Proceedings in Life Sciences

First International Congress on Medicinal Plant Research, held at the University of Munich, Germany, September 6-10, 1976. Section A: N ew Natural Products and Plant Drugs with Pharmacological, Biological or Therapeutical Activity. Eds.: H.K. Wagner/P.M. Wolff. Published by Springer-Verlag Berlin Heidelberg New York 1977.

Plant Tissue Culture and Its Bio-technological Application Proceedings of the First International Congress on Medicinal Plant Research, Section B, held at the University of Munich, Germany September 6-10,1976

Edited by W Barz E. Reinhard M. H. Zenk

With 196 Figures

Springer-Verlag Berlin Heidelberg New York 1977

Professor Dr. W. BARZ Lehrstuhl fUr Biochemie der Pflanzen Westfa]ische Wilhelms-UniversiHit, 4400 Miinster/FRG

Professor Dr. E. REINHARD Lehrstuhl fUr Pharmazeutische Biologie der Universitat Auf der Morgenstelle, 7400 Tiibingen/FRG

Professor Dr. M. H. ZENK Lehrstuhl fUr Pflanzenphysiologie Ruhr-Universitat Bochum, 4630 Bochum-Querenburg/FRG

The cover motive symbolizes a culture vessel with suspended plant cells (left), a vial with a young plant regenerated from a cell culture (center) and a fermenter used for large scale cultivation of plant cells (right).

ISBN-13: 978-3-642-66648-3 DOl: 10.1007/978-3-642-66646-9

e-ISBN-13: 978-3-642-66646-9

Library of Congress Cataloging in Publication Data. International Congress on Medicinal Plant Research, 1st, University of Munich, 1976. Plant tissue culture and its bio-technological application. (Proceedings in life sciences) 1. Plant tissue culture-Congresses. I. Barz, Wolfgang, 1935- II. Reinhard, Ernst, 1926- III. Zenk, M. H., 1933- IV. Title. [DNLM: 1. Plants­Congresses. 2. Tissue culture-Congresses. W 3 IN632M 1976p/OK 725 1976pJ Q K 725.1494.1976.581:07'24.77-7216

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© by Springer-Verlag Berlin Heidelberg 1977.

Softcover reprint of the hardcover 1 5t edition 1977

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Prefu~

The possibility of growing plant cells like micro­organisms offers a basis for the development of new technologies. The scientific and biotechnological as­pects of this new field will be comprehensively dis­cussed in this volume.

The production of the natural ~roducts oj higher plants by means of plant cell suspension cultures seems to be possible. Consequently one can conceive the future industrial production of primary and secondary plant products by fermentation of plant cells.

Prior to the industrial application of plant cell cul­tures, however, several premises need to be investigated.

Cell culture strains have to be selected which produce the desired natural product in the highest possible yields and which simultaneously show high growth rates. Suitable fermentation technologies together with growth and production media for mass cultivation of plant cells have to be developed. Finally it is essential to develop analytical methods which can serve to select suitable cell clones. The radioimmunoassay technique which allows the quantitative analysis of metabolites of single plant cells seems to be a promising approach.

Furthermore tissue culture is becoming increasingly important as a tool for the mass-propagation of desir­able individual plants, the recovery of plants free from specific diseases and the production of somatic hybrids. Research on protoplast formation, recovery of haploid plant cells and the genetic manipulation of such cells are fruitful areas for the advancement of tissue culture application.

In all these fields notable progress has been achieved in the past years. It has been the aim of the organizers of the Munich symposium to offer a worldwide overview of the present situation in the field of technological application of plant cell cultures. Scientists from many countries participated in this effort.

The generous support of the Federal Minister of Research and Technology, Bonn, which is gratefully acknowledged here, made it possible to invite scientists from all parts of the world. They have all made outstanding contributions which have prepared the ground for the basis and development of these new biotechnologies. In comprehensive articles these scientists review the latest development of research in their respective fields.

VI

Much to our regret the colleagues invited from the German Democratic Republic (East-Germany) did not get permission from their government to attend the meet­ing. Unfortunately one of their manuscripts could not be included in this book. We deplore that a free ex­change of peaceful scientific ideas is thus still be­ing hampered.

