IN VITRO HAPLOID PRODUCTION IN HIGHER PLANTS Current Plant Science and Biotechnology in Agriculture VOLUME 29 Scientific Editor RJ. Summerfield, The University o/Reading, Department 0/ Agriculture, P.O. Box 236, Reading RG6 2AT, Berkshire, UK Scientific Advisory Board B.K. Barton, Agracetus Inc., Middleton, Wisconsin, USA F.C. Cannon, University 0/ Massachusetts at Amherst, Amherst, Massachusetts, USA H.V. Davies, Scottish Crops Research Institute, Dundee, Scotland, UK J. Denecke, University o/York, York, UK J. Hamblin, The University o/Western Australia, Nedlands, WA, Australia J. Lyman Snow, Rutgers University, New Brunswick, New Jersey, USA C.P. Meredith, University o/California at Davis, Davis, California, USA J. Sprent, University o/Dundee, Dundee, Scotland, UK D.P.S. Verma, The Ohio State University, Columbus, Ohio, USA Aims and Scope The book series is intended for readers ranging from advanced students to senior research scientists and corporate directors interested in acquiring in-depth, state-of-the-art knowledge about research fmdings and techniques related to all aspects of agricultural biotechnology. Although the previous volumes in the series dealt with plant science and biotechnology, the aim is now to also include volumes dealing with animals science, food science and microbiology. While the subject matter will relate more particularly to agricultural applica tions, timely topics in basic science and biotechnology will also be explored. Some volumes will report progress in rapidly advancing disciplines through proceedings of symposia and workshops while others will detail fundamental information of an enduring nature that will be referenced repeatedly. The titles published in this series are listed at the end 0/ this volume. In Vitro Haploid Production in Higher Plants Volume 5 - Oil, Ornamental and Miscellaneous Plants Edited by S. MOHAN JAIN Plant Production Department, University of Helsinki, Helsinki, Finland S.K. SOPORY School of Life Science, lawaharlal Nehru University, New Delhi, India and R.E. VEILLEUX Department of Horticulture, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, U.S.A. ~. " SPRINGER-SCIENCE+BUSINESS MEDIA, B.Y. Library of Congress Cataloging-in-Publication Data In v!t~o haploid p~oduction in highe~ plants 1 edito~s. S. Mohan ~ain, S.K. Sopo~y. R.E. Veilleux. p. c•. -- <Cu~~ent plant science and b1otechnology in agricultura : v. 23> Includes index. Contents: v. 1. Fundamental aspects ISBN 978-90-481-4683-3 ISBN 978-94-017-1856-1 (eBook) DOI 10.1007/978-94-017-1856-1 1. Mic~op~opagation. 2. Haploidy. 3. Crops--Genetic eng1nee~1ng. 4. Plant b~eeding. I. Jain, S. Mohan. II. Sopo~y. S. K. III. Veilleux, R. E. IV. Se~ies: Cu~~ent plant science and biotechnology in ag~icultu~e : 23. S~123.6.I45 1996 631.5'23--dc20 95-304 ISBN 978-90-481-4683-3 Printed on acid-free paper Ali Rights Reserved © 1997 Springer Science+Business Media Dordrecht Originally published by Kluwer Academic Publishers in 1997 No part of the material protected by this copyright notice may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording or by any information storage and retrieval system, without written permission from the copyright owner. Table of Contents Dedication I.K. Vasil vii General Preface ix Preface to Volume 5 xiii Acknowledgements xiv Section 1 V.1. Haploidy and related biotechnological methods in linseed (Linum usitatissimum L.) R. Bergmann and W. Friedt 1 V.2. Haploid production and application of molecular methods in sunflower (Helianthus annuus L.) W. Friedt, T. Nurhidayah, T. Rocher, H. Kohler, R. Bergmann and R. Horn 17 V.3. Haploidy in niger (Guizotia abyssinica Cass) P.B. Kavi Kishor, T.P. Reddy, A. Sarvesh and G. Venkatesham 37 Section 2 V.4. Haploidy in Petunia S. Mohan Jain and N. Bhalla-Sarin 53 v VI Table of contents V.5. In vitro induction of haploid plants from the gametophytes of lily and tulip R. W .. van den Bulk and J.M. van Tuyl 73 V.6. Anther and microspore culture in Camellia japonica M.C. Pedroso and M.S. Pais 89 Section 3 V.7. Haploidy in asparagus by anther culture H.S. Tsay 109 V.8. Haploidy in Nicotiana spp. A. Cersosimo and C. Sorrentino 135 V.9. In vitro induction of haploidy in cotton Y.P.S. Bajaj and M.S. Gill 165 V.lO. Gynogenic haploids in vitro G. Lakshmi Sita 175 V.11. Haploidy in Citrus M.A. Germana 195 V.12. Haploid plantlet production in tomato W.L. Summers 219 V.13. Somata haploid production by anther culture of interspecific somatic hybrids and their prospects in potato breeding V.-M. Rokka, J.P. T. Valkonen and E. Pehu 233 List of Contributors 245 Species and Subject index 249 Haploid production in higher plants A dedication INDRA K. VASIL The value of haploids in genetic analysis and plant breeding has been known for a long time. Natural haploid embryos and plants, derived from gameto phytic cells, have been described in about 100 species of angiosperms. How ever, haploids occur only rarely in nature. To be useful, they must be produced in large numbers. Therefore, many attempts have been made over the years to increase the efficiency of in ovulo haploid production, but none of these has proven to be of wide practical utility. The early attempts to obtain haploid plants from the male gametophyte of gymnosperms (Tulecke, 1953) and angiosperms (Yamada et at., 1963) resulted only in the production of haploid callus tissues (Vasil, 1980). Embryo-like structures formed in cultured anthers of Datura innoxia were first described by Guha and Maheshwari (1964). They were considered to have originated from the somatic tissues