PK450 (IPPC0959)

Quantitative monitoring of sucrose dynamics for phenotyping of soil moisture-deficit stress tolerance in

rice using hyper-spectral reflectance data

R.N. Sahoo1, Bappa Das1, G. Krishna1, Puja Rai2, Rahul Tiwari2*, R. Verma2, C. Viswanathan2, S.K. Dash4, P. Swain4 and

P.C. Nautiyal3

1Division of Agricultural Physics, 2Division of Plant Physiology, 3Division of Seed Science and Technology, Indian Agricultural ResearchInstitute, New Delhi-110012, India4ICARI-Central Rice Research Institute, Cuttack, Odisha, India


The capacity of rice to store sucrose in the stem is regarded as a promising trait to buffer yield in environments with

limited water availability. Measurement of volume and rate of flow of sucrose to the panicle during grain filling under

water-deficit condition could be one of the reliable tools for measuring soil moisture-deficit stress tolerance than the

currently used tools. Keeping this in view, a study was conducted for non invasive estimation of sucrose in rice stem using

hyper-spectral reflectance data collected in the spectral range of 350 to 2500 nm for different rice genotypes grown under

different soil moisture deficit stress levels. Plant samples were collected synchronizing with spectral measurements for

estimation of sucrose. Spectral data analysis was done to identify suitable spectral indices and models for sucrose

estimation. Two spectral indices, viz., ratio of sucrose index (RSI) and Normalized Difference Sucrose Index (NDSI) in

NIR range were proposed with reliable accuracy of sucrose estimation. Different multivariate spectral models such as

Partial least squares regression (PLSR) and multiple regression model (MLR), Multivariate adaptive regression splines

(MARS), Support Vector Machine (SVM) and random forest (RF) were also evaluated. MARS was found to predict the

sugar content with better accuracy than others, whereas the prediction accuracy of RF was the poorest. The proposed

non invasive in situ method was found to be alternate fast reliable method to labour intensive laboratory methods and a

high throughput field applicable phenotyping technique for water deficit stress studies and rice breeding programs.


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IPPCongress2015


PK451 (IPPC0984)

Application of thermal imaging to monitor the crop stress condition

P. Krishnan*, Kaushik Banerjee and Victor Banerjee

Division of Agricultural Physics, Indian Agricultural Research Institute, New Delhi-110012, India


Monitoring the crop stress condition is an imminent requirement for abiotic stress characterisation. In this study, non-destructive and non-invasive characterisation of crop canopy status of wheat (Triticum aestivum) grown under moistureand temperature stress, was analysed using thermal imaging. When wheat crop is under abiotic stress there is a decreasein latent heat flux at the crop canopy surface, a complementary increase of sensible heat occurs and creates a largertemperature difference between crop canopy and air. Recent thermal imagers can instantly detect minor differences incrop canopy temperatures which correlate to changes in transpiration, thus revealing differences in crop stress condition.The major interest in the application of thermal sensing of field crop was the development of indices of crop stress asguidance for crop management. In this study a crop canopy stress index (CCSI) was derived from the image analysis ofthe thermogram of the crop. A significant correlation was observed between the CCSI derived from the thermogram withthe plant stress indicators like stomatal conductance, Relative Water Content, Water Potential and Leaf Area Index. Inaddition a high correlation was found between yield and CCSI at different growth stages indicate that thermography cancharacterise the crop stress status in wheat. These indices are mostly presumed to reveal variations in stomatal openingand evaporation rate as cellular water becomes the stress regulating factor. Thus thermal imaging as observed in thisstudy can be an efficient tool to characterise the crop responses to abiotic stress conditions.

PK452 (IPPC0989)

Comparative identification of microRNAs and their potential targets between Capsicum chinense and

Capsicum frutescens

Mukesh Nitin1*, Rajnish Kumar1, Onkar Nath2, Nirala Ramchiary1, Sarpras M1, Rashmi Gaur1, Vineet Sharma1, Sushil

Chhapeker1, Toshi Mishra3, Vijaya Bramha2 and Andrew Lynn2

1School of Life Sciences, 2School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India3Department of Bioinformatics, BHU, Varanasi, India


MicroRNAs (miRNAs) are small non-coding RNAs which regulate gene expression involved in various developmentalprocesses of plants and animals. Although many miRNAs and their potential target genes have been identified in variouscrop plants, there is very limited information in Capsicum species. Therefore, here in this study, we used transcriptomedata of Capsicum frutescens and C. chinense to identify miRNAs and their potential target genes. The transcriptome datawere generated from leaf, flower and three fruit stages of each Capsicum species using Illumina High Seq2000 sequencingmachine. Using matured miRNAs of already reported plant miRNAs from Plant Non coding RNAs database (PNRD) andmiRbase against 184975 C. chinense and 179780 C. frutescens transcripts, we identified a total of 747 and 708 potentialmiRNA transcripts in C. chinense and C. frutescens, respectively. Further structure folding and minimum folding freeenergy (MFE) analysis with VieenaRNAfold programe using standard criteria finally gave a total of 127 potential miRNAsin C. chinense and 146 miRNAs in C. futescens, of which 9 and 18 miRNAs showed differential expression in differenttissues of C. chinense and C. frutescens respectively. Out of predicted miRNAs in both the species, majority of miRNAswere conserved but few are found species specific. The prediction of potential targets for differentially expressed miRNAsusing PsRNAtarget programe identified a total of 218 and 360 mRNA target genes in C. chinense and C. frutescens,respectively, which are involved in various cellular functions. Few miRNA families showing differential expression in C.chinense and C. frutescens, were experimentally validated using qRT-PCR. Development of miRNAs based molecularmarkers is being in progress which could be used in Capsicum breeding programe.

Session L : Climate Change and Agriculture

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IPPCongress2015


PL453 (IPPC0073)

Impact of reduced PAR in determinate, semi-determinate and indeterminate soybean (Glycine max L.)

genotypes under climate change scenario

Kiran Bhagat*, Santanu Bal, Yogeshwar Singh, Ratnakumar Pasala, Sunayan Saha and P.S. Minhas

National Institute of Abiotic Stress Management (NIAM), ICAR, Malegaon, Baramati-413115, Pune, Maharashtra


Soybean is an important crop, which is photosensitive in nature and hence will be highly affected by the increasingintensity of atmospheric brown clouds, commonly known as “surface dimming”, and moisture stress condition, which mayprevail in the near future. Under present climatic condition, the average incoming PAR during morning hours is around1300 during kharif season, and it is predicted to decrease by 15-20% in near future. Three different structures were testedto standardize the uniformity of PAR interception within the structures. Perusal of data clearly revealed that as there ishomogeneity in every aspect in semi-circular dome shape structure as compared to rectangular and octagonal domestructures, and therefore, semi-circular dome shape structure was considered most appropriate for conducting such typeof research work. Light saturation point (LSP) calibration revealed that determinate soybean genotype showed LSP nearto 800 μmol m-2 s-1 PAR, with photosynthetic rate of 20.77 μmol m-2 s-1, whereas semi-determinate genotype showed LSPnear to 1100 μmol m-2 s-1 PAR, with photosynthetic rate of 21.95 μmol m-2 s-1, and indeterminate genotype showed LSPnear to 1000 μmol m-2 s-1 PAR, with photosynthetic rate of 28.88 μmol m-2 s-1. Determinate, semi-determinate andindeterminate soybean genotypes tested under control and in semi-circular dome shape structure by using white shadenet with 15-25% PAR reduction factor, revealed that yield of indeterminate soybean genotype was higher than determinateand semi-determinate types under reduced PAR. Hence, indeterminate soybean genotypes may perform better underreduced PAR condition under future climate change scenario.

PL454 (IPPC0148)

Effect of elevated CO2 and temperature on wheat and Phalaris minor

Kamlesh Paraste, P.P. Choudhury, M. Rathore, R. Singh, C. Sarathambal, N. Tripathi and Bhumesh Kumar*

ICAR-Directorate of Weed Research, Jabalpur-482004, India


Effect of elevated CO2 and elevated temperature on wheat and Phalaris minor (an important weed of wheat crop) was

studied in open top chambers (OTCs). Wheat (cv. GW-273) was sown following recommended practices and weedpopulation was maintained 15 plants/m2. Plants of both the species were exposed to different treatments [viz. ambient,elevated CO

2 (550 ± 50 ppm), elevated temperature (ambient + 3.0 ± 0.5 oC) and elevated CO

2 + elevated temperature]

from emergence to physiological maturity. Enrichment of atmospheric CO2 had a positive effect on overall growth of

wheat as well as P. minor, however, elevated temperature alone or in combination of elevated CO2 had adverse effect on

growth and development of both the species and more severely of wheat as compared to P. minor. Rate of photosynthesisincreased in both the species when plants were exposed to elevated CO

2, however, a decrease in photosynthesis was

observed at elevated temperature alone or in combination with elevated CO2.Species-specific as well as treatment-

specific differential expression of genes involved in photosynthesis and defence pathways was evident. As compared toambient conditions, advancement of panicle emergence was observed in both the species when exposed to elevatedtemperature (by 15 days), elevated CO

2 (by 7 days), and combination of elevated temperature and CO

2 (by 10 days).

Efficacy of sulfosulfuron against P. minor was considerably reduced at elevated CO2 and elevated temperature indicating

adverse effects on weed management. Results suggest that climate change may alter crop-weed interaction in favour ofweeds.


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PL455 (IPPC0164)

Lodging: A new face of climate change devastating agricultural productivity

Rinki Khobra*, H.M. Mamrutha, Raj Kumar Gupta and Indu Sharma

Division of Crop Improvement, Indian Institute of Wheat and Barley Research, Karnal-132001, Haryana, India


Lodging is a complex phenomenon that is influenced by interaction of crop plant, rain, wind, soil and result into permanent

displacement of plant stem from its normal position. Introduction of dwarfing gene, has increased lodging resistance in

Indian wheat varieties and hence lodging was not considered as a major problem till date. But, due to untimely rainfall in

the major wheat growing area (North West Plain Zone) during, 2014-15 crop season, significant (5-7%) crop lodging has

been reported. Hence, it is need of hour to explore the effect of lodging for developing lodging resistant wheat varieties for

secured agriculture and food security in India. As an initial attempt to analyze the post harvest effect on wheat crop

lodging, some of the morphological, physiological and quality traits were studied in the grains of lodged and non-lodged

genotypes. The grain samples were taken from IIWBR field trial having 25 genotypes with two replications in two conditions

during 2014-15 crop seasons. The results of initial screening showed a significant reduction in germination percentage,

shoot length and protein content in lodged genotypes but increase in their test weight. Significant visual differences in

grain color, size and luster were also observed. However these results are very preliminary and more study is required to

understand the negative effect of lodging on wheat grains and to develop the non-lodging wheat ideotype.

PL456 (IPPC0313)

Effect of water management practices during post-vegetative stage under system of rice intensification (SRI)

Amod Kumar Thakur*, K.G. Mandal and S. Raychaudhuri

ICAR-Indian Institute of Water Management, Bhubaneswar-751023, Odisha, India


The system of rice intensification (SRI) has been promoted for enhancing yield using less water. SRI recommendation for

water management is to apply a minimum of water during the vegetative stage, and then to maintain a thin layer of water

during post-vegetative stage. Moreover, it is not clear about optimum water management during post-vegetative phase of

rice crop for the best results. The present study was carried out to investigate the impact of continuous vs. alternate

flooding of paddy fields during post-vegetative stage on crop phenotype, physiology and performance under two crop

management systems, namely, SRI and conventional transplanting system (CTS). During vegetative stage of crop growth,

CTS plots were kept flooded and in SRI plots 3-DAD (days after disappearance of ponded water) irrigation method were

followed. Post-vegetative stage, four water management treatments, viz., continuous flooding (CF), 1-DAD, 3-DAD and

5-DAD were imposed both in CTS and SRI. Rice grown with SRI practices showed significant improvements in the plants’

morphology-plant height, tillering, leaves number, and expanded root systems. These were accompanied by improvements

in physiological functions like xylem exudation rate, canopy light interception, leaf chlorophyll content, light utilization,

and photosynthetic rates during flowering. These factors were responsible for increased grain yield as compared with

CTS. Significantly higher water productivity was found under SRI (6.3 kg/ha-mm) than CTS (3.3 kg/ha-mm). Under CTS,

the highest water productivity was obtained with the 1-DAD treatment, while under SRI, it was achieved in the 3-DAD

treatment.


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PL457 (IPPC0339)

Effect of elevated CO2 level on fate of azoxystrobin fungicide and soil microbial parameters in rice soil

Suman Manna1, V.P. Singh2 and Neera Singh1*

1Division of Agricultural Chemicals, 2Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India


A major indirect response to increased CO2 levels in the atmosphere is the greater belowground C allocation through root

exudation, which is likely to lead to change in size and activity of soil microflora and would have direct effect by increasing

temperature. Both may affect the degradation and persistence of pesticides in soil used for crop protection. We report the

effect of elevated CO2 (580±20 ppm) on the degradation of azoxystrobin [methyl (E)-2-{2-(6-(2-cyanophenoxy) pyrimidin-

4-yloxy) phenyl}-3-methoxyacrylate] and soil microbial activity in rice planted soil in open top chambers (OTC). Results

indicated that there was not much difference on the persistence of azoxystrobin in rice planted soil maintained under

different CO2 levels. The half life values for the azoxystrobin in rice soils were 20.3 days in control (rice grown at ambient

CO2 outdoor), 19.3 days in rice grown under ambient CO

2 atmosphere in OTC and 17.5 days in rice grown under elevated

CO2 atmosphere (580±20 μmol mol-1) in OTC. Azoxystrobin acid was recovered as the only metabolite of azoxystrobin,

but it did not accumulate in the soil/water and was further metabolized. Elevated CO2 enhanced soil microbial biomass

carbon (MBC) and alkaline phosphatase activity in soil. Compared with rice grown at ambient CO2 (both outdoors and in

OTC), the soil MBC at elevated CO2 increased by two fold. Elevated CO

2 did not affect dehydrogenase, fluorescein

diacetate (FDA) and acid phosphatase activities. Azoxystrobin application to soils, both under ambient and elevated CO2

levels inhibited alkaline phosphates activity, while no effect was observed on activity of other enzymes.

