Volumes already published

Volume 1: Trees I (1986)

Volume 2: Crops I (1986)

Volume 3: Potato (1987)

Volume 4: Medicinal and Aromatic Plants 1(1988)

Volume 5: Trees II (1989)

Volume 6: Crops II (1988)

Volume 7: Medicinal and Aromatic Plants II (1989)

Volume 8: Plant Protoplasts and Genetic Engineering I (1989)

Volume 9: Plant Protoplasts and Genetic Engineering II (1989)

Volume 10: Legumes and Oilseed Crops I (1990)

Volume 11: Somaclonal Variation in Crop Improvement I (1990)

Volume 12: Haploids in Crop Improvement 1(1990)

Volume 13: Wheat (1990)

Volume 14: Rice (1991)

Volume 15: Medicinal and Aromatic Plants III (1991)

Volume 16: Trees III (1991)

Volume 17: High-Tech and Micropropagation I (1991)

Volume 18: High-Tech and Micropropagation II (1992)

Volume 19: High-Tech and Micropropagation III (1992)

Volume 20: High-Tech and Micropropagation IV (1992)

Volume 21: Medicinal and Aromatic Plants IV (1993)

Volume 22: Plant Protoplasts and Genetic Engineering III (1993)

Volume 23: Plant Protoplasts and Genetic Engineering IV (1993)

Volume 24: Medicinal and Aromatic Plants V (1993)

Volumes in preparation

Volume 25: Maize (1994)

Volume 26: Medicinal and Aromatic Plants VI (1994)

Volume 27: Somatic Hybridization in Crop Improvement

Volume 28: Medicinal and Aromatic Plants VII

Volume 29: Plant Protoplasts and Genetic Engineering V

Volume 30: Somatic Embryogenesis and Synthetic Seed I

Biotechnology in Agriculture and Forestry 22

Plant Protoplasts and Genetic Engineering III

Edited by Y P. S. Bajaj

With 103 Figures

Springer-Verlag Berlin Heidelberg New York London Paris Tokyo Hong Kong Barcelona Budapest

Professor Dr. Y P. S. BAJAJ

A-137 New Friends Colony New Delhi 110065, India

ISBN-13: 978-3-642-78008-0 DOl: 10.1007/978-3-642-78006-6

e-ISBN-13: 978-3-642-78006-6

Library of Congress Cataloging-in-Publication Data. (Revised for volume 3). Plant protoplasts and genetic engineering. (Biotechnology in agriculture and forestry; 8-9, 22) Includes bibli­ographies and indexes. 1. Plant protoplasts. 2. Plant genetic engineering. I. Bajaj, Y.P.S., 1936-II. Series. QK725.P578 1989 581.87'3 88-31197

This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microftlms or in any other way, and storage in data banks. Duplication of this publication or parts thereof is permitted only under the provisions of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer-Verlag. Violations are liable for prosecution under German Copyright Law.

© Springer-Verlag Berlin Heidelberg 1993 Softcover reprint of the hardcover 1st edition 1993

The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use.

Production Editor: Ingrid Samide, Heidelberg Typesetting: Macmillan India Limited, Bangalore 31/3145-5 4 3 2 1 0 - Printed on acid-free paper

Dedicated to Professor Dr. Martin Bopp with whom I had the privilege of working at the University of Heidelberg, Germany, from 1970-1972

Preface

Since the publication of Plant Protoplasts and Genetic Engineering I and Ifin 1989, there has been a steady inflow of new information in the area of genetic engineering of plants. The number of species in which complete plants have been regenerated from isolated proto­plasts has increased considerably. Likewise, numerous genetic trans­formation studies have been conducted successfully on a number of crop species. This is the most compelling reason for compiling the following two new volumes which contain 58 chapters contributed by international experts.

Plant Protoplasts and Genetic Engineering III comprises 29 chapters dealing with the regeneration of plants from protoplasts, and ge­netic transformation in various species of Agrostis, Arabidopsis, Atropa, Brassica, Catharanthus, Cucumis, Datura, Daucus, Digitalis, Duboisia, Eustoma, Festuca, Helianthus, Hordeum, Kalanchoe, Linum, Lobelia, Lolium, Lotus, Lycium, Lycopersicon, Mentha, Nicotiana, Pelargonium, Pisum, Pyrus, Salvia, Scopolia and Solanum.

