D

EXS 46: Experientia Supplementum Vol. 46

Springer Basel AG

Protoplasts 1983 Lecture Proceedings

6th International Protoplast Symposium Basel, August 12-16, 1983

1983

Edited by

I. Potrykus, Basel C. T. Harms, Zürich A. Hinnen, Basel R. Hütter, Zürich P. J. King, Basel R. D. Shillito, Basel

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 1 st 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, Not­tingham (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 sum­maries 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 oth­er 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 deci­ded to publish the posters together with a one page commentary containing detailed informa­tion. 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 inclu­ded 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 org­anizers (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 Proto­plast Symposium will be organized by scientists ofthe Max-Planck-Institut for Plant Breeding, Cologne, FRG, in 1987.

logo Potrykus Secretary General

Contents

Prefaee ........................ . Seientifie Programme ofthe Symposium ..... . Photos of speakers, ehairpersons, and partieipants .

Symposium Lectures and Workshop Summaries Plant Protoplasts

Symposium Lectures

Reeent developments in the eulture and regeneration of plant protoplasts

7

5 11 12

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 . . . . . . . . . . . . . . . . . . . .

Protoplasts and variation from eulture P. J. Larkin, R. BrettelI, S. Ryan, and W. Scowcroft.

Systems for plant protoplast transformation T. Manzara and P. F. Lurquin . . . . . . . .

Reeent developments in plant protoplast fusion and seleetion teehnology G. B. Lazar .................................. .

Somatie hybridization by plant protoplast fusion c. T.Harms .................... .

The segregation of organelles and eytoplasmie traits in higher plant somatie fusion hybrids

43

51

57

61

69

R. Fluhr . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

The first mitotie eycle of mesophyll protoplasts Y. M eyer and L. Aspart . . . . . . . . . . . . . .

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 ......... .

93

101

8

The use of protoplasts in plant virus research H. P. Mühlbach . . . . . . . . . . . . . . . . .

Applications of protoplast technology to agriculture E. C. Cocking . . . . . . . . . . . . . . . . . . . . . .

Workshop Reports

Organelle transfer, sorting out, recombination H. LörzandS. Izhar .............. .

Plant protoplasts as tools for physiological studies T. Boiler and D. W. Galbraith ........... .

Genetic transformation R. D. Shillito and H. H. Steinbiss

Protoplasts as tools in pathology, virology and plant-microbe interactions H. R. Hohl and H. P. Mühlbach . . . . . . . . . . . . . . . . . . . . . . . . .

Microbial Protoplasts

Symposium Lectures

Current questions of gene transfer via protoplast fusion in microorganisms (Opening Adress)

Contents

111

123

129

130

131

132

L. Ferenczy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137

Advances in protoplast fusion and transformation in Streptomyces R. H. Baltz and P. Matsushima. . . . . . . . . . . . . . . . . . . . .

Chromosome interactions and expression in fused Bacillus protoplasts R. D. Hotchkiss and M. Gabor ..................... .

Yeast strain improvement by protoplast fusion and transformation A. J. Morgan . ............................. .

Protoplasts of filamentous fungi in genetics and metabolite production J.Anne . ................................... .

Interspecific somatic hybridisation in Aspergillus H. J. Croft and R. B. G. Dales ........... .

Synthesis and assembly of wall polymers on regenerating yeast protoplasts R. Sentandreu, E. Herrero, M. V. Elorza, H. Rico, and J. Pastor . ...... .

143

149

155

167

179

187

Contents

The theory and practical applications of liposome-protoplast interactions J. F. M akins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Transport of nutrients in yeast protoplast A. Kotyk .................. .

Protoplasts and spheroplasts of gram-negative bacteria - with special emphasis on Proteus mirabilis

9

197

209

H. H. Martin ........................................ 213

Characteristic properties and biological significance of stable protoplast type L­forms J. Gumpert and U. Taubeneck ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227

Advances in microbial protoplasts (Bacillus licheniformis ß-Iactamase and the proto­plast surface) J. o. Lampen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243

Workshop Reports

Fungal protoplasts as genetic tools J. F. Peberdy and L. Ferenczy. . . .

Non-complementing diploids: Prokaryotic microorganism protoplast fusion and chromosome inactivation

251

L. Hirschbein ........................................ 251

Index of Organism . . . . . . . . . . . . . . . . . . . . . . . . . 253 Table of Contents of the Poster Porceedings of Protoplasts 1983 257

SCIENTIFIC PROGRAMME

FRlDAY. AUGUST 12

09.00 Opening Remarks: I. Potrykus 09.15 Opening Address: L. Ferenczy (Szeged, Hungary)

Current quest ions of gene transfer via proto­plast fusion in microorganisms. Chairperson: R. Hütter (Zürich, Switzerland)

10.00 Coffee Break 10.30- Poster Session 1: Microbial Protoplasts. 13.00 Poster Code: MB 1-11 10.30- Lecture Session 1: Plant Protoplasts. 12.00 Chairperson: I.K. Vasil (Gainesville, USA)

10.30 M.R. Davey (Nottingham, U.K.) - Recent developments in the culture and regeneration of plant protoplasts. 11.15 P. Dale (Aberystwyth, U.K.) - Protoplast culture and plant regeneration of cereals and other recalcitrant crops.

