Electron Microscopy of Plant Pathogens

341 Pages·1991·11.417 MB·English

by  H. C. Hoch (auth.), Prof. Dr.rer.nat. Kurt Mendgen, Dr. Dietrich-Eckhardt Lesemann (eds.)

Cover picture
Young pustule with uredospores of Puccinia striiformis (West.) (yellow stripe rust)
emerging through the epidermis of a wheat leaf (1.300 x, SEM-Laboratory
University of Basel in cooperation with Sandoz Agro AG Basel/Switzerland)
K. Mendgen . D.-E. Lesemann (Eds.)
Electron Microscopy
of
Plant Pathogens
With 281 Figures
Springer-Verlag Berlin Heidelberg New York
London Paris Tokyo HongKong Barcelona
Prof. Dr. rer. nat. Kurt Mendgen
Universitat Konstanz
Fakultat fur Biologie
Lehrstuhl Phytopathologie
D-7750 Konstanz, FRG
Dr. Dietrich-Eckhardt Lesemann
Biologische Bundesanstalt
fur Land- und Forstwirtschaft
Institut fur Viruskrankheiten der Planzen
Messeweg 11/12
D-3300 Braunschweig, FRG
ISBN-13: 978-3-642-75820-1 e-ISBN-13: 978-3-642-75818-8
DOl: 10.1007/978-3-642-75818-8
Library of Congress Cataloging-in-Publication Data
Electron microscopy of plant pathogens 1 K. Mendgen, D.-E. Lesemann, eds. p. cm. Results of the International
Symposium Electron Microscopy Applied in Plant Pathology, held at the University of Konstanz in September 1989.
Includes bibliographical references.
1. Plant diseases Congresses. 2. Phytopathogenic microorganisms Congresses. 3. Electron microscopy--Congresses.
I. Mendgen, K. (Kurt). 1944- . II. Lesemann, D.-E. (Dietrich-Eckhardt), 1939- Ill. International Symposium
Electron Microscopy Applied in Plant Pathology (I 989: University of Konstanz)
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, re-use of illustrations, recitation, broadcasting, reproduction on
microfilms or in other ways, and storage in data banks. Duplication ofthis publication or parts thereof is only permitted
under the provisions ofthe German Copyright Law ofSeptcmber 9,1965, in its current version, and a copyright fee must
always be paid. Violations fall under the prosecution act of the German Copyright Law.
© Springer-Verlag Berlin Heidelberg 1991
Softcover reprint of the hardcover 1s t edition 1991
The use of registered names, trademarks, etc. in this publications 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.
2!31/3020-54321O-Printed on acid-free paper
Contributors
You will find the addresses at the beginning of the respective contribution
Ammar, EI-Desouky 133 Kunoh, H. 223
Bailey, J. A. 211 Latge, J. P. 43
Bonfante-Fasolo, P. 265 Lherminier, J. 177
Boudon-Padieu, E. 177 Lesemann, D.-E. 147
Bourett, T. M. 251 Lukacs, N. 161
Brown, J., Mansfield, J. 185 Mansfield, J. 185
Caudwell, A. 177 Martelli, G. P. 103
Chong, J. 235 Meignoz, R. 177
Dollet, M. 307 Mendgen, K. 31
Endo, B. Y. 291 Milne, R. G. 87, 177
Ferrari, M. A. 251 Nicholson, R. L. 223
Gruner, R. 161 O'Connell, R. J. 211
Gubler, F. 197 Perotto, S. 265
Harder, D. E. 235 Read, N. D. 17
Harders, J. 161 Riesner, D. 161
Hardham, A. R. 197 Robert-Nicoud, M. 161
Hippe, S. 317 Santore, U. J. 161
Hoch, H. C. 1 Sanger, H. L. 161
Honegger, R. 277 SchefTold, F. 31
Howard, R. J. 251 van Lent, J. W. M. 119
Jovin, T. M. 161 Verduin, B. J. M. 119
Klotz, G. 161 Welter, K. 31
Knauf-Beiter, G. 31 Wurtz, M. 73
Kobayashi, J. 223 Zeyen, R. J. 59
Preface
Plants, fungi, and viruses were among the first biological objects studied with an
electron microscope. One of the two first instruments built by Siemens was used by
Helmut Ruska, a brother of Ernst Ruska, the pioneer in constructing electron
microscopes. H. Ruska published numerous papers on different biological objects in
1939. In one of these, the pictures by G. A. Kausche, E. Pfankuch, and H. Ruska of
tobacco mosaic virus opened a new age in microscopy. The main problem was then
as it still is today, to obtain an appropriate preparation of the specimen for
observation in the electron microscope. Beam damage and specimen thickness were
the first obstacles to be met. L. Marton in Brussels not only built his own instrument,
but also made considerable progress in specimen preparation by introducing the
impregnation of samples with heavy metals to obtain useful contrast. His pictures of
the bird nest orchid root impregnated with osmium were revolutionary when
published in 1934.
It is not the place here to recall the different techniques which were developed in
the subsequent years to attain the modern knowledge on the fine structure of plant
cells and of different plant pathogens. The tremendous progress obtained with
tobacco mosaic virus is reflected in the chapter by M. Wurtz on the fine structure
of viruses in this Volume. New cytochemical and immunological techniques
considerably surpass the morphological information obtained from the pathogens,
especially at the host-parasite interface. Information on nonpathogenic systems,
such as on mycorrhizae and lichens, is added, since this may help to elucidate the
function of the morphological changes observed during the interaction of two
organisms.