PL458 (IPPC0355)

Physiological and biochemical plasticity of Lepidium latifolium L. suggests it as sleeper weed in western

Himalayas under climate change scenario

Dhiraj Vyas*, Tarandeep Kaur and Hilal Bhat

Biodiversity and Applied Botany, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu-180001, J&K, India


Unlike the aggressive invasive character of Lepidium latifolium in riparian plains of North America, its growth is restricted

in Ladakh Himalayas. In order to understand its future spread under climate change scenario in ecologically sensitive

region of Western Himalayas, native populations of Lepidium latifolium in different microclimates were studied for

physiological and biochemical plasticity. Different populations from sparsly populated site (SPS; 2807 m asl; 34.54 N,

76.13 E), moderately populated site (MPS; 4139 m asl; 33.20 N, 78.64 E) and densly populated site (DPS; 3164 m asl;

34.15 N, 77.57 E) were studied for their fluorescence, photosynthetic and biochemical characteristics. Results showed

that the DPS had higher photosynthetic accumulation and transpiration rate despite lower VpdL and higher relative

humidity. This suggests the regulation of its leaf temperature by evaporative cooling. Apart from its nutritive value, higher

amounts of sinigrin obtained in DPS might play an important role in water transport under environmental stress. The

quantum efficiency of PSII photochemistry (Fv/Fm, NPQ, PSII) and light curve at various PPFD’s suggests better light

harvesting potential and light compensation point at DPS than the other two sites. Concomitantly, plants at SPS had

significantly higher lipid peroxidation, suggesting a stressful environment, and higher induction of antioxidative enzymes.

High light intensities at MPS are managed by specialized contrive of carotenoid pigments and PsBs gene product. It is

suggested that L. latifolium is present as ‘sleeper weed’ that has inherent biochemical plasticity involving multiple processes

and its potential spread is linked to site-specific micro-environment.


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PL459 (IPPC0386)

Physiological basis for development of climate resilient cultivars

R.K. Bhatt*, V.K. Manga, R. Sharma, A. Saxena and Uday Burman

Central Arid Zone Research Institute, Jodhpur-342003, Rajasthan, India


Climate change is experienced by increase in temperature and extreme weather events. It is projected that its impact will

be more significant in the coming years. Research is required to enhance crop resilience and adaptation to climate

change for sustainable production, food and nutrition security. This may be achieved by developing varieties with in-built

developmental plasticity to tolerate high temperature and soil moisture stress particularly at flowering and grain filling

stage, resulting in higher productivity. Screening under varying environments in pearl millet showed that some populations

(CZP2K9, CZP 923, CZP2K3, CZP9802) and hybrids (CZH226, CZH227, CZH225, ICMH 356) were promising climate

resilient genotypes based on yield. CZP2K9, CZP2K11, CZH227, CZH226, GHB538 reflected less than 10% variation in

leaf water content and membrane stability index. CZP2K9 also recorded higher grain yield with higher water use efficiency

over the environments differing in total rainfall, temperature and number of rainy days. In clusterbean genotypes HG884,

RGC1066, RGR-07, RGC1031, CAZG11-1 and RGC936 were selected as promising on the basis of grain yield and less

than 10% variation in leaf water content. In terms of membrane stability index, however, only HGS-03-52 and RGC1031

recorded less than 10% variation. Seed yield significantly correlated with dry weight of root and could be considered as

one of the parameters for screening the germplasm for hot arid environment. Thus, genotypes exhibiting more resilience

with respect to yield and physiological parameters to variable climatic conditions could form the backbone of future

breeding programs in both pearl millet and clusterbean.

PL460 (IPPC0716)

Phenotyping guar germplasm suitable for rainfed ecosystem of Tamil Nadu

N. Sritharan1* and A. Manivannan2

1Department of Crop Physiology, 2Central Institute for Cotton Research, Tamil Nadu Agricultural Univ., Coimbatore-641003, India


Guar (Cyamopsis tetragonoloba L. Taub.) production is greatly affected by climatic factors (rainfall and temperature), soil

factors and cultural practices. Although guar is grown under wide range of annual precipitation, the amount and distribution

of rainfall is the limiting factor for guar production under semiarid regions. In this light, the objectives were formulated to

study the performance of guar germplasm based on morpho-physiological traits as well as to assess the yield attributes

under rainfed ecosystem. A field experiment was conducted at Agricultural Research Station, Kovilpatti during the year

2014 rabi season, and a total of 42 guar genotypes (including vegetable, fodder and gummy types) were screened under

rainfed condition. Plant morphological characters, chlorophyll index, total dry matter production (TDMP), yield and yield

attributes were recorded. Soil moisture status at different depth was also measured. The fodder type, HFG 119 recorded

higher grain yield (824 kg/ha). In the case of gummy types, GAU513 recorded superior seed yield (818 kg/ha) which was

on par with HGS3-2, RGC 1003 and CAZG10. Among the vegetable types, T local recorded greater seed yield (607 kg/

ha). In correlation studies, TDMP, leaf area, leaf area index and number of seeds per pod were positively correlated with

grain yield.


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IPPCongress2015


PL461 (IPPC0800)

Oryza2000 model for effective rice crop management under irrigated ecosystem

B. Sailaja*, S.R. Voleti and D. Subrahmanyam

ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad, India


Biophysical models predict the behaviour of the land use system in physical terms such as crop yields, environmental

effects, and effect on management. Oryza2000 model is an eco-physiological crop model to simulate the growth and

development of rice crop in situations of potential production, water limitations, and nitrogen limitations. This paper

reports on the calibration and validation of the model under irrigated ecosystem using independent data sets of wet

season rice crop. Data are originated from the nitrogen and water limited experiments conducted by the physiology

department of IIRR, Hyderabad 2006. The calibration of the model was performed by deriving the crop growth parameters

for long (BPT 5204), mid (Ajaya) and short (Rasi) duration varieties suitable to the local climatic conditions. Oryza2000

model was able to simulate the site-specific rice production with good performance at high nitrogen level as simulated

values were very close to observed values. On the other, overestimation of leaf area index (LAI) at all nitrogen levels was

consistent indicating a need in this area for further improvement of the model. Model performance was also assessed by

statistical analysis (R2, D-index and NOF). D-index is *0.8–0.9 for two varieties at 3 nitrogen application levels for Ajaya

and BPT varieties. The results indicated applicability of model Oryza2000 can be extended to rice genotypes varying in

their growth periods and situations like fertilizer, water limited conditions.

PL462 (IPPC0972)

Morpho-Physiological efficiency of Soybean (Glycine max L.) cultivars under acid soils of mid-altitudes

of Eastern Himalayas

Krishnappa Rangappa1*, Azad Singh Panwar2, Lotika Kalita2, Meghna Haloi1, Hadienlarisa Syiemlieh2, Savita3,

Anjan Kumar Sarma4, Nishant Anandrao Deshmukh5 Anup Das3 and Shishom Vanao Ngachan6

1Plant Physiology Lab, 2Integrated Farming System Section, 3Agronomy Section, Division of Crop Production, ICAR Research complexfor NEH Region, Umiam-793103, Meghalaya, India4Division of Natural Resource Management, 5Division of Horticulture, 6Director, ICAR Research complex for NEH Region,Umiam-793103, Meghalaya, India


Acid soils predominantly present in EHR (80% total geographical area) limit the productivity of most of agricultural and

horticultural crops to great extent. Increased concentrations of Al, Fe, Mn, Cu and reduced availability of phosphorus in

these acid soils significantly influence physiological, biochemical and molecular plant response. In this connection, present

investigation aimed at characterising the morpho- physiological response of 14 soybean cultivars under severly affected

acid soils of upland conditions in mid-altitudes of Eastern Himalaya. Results show that, soybean cultivars exhibited

significantly varied soil acidity tolerant physiological adaptations in terms of differential root growth traits especially in

rhizosphere acidification, root architecture and R/S ratio, altered leaf characteristics viz., leaf pigmentation, leaf CMS,

stomatal size and density), changed total plant biomass and nutrient accumulation in root and leaves which in turn

influence the yield and yield attributes of the soybean plant in later stages. This altered pattern of morpho-physiological

response of different soybean cultivars might have dependent on the extent of soil nutrient regimes under acid soil. This

physiological characterisation and identification of efficient soybean cultivars can be further used in breeding program for

developing suitable soybean genotypes tolerant to acid soils of Eastern Himalaya.

Session M : Reproductive Biology

350

IPPCongress2015


PM463 (IPPC0294)

OsMADS29: A newly evolved regulator for an ancient metabolic pathway

Vibha Verma1*, Meenu Kapoor2 and Sanjay Kapoor1

1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India2University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi-110075, India


OsMADS29 is a cereal seed-specific transcription factor that has been shown to affect hormone homeostasis, causing

several-fold increase in the levels of active cytokinin (trans-zeatin) and a marked decrease in the levels of auxins. In order

to investigate if (a) there existed a conserved biochemical and/or regulatory pathway for cytokinin metabolism in diverse

taxonomic groups in the plant kingdom and (b) that rice MADS29 could exploit it for the purpose of starch accumulation,

we have generated tobacco BY-2 and Physcomitrella patens transgenic lines expressing rice MADS29 under maize

ubiquitin and 35S CaMV promoters. We had earlier shown that the expression of OsMADS29 could mimic the effects of

exogenously added cytokinins in the heterologous, tobacco BY-2 cell line, system. Our recent work indicates that the

OsMADS29 expression in BY-2 cell lines causes early and enhanced differentiation of proplastids into amyloplasts. This

differentiation is associated with positive modulation of several genes involved in plastid biogenesis and starch biosynthesis.

In moss also, OsMADS29 expression results in several developmental and morphological abnormalities that are suggestive

of excess of cytokinins in the system. These results are suggestive of the existence of a cytokinin metabolism regulatory

system that has largely remained conserved during the course of evolution from lower plants to present day eukaryotes

and OsMADS29 is able to target certain regulatory components of this pathway for shifting the hormonal balance in favor

of cytokinins.

PM464 (IPPC0295)

Rice SKP-like gene, a component of E3 ligase, is involved in regulating pollen development and anther

dehiscence

Mrinalini Chawla1*, Meenu Kapoor2 and Sanjay Kapoor

1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India2University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi-110075, India


Meiosis is an important event in the life of plants, as it marks the time of developmental transition from the sporophytic to

the gametophytic phase. A comprehensive transcriptomics analysis using our in house data representing different stages/

tissues of vegetative and reproductive development in rice had revealed an important role of gene inactivation along with

that of specific genes activation during male gametophyte development. We also found a marked increase in the activity

of genes involved in ubiquitin-mediated protein degradation pathway. Central to this protein turnover machinery is the

Skp1-Cullin-F-box (SCF) class of E3 ligases, which identify and prepare specific proteins for degradation by adding

ubiquitination marks. Our investigations revealed 23 F-box and five OSK encoding genes expressing specifically in

meiotic anthers in rice. Here, we show that silencing of one of these OSKs results in anther indehiscent phenotype and

severe reduction in pollen viability. Silencing of this SKP gene does not affect the process of meiosis as well as mitotic

divisions in the microspores. However, the microspores in OSK-KD lines remain smaller (by ~26%) in size in comparison

to those in the wild type and fail to accumulate starch at the time of maturation, causing loss of viability. Furthermore, the

endothecial layer of anther wall fails to develop the characteristic lignin striations, which have been shown to provide

necessary mechanical pressure/tensile force required during anther dehiscence. These results highlight novel and essential

roles of this gene in the 26S proteasome complex-mediated protein degradation during male gametophyte development.


351

IPPCongress2015


PM465 (IPPC0298)

Transcriptional properties of five seed-high NAC transcription factors

Pinky Agarwal*, Iny Mathew and Akanksha Panwar



Five NAC genes, which encode for transcription factors (TFs), express at high levels during rice seed development. This

is reiterated by promoter-reporter rice transgenic plants. Rice genotypes with variable seed weights show variation in the

expression patterns and levels of these genes during seed development. The promoter sequences of these genes, from

different genotypes, show limited correlation with the expression. In terms of transcriptional properties, only one of the

proteins is nuclear localized. However, many of these proteins dimerize amongst themselves and are subsequently

nuclear localized. They are not only transcriptional activators, but strong repressors as well. Amongst the five NAC

genes, one is alternatively spliced while two others are transcriptionally fused. Interestingly, the multiple forms of these

genes have variable transcriptional properties. Summarily, the five seed-high NAC TFs and their forms have overlapping

as well as distinct transcriptional properties, indicating their multiple roles in rice seed development.

PM466 (IPPC0358)

PROTEIN L-ISOASPARTYL METHYLTRANSFERASE (PIMT) is implicated in seed desiccation tolerance

and seed longevity in rice

Nitin Kamble*, Bhanu Petla and Manoj Majee



PROTEIN L-ISOASPARTYL O-METHYLTRANSFERASE (PIMT) is a protein repairing enzyme and catalyses the conversion

of spontaneously modified isoAsp to Asp in protein. The present study in rice demonstrates that PIMT activity sharply

increases at maturation phase, retains in dry seed and then rapidly decline upon completion of germination. Likewise,

deleterious isoaspartyl accumulation also increases during seed maturation and is highly abundant in dry seed but

decreases upon imbibition. Western blot analyses clearly demonstrated distinct tissue and seed development stage

specific accumulation of these PIMT isoforms, indicating their participation and specific contribution in seed longevity and

germination in rice. For further analysis, we raised transgenic lines for each isoform and the data reveals the distinct roles

of each OsPIMT isoform in restricting deleterious isoAsp accumulation to improve seed vigor and longevity. Collectively,

these data imply that PIMT mediated protein repair mechanism initiates during the course of seed development in rice

and each PIMT isoform plays a distinct yet coordinated role in maintaining seed vigor and longevity by restricting deleterious

isoAsp accumulation in tissue and development stage specific manner.


352

IPPCongress2015


PM467 (IPPC0369)

Differentially expressed galactinol synthase gene (s) of chickpea (Cicer arietinum L.) improves seed

vigor and longevity by limiting the hydroxyl radicals during aging

Prafull Salvi* and Manoj Majee

Plant Molecular Biology and biochemistry, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India


Galactinol synthase (EC: 2.4.1.123) (GolS) catalyzes the rate limiting step in Raffinose Family Oligosaccharide (RFO)biosynthetic pathway. RFOs including galactinol are known as anti-stress agents and suggested to play important role inabiotic stress responses and seed desiccation tolerance. In this study, we have shown galactinol synthase activity as wellas galactinol content is highly concentrated in pod and dry seed and gradually decreased as seed germination progresses.Galactinol synthase activity was also found to be enhanced in response to stresses. In chickpea, galactinol synthaseenzyme is encoded by two divergent genes (CaGolS 1 and CaGolS 2) which produce 5 different types of transcripts.Transcription analysis revealed that the CaGolS1 and CaGolS2 are differentially regulated. Further, all isoforms werebacterially expressed, purified and biochemically characterized. All isoforms exhibit differential enzyme activity. Nativemolecular weight and subunit composition was determined for enzymatically active isoform through gel filtrationchromatography. Confocal microscopy showed that a transiently transfected CaGolS-YFP fusion proteins primarilydistributed to nucleus and membrane in onion epidermal cell. Further we found that seed-specific expression of CaGolS1and CaGolS2 in Arabidopsis play a role in maintaining seed vigor and longevity, which was correlated well with increasedgalactinol and raffinose contents and reduced hydroxyl radicals in seed of transgenic lines. Collectively, our work clearlydemonstrates that CaGolS(s) are differentially regulated in chickpea and play important role in enhancement of seedvigor and longevity.

