The Plant Viruses Volume 3 POLYHEDRAL VIRIONS WITH MONO PARTITE RNA GENOMES

THE VIRUSES

Series Editors HEINZ FRAENKEL-CONRAT, University of California

Berkeley, California

ROBERT R. WAGNER, University of Virginia School of Medicine

Charlottesville, Virginia

THE VIRUSES: Catalogue, Characterization, and Classification Heinz Fraenkel-Conrat

THE ADENOVIRUSES Edited by Harold S. Ginsberg

THE BACTERIOPHAGES Volumes 1 and 2 • Edited by Richard Calendar

THE HERPESVIRUSES Volumes 1-3 • Edited by Bernard Roizman Volume 4 • Edited by Bernard Roizman and Carlos Lopez

THE PAPOVAVIRIDAE Volume 1 • Edited by Norman P. Salzman Volume 2 • Edited by Norman P. Salzman and Peter M. Howley

THE PARVOVIRUSES Edited by Kenneth I. Berns

THE PLANT VIRUSES Volume 1 • Edited by R. I. B. Francki Volume 2 • Edited by M. H. V. Van Regenmortel and Heinz Fraenkel-Conrat Volume 3 • Edited by Renate Koenig Volume 4 • Edited by R. G. Milne

THE REOVIRIDAE Edited by Wolfgang K. Joklik

THE RHABDOVIRUSES Edited by Robert R. Wagner

THE TOGA VIRIDAE AND FLA VIVIRIDAE Edited by Sondra Schlesinger and Milton J. Schlesinger

THE VIROIDS Edited by T. O. Diener

The Plant Viruses Volume 3 POL YHEDRAL VIRIONS WITH MONOPARTITE RNA GENOMES

Edited by

RENATE KOENIG Institute for Plant Viral Diseases Braunschweig, Federal Republic of Germany

PLENUM PRESS • NEW YORK AND LONDON

Library of Congress Cataloging in Publication Data

Polyhedral virions with monopartite RNA genomes.

(The Plant viruses; v. 3) (The Viruses) Bibliography: p. Includes index. 1. Viruses, RNA. 2. Plant viruses. I. Koenig, Renate. II. Series. III. Series: Viruses.

QR357.P58 1985 vol. 3 576'.6483 s 87-32809 [QR3951 [576'.64831

ISBN-13 :978-1-4612-8246-4 e-ISBN-13 :978-1-4613-0921-5 DOl: 10.1007/978-1-4613-0921-5

© 1988 Plenum Press, New York Softcover reprint of the hardcover 1st edition 1988 A Division of Plenum Publishing Corporation 233 Spring Street, New York, N.Y. 10013

All rights reserved

No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher

Con tribu tors

J. C. Carrington, Department of Plant Pathology, University of California, Berkeley, California 94729

R. Casper, Plant Virus Institute, Federal Biological Research Center for Agriculture and Forestry, D-3300 Braunschweig, Federal Republic of Germany

H. Fraenkel-Conrat, Department of Molecular Biology and Virus Labo­ratory, University of California, Berkeley, California 94720

D. Gallitelli, Department of Plant Pathology, University of Bari, Bari, Italy

R. Gamez, Cellular and Molecular Biology Research Center, University of Costa Rica, Costa Rica

R. E. Gingery, United States Department of Agriculture, Agricultural Research Service, Department of Plant Pathology, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster, Ohio 44691

L. Givord, Institute of Cellular and Molecular Biology of CNRS, 67084 Strasbourg Cedex, France

1. Hirth, Institute of Cellular and Molecular Biology of CNRS, 67084 Strasbourg Cedex, France

R. Hull, Department of Virus Research, John Innes Institute, Colney Lane, Norwich NR4 7 UH, United Kingdom

R. Koenig, Plant Virus Institute, Federal Biological Research Center for Agriculture and Forestry, D-3300 Braunschweig, Federal Republic of Germany

P. Leon, Cellular and Molecular Biology Research Center, University of Costa Rica, Costa Rica

v

vi CONTRIBUTORS

G. P. Martelli, Department of Plant Pathology, University of Bari, Bari, Italy

T. J. Morris, Department of Plant Pathology, University of California, Berkeley, California 94720

A. F.Murant, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, Scotland, United Kingdom

M. Russo, Department of Plant Pathology, University of Bari, Bari, Italy

Contents

Chapter 1

Polyhedral Plant Viruses with Monopartite RNA Genomes: Introduction and Summary of Important Properties. . . . . . . . . . . . . . .. 1

