D EXS 46: Experientia Supplementum Vol. 46 Springer Basel AG Protoplasts 1983 Lecture Proceedings 6th International Protoplast Symposium Basel, August 12-16, 1983 Edited by I. Potrykus, Basel C. T. Harms, Zürich A. Hinnen, Basel R. Hütter, Zürich 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) CIP-KurztitelauJnahme der Deutschen Bibliothek {Protoplasts nineteen hundred and eighty-threeJ Protoplasts 1983 / 6th Internat. Protoplast Symposium, Basel, August 12-16, 1983. Ed. by I. Potrykus Basel ; Boston ; Stuttgart : Birkhäuser. NE: Potrykus, Ingo [Hrsg.] ; International Protoplast Symposium< 06, 1983, Basel> Lecture proceedings. - 1983. (Experientia : Supp!. ; Vo!. 46) ISBN 978-3-0348-6777-1 ISBN 978-3-0348-6776-4 (eBook) DOI 10.1007/978-3-0348-6776-4 NE: 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. © 1983 Springer Basel AG Originally published by Birkhäuser Verlag Basel in 1983. Softcover reprint ofthe hardcover 1s t edition 1983 Preface The International Protoplast Symposia are organized at four year intervals by the community of scientists working on the scientific and applied aspects of microbial and plant protoplasts. Following Jena (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. The organization was in the hands of scientists from the Friedrich Miescher-Institut, Basel (I. Potrykus - Secretary General, P. J. King - Treasurer, A. Hinnen and R. D. Shillito - Local Organizers) and from the Swiss Federal Institute ofTechnology (ETH), Zürich (R. Hütter-Secretary, C. T. Harms - Local Organizer). Over 300 scientists from more than 30 countries met in the Basel Trade Fair Building for five days to hear Symposium Lectures reviewing all aspects ofmodern protoplast research, to present recent original results in Poster Sessions, and to discuss specific problems and trends offuture research in Workshops. The manuscripts ofthe Symposium Lectures and sum maries ofthe majority ofthe Workshops are published in this volume together with photos of the speakers and chairpersons and ofthe welcome reception in the Basel Art Museum and oth er social events. Special emphasis in the symposium was placed on the presentation in Poster Sessions ofthe recent work ofmany 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 deci ded to publish the posters together with a one page commentary containing detailed informa tion. These Poster Proceedings were handed out to the participants at the beginning of the Symposium and were a great success. A table ofthe contents ofthe Poster Proceedings is inclu ded in this volume of the Lecture Proceedings. Those who are interested in getting a complete record of the Symposium are recommended to purchase the Poster Proceedings from the org anizers (Friedrich Miescher-Institut, P. O. Box 2543, CH-4002 Basel) or from the Birkhäuser Verlag Basel, for SFr. 112.-. The Symposium was closed with the announcement that the 7th International Proto plast Symposium will be organized by scientists ofthe Max-Planck-Institut for Plant Breeding, Cologne, FRG, in 1987. logo Potrykus Secretary General 7 Contents Prefaee ........................ . 5 Seientifie Programme ofthe Symposium ..... . 11 Photos of speakers, ehairpersons, and partieipants . 12 Symposium Lectures and Workshop Summaries Plant Protoplasts Symposium Lectures Reeent developments in the eulture and regeneration of plant protoplasts M. R. Davey . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 Protoplast eulture and plant regeneration of eereals and other reealeitrant erops P. J. Dale ........................................ 31 Protoplasts and the isolation of plant mutants J. P. Bourgin . . . . . . . . . . . . . . . . . . . . 43 Protoplasts and variation from eulture P. J. Larkin, R. BrettelI, S. Ryan, and W. Scowcroft. 51 Systems for plant protoplast transformation T. Manzara and P. F. Lurquin . . . . . . . . 57 Reeent developments in plant protoplast fusion and seleetion teehnology G. B. Lazar .................................. . 61 Somatie hybridization by plant protoplast fusion c. T.Harms .................... . 69 The segregation of organelles and eytoplasmie traits in higher plant somatie fusion hybrids R. Fluhr . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 The first mitotie eycle of mesophyll protoplasts Y. M eyer and L. Aspart . . . . . . . . . . . . . . 93 Protoplasts for studies of the plasma membrane and associated eell organelles L. C. Fowke, L. R. Griffing, B. G. Mersey, and P. Van der Valk ......... . 