PM468 (IPPC0485)

Overexpression of OsbHLH142 causes complete male sterility in rice

Rajeev Ranjan1*, Reema Khurana2 and Akhilesh Tyagi1

1National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India2Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India


Investigations on male sterility in crop plants are important for the hybrid seed production which provides improved yieldas a result of heterosis. Rice is an important crop plant which is the source of food for almost half of the world population.Anther development in rice has been studied for many years which led to the identification of many molecular players.Yet, detailed mechanism associated with it remains elusive. Previously, we reported a bHLH transcription factor OsbHLH142as an anther specific gene in rice. Now, we aimed to functionally characterize this during anther development in rice.Expression analysis of OsbHLH142 transcripts and polypeptide showed delayed accumulation of its protein compared totranscripts. Overexpression of OsbHLH142 in transgenic rice results in complete sterility and no seed set was observed.Crossing of transgenic plants with wild type pollen results in restoration of seed formation. Detailed observation ofreproductive organs of transgenic plants revealed that anthers did not dehisce and most of the pollen grains were shrunken,abnormal shaped and non-viable. Electron microscopic and paraffin based tissue section analysis revealed detailedhistological reason for these defects. RNA-Seq based transcriptome analysis of early and mature anthers suggestsOsbHLH142 directly or indirectly regulates expression of different set of genes in the two stages of anthers. Moreover,through yeast two-hybrid and BiFC assays we have identified some OsbHLH142 interacting proteins which gives moreinsight on its molecular functions. Therefore, this study has developed new male sterile rice plants and explored novelfunctions of OsbHLH142 during rice anther development which will essentially help in the control of male fertility in cropplants.


353

IPPCongress2015


PM469 (IPPC0508)

Effects of auxins and other hormones on rice OsMADS29 expression

Ridhi Khurana* and Sanjay Kapoor



Rice MADS29 is a seed-specific MADS-box transcription factor that has been implicated in embryo and endosperm

development. The OsMADS29 expression has been found to be tightly regulated at transcriptional, post-transcriptional

as well as post-translational levels. We have previously demonstrated that OsMADS29 regulates cytokinin levels at the

site of its action, not only in rice but also in heterologous tobacco BY-2 cells. In rice, the MADS29 transcripts begin to

accumulate in high amounts soon after pollination, however, molecular basis of this induction is not known. Careful

inspection of rice MADS29 upstream regulatory region (URR) revealed a number of auxin response elements (auxREs),

besides a number of other conserved regulatory motifs. The auxREs are six base pair conserved motifs (TGTCTC) found

in multiple tandem repeats in the promoters of early auxin response genes like GH3 and SAURs. And, the auxREs have

been shown to be the targets of the DNA-binding domain of auxin response factors (ARFs). We have developed a

tobacco BY-2 cell-line based assay system, where the transcriptional activity of OsMADS29URR::GFP under different

hormonal regimes can be analysed. The results of these analyses will be discussed.

PM470 (IPPC0524)

Downregulation of a seed-preferential mediator (MED) subunit affects reproductive development in rice

Naveen Malik*, Pinky Agarwal and Akhilesh K. Tyagi



Mediator, a multi-subunit protein complex in eukaryotes, assists GTFs in RNA polymerase II mediated transcription by

acting as a co-regulator. The whole mediator complex is structurally organized in head, middle, tail and CDK-cyclin C

modules. Direct interaction of the tail module subunits with various transcription factors facilitates relay of signal to RNA

pol II transcription machinery through head and middle modules. There is growing evidence for important roles played by

various mediator subunits in Arabidopsis, but in rice the complex still remains structurally and functionally unexplored. In

past, our group reported several mediator subunit genes having high expression during rice seed development stages

and we chose one of these subunits for functional characterization. To unravel the function of this gene in rice, both

overexpression and knockdown transgenic plants were raised. Overexpression plants were found to behave normally in

most aspects of development and yield. On the other hand, knockdown plants showed various defects during reproductive

development like non-viable pollen, reduced seed set and defective seed filling. Progress of the work on functional

delineation of this MED subunit will be presented.


354

IPPCongress2015


PM471 (IPPC0659)

Various properties of a seed-preferential NAC transcription factor in rice

Iny Mathew* and Pinky Agarwal



NACs are a large group of plant-specific transcription factors (TFs), playing important roles in plant life. All these proteins

contain a highly conserved NAC domain identified initially in NAM, ATAF1/2 and CUC2 TFs, giving rise to the acronym

‘NAC’. In our microarray analysis, one member of this family has been shown to have seed-preferential expression,

indicating its significance in seed development. Further, Q-PCR analysis confirms a high expression of this gene in rice

seed developmental stages of five different genotypes with varying seed weight. The specific expression of the gene in

the seeds can be credited to its 2 Kb promoter region as evident by the promoter-GUS analysis of transgenic plants.

Alternative splicing has resulted in three different forms (A, B and C) of the transcript, which show difference in their C-

terminal transcriptional regulatory region. Although, these proteins by themselves are localized in the cytoplasm, interaction

with another NAC TF, recruit the A form to the nucleus as shown by BIFC experiments. Further, the different forms also

show varying levels of transactivation ability. Over expression of this protein in planta points to the necessity of maintaining

the temporal expression of this gene at an optimal level, as these plants show a slower growth rate with weaker and

thinner aerial parts. The poster will focus primarily on the functional aspects of this gene in rice seed development.

PM472 (IPPC0666)

Stigma and stylar movements vis-a-vis pollination dynamics in two medicinally important threatened

plants

Susheel Verma* and Ankush Khajuria

Centre for Biodiversity Studies, Baba Ghulam Shah Badshah University, Rajouri-185232, J&K, India


Reproduction in flowering plants is a complex process involving several steps of communication between male and

female counterparts. Many a times these interactions are mediated by agents both biotic and abiotic. Obviously the

mechanisms underlying these interactions must be mediated by simple to complex signaling mechanisms operating

either singly or interwoven. Two threatened and prized medicinal angiosperms namely Veleriana wallichii and Incarvellia

emodi accomplish their mating in beautifully controlled ways. At the physical level, the manifestations are interesting and

constitute the matter for this presentation. Flowers of Valeriana wallichii under pollinator limiting conditions fetch pollen

themselves by moving their styles towards the pollen source, either within or in the neighboring flower. This way their

stigmata get pollinated and reproduction is ensured. Stigma of Incarvillea emodi waits for requisite number of pollen to

land on its surface even upto 9 days and then closes its lobes permanently. These mechanisms are responsible for

successful sexual reproduction in these threatened taxa. Their control, however, remains a mystery.


355

IPPCongress2015


PM473 (IPPC0682)

Courtship in plants! What initiates it?

Namrata Sharma* and Veena Sumbria

Department of Botany, University of Jammu, Jammu-180006, J&K, India


Concealment of ovules inside the ovary has made pollen-pistil interaction an inseparable part of sexual reproduction in

angiosperms. In most cases, this interaction initiates with the release of pollen from the anthers and their landing on the

receptive part of pistil, i.e., stigma. This landing may be autonomous because of the close proximity of the two structures

as happens in many autogamous taxa, or may involve a vector in cross-pollinated ones. Pistil in all these cases remains

static structure while the pollen is mobile. During our work on the pollination mechanism in several flowering plants, we

have encountered several cases, which defy these rules (e.g., in-situ pollen germination in Fumaria indica). Herein we

present a recently encountered case in Tephrosia purpurea, wherein pistil exhibits beautiful movements to capture the

pollen, which is being released by the anthers situated below the level of stigma. Although stylar movements have been

reported previously also, the present study shows definitive phases of interaction between pollen and stigma facilitated

by regular movements of both stigma and style. Prior to successful fertilization what stimulates these movements remains

an open question. We speculate some sort of signalling mechanism operative in these movements. It is an endeavour for

us to present these examples before the learned forum of plant physiologists to get a clue to the possible mechanisms

involved in such interactions.

PM474 (IPPC0784)

Phylogeny, expression and interaction of NF-Y TF family in rice

Sweta Das*, Swarup Kumar Parida, Pinky Agarwal and Akhilesh Kumar Tyagi



Nuclear factor-Ys (NF-Ys) are histone-like transcription factors which are present in all eukaryotes. These consist of three

subunits, namely, NF-YA, NF-YB and NF-YC. These are encoded by single genes in yeast and animals but multiple

number of genes in plants. NF-YB and NF-YC share sequence homology with H2B and H2A, respectively. They form

dimers to which, NF-YA binds forming a trimer which in turn recognizes and binds to CCAAT box in the promoters. In

plants, due to the presence of multiple subunits in each sub-class, there are manifold possibilities of these combinatorial

complexes, which eventually lead to the functional diversity of NF-Ys. In rice, a total of 40 genes have been found to

encode NF-Y TFs among which 11 code for NF-YA, 13 for NF-YB and 16 for NF-YC. Phylogenetic analysis shows

homology of some of these genes with certain Arabidopsis NF-Ys and yet a few are unique to rice. Microarray data

suggests that the expression of several of these TFs is constitutive and a few genes express in a seed specific/preferential

manner. SNPs in 10 of these genes have been found to associate with seed size traits. These genes further exhibited

differential expression patterns in seed developmental stages of rice genotypes with contrasting seed features. Study of

transactivation properties of these TFs has revealed that few subunits belonging to each of the sub-class positively

regulate transcription. Yeast two-hybrid interactions amongst OsNF-Ys suggest interaction of certain subunits with each

other. An overview of the OsNF-Y family will be presented here.


356

IPPCongress2015


PM475 (IPPC0788)

Understanding sex expression and modification in Coccinia grandis - A dioecious member of

Cucurbitaceae

Ravi Devani1*, Amita Ghadge1, Kanika Karmakar2, Jayeeta Banerjee1, Sangram Sinha2 and Anjan Banerjee1

1Indian Institute of Science Education and Research, Pune-411008, Maharashtra, India2Department of Botany, Tripura University, Suryamaninagar-799022, Tripura, India*Presenting author: [email protected]

Majority of the angiosperm species are co-sexual (Hermaphrodite or Monoecious). Only about 6% of angiosperm species

are dioecious having separate male and female individuals. From the phylogenetic distribution of dioecious species, it is

clear that dioecy has evolved many times in plant kingdom independently with multiple molecular mechanisms governing

sex expression. Also, a few dioecious species have further evolved sex chromosomes like most of the mammalian

species. Dioecious systems also exhibit a certain degree of lability in sex expression. Chemical treatments like AgNO3

and Ag2S

2O

3, treatment with some phytohormones, and hypomethylating agent 5-azacytidine are known to alter sex

expression in dioecious plants. Coccinia grandis is one such dioecious species having heteromorphic sex chromosomes.

C. grandis belongs to Cucurbitaceae, a family well known for its diverse sexual systems. Availability of four sequenced

genomes in Cucurbitaceae gives C. grandis, an advantage as a model system to study dioecy. Sex modification has

been studied in hypogynous Silene latifolia (Caryophyllaceae) but there are no such reports in epigynous Coccinia grandis.

We have taken a candidate gene approach to understand the role of flowering related genes in the development of

unisexual flowers of C. grandis. Simultaneously, a comparative transcriptomic approach has been initiated to identify

novel players involved in sex expression and modification. Investigations on sexual phenotypes of C. grandis including a

rare gynomonoecious (GyM) form and AgNO3 mediated sex modification will add a new dimension to the understanding

of sex expression in dioecious flowering plants.

PM476 (IPPC0852)

Expression and promoter analysis of seed storage protein encoding genes in rice

Akanksha Panwar*, Iny Elizebeth Mathew and Pinky Agarwal



In monocots such as rice, seed storage proteins (SSPs) are synthesized in the endosperm during seed development,

which are essential source of proteins in human diet. On the basis of solubility, seed storage proteins are classified into

four categories: albumins (soluble in water), globulins (soluble in saline), prolamins (soluble in aqueous alcohol) and

glutelins (soluble in alkali). Glutelin is a major SSP in rice, present in 60-80% by weight of the total protein, whereas,

prolamin contributes to 20-30% of total proteins. Glutelins in rice have been reported to be homologs of 11S globulins of

legumes, while prolamins have been shown to be originated from an alpha-globulin storage protein. Endosperm-specific

expression of SSP genes is regulated by cis-elements present in their promoters, such as GCN4, a prolamin box (P-box),

AACA, ACGT. These motifs act as binding sites for TFs to regulate their expression, of which many are known to be

conserved. We have attempted to identify SSP encoding genes of rice and classify them phylogenetically, along with their

promoter and expression analysis.


357

IPPCongress2015


PM477 (IPPC0886)

Associated bacteria assist in reproduction of seaweed, Gracilariadura

Ravindra Pal Singh*

Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1, Fukuoka,812-8581, Japan


Seaweed associated bacteria play a crucial role in morphogenesis and the growth of seaweeds in direct and/or indirect

manners. Bacteria of phylum proteobacteria and firmicutes are generally most abundant on the seaweed surface. Associated

bacteria produce hormones, quorum sensing signals, bioactive compounds and other effective molecules that are

responsible for normal morphological structure and development in seaweeds. Zoospores of Ulva species respond to N-

acyl-homoserine lactones (AHL) produced by gram- negative bacteria and settle near the centre of AHL production. In

this regards, we isolated several gram-negative bacteria from different seaweeds and out of them seven isolates were

found to produce different types of N-acyl homoserine lactones (AHLs). Interestingly, Shewanella algae produced five

types of AHL: C4-HSL, HC4-HSL, C6-HSL, 3-oxo-C6-HSL and 3-oxo-C12-HSL. Subsequently, the AHLs producing bacterial

isolates were screened for carpospore liberation from G. dura and these isolates were found to positively induce carpospore

liberation over the control. Also, we observed that carpospore liberation increased significantly in C4- and C6-HSL treated

cystocarps. Sodium dodecyl sulfate and native polyacrylamide gel electrophoresis of the total protein of the C4- and C6-

HSL-treated cystocarps showed two specific peptide bands of different molecular weights (50 and 60 kDa) as compared

to the control, confirming their indirect effect on carpospore liberation.