R. Koenig

Chapter 2

Tombusviruses ................................................ 13

G. P. Martelli, D. Gallitelli, and M. Russo

I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 II. Diseases, Host Ranges, and Geographical Distribution. . . . . . 16

A. General Features. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 B. Diseases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

III. Transmission, Ecology, and Control ....................... 23 A. Transmission ........................................ 23 B. Ecology.............................................. 25 C. Control.............................................. 27

IV. Purification.............................................. 28 V. Properties of Virus Particles. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

A. Particle Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 B. Sedimentation........................................ 32 C. Electrophoretic Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 D. Coat Protein Subunits ................................ 33 E. Structure of Virus Particles. . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

VI. Serology and Immunochemistry. .. . .. . . .. . . ... . .. . . . . .. . . . 37 A. General Features ..................................... 37 B. Serology in Virus Identification. . . . . . . . . . . . . . . . . . . . . . . . 38 C. Serology in Virus Classification. . . . . . . . . . . . . . . . . . . . . . . . 41 D. Immunochemical Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

vii

viii CONTENTS

VII. Genome Properties and Replication. . . . . . . . . . . . . . . . . . . . . . . . 43 A. Structure of Genomic RNA ........................... 43 B. Expression of the Genome ............................ 44 C. Viral Replication .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

VIII. Relations with Cells and Tissues. . . . . . . . . . . . . . . . . . . . . . . . . . 46 A. General Cytology of Infected Cells. . . . . . . . . . . . . . . . . . . . . 46 B. Appearance and Intracellular Distribution of Virus

Particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 C. Inclusion Bodies. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

IX. Satellites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 X. Recent Developments .................................... 65

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 65

Chapter 3

Carnation Mottle Virus and Viruses with Similar Properties ...... 73

T. f. Morris and f. c. Carrington

I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 II. Geographical Distribution, Host Range, and Transmission .. 76

A. Diseases of Ornamental Crops. . . . . . . . . . . . . . . . . . . . . . . . . 76 B. Diseases of Cucurbits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79 C. Diseases of Legumes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81 D. Viruses of Other Hosts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83

III. Purification.............................................. 84 IV. Properties of the Virus Particles. . . . . . . . . . . . . . . . . . . . . . . . . . . 85

A. Particle Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 B. Physical and Chemical Properties. . . . . . . . . . . . . . . . . . . . . . 86 C. Virus Particle Structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

V. Serological Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 VI. Genome Properties and Replication. . . . . . . . . . . . . . . . . . . . . . . . 95

A. Genome Structure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 B. Regulation and Expression of the Genome. . . . . . . . . . . . . . 98 C. Viral Replication ..................................... 102

VII. Cytopathology........................................... 103 VIII. Satellites................................................ 104

IX. Conclusions ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 105 X. Recent Developments .................................... 107

References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 107

Chapter 4

The Sobemovirus Group. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 113

KRull

CONTENTS ix

I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 113 II. The Viruses and Diseases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 115

A. Southern Bean Mosaic Virus. . . . . . . . . . . . . . . . . . . . . . . . . .. 115 B. Blueberry Shoestring Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 116 C. Cocksfoot Mottle Virus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 116 D. Lucerne Transient Streak Virus.. . .. . .... . ... .......... 116 E. Rice Yellow Mottle Virus . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 117 F. Solanum Nodiflorum Mottle Virus. . . . . . . . . . . . . . . . . . . .. 117

G. Sowbane Mosaic Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 117 H. Subterranean Clover Mottle Virus ..................... 118 I. Turnip Rosette Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 118 J. Velvet Tobacco Mottle Virus. . . . . . . . . . . . . . . . . . . . . . . . .. 118

K. Cocksfoot Mild Mosaic Virus .................. .' . . . . . .. 118 1. Cynosurus Mottle Virus .............................. 119

M. Ginger Chlorotic Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . .. 119 N. Maize Chlorotic Mottle Virus........... .............. 119 O. Olive Latent Virus-I. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 119 P. Panicum Mosaic Virus ........................... '" .. 119

III. Transmission ............................................ 120 IV. Purification.............................................. 121 V. Particle Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 123