12.10- WOrkshop 1: Plant Protoplasts 13.10 Organelle transfer, sorting-out, recombination.

Organizers: S. Izhar (Bet Dagan, Israel), H. Lörz (Köln, FRG)

13.10- Lunch Break 15.00- Poster Session 2: Plant Protoplasts 18.00 Poster Code: PA 1-8. PB 1-17. PC 1-7 15.00- Lecture Session 2: Microbial Protoplasts 16.30 Chairperson: J. Peberdy (Nottingham, U.K.)

15.00 H.R. Baltz (Indianapolis, USA) - Recent advances in protoplast methodology for antibio­tic-producing Streptomyces. 15.45 R.D. Hotchkiss (Albany, USA) - Chromosome interaction and expression in fused Bacillus protoplasts.

18.00- Dinner Break 20.30- Workshops 23.00 WOrkshop 2: Microbial Protoplasts.

Non-complementing diploids. Organizers and con­tributors: L. Alföldi. K. Fodor. L. Hirschbein. D. Hotchkiss. P. Schaefer. WOrkshop 3: Plant Protoplasts. Protoplast culture and plant regeneration with special emphasis on crop plants. Organizers: I. Potrykus (Basel, Switzerland), I.K. Vasil (Gainesville. USA). WOrkshop 4: Plant Protoplasts. Mutation and stable variation. Org?nizers: P.J. King (Basel, Switzerland). J. io.!dholm (Urbar,a, USA).

SATURDAY. AUGUST 13 09.30- Poster Session 'I: Microbial Protoplasts 12.00 Poster Code: Me 1-5. ME 1-4 09.00- Lecture Se,ssion 3: Plant Protoplasts 11.00 Chairperson: P. Maliga (Szeged, Hungary)

09.00 J.P. Bourgin (Versailies, France) - Proto­plasts and the isolation of mutants. 09.30 P.,J. Larkin (Canberra, Australia) - Proto­plasts and var·iation in culture. 10.00 P. Lurquin (Pul Iman, USA) - Systems for plant protoplast transformation.

11.00- Poster Se~sion 4: Plant Protoplasts. 13.00 Po~ter Code PK 1-16 13.00- Lun/:l Break l!i.dU- Poster Session 5: Plant Protoplasts. 18.00 Poster Code PF 1-6. PH 1-4. PN 1-2. PO 1-2.

PQ 1-8

15.00- lecture Session 4: Microbial Protoplasts. 16.45 Chairperson: F. Kevei (Szeged, Hungary).

15.00 A.J. Morgan (Sunburry-on-Thames, U.K.) -Yeast strain improvement by protoplast fusion and transformation. 15.45 J. AnnA (Louvain, Belgium) - Protoplasts of filamentous fungi in genetics and metabolite production. 16.15 J. Croft (Birmingham, U.K.) - Interspeci­fic somatic hybridization in Aspergillus.

18.00- Dinner Break 20.30 WOrkshop 5: Plant Protoplasts. Protoplasts as

tools for physiological studies. Organizers; T. BoIler (Basel), D. Galbraith (Lineoln. USA). WOrkshOp 6: Plant Protoplasts: Genetic trans­formation. Organizers: R.D.Shillito (Basel). H.H. Steinbiss (Köln. FRG)

11

tIlNDAY. AUGUST 15 09.30- Poster Session 6: Microbial Protoplasts. 12.00 Poster Code NA 1-8. Me 1-4 09.00- lecture Session 5: Plant Protoplasts. 11.00 Chairperson: O. Schieder (Köln. FRG)

09.00 G.B. Lazar (Szeged. Hungary) - Recent De­velopments in fusion and selection technology. 09.30 C.T.Harms (Zürich. Switzerland) - Somatic hybridization by protoplast fusion. 10.30 R. Fluhr (Rehovot. Israel) - Sorting out of organelles and cytoplasmic traits in samatic fusion hybrids.

11.00- Poster Session 7: Plant Protoplasts. 13.00 Poster Code PR 1-7 13.00 Lunch Break 15.00- Poster Session 8: Plant Protoplasts. 18.00 Poster Code PD 1-21. PE 1-6. PG 1-5 15.00- lecture Session 6: Microbial Protoplasts. 17.15 Chairperson: A. Hinnen (Basel, Switzerland)

15.00 R. Sentandreu (Valencia, Spain) - Cytoche­mical and physiological aspects of yeast proto­plasts. 15.45 J. Makins (Syracuse. USA) - The theory and practical application of liposome-protoplast interactions. 16.30 A. Kotyk (Prague. CSSR) - Transport of nutrients in yeast protoplasts.