PM478

Effect of high temperature stress on physiological attributes, antioxidative enzymes and expression of

heat shock proteins in wheat (Triticum aestivum L.)

N.K. Gupta*, Aarif Khan and R. S. Sain

Seed Technology Research, Rajasthan Agricultural Research Institute, Durgapura- Jaipur-302018, Rajasthan, India*Presenting author: [email protected]

Experiment was conducted to study the effect of heat stress on various parameters in five diverse wheat genotypes at

seedling stage. Seeds were sown in plastic pots in laboratory at 25±2°C. For heat treatment, 20 days old seedlings were

exposed to 36°C/42°C for 6 hours. Results showed that the heat stress significantly reduced the dry weight and length of

seedlings in all the genotypes. Chlorophyll content and membrane stability index decreased but proline content increased

in all the genotypes under stress conditions. Among genotypes, Raj 4083 exhibited better heat tolerance, particularly at

42°C. Activities of SOD, CAT, APX and POX increased significantly in all the genotypes but per cent enhancement was

maximum in Raj 4083 and minimum in PBW 502. In SDS-PAGE protein profiling, 15 polypeptides were recovered in

control plants with slight variations in temperature treated plants of most genotypes. Proteins with molecular weights of

85 kDa and 15 kDa were constitutively expressed in all the genotypes but proteins of 45 kDa were expressed only in

tolerant genotypes. Expression analysis of heat shock proteins, particularly hsp 90 further indicated the existence of

tolerance mechanism in Raj 4083. Thus, using a multi parametric approach, Raj 4083 was found most tolerant and PBW

502 most susceptible to heat stress. The study also provides evidence that antioxidant enzymes like SOD, CAT, GPX,

APX and heat shock proteins like HSP 90 have association with heat tolerance in wheat and thus can be used as efficient

markers for heat tolerance at seedling stage.