Plant Protoplasts and Genetic Engineering IV comprises 29 chapters on various species of Actinidia, Armoracia, Beta, Brassica, Cicer, Citrus, Cucumis, Duboisia, Fragaria, Glycine, Ipomoea, Lactuca, Lotus, Lycopersicon, Manihot, Medicago, Nicotiana, Petunia, Phase­olus, Picea, Pisum, Prunus, Psophocarpus, Saccharum, Solanum, Sorghum, Stylosanthes, Vitis and Zea mays.

These studies reflect the far-reaching implications of protoplast technology in agricultural and forest biotechnology. Therefore, these books may be of special interest to advanced students, teach­ers, and research scientists in the field of plant tissue culture, molec­ular biology, genetic engineering, plant breeding, and general bio­technology.

New Delhi, September 1993 Professor Y. P. S. BAJAJ Series Editor

Contents

Section I Regeneration of Plants from Protoplasts

1.1 Regeneration of Plants from Protoplasts of Agrostis alba (Redtop) Y. ASANO (With 5 Figures)

1 Introduction ........................................ 3 2 Induction of Embryogenic Callus and Establishment

of Embryogenic Suspension Culture . . . . . . . . . . . . . . . . . . . . 3 3 Isolation of Pro top lasts .............................. 6 4 Culture of Protoplasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 5 Regeneration of Plants ............................... 8 6 Conclusions ........................................ 9 7 Protocol ........................................... 10 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 11

1.2 Regeneration of Plants from Protoplasts of Atropa belladonna L. (Deadly Nightshade) Y. P. S. BAJAJ (With 5 Figures)

1 Introduction ........................................ 13 2 Isolation and Culture of Protoplasts ................... 13 3 Protoplast Fusion and Somatic Hybridization ........... 19 4 Cryopreservation of Protoplasts ....................... 19 5 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 20 6 Protocol ........................................... 20 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 21

1.3 Regeneration of Plants from Protoplasts of Brassica alboglabra Bailey (Chinese Kale) E.-C. PUA (With 4 Figures)

1 Introduction ........................................ 22 2 Isolation of Protoplasts .............................. 23 3 Culture of Protoplasts ....... : . . . . . . . . . . . . . . . . . . . . . . .. 24 4 Regeneration and Acclimatization of Plants ............. 26 5 Characterization of Protoplast-Derived Plants ........... 27 6 Gene Transfer into Protoplasts Using Electroporation .... 27

x Contents

7 Conclusions and Prospects .. . . . . . . . . . . . . . . . . . . . . . . . . .. 28 8 Protocol ........................................... 29 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 30

1.4 Regeneration of Plants from Protoplasts of Cucumis sativus L. (Cucumber) Z. K. PUNJA and S. H. T. RAHARJO (With 4 Figures)

1 Introduction ........................................ 31 2 Isolation of Pro top lasts .............................. 34 3 Culture of Protoplasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 36 4 Regeneration of Plants ............................... 37 5 Fusion of Protoplasts ................................ 39 6 Conclusions and Prospects. . . . . . . . . . . . . . . . . . . . . . . . . . .. 41 7 Protocol for Isolation, Culture, and Regeneration

of Cucumber Protoplasts ............................. 42 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 43

1.5 Regeneration of Plants from Protoplasts of Lolium (Ryegrasses) and Festuca (Fescues) S. J. DALTON (With 3 Figures)

1 Introduction ........................................ 46 2 Isolation of Protoplasts .............................. 49 3 Culture of Protoplasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 56 4 Culture Media and Conditions ........................ 58 5 Regeneration from Protoplast Colonies ... . . . . . . . . . . . . .. 60 6 Somaclonal Variation in Regenerated Plants . . . . . . . . . . . .. 61 7 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 62 8 Protocol ........................................... 64 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 67

