Recent Advances in

Plant Biotechnology and its Applications

Recent Advances in

Plant Biotechnology and its

Applications

Editors

Ashwani KumarProfessor

Department of Botany

University of Rajasthan,

Jaipur, India

Sudhir K. SoporyHead of Plant Molecular Biology Division

International Centre for Genetic Engineering and Biotechnology,

New Delhi, India

I.K. International Publishing House Pvt. Ltd.New Delhi • Bangalore • Mumbai

Prof. Dr. Karl-Hermann Neumann Commemorative Volume

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Preface

In simple terms, the application of technology to living organisms can be termed as biotechnology.

During the last decade, the advancements in biology have led to the development of newer areas

like cellular engineering, biochips and biomaterial science, stem cells, nanobiotechnology, etc.

The present book is restricted to the area of plant biotechnology, in particular, and attempts

to expand only a few of the topics. Plant biotechnology had its beginning in the early twentieth

century. It was in 1930’s that Philip White and Roger Gautheret established plant cell culture systems.

This was followed by the discovery of hormones, which led to the development of different

techniques for the induction of cell cultures to differentiate into shoots, roots, or embryos in many

plant systems—dicots and monocots. With the advent of recombinant DNA technology, and

knowledge that accumulated related to the mode of transfer of DNA from Agrobacterium to the

plants to induce cell division and gall formation, the field of plant transformation took birth and

genetically engineered plants saw the light of the day. It is hoped that these technologies will

greatly benefit the human kind by improving the yield of crop plants, horticulture produce, and

the compounds having medicinal properties.

The book is divided into five sections. The first section deals with the methodology and

bioresource generation. The techniques related to genetic engineering and gene transfer to the

nuclear genome and chloroplast genome have been dealt with. The new techniques of genome

profiling and gene silencing are also presented in two chapters. The importance and usefulness of

the technology to preserve and utilize useful alleles in the wild germplasm is given in the chapter

contributed by Cho et al.

The second section of the book deals with the classical aspect of plant biotechnology viz.

tissue culture and micropropagation. There are eight chapters in this section, which bring out the

usefulness and application of protoplast culture, microspore culture, in vitro regeneration and

multiplication in cereals, bamboo, fruit, and foliage plants. This section also includes some reviews

and original papers.

As mentioned above, genetic engineering via Agrobacterium has been standardized in a number

of plants including cereals. In addition, direct transfer of DNA via particle bombardment has also

been successful with many plants. Use of this technology to develop transformed plants in Artemesia,

castor and orchids is given in the third section. In addition, a chapter by Soderquist and Lee

reflects work on the production of recombinant proteins in plant cells.

The fourth section deals with the stress tolerance in plants. Both abiotic and biotic stresses put

a very heavy penalty on the yield of many crop plants. Attempts to breed for these traits have

limited success. It seems there is hope to improve tolerance in plants to both biotic and abiotic

stresses using the modern biotechnological tools, and utilizing the knowledge generated via genetics

and genomics. The basic biology of some of the stress responses and designing plants for stress

tolerance is discussed in this section.

The last section deals with medicinal plants. How to increase the alkaloid production, and how

to manipulate pathways to increase the desired metabolite using tissue culture, micropropagation,

genetic engineering etc. are some of the questions that have been discussed in various chapters of

this section.

Although plant biotechnology encompasses many different areas, this book is designed to

include chapters on a few of the selected topics of interest. The authors are given freedom to

express their views and also to present data of their own. However, the editors have taken the

liberty, with the know how of the authors, to review and revise the chapters to avoid presentation

of any unsubstantiated data. We hope this book will be useful for the researchers to update their

information on the topics dealt within this book in the area of Plant Biotechnology.

The present book is dedicated to Professor Dr. Karl-Hermann Neumann, Institut für

Pflanzenernährung, Justus Liebig Universität, Giessen, Germany. One of us (Ashwani Kumar) had

a chance to work with him under Alexander von Humboldt Foundation fellowship. Professor

Dr. Neumann has been pioneer in introducing plant tissue culture in Germany, and has used this

technique for solving various fundamental and applied problems of biology, especially understanding

the mechanism of photosynthesis, hormone metabolism, nutrient uptake and cycling, growth

regulation, DNA metabolism, genetic transformations and production of genetic vaccines.