ted bacteria assist in

Author Index

359

IPPCongress2015


Abass Nazia ...................................... 224

AbdElgawad Hamada ....................... 110

Abdin M.Z. ............................................. 39

Abdul R. ............................................. 151

Abdullah Hesham ............................... 37

Ablordeppey Kenny ............................ 37

Achary Mohan .................................. 239

Achary Mohan Murali ......................... 77

Achary V. Mohan ................ 225,240,315

Acharya Vishal ............................ 57,340

Adhikari Kailash .................................. 65

Adhikari Kedar .................................. 121

Aftab Tariq ........................................ 294

Agarwal Aakrati ..................... 77,129,239

Agarwal Aditya Vikram ...................... 318

Agarwal Gaurav ................................. 216

Agarwal Manu ...........................17,51,176

Agarwal Meenakshi ...........................242

Agarwal P. .......................................... 43

Agarwal Pallavi .................................. 182

Agarwal Parul .................................... 317

Agarwal Pinky ....... 351,353,354,355,356

Agarwal Preeti .................................. 147

Agarwal R.M. .................................... 276

Agarwal Surekha Katiyar ............. 17,176

Aggarwal P.K. ....................................156

Agurla Srinivas ..................................187

Ahamed M. Lal ...................................174

Ahanger Mohammad A. .....................276

Ahlawat Yogesh ...................................214

Ahmad Altaf ..........................................309

Ahmad Syed Sabhi .............................. 83

Ahmad Tauqueer ....................... 335,338

Ahmadi N. ............................................ 99

Ahmadizadeh Mostafa ........................ 103

Ahsan Hafiza .....................................270

Akbari Parisa ........................................ 37

Akula Shalini .................................... .177

Alharbi Najeeb .................................. 121

Alivelu K. ........................................... 142

Aloor Bindu Prasuna .......................... 306

Alvarez-Fernandez Ruben .................. 69

Amardeep ..........................................123

Amaregouda A. ...........................131,264

Amarshettiwar S.B. ............................ 152

Amasino Richard M. ............................ 44

Anand Abhishek ................................. 247

Anand Anjali ......................................145

Anand Saurabh ................................. 319

Ananya Chakrabarty .........................140

Anders Nadine ...................................201

Anderssen Bob .................................... 81

Andreuzza Bindu ................................ 79

Andreuzza S. ..................................... 102

Andrew Lynn ...................................... 344

Angadi Sangamesh .......................... 149

Ansari Mohammad Wahid .................. 118

Ansari Shamim ................................. 316

Antony Edna ....................................... 275

Anuj Abhishek Kumar ....................... 314

Anuradha M. ..................................... 124

Anwar Khalid ............................... 122,226

Archak Sunil ....................................... 156

Argal Surendra ................................. 276

Ariyadasa Thilini ............................... 122

Aro Eva Mari ..................................... 109

Arora Ajay ...36,127,132,143,144,286,287

Arora Priyanka .................................. 181

Arora Sandeep ................................. 166

Aruna C. ........................................... 130

Arya Deepshikha ................................ 35

Arya Gulab ......................................... 190

Arya Preeti ......................................... 340

Arya R.R. .......................................... 160

Asard Han .......................................... 110

Ashish Subba .................................... 122

Ashraf Nasheeman ..................... 223,224

Ashwini N. .......................................... 198

Asif M.H. ........................................... 299

Asif Mehar ................................. 205,317

Asrey Ram ......................................... 335

Aswani Vetcha ..................................... 67

Atkins Peggy Ozark ............................ 199

Atri Anupama .................................... 332

Auguin Daniel ....................................... 9

Augustine Rehna ................................ 41

Avasthi Uday Kiran ........................... 306

Awakale Pramod .............................. 190

Azeez Abdul ..................................... 299

Baba Shoib ....................................... 224

Babita.................................................. 210

Babitha K.C. ......................................... 45

Babu O. Sudhakar ............................. 133

Baburam V. ........................................ 239

Bagri Deepak Singh .................... 262,281

Bahuguna Rajeev K. ............................ 11

Baig K.S. ...........................................132

Bains Gurdeep ................................... 118

Bains Navtej Singh ............................ 149

Bajaj Deepak ...................................... 104

Bakshi Achala ................................... 180

Bal Santanu ....................................... 345

Bala Kiran .......................................... 220

Bala Suman ....................................... 260

Balaji M. ................................................ 17

Balyan Sonia Chhillar ......................... 12

Bamel Kiran ....................................... 285

Banday Zeeshan Zahoor ................... 208

Bandyopadhyay Anindya .................... 70

Banerjee Anjan ........................... 189,356

Banerjee Jayeeta ............................... 356

Banerjee Kaushik ............................... 344

Banerjee Nirupama .......................... 253

Bangar Pooja ..................................... 154

Bannerjee K. ...................................... 102

Bansal K.C. ............................. 7,156,326

Banu Sofia .......................................... 251

Bapatla Ramesh B........................ 67,229

Barchi Boubker ..................................... 8

Bareth S.S. ......................................... 273

Barik Jayanta ..................................... 293

Barman Paramananda ..................... 218

Barnabas A.D. .................................. 336

Barua Praga ...................................... 207

Baruah K.K..................... 62,283,297,305

Baruah Sunitee Gohain .................... 305

Basak Sandip Kumar ......................... 141

Basu P.S. .......................................... 143

Basu Partha ....................................... 272

Basu Saumik ....................................... 47

Basu Urmila ........................................ 312

Basumatary Nabami .......................... 176

Batra V. ............................................. 161

Bechtold Ulrike ..................... 32,123,136

Beckmann Hauke ............................... 201

Bedi Seema ...................................... 332

Beemster Gerrit T.S. ........................ 110

Begam Momtaj .................................. 141

Begna Sultan .................................... 149

Behera Tusarkanti ............................. 315

Bender Kyle ...................................... 114

Bertheau Lucie ..................................... 9

Best Norman ....................................... 75

Bethoju K. .......................................... 102

Author Index

360

IPPCongress2015


Bhaduri Debarati ................................ 159

Bhagat Kiran ..................................... 345

Bhagat Prakash Kumar .................... 185

Bhalla Prem L. ............................... 73,80

Bhambhani Sweta ............................. 317

Bhandari Pinki .................................. 242

Bhandari Purnima ............................. 265

Bhandawat Abhishek ..................221,325

Bharadwaj Chellapilla ......................... 323

Bharali Ashmita ............................ 62,305

Bhardwaj Archana ............................. 210

Bhardwaj Chhavi .............................. 220

Bhardwaj Mansi .................................... 49

Bhardwaj Vasudha ............................ 314

Bhargav R. ........................................ 120

Bharti Amrit ....................................... 134

Bharti Pooja ...................................... 328

Bhaskar Avantika ................................ 186

Bhaskar Yogendra ............................ 202

Bhasker Pattipaka ............................. 177

Bhat Hilal A. .............................. 113, 347

Bhat K.V. .................................... 154,326

Bhat Prabhakar ....................................130

Bhat Ravi............................................. 100

Bhatia Arti ......................................... 278

Bhatia Chitra .................................... 206

Bhatia Sabhyata ................... 200,203,316

Bhatnagar Akanksha .................. 192,193

Bhatt R.K. .................................... 283,348

Bhatt R.M. ......................................... 343

Bhattacharjee Annapurna .................. 211

Bhattacharjee Payel ........................... 259

Bhattacharjee Saikat ......... .. 256,257,258

Bhattacharrya Abhishek ..................... 33

Bhattacharya M.K. ............................ 163

Bhattacharyya Dhriti ........................... 245

Bhave Sanjay ......................... 85,284,324

Bhisth Kavitha ................................... 112

Bhowmick P.K. ............................ 74,320

Bhushan Deepti ................................... 131

Bhyri Priyanka .................................... 79

Birbal......................................................120

Bishi S.K.......................... 42,159,255,302

Bisht Naveen ............................... 41,190

Biswal Basanti ..................................... 71

Biswas Ashish .................................... 268

Biswas Asok Kumar ........................... 225

Bobba Sunil ...................................... 305

Bohra Burhanuddin........................... 280

Boni Rainer ....................................... 243

Boominathan P. .................................. 154

Bora Ranjan Kumar ........................... 176

Borah Leena ..................................... 297

Borah Pratikshya .............................. 218

Bordoloi Nirmali .................................. 283

Borphukan Bhabesh ...... 225,239,240,315

Brahma Vijya ............................. 220,340

Bramley H. .................................... 95,121

Brignolas Franck .................................... 9

Burman Naini ............................. 192,193

Burman Uday ............................. 157,348

Burondkar Murad ........... 85,284,306,324

Calapit-Palao Cecilia Diana O.............103

Carlos Guzman.................................... 318

Carolina Paola Sansaloni.................. 318

Carolina Saint-pierre ........................... 318

Carpin Sabine ....................................... 9

Chaitanya Bharatula ........................... 231

Chakrabarti B. .................................... 277

Chakrabarty Broto............................. 233

Chakraborty Anirban ........................... 155

Chakraborty Koushik...................... 72,159

Chakraborty Navjyoti .................... 55,191

Chakraborty Niranjan..131,133,152,196, 207

Chakraborty Subhra ............. 133,152,207

Chakraborty Supriya .................... 49,245

Challabathula Dinakar ...................... 139

Chamanthula Gayathri ........................ 339

Chand Gurdev .................................... 177

Chand Mehar .................................... 135

Chandra A. ................................. 262,271

Chandra Amaresh ........................ 155,222

Chandra Deepak ............................... 318

Chandran Divya ........................... 256,257

Chandrapal........................................ 296

Chandrasekhar K. ............................. 129

Changwal Chunoti ....................... 286,287

Chatrath Ravish .................................. 140

Chattha Yadvendra Singh ..................186

Chattopadhyay Debasis ........ 207,252,319

Chattopadhyay Sudip .................. 13,320

Chaturvedi Arjit .................................... 227

Chaturvedi Ashish K. ............... 98,159,169

Chaturvedi S.K. ........................... 272,322

Chaudhari Vidhya .............................. 269

Chaudhary Bhupendra ........................ 191

Chaudhary Chanderkant .................... 219

Chaudhary Saurabh ........................... 28

Chaudhury Ashok ...................... 154,220

Chauhan D.A. ................................... 168

Chauhan Harsh ................................... 243

Chauhan P.S. .................................... 182

Chauhan Rajinder Singh .................. 337

Chauhan Rohit .................................... 57

Chaurasia Akhilesh Kumar .................. 331

Chawla Mrinalini ........................... 114,350

Chawla Vandna .................................... 57

Cheema H.S. .................................... 273

Chefdor Françoise ................................. 9

Cheong Yong-Hwa............................... 15

Chetti M.B. .................................... 82,284

Chettry Upasna ................................... 217

Chevala Narayana ............................. 211

Chhajer Sidhika .................................. 283

Chhapekar Sushil.................. 220,328,344

Chhikara Sudesh .................................37

Chhuneja Parveen ............................. 112

Chikani B.M. ...................................... 302

Chinreddy Subramanyam .................. 129

Chitikineni Annapurna .................. 217,322

Chitwood Daniel ................................ 110

Chopra A.K. ...................................... 169

Chopra Preeti ..................................... 272

Chopra Renu Khanna .................... 1,134

Choubey Ami ...................................... 289

Choudhary Chetan Kumar ................ 140

Choudhary Harshawardhan ............... 315

Choudhary R.L. ............................ 120,276

Choudhary Ritika .............................. 140

Choudhur Devapriya ............................. 55

Choudhury P.P. ................................ 345

Choudhury Swarup Roy ...................... 18

Chowdappa P. .................................. 100

Chowdhury A. Roy .............................. 331

Chowdhury Rajojit .............................. 141

Christopher Penfold .............................. 69

Chrungoo N.K. ................. 21,217,322,323

Clerget Benoit ...................................... 264

Clevenger Josh ................................. 199

Courdavault Vincent .............................. 9

Cressman Kurtis ................................. 199

Dadlani Malavika ............................... 230

Dafyd Jenkins ........................................ 69

Dagla M.C. ........................................ 302

Author Index

361

IPPCongress2015


Dahuja Anil ........................................ 338

Dalal Girish ........................................ 280

Dalal Monika ......................... 190,282,324

Dalvi D.G. ......................................... 132

Dalvi Vijay .......................................... 324

Damodarreddy D. .............................. 124

Damor Parthvee R. ........................... 168

Dar M.A. ........................................... 270

Das Amit ............................................. 79

Das Anup ........................................... 349

Das Ayan........................................... 246

Das Bappa ..................................... 92,343

Das M. ................................................. 171

Das Madhumanti .............................. 184

Das Madhurima ................................ 233

Das Priyanka ........................... 11,116,165

Das Rohit .......................................... 259

Das Roma .......................................... 323

Das S. .................................................. 43

Das Sampa ........................................ 246

Das Sandip ........................................319

Das Satyajit ........................................160

Das Shouvik ........................................104

Das Sweta ........................................ .355

Dash S.K. .......................................... 343

Dasmunshi Anilabh............................ 315

Datta Swapan K. ............................... 322

Davda J. ........................................... 102

David Wild ........................................... 69

Dayanandan P. .................................... 336

Debnath Suprokash ............................ 174

Deokate P.P. ...................................... 276

Deosarkar D.B. ................................. 132

Depierreux Christiane ..............................9

Desai B.K. ......................................... 264

Deshmukh Nishant Anandrao ........... 349

Deshpande S.P. ................................. 101

Dev Brihama ........................................ 315

Devadasu Elsin Raju ........................... 309

Devadiga Pooja ................................. 241

Devani Ravi ........................................ 356

Devanna B.N. ........................................ 22

Devi Ayam Gangarani ........................... 21

Devi Sarita ........................................ 177

Devmore Jalindar .............................. 324

Dey Narottam ...................................... 321

Dey Sayani ........................................ 259

Dhakar R. ................................... 127,144

Dhandapani R. .................................... 288

Dhankher Om Parkash ....................... 37

Dhanoji M.M. .............................. 131,277

Dhansu Pooja .................................... 135

Dhanyalakshmi K.H. ......................... 326

Dhar Swatismita ........................... 241,255

Dharanipragada Prashanthi .............. 233

Dharmaraj P.S. .................................... 217

Dhawan Gaurav ................................ 320

Dhepe A.P. ........................................ 311

Dhingra Anuradha .............................. 155

Dhuppar Pooja ............................. 189,191

Dilkes Brian ........................................ 75

Dinesh Rachana................................ 292

Dingkuhn M. ........................................99

Dixit Sandeep ...................................... 246

Djeghdir Inès ......................................... 9

Dkhar Jeremy ...................................... 202

Doddamani Dadakhalandar ......... 216,322

Doddamani M.B. ................................ 275

Doddaraju Pushpa .............................. 234

Dohtdong Lashaihun ........................... 323

Donald James .............................. 240,315

Dutt Som ........................................... 236

Dutta Ayan .......................................... 33

Dutta Tanima ....................................... 114

Dutta Tushar Kanti ............................. 251

Dwivedi Priyanka ............................... 320

Dwivedi S.K. ........................................ 116

Dwivedi Varun ..................................... 235

Dwivedi Vikas ...................................... 319

Ekamber Kariali ................................. 290

Elanchezhian R.................................. 268

Ellur R.K. ............................................. 74

Ewers Brent E. ..................................... 44

Fareed Mohd. Khan .............................. 67

Farhi Moran ........................................ 110

Fartyal Dhirendra .......... 77,129,239,315

Faseela P. ........................................ 303

Ferguson John ...................................... 32

Finnegan Jean....................................... 81

Forster Genevieve M. ........................ 235

Foureau Emilien ..................................... 9

Francis Aleena ............................. 203,314

Fulzele Devanand P. ........................ 139

Gadag R.N. ........................................ 169

Gadi V.K............................................ 172

Gaikwad Kishor ................................. 216

Gajjeraman Prabu ...................... 259,341

Gali S.K. ............................................ 275

Ganapathi Thumballi .................... 266,267

Gandotra Vandit ................................ 209

Ganesh S. Sree ................................ 168

Ganie Showkat Ahmad .............. 128,212

Garai Sampurna .................................. 68

Garg A. .............................................. 172

Garg Neera ... 125,134,173,179,265,266,271

Garg Rashmi ..................................... 301

Garg Rohini ........................... 111,211,212

Garg Vanika .............................. 216,217

Garggi G. ........................................... 226

Gaur Asha .................................. 262,271

Gaur Pooran M. .................... 215,322,323

Gaur Rashmi ....................... 220,328,344

Gautam Anuradha ........................ 74,297

Gautam Janesh Kumar ..................... 320

Gautam Manisha ................................ 171

Gayatri Gunja .............................. 187,188

Geeta R. ............................................ 218

Geetha Gouribidanur A. .................... 106

Gera Rajesh ........................................ 261

Ghadge Amita ................................... 356

Ghangal Rajesh ..................................... 28

Ghanta Anuradha .............................. 217

Gharpure S. ........................................ 102

Ghate Tejashree ............................... 189

Ghazi Irfan A. .................................... 235

Ghosh Ajit ........................................... 64

Ghosh Ananta Kumar .......................... 150

Ghosh Daipayan ........................ 175,177

Ghosh Indira ................................. 91,233

Ghosh Jyotirmoy ............................... 331

Ghosh Shraboni ................................ 137

Ghosh Sourav ..................................... 176

Ghosh Srayan .............................. 243,248

Ghosh Zhumur .................................. 259

Gill Kulvinder Singh .......................... 318

Giri Jitender ........................... 250,263,313

Giri Mrunmay ....................................... 320

Glévarec Gaëlle .................................... 9

Gnanaraj Muniraj ................................ 291

Gnanasekaran Prabu .......................... 245

Godara O.P. ........................................ 144

Goel Parul ...................................... 57,273

Goel Ridhi .......................................... 299

Goel Shailendra ............................. 51,176

Author Index

362

IPPCongress2015


Gohain Bornali .................................. 258

Gohara Elizabeth ................................ 90

Gokhale Nitin ....................................... 284

Golakiya B.A. ........................................ 42

Gopalakrishnan S............................... 272

Goplakrishnamurthy T. ...................... 124

Goswami Nisha .............................. 72,159

Govindasamy V. ................................ 120

Govindjee............................................ 86

Greene Thomas W. .............................. 79

Greenham Kathleen .............................. 44

Grover Anil .................................... 25,155

Grover Khulbhushan ........................... 149

Grover Minakshi ................................. 177

Guerinot Mary Lou .............................. 87

Gujjarlapudi Mariyamma ..................... 252

Gulabani Hitika ................................. 256

Gulati Ranju ........................................ 179

Gunjan............................................... 331

Gupta Aarti .......................................... 246

Gupta Aditi ................................. 280,296

Gupta Anil .......................................... 149

Gupta Asmita ....................................... 301

Gupta Brijesh K............................... 27,121

Gupta Deepti ..................................... 196

Gupta H.S. ...................................... 53,274

Gupta Maitrayee Das .......................... 33

Gupta Megha ....................................... 257

Gupta N.K. ................................. 273,357

Gupta Nakul ........................................ 230

Gupta Parul .................................. 317,318

Gupta Prakash Chandra .................... 279

Gupta Priya ................................ 222,249

Gupta Priyanka ............................ 119,188

Gupta Raj Kumar .............................. 346

Gupta Rajendra ................................. 285

Gupta Ramneek .................................. 51

Gupta Shrish C. ................................... 285

Gupta V.K. ............................ 126,335,338

Gupte Yash ........................................ 241

Gurrapu Ramya ................................... 233

Habib Farhat ........................................ 189

