R EXS 45: Experientia Supplementum Vol. 45 Springer Basel AG Protoplasts 1983 Poster Proceedings 6th International Protoplast Symposium Basel, August 12-16, 1983 Edited by I. Potrykus, Basel c. T. Harms, Ziirich A. Hinnen, Basel R. Hiitter, Ziirich P. J. King, Basel R. D. Shillito, Basel 1983 Springer Basel AG Address oj the Symposium Office 6th International Protoplast Symposium Organization Committee P. O. Box 141 CH-4007 Basel (Switzerland) Front cover illustration Microsphore tetrades of maize (Zea mays L.), the source for microspore protoplasts (see also Poster PB 15, p. 48). Photo by I. Potrykus. CIP-KurztitelauJnahme der Deutschen Bibliothek [Protoplasts nineteen hundred and eighty-three} Protoplasts 1983 : poster proceedings / 6th Internat. Protoplast Symposium, Basel 12-16, 1983. Ed. by I. Potrykus ... - Basel ; Boston ; Stuttgart : Birkhiiuser, 1983. (Experientia : Suppl. ; Vol. 45) NE: Potrykus, Ingo [Hrsg.]; International Protoplast Symposium (06,1983, Basel); Experientia / Supplementum All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, without the prior permission of the copyright owner. ISBN 978-3-0348-6557-9 ISBN 978-3-0348-6556-2 (eBook) DOI 10.1007/978-3-0348-6556-2 © 1983 Springer Basel AG Originally published by Birkhiiuser Verlag Basel in 1983. Softcover reprint of the hardcover 1st edition 1983 Preface The International Protoplast Symposium is organized at four year intervals by the community of scientists working on the scientific and applied aspects of microbial and plant protoplasts. Following Halle (DDR) 1963, Brno (CSSR) 1967, Salamanca (Spain) 1971, Not- tingham (UK) 1975, and Szeged (Hungary) 1979, Basel (Switzerland) was chosen as host city for the 6th International Protoplast Symposium in 1983. Over 300 scientists from more than 30 countries met for five days to hear symposium lectures on all aspects of modern protoplast research, and to discuss in 10 workshops specific problems and trends of future research. These presentations and discussions will be published in the Lecture Proceedings of the Symposium. Special emphasis was placed on the presentation in Poster Sessions of the recent work of many participants. To derive as much benefit as possible from the posters and to allow the information to be distributed to the scientific community the organizers undertook the innova- tive of publishing the posters together with a one page commentary containing additional de- tailed information. The content and the form of presentation of the posters was left entirely up to the authors, and the editors find the experiment to have been a success since the quality of these presentations general-matches that of the lectures. The posters were prepared by the authors in a camera-ready form on A3 sheets and were photo-reduced for inclusion in this volume. (For display purposes, the posters were enlarged to a 70 x 90 cm format.) The organisers feel that the difficulties of sumitting the posters 10 weeks before the Symposium have been well compensated by the opportunity for all participants to take home with them a permanent record of the information presented in their personal copy ofthe Poster Proceedings. Further copies of the Poster Proceedings, and of course of the Lecture Proceedings, are available from the organisers and from the Birkhiiuser Verlag, Basel. ~~ Ingo Potrykus Secretary General VII Contents Plant Protoplasts Protoplast Culture: Preparatory Studies PA 1 Protoplast isolation from morphogenetic cultures of Rice (Oryza sativa L.). Thompson, J. A. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 P A 2 Preliminary studies on isolation and culture of protoplasts from Sandalwood (Santalum album). Sita, G. L., Rani, B. S. ............................... 4 PA3 Isolation and identification of enzymes that are responsible for the isolation of plant protoplasts. Ishii, S., Mogi, Y. 6 PA4 Isolation and culture ofleafprotoplasts from Ulmus sp. Preliminary report. Dorion, N., Godin, B., Bigot, C. ......................... 