1.6 Regeneration of Plants from Protoplasts of Lotus spp. (Birdsfoot trefoil) M. NIIZEKI (With 2 Figures)

1 Introduction ........................................ 69 2 Protoplast Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 70 3 Protoplast Culture and Plant Regeneration . . . . . . . . . . . . .. 73 4 Conclusion and Prospects ............................ 75 5 Protocol ........................................... 76 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 77

1. 7 Regeneration of Plants from Protoplasts of Lycium barbarum L. (Wolfberry) Y 1. RAruSHNYAK, V. A. RUDAS, and N. M. PlVEN (With 4 Figures)

1 Introduction ........................................ 79 2 Plant Material and Callus Culture ..................... 79

Contents XI

3 Mesophyll and Callus Protoplast Isolation .............. 81 4 Mesophyll Protoplast Culture and Plant Regeneration .... 81 5 Culture of Callus-Derived Protoplasts .................. 85 6 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 85 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 86

I.8 Regeneration of Plants from Protoplasts of Pelargonium spp. (Geranium) K. B. DUNBAR (With 2 Figures)

1 Introduction ........................................ 87 2 Isolation of Protoplasts .............................. 89 3 Culture of Pro top lasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 90 4 Regeneration of Plants ............................... 92 5 Plastid Inheritance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 93 6 Conclusion ......................................... 93 7 Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 94 References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 95

I.9 Regeneration of Plants from Protoplasts of Pea (Pisum sativum L.) R. LEHMINGER-MERTENS and H.-I JACOBSEN (With 2 Figures)

1 Introduction ........................................ 97 2 Isolation of Protoplasts .............................. 98 3 Protoplast Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 100 4 Regeneration of Plants ............................... 101 5 Protocol ........................................... 103 References ............................................ 104

1.10 Regeneration of Plants from Protoplasts of Pyrus spp. (Pear) S. I OCHATT (With 1 Figure)

1 Introduction ........................................ 105 2 Isolation of Protoplasts .............................. 109 3 Culture of Protoplasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 110 4 Plant Regeneration from Protoplast-Derived Callus ...... 113 5 Protoplast Fusion and Somatic Hybridization Studies

with Pyrus Protoplasts ............................... 114 6 Miscellaneous Studies with Pyrus Protoplasts ............ 114 7 Summary and Conclusion ............................ 117 8 General Protocols ................................... 117 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 119

XII Contents

Section II Genetic Transformation

II.1 Transformation in Arabidopsis thaliana R. BILANG and 1. POTRYKUS (With 3 Figures)

1 Introduction ........................................ 123 2 Plant Culture and Protoplast Isolation ................. 124 3 Direct Gene Transfer to Protoplasts ... . . . . . . . . . . . . . . . .. 125 4 Selection of Transformed Colonies ..................... 128 5 Plant Regeneration .................................. 131 6 Concluding Remarks ................................. 132 References ............................................ 133

II.2 Transformation in Atropa belladonna Y SUZUKI, Y KURIOKA, T. OGASA W ARA, and H. KAMADA (With 4 Figures)

1 Introduction ........................................ 135 2 Preparation of Agrobacterium ......................... 135 3 Transformation ..................................... 138 4 Elimination of Bacteria and Selection of Transformed Cells 138 5 Regeneration of Plants ............................... 139 6 Transgenic A. belladonna ............................. 140 7 Conclusion ......................................... 141 8 Protocols for Transformation ......................... 142 References ............................................ 142

II.3 Transformation in Catharanthus Species (Madagascar Periwinkle) C. DAVID and J. TEMPE (With 3 Figures)

1 Introduction ........................................ 144 2 Importance of Catharanthus . . . . . . . . . . . . . . . . . . . . . . . . . .. 145 3 Genetic Transformation of Catharanthus Species ......... 146 4 Potential of Hairy Root Cultures ...................... 151 5 Transformation of C. roseus and Plant Pathology ........ 152 6 Genetic Engineering with Genes Involved

in Tropane Alkaloids Synthesis ........................ 152 7 Conclusion ......................................... 153 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 154

II.4 Transformation in Datura Species P. CHRISTEN and M. F. ROBERTS (With 1 Figure)