Hairat Suboot ...................................... 290

Haldankar Parag ................................. 306

Haloi Meghna ..................................... 349

Harikrishna ........................................ 112

Hariprasanna K. .................................. 130

Harish M............................................. 131

Harishukumar P. .............................. 267

Harit R.C. .......................................... 277

Haritha B. ............................................ 74

Hashimoto Kenji .................................... 34

Hassan Mohd ...................................... 240

Hebbar K.B. ....................................... 100

Helliwell Christopher .............................. 81

Hemamalini P. ................................... 343

Hemantaranjan A. ...................... 115,286

Héricourt François ................................ 9

Hermand Victor ..................................... 44

Hickey John M. ................................. 323

Hirt Heribert .......................................... 5

Hooley Richard ...................................... 55

Hossain Firoz ............................... 53,274

Hummel Grégoire .............................. 313

Hussain Aubid ..................................... 224

Ichihashi Yasunori .............................. 110

Igor Chernukhin .................................. 69

Imam Muhammad Ali ........................ 342

Ingole Kishor ....................................... 258

Iqbal M................................................. 309

Islam Md Rofiqul .................................. 251

Ismail Abdelbagi M. .......................... 156

Itabashi Etsuko ...................................... 73

Iyer Ganesh ....................................... 241

Jack Matthews ....................................... 69

Jadon K.S. ......................................... 302

Jadon Kuldeep Singh ............................ 42

Jagadis Kapuganti ..............................194

Jagannath Arun ............................ 51,176

Jain A. .................................................... 52

Jain Ajay ........................................... 272

Jain Ankit ....................................... 28,215

Jain Mukesh ............................ 24,211,212

Jain Neelu ............................ 112,178,262

Jain Nitin ....................................... 162,193

Jain Priyanka ....................................... 330

Jain Radha ..................... 155,222,262,271

Jainender ........................................... 135

Jaldhani V. ................................ 321,307

Jaleel Hassan ...................................... 291

Jalmi Siddhi ....................................... 244

James Beynon ..................................... 69

James Donald ............................ 129,239

Jangir Monikam ................................... 342

Jannink Jean-Luc ................................ 323

Janssens Ivan A. ................................. 110

Jayaswall Kuldip ............................... 325

Jerez Ivone Torres ...............................199

Jethva Minesh .............................. 94,335

Jeyakumar P. ........................ 282,292,295

Jeyapriya S. ....................................... 308

Jez Joseph ........................................... 58

Jha Aditya Nath ................................... 268

Jha Bhavanath .................................. 232

Jha Gopaljee ......................... 243,245,248

Jha Gulshan ....................................... 62

Jha S.K. ....................................... 275,335

Jha Sarita ........................................... 215

Jha Saroj K. ....................................... 113

Jha Shweta ....................................... 250

Jhansi K. ............................................ 311

Jian Jianbo ....................................... .322

JoãoAraújo Jr, ..................................... 304

Jodder Jayanti ..................................... 259

Johal Gurmukh S. ............................... 75

Jondhale Ashish .................................. 324

Jorge Franco....................................... 318

José Crossa....................................... 318

Joshi Alka ......................................... 236

Joshi Babita ....................................... 263

Joshi Chandrashekhar P. .................... 214

Joshi K.A. .......................................... 311

Joshi Raj Kumar ........................... 238,312

Joshi Rohit ..................................... 27,281

Juan Burgueno Ferrera........................ 318

Juergens Gerd ..................................... 201

Julia C. .................................................. 99

Jung Chol-Hee ...................................... 80

Kaiser Werner ..................................... 194

Kakani R.K. ....................................... 170

Kakralya B.L. ....................................... 144

Kalariya K.A. ....................................... 72

Kale Manisha ....................................... 231

Kale Sandip .................................. 215,217

Kalia Rajwant K. .................................. 283

Kalita Lotika ....................................... 349

Kalyani Tripathi .................................... 268

Kambalimath S.K................................ 234

Kamble Nitin ....................................... 351

Kannababu N. ..................................... 130

Kannan K. ......................................... 137

Kant Chandra ............................... 200,203

Kantiwal Usha ..................................... 220

Kanwar Poonam ..................... 15,243,248

Author Index

363

IPPCongress2015


Kanyuka Kostya .................................. 55

Kapare Vishwadeep .......................... 139

Kapoor Meenu ................................ 35,350

Kapoor Sanjay ............ 43, 78,114,350,353

Kapuganti Jagadis ............................. 195

Kar Rup Kumar ..................... 160,213,214

Karaba Nataraja N. .............................. 45

Karan Ratna ....................................... 183

Kariali Ekamber ................................... 293

Karmakar Kanika ................................. 356

Karn Rajni .......................................... 315

Karuppanapandian Thirupathi ........... 291

Karuputhula N.B.................................. 233

Karwa Sourabh ................................. 159

kashyap Jyoti ....................................... 342

Kashyap Lakita ................................... 265

Katiyar Deepmala ................................ 286

Kaul Tanushri .............................. 77,264

Kaul Veenu ........................................ 107

Kaur Amarjot ....................................... 333

Kaur Charanpreet .......................... 64,112

Kaur Gundeep ..................................... 114

Kaur Harsimran .......................... 114,332

Kaur Kanwaljeet ................................ 191

Kaur Navdeep .................................... 167

Kaur Parmeet ...................................... 123

Kaur Preeti ....................................... 164

Kaur Pritam ......................................... 51

Kaur Ranjeet ....................................... 150

Kaur Tarandeep .......................... 113,347

Kaushal Jyoti ....................................... 297

Kav Nataraj ....................................... 312

Kavya Y. ............................................ 102

Kaya Hidetaka ....................................... 34

Kelkar Kaustubh .................................. 306

Keller Beat ........................................ 243

Kemisetti Siva Raju .......................... 267

Kevat Nisha ....................................... 167

Khajuria Ankush ................................ 354

Khan Aamir ....................................... 217

Khan Aamir W. ................................... 216

Khan Aarif .................................. 126,357

Khan Akhtar Hussain .......................... 156

Khan F.A. ............................................ 270

Khan Fareed ....................................... 230

Khan M. Iqbal R. .................................. 279

Khan M. Masroor ............................... 294

Khan M. Masroor A. ...................... 291,298

Khan Nafees A. ................................ 279

Khan Sadaf ....................................... 299

Khan Zaiba Hasan ............................... 209

Khandal Hitaishi ................................ 207

Khandelwal Abha .................................. 90

Khangura Rajdeep S. .......................... 75

Khare Ria .......................................... 205

Kharshiing Gayle ................................. 322

Khatediya Nilesh .................................. 42

Khetarpal Sangeeta ..................... 145,274

Khobra Rinki ....................................... 346

Kholova Jana .................................... 313

Khurana J.P. ............ 6,138,162,186,192,

193,194,218,219,333

Khurana Paramjit...........19,138,147,219,

223,290

Khurana Priyanka .............................. 220

Khurana Ridhi ...................................... 353

Kientz Marika ....................................... 201

Kiep Victoria ....................................... 250

Kim Boem-Gi ....................................... 15

Kiran B.O. ......................................... 264

Kiran K.R. ......................................... 195

Kiranmai K. ....................................... 133

Kirti P.B. ............................................ 180

Kitahata Nobutaka ................................. 34

Kohli Ajay ............................................ 23

Kondamudi Rajesh ............................. 307

Kondhare Kirtikumar .......................... 189

Koppad Appanna V. .......................... 334

Korasick David ....................................... 58

Kotvi Payal ........................................ 298

Krattinger Simon .................................. 243

Krishna Bal .......................................... 331

Krishna G...................................... 92,343

Krishna G.K. .......................................171

Krishnamurthy S. .............................. 314

Krishnamurthy S.L. 20,29,158,161,165,173

Krishnan P. ....................................... 344

Krishnan S. Gopala ............... 74,230,320

Krishnan Victor Banerjee .................. 344

Krishnapriya V.............................. 269,274

Kryukov Lavr ....................................... 60

Kuchitsu Kazuyuki ......................... 34,187

Kudapa Himabindu ............................ 216

Kudla Jörg ........................................... 15

Kujur Alice .......................................... 104

Kulkarni Mahesh .......................... 85,306

Kulshreshtha Neeraj ..................... 29,161

Kumar Abhishek ............................... 262

Kumar Amar ................................. 51,176

Kumar Ambarish ............................... 340

Kumar Amish ....................................... 336

Kumar Amit ....................................... 189

Kumar Anil ........................................ 240

Kumar Arun ....................................... 112

Kumar Ashish ....................................... 28

Kumar Ashwani .......................... 135,316

Kumar Atul ....................................... .118

Kumar Bhumesh ............................... 345

Kumar Dameshwar ............................ 268

Kumar Devendra ............................... 126

Kumar Dinesh .................................... 145

Kumar G. Vijaya ................................. 151

Kumar Gulshan ................................. 210

Kumar H.G. Jalendra .............30,234,327

Kumar Jitender ........................... 118,260

Kumar K. Sumanth ............. 180,327,329

Kumar Kamal ....................... 221,234,258

Kumar Krishan ................................... 177

Kumar Kundan .................................. 162

Kumar Lokesh ................................... 302

Kumar M.B. Arun ................................. 230

Kumar M.V. Mohan ........................ 30,327

Kumar Mahesh .................................... 120

Kumar Mukesh ............................. 12,253

Kumar Mukul ..................................... 279

Kumar Muthappa Senthil............. 246,247

Kumar Narendra ................... 127,128,302

Kumar Neeraj ............................. 177,178

Kumar Pramod ...................... 148,302,316

Kumar Praveen ................................. 157

Kumar Rajeev .............. 235,245,335,338

Kumar Rajnish ................................... 344

Kumar Rakesh ....................................... 92

Kumar Ravi ....................................... 315

Kumar Reddy Kishore.......................... 245

Kumar Roshan .................................... 190

Kumar S. Rajesh ................................. 196

Kumar Sanjay ....................................... 2

Kumar Santosh ................................... 116

Kumar Satish ....................................... 177

Kumar Shiwesh ................................... 101

Kumar Smita ....................................... 205

Kumar Sudhir ............................. 142,147

Kumar Sumit .............................. 231,303

Author Index

364

IPPCongress2015


Kumar Sundeep ............................... 165

Kumar V. ............................................ 277

Kumar V.V. Santosh ............. 16,146,157,

164,171,172,189,191,282,324

Kumar Vandna A. ............................. 240

Kumar Verandra .................................. 210

Kumar Vijay ....................................... 278

Kumar Vinay ....................................... 217

Kumar Vinod .................................. 20,300

Kumar Krishan ................................... 178

Kumara K. Sumanth .......................... 197

Kumara P. Mohana............................ 114

Kumari A.P. Padma ............................ 63

Kumari Anita ....................................... 135

Kumari Archana ................................... 330

Kumari Asha ....................................... 181

Kumari B. Ramana ............................ 321

Kumari Chanchal ................................. 251

Kumari Jyoti ....................................... 165

Kumari Poonam ................................. 260

Kumari Punam .................................... 253

Kumria Ratna ....................................... 79

Kundu Pallob ................................ 246,259

Kundu Siddhartha ............................. 339

Kuriakose Saritha V. .......................... 90

Kushwaha Hemant R. .................... 94,335

Kushwaha N.K.............................. 49,245

Kushwaha S.R. ..................... 172,282,296

Lafarge Tanguy ................................... 99

Laha Debabrata ................................ 109

Lakhwani Deepika ....................... 205,317

Lakra Nita .................................... 122,226

Lakshmamma P. .......................... 142,204

Lakshmi N. Jyothi ......................... 135,151

Lal J.P. ............................................... 118

Lal Milan Kumar ................................... 274

Lal P. ................................................ 262

Lalitha V.S.P. ....................................... 311

Lande Nilesh ....................................... 207

Larcher Mélanie ..................................... 9

Lata Charu ....................................... .125

Lavania Dhruv .................................... 155

Lavanya C........................................... 145

Lavekar Sailee ..................................... 241

Laxman R.H. ....................................... 343

Laxmi Ashverya .....124,186,204,280,296

Leach Jan E. ..................................... 235

Lee Sung-Chul ..................................... 15

Lekshmy S. ................................ 275,324

Liew Lim Chee ....................................... 80

Lohot Vaibhav D. .............................. 331

Lokesh U. .......................................... 133

Lokya Vadthya ..................................... 252

Long Stephanie ................................... 26

Lorenz Aaron ....................................... 323

Lou Ping ................................................ 44

Louis Joe............................................. 47

Luan Sheng ....................................... 4,15

Luiz Fernando Almeida...................... 304

Lymm Andrew .................................... 340

Lynn Andrew .............................. 202,220

Macwan Sunil J. ............................... 288

Madireddi S.K...................................... 309

Mahadik Sandip ................................ 284

Mahajan Ameya ................................. 189

Mahajan Gulshan .............................. 123

Mahatma Lalit ..................................... 168

Mahatma M.K. ........................ 42,255,302

Mahato K.K. ....................................... 195

Mahesh S. ......................................... 102

Mahesh Shashank ............................. 316

Maheswari M. .......................... 96,135,151

Maheswari Sellakumar ....................... 291

Mahindra Anuradha ............................ 90

Mahmood Saquib ............................... 253

Mahto Binod Kumar .................... 241,255

Majee Manoj ................. 137,138,351,352

Maji Ranjan Kumar ............................ 259

Maji Sourobh ....................................... 192

Majumdar Arkajo .............................. 214

Majumdar Rajtilak ................................. 8

Majumder Arun Lahiri .......................... 10

Malick N. ............................................. 43

Malik Aubid ....................................... 223

Malik Naveen ....................................... 353

Mallikarjun N.M. ................................... 327

Mallikarjuna Kuruva ..........................306

Mallikarjuna M.G........................... 53,274

Malpathak N.P. ................................. 146

Malpathak Nutan .......................... 139,228

Mamrutha H.M. .......................... 140,346

Mamta .................................................. 48

Manashi Aditya ................................. 227

Mandal Arunava .......................... 246,259

Mandal K.G. ............................... 137,346

Mane Arun .................................... 85,284

Manga V.K. ........................................ 348

Mangrauthia S.K. .............................. 307

Manivannan A. ................................. 348

Manjunath Manju .................................. 76

Mann Anita ....................................... 269

Manna Mrinalini ..................... 77,239,315

Manna Suman ..................................... 347

Mannully C.T. ............................ 186,204

Manoharan Kumariah ......................... 291

Maramreddy M.K............................... 233

Maria Vera Da Costa .......................... 310

Marik Ananya ...................................... 184

Masand Shikha .................................... 117

Mathew Iny ................................ 351,354

Mathew Iny Elizebeth ..........................356

Mathew Reynolds............................... 318

Mathur P.N. ....................................... 156

Mathur Saloni ................................ 12,215

Mathur Supriya .................................... 203

Maurya Sadhana .............................. 169

Max K.F. ........................................... 102

Mayee Pratiksha .................................. 332

Mayer Ulrike ....................................... 201

McClung C. Robertson ......................... 44

Meena Hari Singh .......................... ....169

Meena M.K. ....................................... .82

Meena Mukesh Kumar ....................... 277

Meena R.C. ....................................... 178

Meena Rajendra Prasad ................... 216

Meena Seema ..................................... 235

Meena Shashi ............................. 159,169

Megha Swati ....................................... 312

Mehra Poonam .......................... 263,313

Mehrotra Rajesh .......................... 205,209

Mehrotra Sandhya ....................... 205,209

Mekala Nageswara Rao ...................... 109

Meshram Jayant ................................ 153

Michael Rahul .................................... 318

Mina U. ............................................. 277

Minhas J.S. ....................................... 126

Minhas P.S. .................................. 120,345

Minocha Rakesh .............................. 8,26

Minocha Subhash ............................. 8,26

Mishra Avinash ................................ 232

Mishra B.K. ....................................... 118

Mishra Divya ....................................... 133

Mishra J.K. ....................................... 116

Mishra J.S. ....................................... 130

Author Index

365

IPPCongress2015


Mishra Kumud B. ................................ 86

Mishra Manjari .......................... 165,261

Mishra Neeti Sanan.......................... 247

Mishra P.C. ....................................... 112

Mishra Rukmini ................................. 238

Mishra Satish ....................................... 140

Mishra Sefali ....................................... 20

Mishra Sonal .............................. 195,229

Mishra Sushma ................................. 194

Mishra Toshi .............................. 220,344

Mishra Upama .................................... 169

Misra S.C. ......................................... 112

Mithöfer Axel ....................................... 250

Mithra Amitha C.R. ............................ 112

Mithra S.V. Amitha .............................. 20

Mitra Abhijit ................................... 91,233

Mittal Shikha ................................. 53,274

Mittra Sarika ....................................... 156

Mockler Todd C. .................................. 44

Mog Babli .......................................... 341

Mohammad Naseer ............................. 316

Mohan Amita ..................................... 318

Mohanasundaram A. .......................... 331

Mohanty Amitabh ................................ 79

Mohapatra P.K.................................84,293

Mohapatra Sridev ......................... 175,177

Mohapatra T. ..........................88,197,329

Mohiudin Tabassum .......................... 224

Moin Mazahar ................................... 180

Mondal Tapan Kumar ................ 128,212

Morabito Domenico ............................... 9

Mrinalini Chawla .................................. 78

Mudalkar Shalini ............................... 150

Mudgil Yashwanti ............................... 129

Muhammed Jamsheer K.............. 186,204

Mukesh Nitin................................ 220,344

Mukherjee Abhishek ............................ 10

Mukherjee Chandan .......................... 141

Mukherjee Gairik .......................... 129,184

Mukherjee Sritama .............................. 10

Mukhopadhyay Pradipto ..................... 153

Mullineaux Philp ............... 32,69,123,136

Muniswamy S. ..................................... 217

Munnam Suchandranath B. ............... 307

Mur Luis ........................................... 194

Murtaza Malik....................................... 314

Mushtaq Ruqia .................................... 285

Mustafiz Ananda ................................. 64

Muthulakshmi S. ................................. 164

Muthusamy A. ..................................... 195

Mutum Roseeta Devi .......................... 12

Naeem M. .......................................... 294

Nag T.N. ....................................... 227,228

Nagar Shivani ............................. 127,144

Nagarajan M. .......................... 74,230,320

Nagarajan V.K. ...................................... 52

Nagegowda Dinesh A. ........................ 235

Nagle Vinod ....................................... 241

Naika Mahantesha ............................ 334

Naithani S.C........................... 61,117,183

Naiya Haraprasad ............................. 184

Nakar R.N. .......................................... 72

Nalli A. ................................................. 102

Nama Srilatha ..................................... 309

Nanda Satyabrata ............................. 238

Nandhitha K. ....................................... 292

Nandi Ashis K. ...59,208,222,249,254,320

Nandwal A.S................. 168,135,177,178

Narasimha Swamy Konduri ............... 307

Narayan S. ....................................... 270

Narayankutty M.C. ............................ 143

Narendra Gupta................................ 126

Nareshkumar A. ................................ 133

Narwade Ajay V. ............................... 168

Nataraja Karaba N. .......................... 326

Natarajan S. ....................................... 288

Nath Onkar ....................................... 344

Nath Pravendra .......................... 299,301

Nathawat N.S. ................................... 120

Nautiyal C.S. .............................. 182,300

Nautiyal P.C. ......................... 147,327,343

Nayyar Harsh .......................................