We hope that the present volume comprehensively shows the latest developments in this fascinating field.

April, 1977 W. BARZ E. REINHARD M.H. ZENK

Contents

Cell Cultures and Secondary Products

Recent Advances in the Production of Medicinal Substances by Plant Cell Cultures M. TABATA •.•.....•..........•.•........•.•.••.•. 3

Production of Natural Substances by Plant Cell Cultures Described in Japanese Patents M. MISAWA •..............••.......••..........•.. 17

Formation of the Indole Alkaloids Serpentine and Ajmalicine in Cell Suspension Cultures of Catharan­thus rose us M.H. ZENK, H. EL-SHAGI, H. ARENS, J. ST5CKIGT, E.W. WEILER, and B. DEUS (With 10 Figures) ..•.... 27

Cardiac Glycosides in Cell Cultures of Digitalis T. KARTNIG (With 3 Figures) .................•... 44

Biochemistry, Physiology and Regulatory Aspects

The Common and Unusual Lipids of Plant Cell Cul­tures H.K. MANGOLD (With 3 Figures) .......•......•.... 55

Biosynthesis of Mevalonoid-Derived Compounds in Cell Cultures K.H. OVERTON (With 7 Figures) ..........•..•..... 66

Current Problems in the Regulation of Nitrogen Metabolism in Plant Cell Cultures D. K . DOUGALL ..•......•..........•....•...•....•.. 76

Improvement of Growth Rates of Plant Cell Cultures M. NOGUCHI, T. MATSUMOTO, Y. HIRATA, K. YAMAMOTO, A. KATSUYAMA, A. KATO, S. AZECHI, and K. KATO (With 6 Figures) ..••••••.......•.•....•....••••. 85

Regulatory Aspects of Phenylpropanoid Biosyn­thesis in Cell Cultures K. HAHLBROCK (With 7 Figures) •.........•...•...• 95

Selection and Characterization of Biochemical Mutants J. M. WIDHOLM (With 2 Figures) •....•.•...•.•..... 112

VIII

Biotransformation

Biotransformation of Cardiac Glycosides by Plant Cell Cultures A.W. ALFERMANN, H.M. BOY, P.C. D6LLER, W. HAGEDORN, M. HEINS, J. WAHL, and E. REINHARD (With 18 Fig-ures) •........•..••.......•.........•.......••.. 125

Metabolism of Steroids in Plant Tissue Cultures S.J. STOHS (With 5 Figures) ..•.....••........... 142

Catabolism

Catabolism of Endogenous and Exogenous Compounds by Plant Cell Cultures W. BARZ (With 13 Figures) .......•.•......•...... 153

Enzymological Aspects of Flavonoid Catabolism in Plant Cell Cultures W. H6sEL, G. BURMEISTER, P. KREYSING, and E. SURHOLT (With 5 Figures)...................... 172

Metabolism of Xenobiotics by Plant Cell Cultures H. SANDERMANN, Jr., H. DIESPERGER, and D. SCHEEL (With 6 Figures) ......•.......•................. 178

Uptake and Metabolism of Aldrin in Plant Tissue Cultures K.R. BRAIN and D.S. LINES (With 5 Figures) ....•. 197

Catabolism of y-Hexachlorocyclohexane (Lindane) by Plants and Cell Cultures - a Comparison J. STOCKIGT and B. RIES ..•.•.................... 204

General and Analytical Techniques

Plating of Plant Cells L. BERGMANN (With 7 Figures) 213

The Freeze-Preservation of Plant Cell Cultures L.A. WITHERS and H.E. STREET (With 9 Figures) ... 226

Cultivation of Plant Tissue Cultures in Bioreac­tors and Formation of Secondary Metabolites F. WAGNER and H. VOGELMANN (With 5 Figures) ...•• 245

Growth of Cell Cultures Under Chemos tat Conditions M.W. FOWLER (With 4 Figures) .................... 253

Radioirnrnuno-Screening Methods for Secondary Plant Products E.W. WEILER (With 8 Figures) •...•.....•.••.•.•.• 266

RIA-Test of Digitalis Plants and Tissue Cultures S. NICKEL and E.J. STABA (With 1 Figure) .......• 278

Somatic Hybridization, Fusion and Haploids

Somatic Cell Hybridization by Protoplast Fusion and Morphogenesis

IX

O.L. GAMBORG (With 7 Figures) .......•..•.•...•.. 287.

