biotech research at tamil nadu agricultural university 2011

GenesisA manpower development programme in Biotechnology

initiated during 1988 through PG-Biotechnology programme with the support of DBT, GoI.

Plant Molecular Biology and Biotechnology

Major activitiesEducationResearch

EducationUnder Graduate Education

B. Tech. (Biotechnology)B. Tech. (Bioinformatics)

Post Graduate EducationM.Sc. (Biotechnology)M.Tech (Biotech & Business

Management)PhD (Biotechnology)

Major Areas of Biotech Research Isolation of genes conferring agronomic traits

Novel genes to overcome IPR issues Genetic transformation of crops for improved

agronomic traits Wider choice of genes for incorporation into crops

Marker Assisted Breeding for improving agronomic traits in crop plants

Improves the efficiency with which breeders can select plants with desirable combinations of genes

Major Target TraitsAbiotic stress resistance

Drought resistanceSubmergence toleranceSalt tolerance

Biotic stress resistanceInsect resistanceDisease resistance

Nutritional quality

Abiotic Stress Resistance

• Duration– 100 -105 days • Season– September-October (NE Monsoon)• Yield (from 40 OFTs) – 3900 kg/ha (rainfed)

– 6600 kg/ha (semi dry)• Grain type – Long slender white rice • Increase in yield over PMK (R) 3 - 14.40 %• Suitable for cultivation under rainfed

condition in Ramnad and Sivagangai Districts• Moderately resistant to sheath rot and blast

diseases

Submergence ToleranceAbout 3 lakh ha paddy is affected frequently by submergence

in cauvery delta zoneNecessitates the development of submergence tolerant rice

varietiesMarker Assisted Introgression of Sub1 locus conferring

tolerance to flooding into elite rice varieties of Tamil Nadu

Submergence ToleranceFR13A Sub1 locus was introgressed into a widely grown Asian

rice cultivar “Swarna” using marker-assisted selectionSubmergence tolerant versions of CR1009, Savithri, Samba

Mahsuri and IR 64 are already available for cultivationSubmergence tolerant versions of CO 43 and White Ponni are

being developed to target flooding prone areas

White Ponni Co43 FR13A

Recovery after 13 days of submergence

Salt ToleranceIntrogression of salT locus conferring tolerance against salinity

stress into elite rice varieties of Tamil NaduA major QTL called salT located on chromosome 1 of a salt

tolerant variety “FL 478” controlling salinity tolerance will be targeted for introgression

Development of salinity tolerant versions of ADT 36 and White Ponni

White Ponni FL 478White Ponni FL 478

Control 80mM 150 mM Control 80 mM 150 mM 10 days after treatment10 days after treatment

Genetic engineering of sugarcane for abiotic stress toleranceAtDREB1a gene from Arabidopsis thalianaBcZAT12 gene from Brassica carinata

Co86032 transformed with AtDREB1a 20 transgenic lines obtainedCoC671 transformed with BcZAT12 11 transgenic lines obtained

Cotton Varieties with Drought Tolerance and Extra Long Staple FibreObjectives

To characterize cotton germplasm for drought tolerance using biochemical, physio-morphological traits and molecular markers

To identify DNA markers linked to drought tolerance and fibre quality traits

Development of mapping populations in progressMCU5 X KC3 MCU5 X TCH1218 KC3 X Suvin and TCH1218 X Suvin

Insect & Disease Resistance

Development of DNA markers linked toplant-hopper resistance in rice A Mapping population involving from IR50 and

Rathu-Heenathi (a genotype resistant to BPH) developed and now in F8 stage

Two SSR markers viz., RM3180 (18.22 Mb) and 25 RM2453 (20.19 Mb) showed significant association with BPH resistance in the Bulked Segregant Analysis at F7 stage

Developing bacterial blight resistant riceXa genes conferring resistance to bacterial blight are

routinely transferred to elite rice cultivars through maker assisted breeding

Pyramiding resistance genes Xa4,xa5, xa13 and Xa21 into high yielding adopted rice varieties (ADT 43 and ADT 47) through MAS using IRBB60-1-5 (resistant) as donor is in progress

30 genotypes showing higher level of resistance were identified with three genes (xa5, xa13 and Xa21) in homozygous state.

A BResistance reaction of susceptible (A) and

pyramided (B) lines against Xoo isolates

Development of gall midge, blast and bacterial blight resistant rice varietiesGall midge resistant version of Co43

pyramided with GM1 and GM4 developed through marker assisted breeding

Bacterial blight resistant version of ASD16 pyramided with xa13 and Xa21 developed through markers assisted selection

Gall midge resistant Co43 and bacterial blight resistant ASD16 will be crossed with a DRR line (BPT5204 X Tetap) and VRP1 for incorporation of blast resistant genes Pikh and Pi9 respectively

Silver shoot in ratoon crop

Bt Brinjal• Four elite genotypes, CO2, MDU1, KKM1 and PLR1 were

converted into Bt versions by introgression of cry1Ac gene from Mahyco’s elite event through back-cross breeding programme

• All Bt genotypes had significantly higher fruit yield as compared to their non-Bt counterparts.