8 PA 5 Isolation of high yields of viable protoplasts from Quaking Aspen seedlings and cultured loblolly pine cell suspensions. Verma, D. c., Wann, S. R. ............................ 10 P A 6 Approaches to cereal protoplast culture: morphogenic cultures in Wheat. Potrykus, 1., Petruska, J. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 PA 7 Towards a protoplast culture system for Wheat (Triticum aestivum). Maddock, S. E. . ............................ . 14 PA 8 Approaches to cereal protoplast culture: analysis of U stilago maydis induced cell divisions in Zea mays. Brown, P. T. H., Potrykus, 1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 Protoplast Culture: Induction o/Sustained Divisions PB 1 Mix-culture of Nicotiana and Capsicum mesophyll protoplasts. Hsia, C. A., Wang, G. Y., Wang, L. F. . ............ . 20 PB 2 Study ofhypocotyl and mesophyll protoplasts. Roy, S. c., De, K. K. .............. . 22 PB 3 Isolation and culture of cotyledon protoplasts of Cotton (Gossypium barba- dense cv. GIZA 70). El-Shihy, O. M., Evans, P. K.. . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 VIn Contents PB 4 Studies on isolation of prot op lasts from some oil-yielding plants: Niger (Gui- zotia abysssinica) and Safflower (Carthamus tinctorius L.) Pandey, R. K., Cocking, E. C. ........................... 26 PB5 Developmental potential of mega and normal protoplasts in Populus Ahuja, M. R . ............................... . 28 PB 6 A study of protoplast regeneration in ornamental species within the Composi- tae. Rashid., A., Power, J. B. .............................. 30 PB 7 Callus formation from cotyledon protoplasts of Browallia, Hyoscyamus and Physalis species. Rybczinski, J. J., Power, J. B. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 PB 8 Culture of apical protoplasts from shoot cultures in the orders Fabales, Rosa- les, and Caryophyllales. Binding, H., Joergensen, J., Krumbiegel-Schoeren, G., Finger, J., Mordhorst, G., Suchowiat, G. ................................. 34 PB 9 Production, culture and callus formation from leaf protoplasts of Hedysarum coronarium L. Lakhoua, L. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 PB 10 Sustained division of Sugarbeet (Beta vulgaris L.) protoplasts: Stimulating ef- fect of conditioned media. Szabados, L., Gaggero, C. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 PB 11 Plant regeneration from suspension culture and mesophyll protoplasts of Alfalfa. Atanassov, A. I., Brown, D. C. W. . ................. . 40 PB 12 Protoplast isolation, culture and plant formation of Physcomitrella. Batra, A., Abel, W. O . ......................... . 42 PB 13 Optimisation of Turnip (Brassica rap a) protoplast culture for Cauliflower Mo- saic Virus Transformation. Pisan, B., Potrykus, I., Paszkowski, J. ...................... 44 PB 14 Lycopersicon esculentum: globular embryos from microspores and calli from diploid protoplasts. Kruger-Lebus, S., Potrykus, I., Imamura, J. . . . . . . . . . . . . . . . . . . . . 46 PB 15 Isolated tetrade protoplasts develop to the binucleate stage in Tobacco (Nico- tiana tabacum cv Havana). Imamura, J., Portykus, I. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 PB 16 Protoplast regeneration of some legume species. Oelk, M. M., Rao, P. S., Ozias-Akins, P., Schieder, O. 50 PB 17 Isolation and culture of Coffee leaf protoplasts. Orozo, F. J., Schieder, O. ........... . 52 Contents IX Protoplast Culture: Plantsfrom Protoplast-Derived Clones PC 1 Viability and development of Potato protoplast culture. Kikuta, Y., Saito, W., Okazawa, Y. . .......... . 56 PC 2 Somatic embryos and plants from cultured protoplasts of Pennisetum pur- pureum Schum. (Napier grass). Vasil, 1. K., Vasil, v., Wang, D. Y . ........................ . 58 PC 3 Plant regeneration from mesophyll protoplasts of White Clover (Trifolium re- pens L.) White, D. W. R . .................................. . 60 PC 4 The isolation, purification and culture of Brassica napus cv. LINGOT cotyle- don protoplasts. Alexander, R., Dubert, F . ............................. . 