1 Introduction ........................................ 157 2 Tropane Alkaloid Production ......................... 157 3 Transformation ..................................... 159 4 Summary and Conclusion ............................ 169 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 169

Contents

II.5 Transformation in Daucus carota L. (Carrot) R. BOWER and R. G. BIRCH (With 2 Figures)

XIII

1 Introduction ........................................ 172 2 Stable Genetic Transformation of Carrot ............... 172 3 Applications of Electroporation of Carrot Protoplasts .... 173 4 Isolation and Culture of Carrot Pro top lasts ............. 175 5 Electroporation Parameters ........................... 175 6 Protocol for the Establishment of an Electroporation System 179 7 Conclusion ......................................... 180 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 180

II.6 Transformation in Digitalis purpurea L. (Foxglove) K. SAITO, M. YAMAZAKI, K. SHIMOMURA, K. YOSHIMATSU, and I. MURAKOSHI (With 6 Figures)

1 Introduction ........................................ 182 2 Transformation ..................................... 182 3 Expression of Chimeric Genes in Transgenic Digitalis Roots . 185 4 Production of Cardioactive Glycosides ................. 186 5 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 186 6 Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 187 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 188

II. 7 Transformation in Duboisia spp. y. MANO (With 4 Figures)

1 Introduction ........................................ 190 2 Genetic Transformation in Duboisia by the Ri T-DNA

of Agrobacterium rhizogenes ........................... 190 3 Somatic Hybrids Between Duboisia and Nicotiana

by Protoplast Fusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 196 4 Summary and Conclusions ............................ 199 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 199

II.8 Transformation in Eustoma grandiflorum (Lisianthus) S. C. DEROLES, S. E. LEDGER, R. M. MILLER, K. M. DAVIES, and N. K. GIVEN (With 3 Figures)

1 Introduction ........................................ 202 2 Breeding and In Vitro Propagation .................... 202 3 Agrobacterium-Mediated Transformation ............... 203 4 Analysis of Early Transformation Events ............... 204 5 Production of Kanamycin-Resistant Shoots ............. 206 6 Southern Hybridization Analysis ...................... 209 7 Future Directions: Manipulation of Color in Lisianthus .. 210 References ............................................ 212

XIV

11.9 Transformation in Helianthus annuus L. (Sunflower) B. SCHRAMMEIJER, A. HOEKEMA, and P. C. SUMONS (With 3 Figures)

Contents

1 Introduction ........................................ 213 2 Methodology and Experimental Results ................ 214 3 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 218 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 219

II.10 Transformation in Hordeum vulgare (Barley) H. TOYODA (With 7 Figures)

1 Introduction ........................................ 220 2 Culture of Barley Coleoptile Tissues ................... 220 3 Transient Expression of Foreign Genes Introduced

into Barley Coleoptile Cells by Microinjection ........... 221 4 Assessment of Transformation Strategy with Respect

to Disease Resistance ................................ 225 5 Promoter Sequences of Barley Responsible for Infection

with Powdery Mildew Pathogen ....................... 228 6 Conclusion and Prospects ............................ 231 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 232

11.11 Transformation in Kalanchoe S.-R. JIA (With 5 Figures)

1 Introduction ........................................ 234 2 Transient Gene Expression . . . . . . . . . . . . . . . . . . . . . . . . . . .. 235 3 Stable Transformation ............................... 237 4 Transformation Protocol ............................. 240 5 Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 241 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 242

11.12 Transformation in Linum usitatissimum L. (Flax) M. C. JORDAN and A. McHuGHEN (With 1 Figure)

1 Introduction ........................................ 244 2 Agrobacterium-Mediated Flax Transformation ........... 244 3 Genetic Engineering for Flax Crop Improvement ........ 246 4 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 250 References ............................................ 251

II.13 Transformation in Lobelia injlata M. TANAKA, H. YONEMITSU, K. SHIMOMURA, K. ISHIMARU, S. MOCHIDA, T. EDNO, and A. KAJI (With 7 Figures)