101 8 Contents The use of protoplasts in plant virus research H. P. Mühlbach . . . . . . . . . . . . . . . . . 111 Applications of protoplast technology to agriculture E. C. Cocking . . . . . . . . . . . . . . . . . . . . . . 123 Workshop Reports Organelle transfer, sorting out, recombination H. LörzandS. Izhar .............. . 129 Plant protoplasts as tools for physiological studies T. Boiler and D. W. Galbraith ........... . 130 Genetic transformation R. D. Shillito and H. H. Steinbiss 131 Protoplasts as tools in pathology, virology and plant-microbe interactions H. R. Hohl and H. P. Mühlbach . . . . . . . . . . . . . . . . . . . . . . . . . 132 Microbial Protoplasts Symposium Lectures Current questions of gene transfer via protoplast fusion in microorganisms (Opening Adress) L. Ferenczy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137 Advances in protoplast fusion and transformation in Streptomyces R. H. Baltz and P. Matsushima. . . . . . . . . . . . . . . . . . . . . 143 Chromosome interactions and expression in fused Bacillus protoplasts R. D. Hotchkiss and M. Gabor ..................... . 149 Yeast strain improvement by protoplast fusion and transformation A. J. Morgan . ............................. . 155 Protoplasts of filamentous fungi in genetics and metabolite production J.Anne . ................................... . 167 Interspecific somatic hybridisation in Aspergillus H. J. Croft and R. B. G. Dales ........... . 179 Synthesis and assembly of wall polymers on regenerating yeast protoplasts R. Sentandreu, E. Herrero, M. V. Elorza, H. Rico, and J. Pastor . ...... . 187 Contents 9 The theory and practical applications of liposome-protoplast interactions J. F. M akins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197 Transport of nutrients in yeast protoplast A. Kotyk .................. . 209 Protoplasts and spheroplasts of gram-negative bacteria - with special emphasis on Proteus mirabilis H. H. Martin ........................................ 213 Characteristic properties and biological significance of stable protoplast type L forms J. Gumpert and U. Taubeneck ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227 Advances in microbial protoplasts (Bacillus licheniformis ß-Iactamase and the proto plast surface) J. o. Lampen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243 Workshop Reports Fungal protoplasts as genetic tools J. F. Peberdy and L. Ferenczy. . . . 251 Non-complementing diploids: Prokaryotic microorganism protoplast fusion and chromosome inactivation L. Hirschbein ........................................ 251 Index of Organism . . . . . . . . . . . . . . . . . . . . . . . . . 253 Table of Contents of the Poster Porceedings of Protoplasts 1983 257 11 SCIENTIFIC PROGRAMME tIlNDAY. AUGUST 15 09.30- Poster Session 6: Microbial Protoplasts. FRlDAY. AUGUST 12 12.00 Poster Code NA 1-8. Me 1-4 09.00- lecture Session 5: Plant Protoplasts. 09.00 Opening Remarks: I. Potrykus 11.00 Chairperson: O. Schieder (Köln. FRG) 09.15 Opening Address: L. Ferenczy (Szeged, Hungary) 09.00 G.B. Lazar (Szeged. Hungary) - Recent De Current quest ions of gene transfer via proto velopments in fusion and selection technology. plast fusion in microorganisms. 09.30 C.T.Harms (Zürich. Switzerland) - Somatic Chairperson: R. Hütter (Zürich, Switzerland) hybridization by protoplast fusion. 10.00 Coffee Break 10.30 R. Fluhr (Rehovot. Israel) - Sorting out 10.30- Poster Session 1: Microbial Protoplasts. of organelles and cytoplasmic traits in samatic 13.00 Poster Code: MB 1-11 fusion hybrids. 10.30- Lecture Session 1: Plant Protoplasts. 11.00- Poster Session 7: Plant Protoplasts. 12.00 Chairperson: I.K. Vasil (Gainesville, USA) 13.00 Poster Code PR 1-7 10.30 M.R. Davey (Nottingham, U.K.) - Recent 13.00 Lunch Break developments in the culture and regeneration 15.00- Poster Session 8: Plant Protoplasts. of plant protoplasts. 18.00 Poster Code PD 1-21. PE 1-6. PG 1-5 11.15 P. Dale (Aberystwyth, U.K.) - Protoplast 15.00- lecture Session 6: Microbial Protoplasts. culture and plant regeneration of cereals and 17.15 Chairperson: A. Hinnen (Basel, Switzerland) other recalcitrant crops. 15.00 R. Sentandreu (Valencia, Spain) - Cytoche 12.10- WOrkshop 1: Plant Protoplasts mical and physiological aspects of yeast proto 13.10 Organelle transfer, sorting-out, recombination. plasts. Organizers: S. Izhar (Bet Dagan, Israel), 15.45 J. Makins (Syracuse. USA) - The theory and H. Lörz (Köln, FRG) practical application of liposome-protoplast 13.10- Lunch Break interactions. 