18.00 Dinner Break 20.30- Workshops 23.00 Workshop 7: Microbial Protoplasts.

Fungal protoplasts. Organizers: l. Ferenczy (Szeged.Hungary). J.F. Peberdy (Nottingham. UK) Workshop 8: Plant protoplasts. Protoplasts as tools in pathology. virology.and plant-microbe interactions. Organizers: R. Hohl (Zürich. Switzerland), H.P. Mühlbach (München. FRG). WOrkshop 9: Plant protoplasts. Somatic hybridi­zation. Organizers: P. Maliga (Manhattan. USA). O. Schieder (Köln. FRG).

TUESDAY, AUGUST 16 09.00- Lecture Session 7: Plant Protoplasts 11.00 Chairperson: E. Galun (Rehovot, Israel).

09.00 Y. Meyer (Perpignan. France) - The first mitotic cycle of mesophyll protoplasts. 09.30 L. Fowke (Saskatoon. Canada) - Protoplasts for studies of plasma membrane and associated organelles. 10.00 H.P. Mühlbach (München. FRG) - The use of protoplasts in plant virus research.

11.00 Workshop 10: Plant protoplasts. Protoplasts and plant breeding. Organizers: D. Evans (Cinnaminson. USA), E. Galun (Rehovot)

13.00 Lunch break 14.00- Lecture Session 8: Microbial Protoplasts. 15.00 Chairperson: R. Hütter (Zürich. Switzerland)

14.00 H.H. Martin (Darmstadt. FRG) - protoplasts and spheroplasts in the gram-negative bacterium Proteus mirabilis. Physiological and biochemi­cal properties. 14.30 U. Taubeneck (Jena. DDR) - The stable pro­toplast type L-form of bacteria - characteristic properties and biological significance.

14.00- Poster Session 9: Plant Protoplasts 16.00 Poster Code PI 1-2, PJ 1-2, Pl 1-6, PM 1-3,

PP 1-4 16.00- Closing Lectures 18.00 16.00 O. lampen (Piscataway. USA) - Advances in

microbial protoplasts. Chairperson: R. Hütter (Zürich, Switzerland) 17.00 E.C. Cocking (Nottingham, U.K.) - Appli­cations of protoplast technology in agriculture. Chairperson: G. Melchers (Tübingen. FRG)

18.00 Closing remarks: I. Potrykus

ORGAN I ZERS I. Potrykus (Basel), R. Hütter (Zürich). P.J. King (Basel) C.T. Harms (Zürich). A. Hinnen (Basel). R.D. Shillito (Basel), R. Forlen (B3sel). M. Marti (Basel),

Secretary General Secretary Treasurer Local Organizer Loca I Organizer Local Organizer Congress Office MUBA service

12

Welcome Reception offered by the City Council of Basel in the court yard of the Basel Art Museum, where the participants met beside Rodin's 'Citisens of Calais' (left) and under Borofsky's 'Flying Man' (right)

to listen to the Welcome of the Dean of the Faculty for Natural Sciences of the University of Basel (left), to taste Swiss wine (right), to exchange brand-new ideas or gossip, to revive friendships or create contacts.

M.R. Davey (Nottingham) and R.D. Shillito (Basel)

M. Gabor and R.D. Hotchkiss (Albany, New York) and K. Fodor (Szeged).

L. Alföldi (Szeged) and C.J. Bos (Wageningen)

A. Maretzki (Hawaii) and T. BoIler (Basel)

Evening Workshops: Participants from China, Japan, India, Germany, Switzerland, United States, Nigeria, France, Belgium, Israel and other countries folIowand participate in discussions often beyond 11.00 p.m.

Rhine swimming: The weather was warm enough and the water clean enough to allow for refreshment between afternoon sessions and evening workshops by floating down the Rhine through the old City of Basel.

13

Symposium Party: Organized by the 'Männerchor of a nearby village (left, fourth fram left is P.J.King), in the gymnasium of a local sChool in the form of a Swiss 'Heimatabend' with typical folk music and dancing.

14

M.R. Davey (speaker) I.K. Vasil (chairman) P. Dale (speaker)

P.J. Larkin (speaker) J.P. Bourgin (speaker) P. Lurquin (speaker)

G.B. lazar (speaker) C.T. Harms (speaker)

o. Schieder (chairman) R. Fluhr (speaker) Y. Meyer (speaker)

E.C. Cocking (speaker) L. Fowke (speaker) E. Galun (chairman)

15

F. Kevei (chairman) J. troft (soeaker) H.H. Martin (speaker)

O. Lampen (speaker) L. Ferenczy (speaker) A.J. Morgan (speaker),

J. Makins (speaker) R. Sentandreu (speaker) J. Peberdy (chairman)

J. Anne (speaker) R. Hütter (chairman) U. Taubeneck (speaker)

G. Melchers (chairman)