Protoplasts in Studies of Cell Genetics and Mor­phogenesis E. GALUN, D. AVIV, D. RAVEH, A. VARDI, and A. ZELCER (With 6 Figures) .......•.•.••.••..•..•. 302

Technical Advances in Protoplast Isolation and Cultivation T. ERIKSSON (With 2 Figures) .•..•...•.........•. 313

On Some Selected Problems and Results Concerning Culture and Genetic Modification of Higher Plant Protoplasts I. POTRYKUS, H. L6RZ, and C.T. HARMS (With 1 Figure) .•.......••...•...•....•...•...•.. 323

Theoretical Aspects of Haploid Techniques J. STRAUB ...........••...•.•.......••...•.•.•... 334

Potential from Haploid and Possible Use in Agriculture R. DEPAEPE, C. NITSCH, M. GODARD, and J. PERNES (With 3 Figures) ............•.•.......•......... 341

Regeneration and Organogenesis

Basic Aspects of Differentiation and Plant Regen­eration from Cell and Tissue Cultures H.W. KOHLENBACH (With 4 Figures) .....•.•....••.. 355

Regulation of Morphogenesis K. TRAN THANH VAN (With 26 Figures)

Regeneration of Virus-free Potato from Tissue Culture

367

P.J. WANG (With 12 Figures) ..•.................. 386

Clonal Crops Through Tissue Culture T. MURASHIGE (With 11 Figures) ..........•.•...•. 392

Application of Tissue Culture for Plant Improve­ment P. SPIEGEL-ROY and J. KOCHBA •.........•....•.... 404

Subject Index ...•....•.........•......•.•..•.... 415

List of Contributors

ALFERMANN, A.W., Dr., Lehrstuhl fUr Pharmazeutische Biologie, Universitat TUbingen, Auf der Morgen­stelle 8, 7400 TUbingen / FRG

ARENS, H., Lehrstuhl fUr Pflanzenphysiologie, Ruhr­Universitat Bochum, 4630 Bochum / FRG

AVIV, D., Department of Plant Genetics, The Weizmann Institute of Science, Rehovot, Israel

AZECHI, S., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

BARZ, W., Prof. Dr., Lehrstuhl fUr Biochemie der Pflan­zen, Westfalische Wilhelms-Universitat, 4400 MUn­ster / FRG

BERGMANN, K., Prof. Dr., Botanisches Institut der Uni­versitat, GyrhofstraBe 15, 5000 Koln / FRG

BOY, H.M., Lehrstuhl fUr Pharmazeutische Biologie der Universitat, Auf der Morgenstelle, 7400 TUbingen/ FRG

BRAIN, K., Dr., The Welsh School of Pharmacy, UWIST, Cardiff CF1, 3NU, Wales / Great Britain

BURMEISTER, G., Lehrstuhl fUr Biochemie der Pflanzen, Westfalische Wilhelms-Universitat, 4400 MUnster / FRG

DEPAEPE, R., Dr., Laboratoire de Genetique et Physio­logie du Developpement des Plantes, CNRS, 91190 Gif-sur-Yvette, France

DEUS, B., Lehrstuhl fUr Pflanzenphysiologie, Ruhr­Universitat Bochum, 4630 Bochum / FRG

DIESPERGER, H., Institut fUr Biologie II, Biochemie der Pflanzen, Universitat Freiburg, 7800 Freiburg/ Brsg. / FRG

DOELLER, P.C., Lehrstuhl fUr Pharmazeutische Biologie der Universitat, Auf der Morgenstelle, 7400 TUbingen/ FRG