15.00- Poster Session 2: Plant Protoplasts 16.30 A. Kotyk (Prague. CSSR) - Transport of 18.00 Poster Code: PA 1-8. PB 1-17. PC 1-7 nutrients in yeast protoplasts. 15.00- Lecture Session 2: Microbial Protoplasts 18.00 Dinner Break 16.30 Chairperson: J. Peberdy (Nottingham, U.K.) 20.30- Workshops 15.00 H.R. Baltz (Indianapolis, USA) - Recent 23.00 Workshop 7: Microbial Protoplasts. advances in protoplast methodology for antibio Fungal protoplasts. Organizers: l. Ferenczy tic-producing Streptomyces. (Szeged.Hungary). J.F. Peberdy (Nottingham. UK) 15.45 R.D. Hotchkiss (Albany, USA) - Chromosome Workshop 8: Plant protoplasts. Protoplasts as interaction and expression in fused Bacillus tools in pathology. virology.and plant-microbe protoplasts. interactions. Organizers: R. Hohl (Zürich. 18.00- Dinner Break Switzerland), H.P. Mühlbach (München. FRG). 20.30- Workshops WOrkshop 9: Plant protoplasts. Somatic hybridi 23.00 WOrkshop 2: Microbial Protoplasts. zation. Organizers: P. Maliga (Manhattan. USA). Non-complementing diploids. Organizers and con O. Schieder (Köln. FRG). tributors: L. Alföldi. K. Fodor. L. Hirschbein. TUESDAY, AUGUST 16 D. Hotchkiss. P. Schaefer. WOrkshop 3: Plant Protoplasts. 09.00- Lecture Session 7: Plant Protoplasts Protoplast culture and plant regeneration with 11.00 Chairperson: E. Galun (Rehovot, Israel). special emphasis on crop plants. 09.00 Y. Meyer (Perpignan. France) - The first Organizers: I. Potrykus (Basel, Switzerland), mitotic cycle of mesophyll protoplasts. I.K. Vasil (Gainesville. USA). 09.30 L. Fowke (Saskatoon. Canada) - Protoplasts WOrkshop 4: Plant Protoplasts. for studies of plasma membrane and associated Mutation and stable variation. organelles. Org?nizers: P.J. King (Basel, Switzerland). 10.00 H.P. Mühlbach (München. FRG) - The use of J. io.!dholm (Urbar,a, USA). protoplasts in plant virus research. 11.00 Workshop 10: Plant protoplasts. SATURDAY. AUGUST 13 Protoplasts and plant breeding. Organizers: 09.30- Poster Session 'I: Microbial Protoplasts D. Evans (Cinnaminson. USA), E. Galun (Rehovot) 12.00 Poster Code: Me 1-5. ME 1-4 13.00 Lunch break 09.00- Lecture Se,ssion 3: Plant Protoplasts 14.00- Lecture Session 8: Microbial Protoplasts. 11.00 Chairperson: P. Maliga (Szeged, Hungary) 15.00 Chairperson: R. Hütter (Zürich. Switzerland) 09.00 J.P. Bourgin (Versailies, France) - Proto 14.00 H.H. Martin (Darmstadt. FRG) - protoplasts plasts and the isolation of mutants. and spheroplasts in the gram-negative bacterium 09.30 P.,J. Larkin (Canberra, Australia) - Proto Proteus mirabilis. Physiological and biochemi plasts and var·iation in culture. cal properties. 10.00 P. Lurquin (Pul Iman, USA) - Systems for 14.30 U. Taubeneck (Jena. DDR) - The stable pro plant protoplast transformation. toplast type L-form of bacteria - characteristic 11.00- Poster Se~sion 4: Plant Protoplasts. properties and biological significance. 13.00 Po~ter Code PK 1-16 14.00- Poster Session 9: Plant Protoplasts 13.00- Lun/:l Break 16.00 Poster Code PI 1-2, PJ 1-2, Pl 1-6, PM 1-3, l!i.dU- Poster Session 5: Plant Protoplasts. PP 1-4 18.00 Poster Code PF 1-6. PH 1-4. PN 1-2. PO 1-2. 16.00- Closing Lectures PQ 1-8 18.00 16.00 O. lampen (Piscataway. USA) - Advances in microbial protoplasts. 15.00- lecture Session 4: Microbial Protoplasts. Chairperson: R. Hütter (Zürich, Switzerland) 16.45 Chairperson: F. Kevei (Szeged, Hungary). 17.00 E.C. Cocking (Nottingham, U.K.) - Appli 15.00 A.J. Morgan (Sunburry-on-Thames, U.K.) - cations of protoplast technology in agriculture. Yeast strain improvement by protoplast fusion Chairperson: G. Melchers (Tübingen. FRG) and transformation. 18.00 Closing remarks: I. Potrykus 15.45 J. AnnA (Louvain, Belgium) - Protoplasts of filamentous fungi in genetics and metabolite production. 16.15 J. Croft (Birmingham, U.K.) - Interspeci ORGAN I ZERS fic somatic hybridization in Aspergillus. I. Potrykus (Basel), Secretary General 18.00- Dinner Break R. Hütter (Zürich). Secretary 20.30 WOrkshop 5: Plant Protoplasts. Protoplasts as P.J. King (Basel) Treasurer tools for physiological studies. Organizers; C.T. Harms (Zürich). Local Organizer T. BoIler (Basel), D. Galbraith (Lineoln. USA). A. Hinnen (Basel). Loca I Organizer WOrkshOp 6: Plant Protoplasts: Genetic trans R.D. Shillito (Basel), Local Organizer formation. Organizers: R.D.Shillito (Basel). R. Forlen (B3sel). Congress Office H.H. Steinbiss (Köln. FRG) M. Marti (Basel), MUBA service