1 Introduction ........................................ 253 2 Establishment of Hairy Root Culture and Production

of Secondary Metabolites . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 253

Contents xv

3 Results and Discussion ............................... 256 4 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 263 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 264

II.14 Transformation in Lycopersicon esculentum L. (Tomato) C. BELLINI (With 5 Figures)

1 Introduction ........................................ 265 2 Parameters Important for the Preparation and Culture

of Tomato Protoplasts ............................... 266 3 Direct Gene Transfer into Mesophyll Protoplasts

of L. esculentum and L. peruvianum by Electroporation ... 270 4 Conclusion ......................................... 274 References ............................................ 276

11.15 Transformation in Mentha Species (Mint) A. SPENCER, J. D. HAMILL, and M. J. C. RHODES (With 1 Figure)

1 Introduction ........................................ 278 2 Procedures for the Genetic Manipulation of Mentha . ..... 279 3 Phenotypes of Mentha Galls Transformed

by Different Strains of A. tumefaciens .................. 284 4 Analysis of Transformed Shoot Cultures .............. .. 288 5 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 291 References ............................................ 291

II.16 Transformation in Nicotiana edwardsonii J. M. KIERNAN, F. C. Wu, K.-B. GOLDBERG, S. GOWDA, and R. J. SHEPHERD (With 6 Figures)

1 Introduction ........................................ 294 2 Leaf Disc Transformation and Regeneration ............ 295 3 Transient Expression of Introduced Genes

in Nicotiana edwardsonii .............................. 301 4 Conclusion ......................................... 304 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 305

II.17 Transformation in Salvia chinensis ZHI-HONG Xu and LI-JUN YANG (With 4 Figures)

1 Introduction ........................................ 308 2 Plant Regeneration System ........................... 308 3 Transgenic Plants and Shoots Produced

by Agrobacterium-Mediated Transformation ............. 309 4 Protocol ........................................... 312 5 Conclusions ........................................ 313 References ............................................ 313

XVI

II.18 Transformation in Scopolia Y L. ZHANG (With 4 Figures)

Contents

1 Introduction ........................................ 314 2 Cell Culture of Scopolia .......................... ,... 315 3 Ri Plasmid Transformation and Hairy Root Culture ...... 315 4 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . .. 318 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 318

II.19 Transformation in Solanum melongena L. (Eggplant) M. LEONE, E. FILIPPONE, and P. F. LURQUIN (With 2 Figures)

1 Introduction ........................................ 320 2 General Transformation Strategies ..................... 321 3 Eggplant Transformation by Cocultivation .............. 321 4 Transgenic Somatic Embryos . . . . . . . . . . . . . . . . . . . . . . . . .. 324 5 Conclusions ........................................ 326 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 327

SUbject Iudex ......................................... 329

List of Contributors

ASANo, Y, Faculty of Horticulture, Chiba University, 6-2-1 Kashiwa-no-ha, Kashiwa city, Chiba Pref., Japan 277

BAJAJ, Y P. S., Former Professor of Tissue Culture, Punjab Agricultural University, Ludhiana, India. (Present address: A-137 New Friends Colony, New Delhi 110065, India)

BELLINI, C., Laboratoire de Biologie Cellulaire, I.N.R.A., Route de Saint-Cyr, 78026 Versailles Cedex, France

BILANG, R., Institute of Plant Sciences, Swiss Federal Institute of Technology, ETH Zentrum, 8092 Zurich, Switzerland

BIRCH, R. G., Department of Botany, The University of Queensland, Brisbane, 4072, Australia

BOWER, R., Department of Botany, The University of Queensland, Brisbane, 4072, Australia

CHRISTEN, P., Dept. de Pharmacognosie, Ecole de Pharmacie, Universite de Geneve, 30, Quai E.-Ansermet, 1211 Geneve 4, Switzerland

DALTON, S. 1., Institute of Grassland and Environmental Research, WPBS, PI as Gogerddan, Aberystwyth, Dyfed, SY23 3EB, United Kingdom

DAVID, c., Institut des Sciences Vegetales, CNRS, Avenue de la Terrasse, 91198 Gif sur Yvette, France