105

Neeraj Kumar.......................................168

Neeraja C.N. ................................. 54,321

Negi Bhawna ....................................... 166

Negi Manisha ..................................... 272

Negi Neelam Prabha ..........................148

Ngachan Shishom Vanao .................. 349

Nijhawan Aashima ............................. 333

Nikalje Ganesh .................................... 202

Nikam T.D. ................................ 202,293

Nirala Ramchiary.................................. 344

Niranjana P......................................... 329

Nisarga K.N. ........................................ 198

Nithia S. Jannita .................................. 304

Nitin Mukesh ....................................... 328

Nivedita D. ....................................... 141

Nogia Panchsheela ........................... 205

Nongpiur Ramsong .......................... 183

Noor Jewel Jameeta .......................... 256

Nutan Kamlesh Kant .................. 111,333

Oudin Audrey ........................................ 9

Padaria Jasdeep Chatrath ................ 216

Padmasree Kollipara .................. 252,253

Pakrasi Himadri B. .............................. 89

Pal Madan ........... 98,148,159,169,174,

216,302,320

Pal Shaifali ......................................... 235

Pal Tarun .......................................... 337

Palta J.A. ............................................. 95

Pan Yong-Bao ..................................... 222

Panda Ashok ....................................... 181

Panda Binay ....................................... 84

Panda S.K. ....................................... 163

Pandey A. .......................................... 175

Pandey Amita ....................................... 15

Pandey Ashutosh ......................... 206,317

Pandey Bipin ............................. 263,313

Pandey Daya K. ................................ 238

Pandey Dinesh .................................... 240

Pandey Garima ................................. 206

Pandey Girdhar K. .......................... 15,113

Pandey Prachi ................................... 247

Pandey Rakesh ............. 178,233,335,338

Pandey Renu ....................... 269,274,278

Pandey S. .......................................... 102

Pandey S.P. ....................................... 301

Pandey Saurabh ............................... 200

Pandey Sona ....................................... 18

Pandey Sonika ................................. 232

Pandey Veena ..................................... 118

Pandey Vinod ..................................... 178

Pandiyan M. ....................................... 130

Pandurangam Vijai ............................ 166

Pangaan Iris Dawn ........................... 103

Panwar Akanksha ........................ 351,356

Panwar Azad Singh .......................... 349

Paramanantham Anjugam ................ 247

Parashar Renu ................................. 287

Paraste Kamlesh .............................. 345

Pareek Ashwani .............11,27,111,

114,116,1 19,121,122,136,165,183,184,

188,199,202,226,261,281,314,323

Author Index

366

IPPCongress2015


Pareek Sneh L. Singla .............11,27,64,

111,112,116,119,121,122,136,183,184,

188,199,226,261,281

Parekh Nita .................................. 91,233

Parida Adwaita .................................. 201

Parida Asish Kumar ............................. 181

Parida S.K. ....................... 43,104,319,355

Parihar Vimala ..................................... 35

Park Se Won ....................................... 115

Parmar Hemangini ............................. 297

Parmar Rajni ....................................... 325

Parry Martin A.J. ................................. 66

Partheeban C. .................................... 130

Parupalli Swathi ................................ 217

Parveen Shaista ................................ 131

Pasala Ratnakumar ..................... 276,345

Patel C.B. .......................................... 269

Patel Kiran ........................................ 159

Patel Manish Kumar .......................... 232

Patel Prashanti Devendra ................. 266

Pathak A.D......................................... 155

Pathak Garima .................................... 299

Pathak H. ....................... 145,169,277,278

Pathak Ravi ........................................ 55

Pathak Sumya ..................................... 317

Pathania Shivalika .............................. 57

Pati Pratap Kumar ............................ 167

Patil Amol .......................................... 135

Patil Hemant ....................................... 331

Patil Mahesh ....................................... 224

Patil R.P. ............................................ 131

Patil Supriya ....................................... 264

Patil Suyash ....................................... 217

Patil Vidya ......................................... 146

Patra Ashok ....................................... 268

Pattiwala Yasir Umar .................. 259,341

Paul Asosii ........................................ 215

Paul Priyanka ...................................... 123

Paul Vijay ....................... 178,233,335,338

Pazhamala Lekha ........................ 216,217

Pednekar Suhas ................................ 241

Penna Suprasanna ............................ 202

Peter Wenzl....................................... 318

Petla Bhanu ....................................... 351

Phazang Paomipem .......................... 123

Phukan Ujjal J. .................................. 229

Pieter Edgar ....................................... 250

Pillai M. Arumugam .......................... 326

Pitagi Anita ....................................... 334

Podile Appa Rao .................................. 187

Pooja B. ............................................ 196

Poolman Mark ....................................... 65

Poonia Vijay ....................................... 272

Poonia Shefali .................................... 171

Prabhu K.V. ................................ 112,142

Pradeep T. ....................................... 114

Prasad Ch. Ram ............................... 135

Prasad Manoj ................. 56,206,209,244

Prasad P.V.V. ..................................... 130

Prasad S.R. ....................................... 288

Prasad Sai ....................................... 140

Prasad Subhash Chandra ................ 136

Prasad T.G. ................................ 224,341

Prashant Vikram ............................... 318

Prathibha M.D. ................ 30,234,327,329

Pratibha M. ........................................ 196

Prayaga Lakshmi ........................ 142,204

Preethi N.V. ....................................... 180

Priyanka A.B. .............................. 196,234

Pudi Vikram .................................. 91,233

Puli Mallikarjuna Rao .......................... 187

Pundir P. ................................ 29,158,161

Punitha S. ......................................... 282

Punyapwar Swapnil .......................... 162

Purayannur Savithri ............. .......221,234

Purty Ram Singh ............................... 268

Purushottam....................................... 171

Pushkar Suchitra .............................. 172

Pushpa D. ................................... 196,327

Puskar Suchitra .......................... 146,296

Puthur Jos ......................................... 303

Rachel Jacob Sherry .......................... 230

Radadiya Nidhi .................................... 302

Radha Beena ...................................... 143

Raghava Nisha ................................. 237

Raghava Ravindra Pratap ................. 237

Raghavendra A.S. ............. 67,187,188,

229,230,232,305,306

Raghothama K.G. ................................ 52

Raghuram Nandula ...................... 55,191

Raghuvanshi Saurabh .......................... 12

Raghuvanshi Utkarsh .......................... 12

Raghuveer Rao Puskur........................ 307

Rai Krishan Mohan ............................ 210

Rai Neha ........................................... 178

Rai Puja ............................... 147,327,343

Rai Ramakant ..................................... 222

Rai Vandna ............................ 20,272,312

Rai Yogita .......................................... 196

Raigond Pinky ..................................... 236

Raina Aparna Shree .......................... 325

Raina Susheel ..................................... 120

Raj A. ................................................ 277

Raj Lakshmi ....................................... 274

Raj S.R. Kiran ...................................... 233

Rajabhoj Mohit .................................. 189

Rajam M.V.............................. 48,248,289

Rajan S. ............................................ 158

Rajanna M.P. ............ 30,197,327,328,329

Rajeev Ranjan .................................. 352

Raju B.R. .. 30,180,196,327,328,329,334

Raju M.B. ........................................... 198

Rakesh K. ............................................ 146

Rakesh Verma .................................. 164

Ram Phool Chand ............................. 156

Ramachandra R. .......................... 180,328

Ramakrishnan Shiv............................ 144

Ramchiary Nirala .......................... 220,328

Ramegouda H.V. ................................ 45

Ramesh Kulasekaran .......................... 268

Ramesha............................................ 102

Ramteke S.D. ...................................... 295

Ramu S.V. ...................................... 45,198

Rana Dheeraj ...................................... 140

Ranawana S.R.W.M.C.J.K. ................. 95

Rane Jagadish .................................. 120

Rangani Jaykumar ............................. 181

Rangappa Krishnappa ....................... 349

Rani P. Prasuna ................................ 174

Rani Y. Ashoka .................................... 174

Ranjan Aashish ................................... 110

Rao A.R. ..................................... 112,216

Rao Ch. Srinivasa .............................. 135

Rao G. Lokanadha .............................. 13

Rao I. Subhakara ................................ 54

Rao K.L. Narasimha .......................... 174

Rao K.V. ............................................ 180

Rao M.V. ...................................... 146,157

Rao P.R. ....................................... 63,321

Rao S.S. ........................................... 130

Rao Sombir ....................................... 215

Rao T.G.N. ....................................... 130

Rao Uma ............................................ 251

Ratcliffe R. .......................................... 194

Author Index

367

IPPCongress2015


Rathanakumar A.L. .......................... 302

Rathi Divya ....................................... 152

Rathi Iti ....................................... .......287

Rathod Khyati J. .................................... 42

Rathod T.H. .......................... .............152

Rathor Ray Singh ............................. 116

Rathore Abhishek ........................ 322,323

Rathore M. ....................................... 345

Rathore Ray Singh .............................. 11

Rathore S.S. ....................................... 170

Rathore V.S. ....................................... 120

Raveendran M. .................... 163,164,329

Ravi Iyyakutty .................................... 333

Ravichandran V. .......................... 154,308

Ravikanth G. ....................................... 114

Ravikiran K.T. ...................................... 173

Ravindran Nevedha .......................... 189

Rawal S. ............................................ 126

Raxwal Vivek Kumar .......................... 176

Ray Krishna ....................................... 141

Raychaudhuri S. ............................... 346

Raza S.K. .......................................... 242

Reddy A.R.150,151,180,231,307, 303,308

Reddy C.S.......................................... 264

Reddy Chandrashekar ...................... 224

Reddy D. Damodar ........................... 267

Reddy Kanubothula Sita Rami.............151

Reddy M.K...77,129,200,225,239,240,315

Reddy V. Amarnatha .......................... 133

Reddy V.R. ....................................... ...97

Reddy Vijay ................................. 240,315

Reema Khurana................................. 352

Rey Soham .......................................... 22

Rice Elena........................................... 76

Richter Sandra ................................... 201

Robertson Masumi .............................. 81

Robin S. .............................. 197,308,329

Roopendra Kriti ................................. 222

Roorkiwal Manish ................ 215,322,323

Rout Ellojita ....................................... 238

Roy Amit ............................................ 246

Roy Shweta ................................ 222,249

Roy Suchismita .......................... 119,314

Roy Suman .......................................132

Roy Tapas K........................................ 106

Ruhil Kamal ....................................... 309

Ryan Elizabeth P. .............................. 235

Rydquist Shane W. ........................... 218

Sablok Gaurav ..................................... 202

Sacaria Vinod ...................................... 176

Sadashiva A.T. .................................... 343

Saeed Bushra ..................................... 223

Saha Chinmay ..................................... 184

Saha Jayeeta ...................................... 225

Saha Sudip .......................................... 33

Saha Sunayan..................................... 345

Sahi Shivendra .................................... 272

Sahoo K.K. ............. 27,64,121, 281, 289

Sahoo L. ........................................... 172

Sahoo R.N. ............. 92,118,335,338,343

Sahu Alok Kumar ............................. 183

Sahu Balram ....................................... 117

Sahu Pranav Pankaj ............................ 244

Sahu Sarika ....................................... 216

Sai Prasad S.V. ................................... 287

Sailaja B. ............................. 307,339,349

Sain R.S. ............................. 126,273,357

Saini Niveditha .................................... 112

Sairam R.K. ............................... 142,275

Sajeevan R.S. .................................... 326

Sakai Hajime ....................................... 79

Saketi Ananda Rao............................ 314

Sakhare Akshay .................... 142,147,327

Salini K. ............................................. 135

Salvi Prafull ................................ 138,352

Samanta Subhasis ............................ 213

Sameer C. ......................................... 217

Samundeswari R. ......................... 163,164

Sanagala Raghavendrarao ............... 272

Sane A. .............................................. 331

Sane A.P. .............................. 299,300,301

Sane P.V. ......................................... 331

Sane V.A. ........................... 182,299,300

Sangita Sushree ............................... 203

Sangwan Chetna .............................. 209

Sangwan Rajender Singh .................... 38

Sanyal Sibaji K. ............................ 15,113

Sarada C. ......................................... 204

Saradhi A.V. Pardha .......................... 102

Sarala N. ...................................... 197,329

Saraswati Nayar ................................. 78

Sarathambal C. ................................... 345

Sardar Atish ....................................... 252

Sarin Neera Bhalla .......123,136,148,253

Sarkar Arijita ....................................... 259

Sarkar B. ..................................... 135,151

Sarkar Deepti ..................................... 259

Sarkar Poulami ................................. 237

Sarla N............................................... 307

Sarma Anjan Kumar ......................... 349

Sarma C.M. ....................................... 176

Saroj P.L. ........................................... 341

Saroop Shveta .................................... 107

Sarpras M. ........................... 220,328,344

Sastry P.S. ........................................... 231

Satdive Ramesh .................................. 139

Sathish P. .................................... 135,151

Satyamoorthy K. .................................. 195

Satyavathi C. Tara ............................ 216

Savita ................................................ 349

Sawant S.V........................................ 210

Saxena A. .......................................... 348

Saxena D.C. ............................... 140,287

Saxena Rachit K. .......................... 217,322

Saxena S.N. ....................................... 170

Schaaf Gabriel .................................. 109

Scheel Dierk ........................................ 31

Scheffler Brian .................................... 199

Schnell Danny ....................................... 37

Scholz Sandra .................................... 250

Seal Anindita ................................ 129,184

Sekhar K.M........................................ 308

Sekhar Sudhanshu ............................ 247

Sekhwal Manoj Kumar ........................ 233

Selter Liselotte .................................. 243

Semwal D.P. ....................................... 156

Semwal Vimal ................................... 134

Sen Soumitra Kumar .......................... 150

Sen Sunetra ....................................... 177

Senapati Dhirodatta ............................ 13

Sengupta Sonali .................................... 10

Senthil A. .......................................... 282

Seth Romit ....................................... 325

Shaanker R. Uma .............................. 114

Shah Diti ........................................... 177

Shah Divya .................................... 98,159

Shamim M.Z. .................................... 175

Shamkuwar G.R. ............................... 152

Shankar Rama .................................... 211

Shankar Ravi ....................................... 57

Shanker Arun K. ................................. 135

Shankhdha Shailesh Chandra ...........263

Shankhdhar Deepti .............. 127,128,263

Shankhdhar S.C. ..........................127,128

Author Index

368

IPPCongress2015


Shanthi N. ........................................... 304

Sharan Ashutosh .......................... 183,184

Sharma Abhishek ............................... 271

Sharma Amrapali ................................ 201

Sharma Amrita .................................. 271

Sharma Arun ..................................... 201

Sharma Chhavi .................................. 289

Sharma D.K. .......................... 29,158,161

Sharma Deepika ................. 127,128,205

Sharma Eshan .................................. 186

Sharma Gaurav ................................... 233

Sharma Indu ............................... 140,346

Sharma Isha ....................................... 167

Sharma Kamal Dev............................ 105

Sharma Kamal Dutt .......................... 261

Sharma Kewal K. ............................... 331

Sharma Laxmi .................................. 282

Sharma Madhu ................................... 289

Sharma Mamta ................................. 217

Sharma Manoj K. ................ 203,314,342

Sharma Manvi ................................... 204

Sharma Meenakshi ........................... 250

Sharma Mohan A. .............................. 124

Sharma Namisha ............................... 244

Sharma Namrata ............................... 355

Sharma Naveen ................................ 219

Sharma P.C. ................................. 158,161

Sharma P.K. ............................... 167,310

Sharma Parbodh C. ........ 29,161,165,173

Sharma Prakash C. ............................. 28

Sharma R. ................................. 283,348

Sharma R.P. ............................... 197,329

Sharma Ram Kumar ............210,221,325

Sharma Rameshwar Prasad ................ 3

Sharma Rinku ............................... 53,274

Sharma Rita .................. 203,314,339,342

Sharma Sadan .................................. 208