DOUGALL, O.K., Dr., W. Alton Jones Cell Science Center, Lake Placid, 12946 New York, USA

EL-SHAGI, H., Lehrstuhl fUr Pflanzenphysiologie, Ruhr­Universitat Bochum, 4630 Bochurn / FRG

XII

ERIKSSON, T., Prof. Dr., Institute of Physiological Botany, University of Uppsala, 75121 Uppsala, Sweden

FOWLER, M.W., Dr., Department of Biochemistry, The University, Sheffield S10 2TN / Great Britain

GALUN, E., Prof. Dr., Department of Plant Genetics, The Weizmann Institute of Science, Rehovot, Israel

GAMBORG, O.L., Prof. Dr., Prairie Regional Laboratory, National Research Council, Saskatoon, Saskatchewan S7M OW9, Canada

GODARD, M., Laboratoire de Genetique et Physiologie du Developpement des Plantes, CNRS, 91190 Gif-sur­Yvette, France

HAGEDORN, W., Lehrstuhl fur Pharmazeutische Biologie, Universitat Tubingen, Auf der Morgenstelle, 7400 Tubingen / FRG

HAHLBROCK, K., Prof. Dr., Biologisches Institut II der Universitat, SchanzlestraBe 9-11, 7800 Freiburg/ Brsg. / FRG

HARMS, Chr.-T., Dr., Max-Planck-Institut fur Pflanzen­genetik, Projektgruppen Haploide in der Pflanzen­zuchtung, 6802 Ladenburg/Heidelberg / FRG

HEINS, M., Lehrstuhl fur Pharmazeutische Biologie der Universitat, Auf der Morgenstelle, 7400 Tubingen / FRG

HIRATA, Y., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

HOESEL, W., Dr., Lehrstuhl fur Biochemie der Pflanzen, Westfalische Wilhelms-Universitat, 4400 Munster / FRG

KARTNIG, T., Prof. Dr., Institut fur Pharmakognosie der Universitat Graz, 8010 Graz, Universitatsplatz 4/1, Austria

KATO, A., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

KATO, K., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

KATSUYAMA, A., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

KOCHBA, J., Dr., Division of Fruit· Breeding and Genetics, Volcani Center A.R.O., Bet-Dagan/Israel

XIII

KOHLENBACH, H.W., Prof., Fachbereich Biologie, Bota­nisches Institut, Universitat Frankfurt, 6000 Frank­furt/Main / FRG

KREYSING, P., Lehrstuhl fur Biochemie der Pflanzen, Westfalische Wilhelms-Universitat, 4400 Munster / 'FRG

LINES, D.St., Welsh School of Pharmacy, UWIST, Cardiff CF1 3NU, Wales / Great Britain

LOERZ, H., Dr., Max-Planck-Institut fUr Pflanzengenetik, Projektgruppen Haploide in der PflanzenzUchtung, 6802 Ladenburg/Heidelberg / FRG

MANGOLD, H.K., Prof. Dr., Bundesanstalt fUr Fettfor­schung, Institut fUr Biochemie und Technologie, H.P. Kaufmann-Institut, 4400 MUnster / FRG

MATSUMOTO, T., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

MISAWA, M., Dr., Tokyo Research Laboratory, Kyowa Hakko Kogyo Co. Ltd., Michida-shi, Tokyo, Japan

MURASHIGE, T., Prof. Dr., Department of Plant Sciences, University of California, Riverside, California 92502, USA

NICKEL, St., College of Pharmacy, University of Minne­sota, Minneapolis, MN 55455, USA

NITSCH, C., Prof. Dr., Laboratoire de Genetique et Physiologie du Developpement des Plantes, CNRS, 91190, Gif-sur-Yvette, France

NOGUCHI, M., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

OVERTON, K.H., Prof. Dr., Department of Chemistry, The University of Glasgow, Scotland

PERNES, J., Dr., Laboratoire de Genetique et Physiologie du Developpement des Plantes, CNRS, 91190 Gif-sur­Yvette, France