DAVIES, K. M., Levin Research Centre, Crop and Food Research Ltd, Private Bag, Levin, New Zealand

DEROLES, S. C., Levin Research Centre, Crop and Food Research Ltd, Private Bag, Levin, New Zealand

DUNBAR, K. B., Division of Natural Science and Mathematics, Dalton College, 213 North College Drive, Sequoya 127A,Dalton, GA 30720, USA

EDNO, TOHRU, Research Institute for Molecular Genetics, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki, Ibaraki 300-11, Japan

XVIII List of Contributors

FILIPPONE, E., Dipartimento Scienze Agronomiche e Genetica Vegetale, Universita di Napoli, 80055 Portici, Italy

GIVEN, N. K., died in June 1991

GOLDBERG, K.-B., Department of Plant Pathology, University of Kentucky, Lexington, KY 40546, USA

GOWDA, S., Department of Plant Pathology, University of Kentucky, Lexington, KY 40546, USA

HAMILL, I D., Department of Genetics and Developmental Biology, Monash University, Melbourne, Australia

HOEKEMA, A., MOGEN Int. nv, Einsteinweg 97, 2333 CB Leiden, The Netherlands

McHuGHEN, A., National Research Council of Canada, Plant Biotechnology Institute, Saskatoon, Sask. S7N OW9, and Crop Development Centre, University of Saskatchewan, Saskatoon, Sask. S7N OWO, Canada

ISHIMARU, KANJI, Genetic Engineering Laboratory, Faculty of Agriculture, Saga University, Honjo, Saga 840 Japan

JACOBSEN, H.-I, Lehrgebiet Molekulargenetik, Universitat Hannover, Herrenhauserstr. 2, 30419 Hannover, Germany

JIA, SHI-RoNG, Biotechnology Research Center, Chinese Academy of Agricultural Sciences, 30 Baishiqiao Road, Beijing 100081, China

JORDAN, M. c., National Research Council of Canada, Plant Biotechnology Institute, Saskatoon, Sask. S7N OW9, and Crop Development Centre, University of Saskatchewan, Saskatoon, Sask. S7N OWO, Canada

KAJI, AKIRA, Department of Microbiology, School of Medicine, University of Pennsylvania, Philadephia, PA 19104-6076, USA

KAMADA, HIROSHI, Gene Experiment Center, University of Tsukuba, Tsukuba, Ibaraki 305, Japan

KIERNAN, I M., Department of Plant Pathology, University of Kentucky, Lexington, KY 40546, USA

KURIOKA, YURIKO, Gene Experiment Center, University of Tsukuba, Tsukuba, Ibaraki 305, Japan

LEDGER, S. E., Levin Research Centre, Crop and Food Research Ltd, Private Bag, Levin, New Zealand

LEHMINGER-MERTENS, R., Institut f. Genetik, Universitat Bonn, Kirschallee 1, 53115 Bonn, Germany. (present address: Reiherweg 4, 50374 Erftstadt-Liblar, Germany)

List of Contributors XIX

LEONE, M., Centro di Studio per il Miglioramento Genetico degli Ortaggi, CNR, 80055 Portici, Italy

LURQUIN, P. E, Department of Genetics and Cell Biology, Washington State University, Pullman, WA 99164-4234, USA

MANO, YOSHIHIRO, Department of Biological Science and Technology, Tokai University, 317 Nishino, Numazu, Shizuoka 410-03, Japan

MILLER, R. M., Levin Research Centre, Crop and Food Research Ltd, Private Bag, Levin, New Zealand

MocHIDA, SHUNJI, Research Institute for Molecular Genetics, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki, Ibaraki 300-11, Japan

MURAKOSHI, ISAMU, Faculty of Pharmaceutical Sciences, Chiba University, Yayoi-cho 1-33, Inage-ku, Chiba 263, Japan

NIIZEKI, M., Plant Breeding Laboratory, Faculty of Agriculture, Hirosaki University, Hirosaki, Aomori-ken 036, Japan