Sharma Shivam ................................. 298

Sharma Shweta ..................................... 64

Sharma Surinder Kumar ................... 170

Sharma T.R. ................................. 22,330

Sharma Vasundhara .......................... 270

Sharma Veerendra Kumar ................ 245

Sharma Vijay K. .................................. 76

Sharma Vinay .................................. 148

Sharma Vineet .................... 220,328,344

Sharma Vishakha ............................... 16

Shasany Ajit Kumar .......................... 235

Shashibhushan N.B. .......................... 334

Sheeba............................................... 309

Sheeba J. Annie ............................... 153

Shekhar Shubhendu .......................... 133

Shekharappa Gurumurthy ................ 143

Shembekar W.K. ................................ 152

Sheoran Promila ............................... 220

Sheoran Sonia .................................. 140

Sheri Vijay ...................................... 77,225

Sherp Ashley ...................................... 58

Sheshshayee M.S......... 30,180,196,197,

234,327,328,329

Shine M.B. ......................................... 178

Shiriga Kaliyugam .......................... 53,274

Shirokov Aleksandr.............................. 60

Shivaraj S.M. ....................................... 332

Shivashankara K.S. .......................... 106

Shome S. ............................................ 163

Shrivastava Madhup .......................... 276

Shrivastava Sanya ......... 16,146,189,191

Shrivastva Ruchi Nag .......................... 227

Shukla Alok ................................ 118,263

Shukla Brihaspati N. .......................... 124

Shukla Neha ....................................... 51

Shukla Rakesh K. ............................... 229

Shukla S.K. ................................ 262,271

Shukla Tapsi ....................................... 205

Shukla Tushita ........................... 286,287

Shukla Yogesh M. ............................. 288

Siddappa Sundaresha ........................ 189

Siddiqi E.A. ....................................... 180

Siddiqi I. ........................................... 102

Siddique K.H.M. .................................. 95

Sidhu Gurpreet Kaur .......................... 205

Sikdar Samir Ranjan .......................... 237

Silas Wungrampha ............................ 199

Silvester Naveen ................................. 90

Sindhuja Vajravel............................... 291

Sindhumol P. ....................................... 143

Singal Rajesh Kumar ................. 197,329

Singh A. ............................................. 102

Singh A.K. ............ 43,74,197,273,320,329

Singh A.L. ............................. 72,159,269

Singh Ajay ........................................ 120

Singh Amar Pal ................................ 300

Singh Amarjeet ...................................... 15

Singh Amit Kumar ............................. 165

Singh Anandita ................................... 332

Singh Anil .......................................... 210

Singh Anil Kumar .................................. 57

Singh Anoop ....................................... 220

Singh Anshu .......................... 155,262,271

Singh Anuj .........................................156

Singh Anup Kumar ........................... 235

Singh Anupriya ................................. 249

Singh Ashok K. ...................................... 20

Singh Atar ...................... 178,233,335,338

Singh Balwant ....................................... 20

Singh Bharti ....................................... 286

Singh Bhupinder .................................. 270

Singh Brajesh .............................. 126,236

Singh Brinderjit ................................. 138

Singh D.K. ......................................... 102

Singh D.V. ......................................... 296

Singh Deepika..................................... 300

Singh Deepjyoti ................................... 208

Singh Dhriti ........................................ 186

Singh Dipali ......................................... 65

Singh G.P. ................................. 112,178

Singh Gagandeep ................ 210,221,325

Singh Geeta ....................................... 181

Singh Harpreet .................................... 114

Singh Ishwar ............................... 174,297

Singh Jagdev ....................................... 261

Singh Jagdish ..................................... 143

Singh Jogendra ............................. 29,161

Singh Karmal ....................................... 261

Singh Khangembam .......................... 213

Singh Kushal ....................................... 332

Singh Manjul .............................. 280,296

Singh Manoj Kumar .......................... 201

Singh Mohan ..................................... 272

Singh Mohan B. ................................ 73,80

Singh Muneendra ............................. 323

Singh N.K. ......................................... 112

Singh N.P. ......................................... 143

Singh Nagendra K. ............................... 20

Singh Name ....................................... 126

Singh Namita ....................................... 337

Singh Narendra ................................... 322

Singh Narpinder ................................ 123

Singh Navneet ................................... 191

Singh Neer ....................................... 319

Singh Neera .......................... 127,144,347

Singh Nidhi ................................. 249,254

Singh P.K. ......................................... 112

Author Index

369

IPPCongress2015


Singh Pawan Kumar .......................... 318

Singh Prabhjeet .......................... 114,138

Singh Preeti ....................................... 166

Singh Priya ....................................... 300

Singh R. ............................................ 345

Singh R.K.............................. 155,262,271

Singh Rakesh ...................................... 165

Singh Rakesh K. ............................... 103

Singh Ravindra Pal ............................ 357

Singh Renu ....................................... 278

Singh S. ............................................ 116

Singh S.K. ......................................... 116

Singh S.P. ............................ 155,262,271

Singh Sandeep .................................. 266

Singh Shardendu K. ............................. 97

Singh Shreenivas K. .......................... 234

Singh Smita .......................... 155,262,271

Singh Sudhanshu ............................. 156

Singh Sukhbir ................................... 149

Singh Sukhwinder .............................. 318

Singh Sunil Kumar ............................. 210

Singh Supreet ................................... 114

Singh Surendra Pratap ..................... 210

Singh V.K. ......................................... 158

Singh V.P. ............. 127,132,144,286,347

Singh Vijayata ................................... 208

Singh Vijendra .................................... 165

Singh Vikas K. ................................... 217

Singh Vikash ....................................... 212

Singh Vinay....................................... 342

Singh Yogeshwar ............................... 345

Singla Priyanka ................................. 125

Sinha Alok Krishna .................14,185,244

Sinha Neelima ..................................... 110

Sinha Pallavi ....................................... 217

Sinha Sangram ................................... 356

Sinha Somya ....................................... 176

Sinha Veena ....................................... 247

Sinharoy Senjuti .................................. 199

Sirhindi Geetika .......................... 179,285

Sivakumar R. ............................... 154,292

Sivaprakasam J.V.............................. 295

Sivaraju K. ......................................... 124

Skottke Kyle ....................................... 90

Slewinski Tom ....................................... 76

Smitharani J. ....................................... 327

Smitharani J.A. .................................... 329

Snedden W.A. .................................... 114

Somasundaram G. ............................ 288

Soni Dharamvir ................................. 178

Soni Praveen ...................... 183,184,188

Soni Sweta ....................................... 220

Sonti Ramesh V. ................................. 46

Sood Hemant .................................... 298

Sopory S.K. .......................... 64,112,121

Soumen Bhattacharjee .........140,141,227

Soumya P.R. ....................................... 302

Sowmya H.R. .............................. 197,329

Spall Kaur Nirmal K. .......................... 179

Sreedeep S. ....................................... 172

Sreeharsha R.V................................. 307

Sreevathsa Rohini ............................. 334

Sri Tanu ............................................. 332

Srikanth B. ............................................. 54

Srimany Amitava ................................ 114

Srinivas Agurla .................................... 188

Srinivasan Samineni .................... 215,323

Sritharan N. ....................................... 348

Srivani V.P. ....................................... 233

Srivastava Anjil Kumar .......................... 13

Srivastava Ashish Kumar ............. 156,202

Srivastava J.P. .................................... 118

Srivastava Jayanti ............................. 237

Srivastava M.K .................................... 271

Srivastava Monika ............................ 278

Srivastava S. ....................................... 171

Srivastava Shruti .............................. 301

Srivastava Smriti ............................... 299

Srivastava V.K. .................................... 155

Srivastava Vikas .......................... 221,234

Steinhorst Leonie ............................... 15

Stephen Roy ....................................... 226

Stierhof York ....................................... 201

Strader Lucia ....................................... 58

Subbarayaudu B. .............................. 130

Subbiah V. ....................................... .102

Subrahmanyam D.... 54,63,174,307,339, 349

Subramaniam V.R. ............................ 331

Subramanyam R................................ 309

Suchendranathbabu M. ...................... 321

Sudhakar Chinta .......................... 133,254

Sujatha T. ......................................... 311

Sumbria Veena ................................... 355

Sunder Singh ....................................... 168

Suneja Yadhu ...................................... 149

Sunil Bobba ......................................... 67

Surekha K. .......................................... 54

Surendra S. ....................................... 102

Suryanarayana V. ............................ 217

Sutradhar Tapan.................................. 141

Sutton Tim ....................................... ..323

Swain P. .......................................... 343

Swamy K.N. ....................................... 321

Swapna M. ....................................... 155

Swarnalakshmi K............................... 272

Swarnkar Mohit Kumar ................ 57, 210

Swaroop Vanka ................................. 252

Swathy P.S. ....................................... 195

Syiemlieh Hadienlarisa ........................ 349

Tahir Inayatullah ................................. 83

Taj Gohar .......................................... 240

Talwar H.S. ..................................101,130

Tanushri Kaul .................................... 314

Tasleem Mohd ................................... 240

Tavva Sresty ....................................... 79

Teja V. Dharma ................................... 233

Tetali Sarada .................................... 232

Tewari S. ........................................... 163

Thakur A.K. ................................ 137,346

Thakur Jitendra K. .......... 192,211,213,338

Thamilarasi K...................................... 331

Thawait Lokesh Kumar ........................ 42

Thayssa Schley.................................. 304

Thirunavukkarasu Nepolean .......... 53,274

Thiruvettai Vivek ................................. 326

Thomas Payne .................................. 318

Thomas S. ......................................... 171

Thombare Nandkishore .................... 331

Thudi Mahendar ................................. 215

Thulasiram H.V. ................................. 234

Tikaniya Divya ................................... 220

Tikkanen Mikko .................................. 109

Tittal Megha ........................................ 276

Tiwari Archana .................................. 181

Tiwari Manish .................................... 316

Tiwari Rahul ............................ 92,172,343

Tiwari Shalini ..................................... 125

Tiwari Sneha ....................................... 190

Tiwari Sushma .................................... 20

Tokas Indu ........................................... 15

Tomar Ajeet Singh ............................ 280

Tomar Vandana ................................. 205

Townsley Brad .................................. 110

Tracy Lawson ..................................... 69

Author Index

370

IPPCongress2015


Tripathi A.K. ........................... 27,136,281

Tripathi N. .......................................... 345

Tripathi Shailesh .......................... 104,323

Tripathy B.C. ............................... 68,309

Tripathy Jayant ................................. 199

Trivedi A.K. ....................................... 160

Trivedi P.K............... 40,205,206,317,318

Trivedi S.M. ........................................ 145

Tula Suresh ........................................ 118

Turlapati Swathi ................................ 8,26

Tuteja Narendra ................................. 118

Tuteja Omprakash .............................. 140

Tyagi A.K. ............. 43,153,104,250,352,

353,355

Tyagi Isha .......................................... 245

Tyagi Shikha ........................................ 332

Udayakumar M........ 30,45,180,196,198,

224,234,254,327,329,334

Uddin Moin ............................ 291,294,298

Udhayakumar Nagan ......................... 291

Udvardi Michael ................................ 199

Ulrike Bechtold ...................................... 69

Umesh M.R. ........................................ 131

Upadhyay Natvarlal V. ....................... 288

Upadhyay Rakesh .............................. 301

Upadhyaya C.P. .................... 115,262,281

Upadhyaya D.C. ................................ 262

Upadhyaya H. ................................... 163

Upadhyaya H.D. ................................ 104

Upadhyaya Hari Deo ........................ 272

Vadassery Jyothilakshmi .................... 250

Vadez V. ............................................ 101

Vadez Vincent ................................... 313

Vaganan M. Mayil .............................. 333

Vagheera P. ........................................ 151

Vaishnav Piyush R. ........................... 288

Vaishnaw Vivek ................................. 316

Vakharia Dinesh N. ........................... 255

Vanaja M. ................................... 135,151

Vanvari Rythm ..................................... 195

Varsani Suresh ................................... 47

Varsheny Rajeev K. ............................ 322

Varshney Lalit .............................. 291,298

Varshney Rajeev K. .......215,216,217,323

Vashisht Ira ........................................ 337

Vashist Enesh..................................... 298

Vemanna Ramu S. ............................ 254

Venkatesh B. .................................... 133

Vennapusa Amaranatha .................... 254

Vergish Satyam ........................... 162,193

Verma Baburam .................... 77,239,240

Verma Deepanjali ................................ 185

Verma Indu ........................................ 222

Verma Monika ................................... 220

Verma Pooja ........................................137

Verma Praveen K......50,221,247,234,258

Verma Rakesh .........16,157,171,172, 191,

282,296,324,343

Verma S.K. ........................................160

Verma Sandhya................................. 258

Verma Subodh .................................... 200

Verma Susheel .................................... 354

Verma Vibha ....................................... 350

Veronica N. ....................................... 174

Vetcha Aswani................................... 187

Vigneswaran Veena .......................... 143

Vijay Priyanka ..................................... 112

Vijaya Bramha................................... 344

Vijayakumar H.P. .............................. 288

Vijayalakshmi D. .......................... 163,164

Vijayalakshmi P. ................................ 321

Vijayaraghavan H. ............................ 130

Vijaybhaskar V. ................................ 102

Viji Maluventhen ................................ 291

Vimalakumari N.K............................... 255

Vineeth T.V........................................ 165

Vinod K.K. ............................................ 74

Vinod Kumar ....................................... 278

Vinya V. ............................................. 102

Virdi Singh ........................................ 123

Vishnukiran T. ..................................... 321

Vishwakarma C. .................. 171,172,327

Vishwakarma Harinder ..................... 216

Vispo Naireen A. ............................... 103

Viswanathan C. ......16,92,146,147,157,

164,171,172,189,191,216,230,275,282,

296,324,326,327,341,343

Voleti S.R....... 54,63,143,174,307,321,

339,349

Vyas Dhiraj ................................. 113,347

Wakchaure G.C. ............................... 276

Wang Jun .......................................... 322

Wani Zahoor ....................................... 224

Wany Aakanksha ............................... 195

Watt Mitchelle ..................................... 140

Weiming He....................................... 322

Weinig Cynthia ..................................... 44

Westall Corey ....................................... 58

Wicker Thomas ................................... 243

Wong Chui E. ....................................... 80

Woody Scott T. ..................................... 44

Xie Qiguang ....................................... 44

Yadav A.K. .................................... 15,113

Yadav Amita ....................................... 209

Yadav Archana ..................... 145,220,338

Yadav Chandrabhan .......................... 206

Yadav D.K. ....................................... 115

Yadav Gitanjali ..................... 93,336,338

Yadav Karuna .................................. 267

Yadav Mahesh .................................... 333

Yadav Mohit ....................................... 220

Yadav N.D. ....................................... 120

Yadav P. ............................................ 171

Yadav Pallavi ....................................... 228

Yadav Praduman ............................... 145

Yadav Pragya ............................... 164,191

Yadav Rachita ..................................... 51

Yadav Rakesh .................................... 337

Yadav S. ............................................ 171

Yadav S.K.................................... 135,151

Yadav Sanjay ..................................... 148

Yadav Saurabh ............................ 94,335

Yadav Shashank K ..........16,146,157,164,

172,189,191,324

Yadav Sonam ................................... 220

Yadav Sudesh Kumar ......................... 117

Yadava Pranjal .......................... 174,297

Yadukrishnan P.................................. 166

Yamini K.N. ....................................... 217

Yasin Jeshima Khan .......................... 326

Yogindran Sneha ............................... 248

Yongle Li ........................................... 323

Zala P.V. ...................................... 159,269

Zhang Gengyun .................................. 322

Zhou W.H. ......................................... 172

Zinta Gaurav ....................................... 110

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