We take pleasure to dedicate this book on his 70th birthday, and wish him a long and prosperous

life.

Ashwani Kumar

Sudhir K. Sopory

vi Preface

Prof. Dr. Karl-Hermann Neumann

I was born on 22 May, 1936, at a farm in Morgendorf, near Leitmeritz (Sudetenland), which is

now a part of the Czech Republic. After the end of the Second World War in 1945, my family

was transferred to a place near Bernburg in Saxonia-Anhaltinia (later German Democratic Republic,

GDR). In this place my father acquired another farm, where I grew up. In 1956, following some

problems with the Communist administration of the GDR, I had to move to the Federal Republic

of Germany, where I finished my schooling. In 1957, I entered Justus Liebig University at Giessen,

Germany, to study Agriculture. After having spent one semester as a foreign student with a stipend

at “Den Kgl.Veterinaer og Landbohojskole” in Copenhagen, Denmark, majoring in agricultural

chemistry, I completed my studies, again in Giessen, as “Diplomlandwirt” in 1960. In the same

year I received a scholarship from Cornell University in Ithaca, NY, USA. Here, I enrolled in a

graduate school, with Botany as major and Biochemistry and Physical Chemistry as minor. I also

started working for Ph.D. under the supervision of Prof. F. C. Steward, FRS. The topic of my

study was “Function of some heavy metals (iron, manganese, and molybdenum) on the growth

and metabolism of carrot tissue cultures” (mainly protein metabolism and photosynthesis), which

also became a major topic of my Ph.D. thesis later on. Since the scholarship was given to me for

only one year, I returned to Germany and completed my Ph.D. under the supervision of

Prof. H. Linser in 1962 from the University of Giessen. After spending few years with Prof.

Linser while doing postdoctorate, I finished my habilitation studies in 1969. I was promoted as

“Privat- Dozent” at the agricultural faculty of Justus Liebig University, Giessen.

While working with Prof. H. Linser at the Institut für Pflanzenernährung of Justus Liebig

University, Giessen, I had a chance to establish one of the first cell and tissue culture laboratories

in Germany. Starting from a very modest basis, the protocol of the work, which would be

performed for the next thirty years, was developed. The first problem taken up was the replacement

of White’s Basal Medium with coconut milk, as used in Steward’s laboratory at Cornell, by an

artificial nutrient medium of defined chemical composition. All later investigations can be traced

back to the three experiments performed in the 1960’s. One of the experiments focussed on the

photosynthesis of cultured carrot explants, which was based on work done at Cornell ( Exp. 39);

a second on nucleic acid metabolism of carrot cultures (Exp. 40); and a third on somatic

embryogenesis in carrot cells in a defined nutrient media ( Exp.73 ). The work on photosynthesis

was accelerated after L. Bender and A. Kumar joined our group in the 1970’s; both are now

viii Contents

holding a Professorship. The results, covering cytology as well as biochemistry aspects, were published

in a number of papers. This line of work came to a close with studies on somatic embryogenesis

of autotrophic cultures under normal atmosphere (Dr. E. Plescka).

The work on nucleic acids started with studies on metabolic turnover of both DNA and

RNA, followed by comparative studies of DNA organization of several plant species by cot

hybridization, indicating about 15% identity of unique and repeated DNA. This represents the

basic genetic information, which can be used to distinguish higher plants from other biological

systems. Here also first results turned up on the occurrence of metabolic DNA localized in repeated

fractions and broadly associated with differentiation. This work was done in cooperation with

Dr. A. Schäfer, Dr. E. Duerssen, and Prof. Savedra of Chile. Later, Dr B. Arnholdt, now a