of the anther. In a subsequent study, it was determined that the somatic embryos and the resulting plantlets were indeed derived from the developing microspores and were haploid in nature (Guha and Maheshwari, 1966). As is true of most pioneering studies, these first androgenic haploids were neither grown to maturity, nor were the experimental conditions for their production clearly defined. Their real value was in demonstrating the feasibility of the experimental production of ha ploids. Haploid plants were soon obtained from cultured anthers of Nicotiana sylvestris and N. tabacum by Bourgin and Nitsch (1967). These and subse quent studies by Nitsch and Nitsch (1969) clearly established that the culture of excised anthers at a precise stage of development was the most important requirement for switching the development of pollen from a gametophytic to a sporophytic phase, resulting in the formation of haploid embryos and/or plants. They also described a simple nutrient medium for the culture of anthers, and an easy procedure for obtaining dihaploid homozygous plants. The elegant, simple and reliable method of haploid production invented by Jean Pierre Nitsch and his associates provided much stimulus for future studies by many others. During the past three decades many improved methods as well as nutrient media have been developed to increase the efficiency of production of andro- vii viii A dedication Prot. s.c. Maheshwari Dr. J.P. Nitsch genic haploids, from cultured anthers as well as isolated microspores, in a wide variety of species. Success has also been achieved in obtaining gynogenic haploids from cultured ovaries or ovules. As a result, haploids are being used increasingly and profitably in breeding programmes for the development of new and improved cultivars. The various chapters in this and the com panion volumes describe in detail the basic as well as many applied aspects of haploid production and utilization. It has been my pleasure and privilege to have known the late Jean Pierre Bourgin, Sipra Guha-Mukherjee, Satish C. Maheshwari, Colette Nitsch and the late Jean Pierre Nitsch, all pioneers in haploid research. These volumes are dedicated to them for their seminal contributions to the experimental production of haploids and for creating a whole new field of basic and applied plant research. References Bourgin, J.P. and J.P. Nitsch, 1967. Obtention de Nicotiana haploides II partir d'etamines cultivees in vitro. Ann. Physio!. Veg. 9: 377-382. Guha, S. and S.C. Maheshwari, 1964. In vitro production of embryos from anthers of Datura. Nature 204: 497. Guha, S. and S.C. Maheshwari, 1966. Cell division and differentiation of embryos in the pollen grains of Datura in vitro. Nature 212: 97-98. Nitsch, J.P. and C. Nitsch, 1969. Haploid plants from pollen grains. Science 163: 85--87. Tulecke W., 1953. A tissue derived from the pollen of Ginkgo bi/oba. Science 117: 599-600. Yamada, T., T. Shoji and Y. Sinoto. 1963. Formation of calli and free cells in the tissue culture of Tradescantia reflexa. Bot. Mag. Tokyo 76: 332-339. Vasil, I.K., 1980. Androgenetic haploids. Int. Rev. Cyto!. Supp!. 11A: 195-223. General Preface Since the beginning of agricultural production, there has been a continuous effort to grow more and better quality food to feed ever increasing popula tions. Both improved cultural practices and improved crop plants have allowed us to divert more human resources to non-agricultural activities while still increasing agricultural production. Malthusian population predictions continue to alarm agricultural researchers, especially plant breeders, to seek new technologies that will continue to allow us to produce more and better food by fewer people on less land. Both improvement of existing cultivars and development of new high-yielding cultivars are common goals for breeders of all crops. In vitro haploid production is among the new technologies that show great promise toward the goal of increasing crop yields by making similar germplasm available for many crops that was used to implement one of the greatest plant breeding success stories of this century, i.e., the development of hybrid maize by crosses of inbred lines. One of the main applications of anther culture has been to produce diploid homozygous pure lines in a single generation, thus saving many generations of backcrossing to reach homozygosity by traditional means or in crops where self-pollination is not possible. Because doubled haploids are equivalent to inbred lines, their value has been appreciated by plant breeders for decades. The search for natural haploids and methods to induce them has been ongoing since the beginning of the 20th century. Blakeslee (1921) first identified naturally occurring hap loids of Datura stramonium and subsequently, natural haploids of many other plants were reported by various researchers. However, naturally occurring haploids could not be produced in sufficient numbers by reliable techniques for their extensive use in breeding programmes. In 1964, the research group headed by Prof. S.C. Maheshwari, Department of Botany, Delhi University, India, reported haploid production in Datura innoxia for the first time by anther culture. Since this discovery, many of the limitations of the technique have been overcome such that it is currently employed for the production of haploids and doubled haploids of many crop species throughout the world. The early contributions of Drs. C. Nitsch and J.-P. Nitsch (France), G. Melchers (Germany), M.S. Swaminathan (India), I.K. Vasil (USA), N. ix