A. Size and Shape of Particles. . . . . . . . . . . . . . . . . . . . . . . . . . .. 123 B. Sedimentation........................................ 124 C. Isopycnic Banding. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 124 D. Composition of Particles. . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 125 E. Coat Proteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 125 F. Capsid Stabilization and Structure. . . . . . . . . . . . . . . . . . . .. 126

VI. Serology................................................. 130 VII. Genome Properties, Expression, and Replication. . . . . . . . . . .. 131

A. Virion RNA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 131 B. Nucleic Acid Homologies . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 134 C. Replication and Expression. . . . . . . . . . . . . . . . . . . . . . . . . . .. 134

VIII. Cytopathology ........................................... 138 IX. Conclusions ............................................. 139

References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 139

Chapter 5

Tobacco Necrosis, Satellite Tobacco Necrosis, and Related Viruses ....................................................... 147

H. Fraenkel-Conrat

I. Introduction............................................. 147 II. Geographical Distribution, Host Range, Diseases, and

Epidemiology ........................................... , 148

x CONTENTS

III. Transmission............................................ 148 IV. Purification and Separation of TNV and TNSV . ... ... ... ... 149 V. Properties of the Virions, Their RNAs, and Their Coat

Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 150 VI. Serology of TNV and TNSV. . . . . . ..... . ... ... . ... ... ... . .. 154

VII. Translation and Replication of the Genomes of TNV and TNSV ................................................... 154

VIII. Cytopathology ........................................... 157 IX. Other N ecroviruses ...................................... 157 X. Conclusions ............................................. 157

References .................................................... 158

Chapter 6

Tymoviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 163

L. Hirth and L. Givord

I. Introduction............................................. 163 II. Geographical Distribution, Host Ranges, Diseases, and

Symptomatology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 164 III. Transmission, Epidemiology, and Control. . . . . . . . . . . . . . . . .. 170 IV. Purification.............................................. 173

A. General Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 173 B. Diversity of Purification Products. . . . . . . . . . . . . . . . . . . . .. 174

V. Properties and Constituents of Particles. . . . . . . . . . . . . . . . . . .. 175 A. TheRNAs ........................................... 175 B. The Capsid Protein. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 176 C. Polyamines .......................................... 176 D. Structure of the Capsids of TYMV ... . ... .... ..... ... .. 177 E. Protein-RNA Interactions inside TYMV . . . . . . . . . . . . . . .. 178 F. Formation and Properties of Artificial Capsids . . . . . . . . .. 178

G. Stability and Structure of Tymoviruses Other than TYMV ............................................... 181

H. Decapsidation of TYMV .............................. 182 VI. Serology and Immunochemistry . . . . . . . . . . . . . . . . . . . . . . . . . .. 184

VII. Genome Properties and Replication.. . .... ...... ... ...... .. 189 A. In vitro Messenger Properties of TYMV RNAs.... ... ... 189 B. tRNA-like Structure at the 3'-OH Extremity of TYMV

RNA ................................................ 192 C. Replication .......................................... 196

VIII. Relations with Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 198 A. Cellular Symptomatology. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 198 B. Modifications in Protoplasts infected by TYMV. . . . . . . .. 199

IX. Conclusions ............................................. 200 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 201

CONTENTS xi

Chapter 7

Maize Rayado Fino and Related Viruses ......................... 213

R. Gamez and P. Le6n

I. Introduction............................................. 213 II. Geographical Distribution, Host Range, and Diseases. . . . . .. 214

A. Geographical Distribution. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 214 B. Host Range and Symptomatology. . . . . . . . . . . . . . . . . . . . .. 215

III. Transmission, Epidemiology, and Control. . . . . . . . . . . . . . . . .. 217 A. Transmission ........................................ 217 B. Epidemiology ........................................ 219 C. Control.............................................. 220

IV. Purification.............................................. 220 A. Purification of the Viruses ............................ 220 B. Purification of RNA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 221

V. Particle Properties ....................................... , 221 A. Physicochemical Properties ........................... 221 B. Capsid Protein Studies. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 222 C. Electron Microscopy Studies of Isolated Particles ....... 224 D. Capsid Structure of MRFV ............................ 224

VI. Serology and Immunochemistry. . . . . . . . . . . . . . . . . . . . . . . . . .. 224 A. Gel Double-Immunodiffusion Tests. . . . . . . . . . . . . . . . . . .. 224 B. Blot Transfers and Immunological Detection of Viral

Proteins ............................................. 225 VII. Genome Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 226

A. Properties of the RNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 226 B. In vitro Expression of MRFV-RNA ............... , ..... 226