OCHATT, S. J., LN.R.A., Station d'AmeIioration des Especes Fruitieres et Ornementales, Centre d'Angers, B.P. 57,49071 Beaucouze Cedex, France

OGASAWARA, TAKESHI, Gene Experiment Center, University of Tsukuba, Tsukuba, Ibaraki 305, Japan

PIVEN, N. M., Institute of Cell Biology and Genetic Engineering, Ukrainian Academy of Sciences, Acad. Lebedev Str., 1, 252650, Kiev-GSP-22, Ukraine

POTRYKUS, 1., Institute of Plant Sciences, Swiss Federal Institute of Technology, ETH Zentrum, 8092 Zurich, Switzerland

PUA, ENG-CHONG, Institute of Molecular and Cell Biology, National University of Singapore, 10 Kent Ridge Crescent, Singapore 0511, Republic of Singapore. (Present address: Department of Botany, National University of Singapore, Kent Ridge, Singapore 0511)

PUNJA, Z. K., Department of Biological Sciences, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6

RAHARJO, S. H. T., Department of Biological Sciences, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6

RATUSHNYAK, Y 1., Institute of Cell Biology and Genetic Engineering, Ukrainian Academy of Sciences, Acad. Lebedev Str., 1, 252650, Kiev-GSP-22, Ukraine

xx List of Contributors

RHODES, M. J. C., Genetics and Microbiology Department, AFRC Institute of Food Research, Norwich Laboratory, Norwich, United Kingdom

ROBERTS, M. E, Department of Pharmacognosy, The School of Pharmacy, University of London, London WC1N 1AX, United Kingdom

RuoAS, V. A., Institute of Cell Biology and Genetic Engineering, Ukrainian Academy of Sciences, Acad. Lebedev Str., 1,252650, Kiev-GSP-22, Ukraine

SAITO, KAZUKI, Faculty of Pharmaceutical Sciences, Chiba University, Yayoi-cho 1-33, Inage-ku, Chiba 263, Japan

SCHRAMMElJER, B., MOGEN Int. nv, Einsteinweg 97, 2333 CB Leiden, The Netherlands

SHEPHERD, R. J., Department of Plant Pathology, University of Kentucky, Lexington, KY 40546, USA

SmMoMURA, KOIClflRO, Tsukuba Medicinal Plant Research Station, National Institute of Hygienic Sciences, 1 Hachimandai, Tsukuba, Ibaraki 305, Japan

SIJMONS, P. C., MOGEN Int. nv, Einsteinweg 97, 2333 CB Leiden, The Netherlands

SPENCER, A., Department of Biological Sciences, University of East Anglia, Norwich, Norfolk, United Kingdom

SUZUKI, YOSHIlllTO, Faculty of Agriculture, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan

TANAKA, MASAlflKO, Research Institute for Molecular Genetics, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki, Ibaraki 300-11, Japan

TEMPE, J., Institut des Sciences Vegetales, CNRS, Avenue de la Terrasse, 91198 Gif sur Yvette, France, and Institut National Agronomique Paris Grignon, 75231 Paris, France

TOYODA, HIDEYOSm, Faculty of Agriculture, Kinki University, Nakamachi 3327-204, Nara 631, Japan

Wu, E G., Department of Plant Pathology, University of Kentucky, Lexington, KY 40546, USA

Xu, Zm-HONG, Shanghai Institute of Plant Physiology, Academia Sinica, Shanghai 200032, China

YAMAZAKI, MAMI, Faculty of Pharmaceutical Sciences, Chiba University, Yayoi-cho 1-33, Inage-ku, Chiba 263, Japan

List of Contributors XXI

YANG, LI-JUN, Shanghai Institute of Plant Physiology, Academia Sinica, Shanghai 200032, China

YONEMITSU, HIROSIll, Research Institute for Molecular Genetics, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki, Ibaraki 300-11, Japan

YOSIllMATSU, KAyO, Tsukuba Medicinal Plant Research Station, National Institute of Hygienic Sciences, 1 Hachimandai, Tsukuba, Ibaraki 305, Japan

ZHANG, Y. L., Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Hai Dian District, Dong Bei Wang, Beijing 100094, China