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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
Published by
I.K. International Publishing House Pvt. Ltd.S-25, Green Park Extension
Uphaar Cinema MarketNew Delhi - 110 016 (India)
E-mail: [email protected]
Branch Offices:
A-6, Royal Industrial Estate, Naigaum Cross Road,
Wadala, Mumbai - 400 031 (India)E-mail: [email protected]
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ISBN 81-89866-09-5
© 2008 I.K. International Publishing House Pvt. Ltd.
All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or
transmitted in any form or any means: electronic, mechanical, photocopying, recording, or otherwise,without the prior written permission from the publisher.
Published by Krishan Makhijani for I.K. International Publishing House Pvt. Ltd., S-25, Green Park
Extension, Uphaar Cinema Market, New Delhi - 110 016 and Printed by Rekha Printers Pvt. Ltd.,Okhla Industrial Area, Phase II, New Delhi - 110 020.
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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