POTRYKUS, I., Dr., Friedrich-Miescher-Institut, P.O. Box 273, 4002 Basel, Switzerland

RAVEH, D., Dr., Genetics Laboratory, Department of Bio­chemistry, Oxford University / Great Britain

REINHARD, E. " Prof. Dr., Lehrstuhl fUr Pharmazeutische Biologie der Universitat, Auf der Morgenstelle, 7400 TUbingen / FRG

RIES, B., Lehrstuhl fUr Pflanzenphysiologie der Uni­versitat Bochum, 4630 Bochum / FRG

XIV

SANDERMANN, H., Prof. Dr., Biologisches Institut II der Universitat Freiburg, Lehrstuhl fur Biochemie der Pflanzen, SchanzlestraBe 9-11, 7800 Freiburg/ Brsg. / FRG

SCHEEL, D., Institut fur Biologie II, Lehrstuhl fur Biochemie der Pflanzen, Universitat Freiburg, 7800 Freiburg/Brsg. / FRG

SPIEGEL-ROY, P., Prof. Dr., Division of Fruit Breeding and Genetics, Volcani Center A.R.O., Bet Dagan, Israel

STABA, E.J., Prof. Dr., College of Pharmacy, University of Minnesota, Minneapolis, MN 55455, USA

STOECKIGT, J., Dr., Lehrstuhl fur Pflanzenphysiologie, Ruhr-Universitat Bochum, 4630 Bochum / FRG

STOHS, S.J., Prof., Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Nebraska, Medical Center, Omaha, Nebraska 68105, USA

STRAUB, J., Prof. Dr., Max-Planck-Institut fur Zuch­tungsforschung, 5000 Koln-Vogelsang / FRG

STREET, H.E., Prof., Botanical Laboratories, University of Leicester, Leicester, LE1 7RH / Great Britain

SURHOLT, E., Lehrstuhl fur Biochemie der Pflanzen, Westfalische Wilhelms-Universitat, 4400 Munster / FRG

TABATA, M., Prof. Dr., Faculty of Pharmaceutical Sci­ences, Kyoto University, Shimodachicho, Sakyo-ku, Kyoto 606, Japan

TRAN THAN VAN, K., Dr., Laboratoire du Phytotron, CNRS, Gif-sur-Yvette, France

VARDI, A., Dr., Agricultural Research Organization, Volcani Center, Department of Plant Genetics, The Weizmann Institute of Science, Rehovot, Israel

VOGELMANN, H., Dr., Lehrstuhl fur Biochemie und Bio­technologie der Technischen Universitat Braunschweig, Gesellschaft fur Biotechnologische Forschung mbH, 3300 Braunschweig / FRG

WAGNER, F., Prof. Dr., Lehrstuhl fur Biochemie und Bio­technologie der TU Braunschweig, Gesellschaft fur Biotechnologische Forschung mbH, 3300 Braunschweig / FRG

WAHL, J., Lehrstuhl fur Pharmazeutische Biologie der Universitat, Auf der Morgenstelle, 7400 Tubingen / FRG

WANG, P.J., Dr., Institute of Botany, Academia Sinica, Nankong, Taipei, Taiwan, Free China

xv

WEILER, E.W., Dr., Lehrstuhl fur Pflanzenphysiologie, Ruhr-Universitat Bochum, 4630 Bochum / FRG

WIDHOLM, M., Prof. Dr., Department of Agronomy, Uni­versity of Illinois, Urbana, Illinois 61801, USA

WITHERS, L.A., Dr., Botanical Laboratories, University of Leicester, Leicester LEl 7RH / Great Britain

YAMAMOTO, K., Dr., Central Research Institute, The Japan Tobacco and Salt Public Corporation, 6-2 Umegaoka, Midori-ku, Yokohama, Kanagawa 227, Japan

ZELCER, A., Dr., Department of Plant Genetics, The Weizmann Institute of Science, Rehovot, Israel

ZENK, M.H., Prof. Dr., Lehrstuhl fur Pflanzenphysiolo­gie, Ruhr-Universitat Bochum, 4630 Bochum / FRG