Plant Biotechnology and Molecular Markers Plant Biotechnology and Molecular Markers Edited by P.S. Srivastava Alka Narula Centre for Biotechnology, Faculty of Science Jamia Hamdard, New Delhi, India Sheela Srivastava Department of Genetics University of Delhi South Campus New Delhi, India KLUWER ACADEMIC PUBLISHERS NEW YORK, BOSTON, DORDRECHT, LONDON, MOSCOW Anamaya Publishers NEW DELHI eBook ISBN: 1-4020-3213-7 Print ISBN: 1-4020-1911-4 ©2005 Springer Science + Business Media, Inc. Print ©2004 Anamaya Publishers, New Delhi, India All rights reserved No part of this eBook may be reproduced or transmitted in any form or by any means, electronic, mechanical, recording, or otherwise, without written consent from the Publisher Created in the United States of America Visit Springer's eBookstore at: http://ebooks.springerlink.com and the Springer Global Website Online at: http://www.springeronline.com Professor Sant Saran Bhojwani Sant Saran Bhojwani was born to Mrs. Nam Adhari and Mr. Parmanand on 20th November, 1940 in the serene and tranquil environment of Dayalbagh, about 3 km from the hustle-bustle of the Agra city. He had his early education in Dayalbagh and graduated and postgraduated from Agra University. Soon after finishing M.Sc. (Botany), Dr Bhojwani served his alma-mater (R.E.I. Dayalbagh, Agra) as lecturer for one year before joining the University of Delhi as a doctoral student. His supervisor, late Professor B.M. Johri assigned him a challenging research problem, with the warning that his Ph.D. degree would depend on his demonstrating the cellular totipotency of endosperm, a completely unorganized, short-lived, triploid tissue. Earlier, many students of Professor Johri and scientists elsewhere in the world could establish tissue cultures of endosperm but failed to induce the organogenic differentiation. It was remarkable that within six months of his joining Delhi University, Dr Bhojwani achieved differentiation of normal shoot buds from the endosperm of Exocarpus cupressiformis; a parasitic flowering plant (Nature, 1965). At this stage a very renowned American plant physiologist, Prof. F.C. Steward, visited the University of Delhi who even after observing the cultures could not believe that the endosperm tissue could form shoots and remarked, “Young man, take a bet with me. All the shoots in the cultures are diploid. If that is the case remember me or else forget me”. However, when the shoots of endosperm origin were cytologically analysed, all of them were found to be triploid, which is of considerable practical importance in plant genetics and improvement. Subsequently, Dr Bhojwani established the cellular totipotency of endosperm cells by reporting regeneration of triploid shoots and/or plants in Scurrula pulverulenta, Acacia nilotica (Garg et al., 1996), Morus alba vi PROFESSOR SANT SARAN BHOJWANI (Thoma et al., 2000) and Azadirachta indica (Chaturvedi et al., 2003). In the meantime, many other scientists confirmed the observations of Bhojwani. Dr. Bhojwani and his students worked on a range of basic and applied aspects of in vitro plant morphogenesis. During 1971-1972 he worked with Dr Norman Sunderland at the John Innes Institute, Norwich, U.K. under the British Council Fellowship Programme and reported quantitative changes in nucleic acid and protein contents of microspores during the induction of androgenesis in tobacco using histochemistry and cytophotometry (J. Exp. Bot. 1973). In 1972, he spent three months in the laboratory of Professor Edward C. Cocking, FRS, at the University of Nottingham, U.K. and reported for the first time isolation of microspore protoplasts using helicase enzyme. The report appeared in Nature, New Biology (1972). Dr Bhojwani had another opportunity to work in the U.K. for a year during 1975-1976 under the Royal Society Commonwealth Bursary. This time he spent the whole year with Professor Cocking and worked on wheat tissue culture (Z. Planzenphysiol., 1977) and protoplast isolation and culture in cotton (Plant Sci. Lett. 1977). At this point of time there was considerable interest in the application of biotechnological techniques to crop improvement. However, a major limitation in achieving this goal was the recalcitrance of legumes, cereals and other major crop plants for plant regeneration from cultured cells, an essential step in genetic engineering and somatic hybridization. This prompted Dr Bhojwani to critically review the literature on tissue culture of crop plants which was presented as an invited lecture in a meeting organised by the Agricultural Research Council, London and later published in Euphytica (1977). The review, discussing the progress and problems of tissue culture of major crop plants and emphasizing the need for extensive further research in the area, was a highly cited publication which paved the way for a fresh spurt of research to achieve high frequency regeneration in tissue cultures of these plants. In 1978 Dr Bhojwani was awarded the prestigious Senior Fellowship of the National Research Advisory Council of New Zealand, and the family moved to Palmerston North to join the Plant Physiology Division of the D.S.I.R., New Zealand. Before the expiry of the term of the Fellowship, the D.S.I.R. offered Dr Bhojwani a position of Senior Scientist (Scientist 105) and the Government of New Zealand granted Permanent Residence to him and his family. In 1980 he was confirmed in the job. The stay of Dr Bhojwani in New Zealand was very productive. He published numerous papers on the micropropagation of Willow (N.Z.J. Bot., 1980), Garlic (Sci. Hortic., 1980), Clover (Physiol. Plant. 