VIII. Relations with Cells and Tissues. . . . . . . . . . . . . . . . . . . . . . . . .. 227 A. In Plant Hosts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 227 B. In Insect Hosts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 228

IX. Evolutionary Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 229 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 230

Chapter 8

Luteoviruses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 235

R. Casper

I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 235 II. Taxonomy and Properties of Individual Viruses. . . . . . . . . . . .. 236

A. Beet Western Yellows Virus. . . . . . . . . . . . . . . . . . . . . . . . . .. 240 B. Rice Giallume Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 241 C. Carrot Red Leaf Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 241 D. Barley Yellow Dwarf Virus ............................ 242 E. Bean Leaf Roll Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 242 F. Soybean Dwarf Virus ................................. 243

xii CONTENTS

G. Potato Leaf Roll Virus ................................ 243 H. Tomato Yellow Top Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 243

I. Tobacco Necrotic Dwarf Virus ........................ 243 J. Solanum Yellows Virus ............................... 244

K. Groundnut Rosette Assistor Virus ..................... 244 L. Strawberry Mild Yellow-Edge Virus. . . . . . . . . . . . . . . . . . .. 244

M. Banana Bunchy Top Virus. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 244 III. Transmission and Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 245 IV. Purification.............................................. 247 V. Particle Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 248

VI. Serology................................................. 250 VII. Genome Properties and Replication. . . . . . . . . . . . . . .. . . . . . . .. 250

VIII. Cytopathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 251 IX. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 251

References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 252

Chapter 9

Maize Chlorotic Dwarf and Related Viruses. . . . . . . . . . . . . . . . . . . .. 259

R. E. Gingery

I. Introduction and History ................................. 259 II. Geographical Distribution, Diseases, and Host Ranges ...... 260

A. Geographical Distribution ............................ , 260 B. Main Diseases ....................................... , 260 C. Host Range. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 261 D. Losses ............................................... 262

III. Transmission, Epidemiology, and Control. . . . . . . . . . . . . . . . .. 262 A. Transmission ........................................ 262 B. Epidemiology ........................................ 264 C. Control.............................................. 265

IV. Purification.............................................. 266 V. Particle Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 267

VI. Serology and Immunochemistry. . . . . . . . . . . . . . . . . . . . . . . . . .. 268 VII. Genome Properties and Replication. . . . . . . . . . . . . . . . . . . . . . .. 268

VIII. Relations with Cells and Tissues. . . . . . . . . . . . . . . . . . . . . . . . .. 268 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 268

Chapter 10

Parsnip Yellow Fleck Virus, Type Member of a Proposed New Plant Virus Group, and a Possible Second Member. Dandelion Yellow Mosaic Virus .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 273

A. F. Murant

I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 273 II. Hosts, Diseases, and Geographical Distribution. . . . . . . . . . . .. 274

CONTENTS xiii

A. Parsnip Yellow Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 274 B. Dandelion Yellow Mosaic Virus ....................... 276

III. Transmission, Epidemiology and Control. . . . . . . . . . . . . . . . .. 277 A. Parsnip Yellow Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 277 B. Dandelion Yellow Mosaic Virus ....................... 280

IV. Purification .............................................. 281 A. Parsnip Yellow Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 281 B. Dandelion Yellow Mosaic Virus ............. . . . . . . . . .. 282

V. Particle Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 282 A. Parsnip Yellow Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 282 B. Dandelion Yellow Mosaic Virus ....................... 283

VI. Serology................................................. 283 A. Parsnip Yellow Fleck Virus.. . . . . . . . . . . . . . . . . . . . . . . . . .. 283 B. Dandelion Yellow Mosaic Virus ....................... 284

VII. Genome Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 284 A. Parsnip Yellow Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 284 B. Dandelion Yellow Mosaic Virus ... . . . . . . . . . . . . . . . . . . .. 284

VIII. Relations with Cells and Tissues. . . . . . . . . . . . . . . . . . . . . . . . .. 284 A. Parsnip Yellow Fleck Virus. . . . . . . . . . . . . . . . . . . . . . . . . . .. 284 B. Dandelion Yellow Mosaic Virus .......... . . . . . . . . . . . .. 285

IX. Possible Affinities of Parsnip Yellow Fleck Virus and Dandelion Yellow Mosaic Virus. . . . . . . . . . . . . . . . . . . . . . . . . .. 285

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 287

Index ......................................................... 289