• Bt brinjal offer adequate level of resistance to BFSB and show potential for significantly higher marketable yield

Co2 MDU1 KKM1 PLR1

Indigenous Bt genesLocal isolates of Bacillus thuringiensis collected and

evaluated against major lepidopteran pestsNovel Bt genes isolated from promising isolatesProteins from cloned Bt genes were tested by

bioassayA novel chimeric cry2A gene madePromising Bt genes (belonging to

cry2A group) being introduced in crops

cry2A for leaf folder in rice

S. No Gene Year Country

1 cry2Aa 1989 USA

2 cry2Ab 1989 USA

3 cry2Ac 1991 USA

4 cry2Ad 1999 Korea

5 cry2Ae 2003 USA

6 cry2Af 2007 Australia

7 cry2Ag 2008 China

8 cry2Ah 2008 China

9 cry2Ai 2009 TNAU, India

New Bt gene discovery

http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/intro.html

Bt CottonInsect Resistant CottonStandardization of regeneration and

transformation protocols for Coker genotypes

Agrobacterium-mediated Transformation of Coker genotypes with a chimeric cry2A gene

In planta transformation of MCU5, MCU12 and MCU13

Regeneration of transformed plants

Bt BrinjalIndigenously isolated, synthetic cry2A genes to

control fruit and shoot borerTarget genotypes: CO2, Elavambadi LocalTransgenic plants generated using Agrobacterium

method

Regeneration of plants from cotyledons

Regenerated of plants Transgenic plant

Engineering resistance against banana bunchy top virusThe replication of virus is targeted

through RNA interference approachRNAi vector targeting the BBTV Rep

gene constructedAgrobacterium- mediated

transformation of hill banana with developed RNAi construct to impart BBTV resistance

Engineering resistance against banana bunchy top virusMore than 75 putative

transgenic hill banana plants were generated and established in transgenic greenhouse.

Bioassay results showed that BBTV resistance in all eight transgenic hill banana lines tested

Engineering resistance in papaya against the papaya ring spot virusViral coat protein gene and replicase genes are suppressed

through RNA interference approach to contain virus replication

RNAi vector targeting these genes madeBiolistic and Agrobacterium- mediated transformation of Co7

papaya with the RNAi construct is on-going

Genetic engineering of Cassava for resistant to Cassava Mosaic Disease

RNAi construct targeting the ICMV and SLCMV Replicase gene

Agrobacterium-mediated transformation of cassava with RNAi construct is on-going

Nutritional Quality

Golden Rice• Vitamin A deficiency is prevalent

in the developing world – responsible for the deaths of

two million children annually – deficiency is a leading cause of

blindness• β-carotene expressed in many

plants but not in cereal grains• Introduction of correct metabolic

steps into rice endosperm to facilitate β-carotene synthesis

• ASD16 and ADT43 converted into GR lines

IPP

Geranylgeranyl diphosphate

Phytoene

-carotene(vitamin A precursor)

Phytoene synthase

Phytoene desaturase

Lycopene-beta-cyclase

ξ-carotene desaturase

Maize psy

Erwinia crtl

Iron riceFerritin is a ubiquitous

protein that stores iron and releases it in a controlled fashion

Over-expression of Avecennia marina ferritin in rice using an endosperm-specific promoter

ASD16

ADT43

Low phytate maizeTwenty nine maize inbred lines were selected based on the

percent of phytate phosphorus content in total phosphorus content in the maize kernels, as low and high and were included in the crossing programme

SSR Polymorphism survey among the low and high phytate maize inbreds for the lpa1 (10 SSR markers) and lpa2 (10 SSR markers) regions in chromosome1 is under progress

Content (mg/g) Reported Variability ( Raboy et al.2002)

Observed variability in UMI inbreds

lpa1 / lpa2 mutants

Low High Low High lpa1 lpa2

Phytate P 2.50 3.50 1.12 4.30 1.10 2.60

Total P 3.50 4.50 2.04 5.07 4.60 4.70

Phytase maizeExpression of fungal phytase in maize grain is attempted with

a view to make more micronutrients bioavailable to the poultry birds

Phytase gene from Aspergillus niger isolated, expressed in E. coli and characterized

Gene construct made to express the phytase in maize endosperm in tissue specific manner

Genetic transformation resulted in development of 26 events

Shoots on rooting medium

Callus on shooting media

Somatic embryos from Immature embryos

Tissue cultureMicro-propagation of vegetatively propagated

cropsSecondary metabolite production in medicinal

plants through suspension culture