1981), Japanese Pear (Sci. Hortic, 1984), and Feijoa (Acta Hortic. 1987). He also worked on Trifolium spp and reported, for the first time, regeneration of full plants from mesophyll protoplasts of white clover (Plant Sci. 1982, Euphytica, 1984). Virus-free garlic plants of a Japanese variety imported into New Zealand were produced by shoot tip culture to facilitate its release through quarantine (Sci. Hortic 1982/83). Impressed by the work and publications of Dr Bhojwani the D.S.I.R. decided to promote him to Scientist 106, an opportunity which was pre-empted by his decision to return to India in 1981. However, his post in the D.S.I.R. was not filled for at least two years expecting that Dr Bhojwani might decide to return to New Zealand. He did return to New Zealand in 1983 but only as a Visiting Scientist for three months to finish some experiments which remained incomplete in 1981 and process the data for publication. After his sojourn in New Zealand, Dr Bhojwani made a modest beginning as a Research Associate at the University of Delhi and started guiding Ph.D. students in 1981. Fortunately, a PROFESSOR SANT SARAN BHOJWANI vii major research project on “Micropropagation of Important Horticultural and Silvicultural species of India” was sanctioned to him by the UGC, under which he and his students developed an efficient protocol for clonal propagation of the leguminous tree species, Leucaena leucocephala, and in vitro nodulation of micropropagated plants by Rhizobium to enhance their field survival. He also demonstrated that sugar cubes, produced by Daurala Sugar Mills, was a fair substitute of ‘Analar’ Grade Sugar used in Plant tissue culture media. The sugar cubes were more than 10 times cheaper than the ‘Analar’ Grade sucrose. In 1985 the Department of Environment and Forests, Government of India, awarded another major research project to Dr Bhojwani to work on “In Vitro Conservation of Endangered Plants”. It led to the development of protocols for micropropagation and cold storage of Himalayan Species of three medicinally important plants, viz., Picrorhiza kurroa, Podophyllum hexandrum and Saussurea lappa. In collaboration with the scientists at the Biochemical Engineering and Biotechnology Department of IIT Delhi, Dr Bhojwani studied the kinetics of cell growth in suspension cultures of Podophyllum hexandrum and in vitro production of Podophyllotoxin, an anticancerous drug (Biotechnol. Lett. 2001, J. Biosci. Bioengg., 2002). Dr Bhojwani guided six Ph.D.’s on plant regeneration alone from somatic and gametic cells of Brassica spp and published several papers (Plant Cell Tissue Organ Cult. 1985, 1991; Biol. Plant., 1989; Plant Sci. 1990a,b; Euphytica 1993). A detailed investigation on direct shoot regeneration from excised cotyledons of B. juncea proved a viable system for genetic transformation of this important oleiferous crop of India. His group also achieved high frequency androgenesis and selection of agronomically useful androclones in B. juncea. This work was supported by funds from MOMBUSHO, Japan and European Commission, Brussels. Dr Bhojwani undertook two major projects on mulberry biotechnology and investigated micropropagation of some elite clones and production of gynogenic haploids (Euphytica, 1999) and endosperm derived triploids (Pant Cell Rep. 2000) of this invaluable tree for silk industry, the sole source of feed for silkworms. Recently, he has reported the production of gynogenic haploids (Plant Cell Rep. 2003) and triploids (J. Plant Physiol., 2003) of Neem. Dr Bhojwani has published 75 research papers in journals of international repute, 10 critical reviews and 19 invited chapters in books published from India and abroad. In addition, he has authored and edited several books. His first book “The Embryology of Angiosperms” (Vikas Publishers, New Delhi) has been a popular text book for graduate and post-graduate students in India and many other countries. Running into its 5th edition, the book has been translated into Japanese (1995) and Korean (2001). In 1983, Dr Bhojwani brought out another book titled “Plant Tissue Culture : Theory and Practice”, published by Elsevier, The Netherlands. This has been regarded as the first standard text book on the subject and became so popular worldwide that the publishers brought out its paperback edition in 1986. It was translated into Korean in 1986. Under a project funded by the Department of Biotechnology, Dr Bhojwani completed a mammoth task of compiling ‘A Classified Bibliography of Plant Tissue Culture’, covering the entire literature on the subject up to 1984.