62 PC 5 Improved culture ability of the genus Brassica by using hypocotyls as the sour- ce for protoplasts. Glimelius, K., Ottosson, A. 64 PC 6 Somatic embryogenesis and plant regeneration from protoplasts of Eggplant (Solanum melongena L.). Gleddie, S. c., Keller, W. A., Setterfield, G. . ................. . 66 PC 7 Plants regenerated from isolated protoplasts of Solanum brevi dens. Nelson, R. S., Karp, A., Greissen, G. P., Bright, S. W. J. . ..... . 68 Protoplast Fusion and Somatic Hybridisation PD 1 Factors influencing the frequency of heterokaryon formation in forage legume protoplasts. Ahuja, P. S., Hadiuzzaman, S., Rybczynski, J. J., Davey, M. R. . . . . . . . . . 72 PD 2 Intergeneric fusion of terrestrial Orchid protoplasts induced by different fu- sion promoting agents. Pais, M. S., Barroso, J., Pevereiro, P., Oliveira, M. . . . . . . . . . . . . . . . . 74 PD 3 Complementation of nitrate reductase deficient mutants in somatic hybrids between Nicotiana species. Xuan, L. T., Grafe, R., Milller, A. J. . . . . . . . . . . . . . . . . . . . . . . . . 76 PD 4 Towards somatic hybridisation in the genus Linum (Flax). Barakat, M. N., Cocking, E. C. . . . . . . . . . . . . . . . . 78 PD 5 Assessment of somatic hybridisation between Lycopersicon esculentum and Lycopersicon peruvianum. Kinsara, A., Cocking, E. C. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 PD 6 Factors affecting hybridisation between sexually incompatible species of Pet- unia. Berry, S. F.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82 X Contents PD 7 Ultrastructure and isoenzyme analysis of cultured Soybean-Nicotiana fusion products. Chien, Y.-c., Fowke, L. c., Rennie, P. J., Wetter, L. R. . . . . . . . . . . . . . . 84 PD 8 The transfer of LpDH activity as marker in somatic hybrid plants between To- bacco tumor B6S3 and normal Tobacco Xanthi. Li, x., Schieder, 0., Huang, M., Li, W. ...................... 86 PD 9 Production and characterisation of albino mutants for the use in protoplast culture. Hanson, M. R.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88 PD 10 Increasing the variability of Lycopersicon Mill. by protoplast fusion with Pet- unia L. Tabaeizadeh, z., Bergounioux, c., Perennez, C. ................. 90 PD 11 Carrot x Tobacco somatic cell hybrids selected by amino acid analog resistance complementation. Hauptmann, R., Kumar, P., Widholm, J. ..................... 92 PD 12 Molybdenum-cofactor gene transfer into nitrate reductase deficient plant cells through intergeneric protoplast fusion. Gupta, P. P., Schieder, O. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 PD 13 Developmentally regulated fusion of carrot protoplasts. Boss, W. F., Allen, N. S., Grimes, H. D. . ........ . 96 PD 14 Genetic behavior of somatic hybrids in the genus Nicotiana: N.otophora + N.tabacum and N.sylvestris + N.tabacum. Evans, D., Bravo, J. E. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98 PD 15 Nitrate-reductase deficients in the progeny after selfing complemented soma- tic hybrids of Nicotiana plumbaginifolia mutants. Marton, L., Biasini, G., Sidorov, v., Maliga, P. ................. 100 PD 16 Fusion of protoplasts isolated from Chlorella saccharophila (KrUger) NAD- SON 211-1a Gobel, E., Aach, H. G.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 PD 17 Production of somatic hybrids using auxotrophic nurse culture. Hein, T., Schieder, O . ....................... . 104 PD 18 Confirmation of chloroplast segregation in somatic hybrids of Datura by DNA-DNA hybridisation. Muller-Gensert, E., Landsmann, J., Eckes, P., Schieder, O. ........... 106 PD 19 Species specific DNA used to identify interspecific somatic hybrids. Saul, M., Potrykus, /. ......................... . 108 PD 20 Hyoscyamus-Nicotiana fusion hybrids selected via auxotroph complementa- tion and verified by species-specific DNA hybridisation. Jia, J., Potrykus, I., Lazar, G. B., Saul, M.. . . . . . . . . . . . . . . . . . . . . 110