Professor, joined our group and was mainly concerned with DNA methylation and amplification

as related to differentiation. Based on these early studies in the 1990s, gene technology was taken

up resulting in the insertion of the information of a coat protein of hepatitis B virus into the

carrot genome, which was also expressed in mature carrot roots at harvest. Dr. J. Imani took the

lead with the cooperation of medical virologists (Prof. W. Gerlich) of our university. Clinical

studies with respect to immunization after oral application could not be performed till now. Here

also the results and experience of many studies on the cell cycle and its synchronization of haploid

and diploid Datura cultures and others ( Dr. J. Blaschke, Dr. R. Kibler) was utilized by using

synchronized cultures for insertion of foreign DNA into carrot cells, preferably during S-phase.

Many studies were performed on somatic embryogenesis, mainly with petiole explants of

carrots, including histology, protein, and nucleic acid organization and metabolism. These studies

resulted from the cooperation of Dr. B. Grieb and Professor Li of the University of Huehot,

P. R. China. Carbon metabolism and hormonal system were also studied by Dr. E. Pleschka and

F. Schaefer, and published in a number of papers. A broad research program on the ploidy level

and its significance on development and secondary metabolism was initiated by Dr. E. Forche and

Dr. B. Zeppernick; both associated with our group for several years. The results of these exciting

studies were published in several papers.

Quite interesting were the comparative investigations on diurnal variations of the concentration

of several phytohormones in intact plants, in pot experiments, as well as in cultured cells. In

cultured cells, in constant environment (including continuous illumination), clear maxima of IAA as

well as of several cytokinins was observed 24 hours a day, for several days.

All this work would not have been possible without the dedicated help of my associates,

especially Frau Christa Lein, who had the same position and function in my laboratory as

Mrs. M. Mapes had in F. C. Steward’s laboratory at Cornell.

In 1995, a small book on cell and tissue culture was published in German by Ulmer Verlag,

Stuttgart ( Pflanzliche Zell- und Gewebekulturen). In this book, our results obtained till then were

discussed in context to ideas of the time. A comparable book in English (Springer Verlag,

Heidelberg) is presently in developmental stage (Prof. Dr. Kumar and Dr. Imani).

In 1972, I became Professor in the faculty of nutrition at the University of Giessen (Plant

Nutrition, Biochemistry, and Cell Biology of Plants), and I worked there till my retirement in

2001. During this period, I spent some time abroad mainly as a Visiting Professor or in a similar

position in several countries of Asia and Africa, from where students came to my laboratory to

Contents ix

work for a Ph.D. or Postdoctorate. My longest cooperation was with Prof. Ashwani Kumar,

University of Rajasthan, India (since 1977 till now).

One great challenge before me was to establish a research farm in the south of Frankfurt in

1979 to pursue investigations mainly concerned with irrigation and the quality of irrigation water.

What a change! After more than 20 years doing basic research, I had to turn to practical problems

to continue studies on biochemical and cell biological problems as before! Here Dr. B. Pauler, a

research associate, was a great help, especially in the statistical evaluation of the data obtained from

the experimental work of about 15 years. The work on irrigation and salinity was extended to

studies on sugar beet cultivation in saline conditions in Egypt. This work was done together with

Prof. A. Raafat and Dr. Eisa, Ain Shamps University, including work on biological remediation of

saline fields in cooperation with Prof. Kumar and Dr. Shekhawat, University of Rajasthan, Jaipur,

India. In Egypt, some work was concerned with the control of Orobanche infection of faba

beans based on hormonal studies (Prof. N. Al Gamrawy and Dr. Salem, Cairo University); and

mango malformation, (Prof. A. Raafat and Dr El Deep), concentrating on the hormonal system.

At the university, I occupied the chair of dean several times, and was also the director of the

department as well as the chief of examinations.

KARL-HERMANN NEUMANN

Institut für Pflanzenernährung,

Justus Liebig Universität, Giessen,

Germany

Recent Advances In Plant BiotechnologyAnd Its Applications

Publisher : IK International ISBN : 9788189866099 Author : Ashwani KumarAnd Sudhir K Sopory

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