The results published in the book were presented to a great part as reviews
during the International Symposium Electron Microscopy Applied in Plant
Pathology, held at the University of Konstanz in September 1989. Further
information has been added where it seemed appropriate in order to give a general
account of the fine structure of different plant pathogens. This book is the result of
many efforts of collaborators and students in the department of Plant Pathology at
the University of Konstanz. The authors of the different chapters also referred the
articles of their colleagues. In addition, Bill Bushnell, Wolfram Koller, Gerard de
Leeuw, Klaus Rudolph, Roland Rohringer, and Urs Wyss helped with reviewing.
VIII Preface
Last but not least, we would like to thank the Deutsche Forschungsgemeinschaft,
the Deutsche Phytomedizinische Gesellschaft, and the Freunde and Forderer der
Universitiit Konstanz for financial help in organizing the Symposium and in editing
this book.
Konstanz and Braunschweig K. Mendgen
Summer 1990 D.-E. Lesemann
Contents
Chapter 1 Preservation of Cell Ultrastructure by Freeze-Substitution
H. C. HocH (With 25 Figures) . 1
1 Introduction . . .. 1
2 The Cryofixed Cell . . 3
3 Freezing Methods. . . 6
4 Specimen Preparation 7
5 Substitution . . . . . . 8
6 Problems and Artifacts Associated
with Freeze-Substitution. 12
7 Conclusions 13
References . . . . . . . . . 14
Chapter 2 Low-Temperature Scanning Electron Microscopy of Fungi
and Fungus-Plant Interactions
N. D. READ (With 16 Figures) 17
1 Introduction . . . . . . . . . . 17
2 Specimen Preparation . . . . 17
3 Rapid Immobilization and Stabilization. 19
4 Retention of Water. . . . . . . . . . . . . 19
5 No Exposure to Chemicals, Fixatives or Solvents. 23
6 Artifacts Specific to Low-Temperature Scanning Electron
Microscopy. . . . . . . 27
7 Future Developments. 27
References . . . . . . . 28
Chapter 3 High Pressure Freezing of Rust Infected Plant Leaves
K. MENDGEN, K. WELTER, F. SCHEFFOLD, and G. KNAUF-BEITER
(With 5 Figures) . . . . . . . . . 31
1 Introduction . . . . . . . . . . . . . . . . . . . 31
2 The Specimen Sandwich . . . . . . . . . . . . 32
3 Plunge Freezing vs. High Pressure Freezing 33
4 Infiltration of Leaves ......... . 36
5 Freeze-Substitution and Embedding. . . . . 38
x
Contents
6 A Re-Evaluation of the Haustorial Host-Parasite Interface. 38
7 Conclusions . . . . . . . . . . . . 40
References . . . . . . . . . . . . . . 41
Chapter 4 Cytochemistry of Fungal Surfaces:
Carbohydrate Containing Molecules
1. P. LATGE (With 22 Figures) ..... . 43
1 Introduction . . . . . . . . . . . . . . 43
2 EM Probes for External Carbohydrates. 43
3 EM Methods. 53
4 Conclusion. . . . . . . . . . . . . 54
Refureoc~ . . . . . . . . . . . . . 54
Chapter 5 Analytical Electron Microscopy in
Plant Pathology: X-Ray Microanalysis
and Energy Loss Spectroscopy
R. 1. ZEYEN (With 2 Figures) . 59
1 Introduction . . . . 59
2 XRMA Principles. . . . . 61
3 XRMA Detectors . . . . . 62
4 Applications to Plant Pathology-XRMA . 63
5 Electron Energy Loss Spectrometry (EELS) Principles . 68
6 EEL Spectrometer Systems . . . . . . . . . 68
7 Applications to Plant Pathology-EELS. 69
8 Summary. . . . . . . . . . . . . . . . 69
References . . . . . . . . . . . . . . . 70
Chapter 6 The Fine Structure of Virus Particles
M. WURTZ (With 18 Figures) 73
1 Introduction . . . . . . . . . 73
2 Spherical Plant Viruses ... 73
3 Rod-Shaped Plant Viruses. 77
4 Filamentous Plant Virus. 81
5 Discussion . 81
6 Conclusion. 83
References . 85
Chapter 7 Immunoelectron Microscopy for Virus Identification
R. G. MILNE (With 14 Figures) .. . 87
1 Introduction ................... . 87
2 Leaf-Dip Serology. . . . . . . . . . . . . . . . . 88
3 Immune Complexes from Ouchterlony Plates 89
4 ISEM .............. . 89
5 Decoration. . . . . . . . . . . . 94
6 Mycoplasma-Like Organisms. 98
7 Conclusion . 99
References . . . . . . . . . . . . 99
Contents XI
Chapter 8 Cytochemistry of Virus-Infected. Plant Cells
G. P. MARTELLI (With 4 Figures) ....... . 103
1 Introduction . . . . . . 103
2 Light Microscopy. . . 104
3 Electron Microscopy . 105
4 Concluding Remarks . 114
References . . . . . . . 114
Chapter 9 Immunolabeling of Viral Antigens in Infected Cells
1. W. M. V AN LENT and B. 1. M. VERDUIN (With 9 Figures) 119
1 Introduction. . . . 119
2 Labeling . . . . . . . . . . . . . . . . . . . . . . . 121
3 Initial Interaction. . . . . . . . . . . . . . . . . . 123
4 Site of Coat Protein SynthesisjVirus Assembly. 123
5 Translocation of CCMV Genome 125
6 Detection of RNA. 129
References . . . . . . . . . . . . . . 130
Chapter 10 Mechanisms of Plant Virus Transmission by
Homopteran Insects
EL DESOUKY AMMAR (With 10 Figures). . . 133
1 Introduction . . . . . . . . . . . . . . . . 133
2 The Role of EM in the Elucidation of
Transmission Mechanisms. . . . . . . . 133
3 Studies on Insect Structures Involved in Transmission 142
4 Special Problems in Studying Virus-Vector Relations
by EM . . 142
References . . . . . . . . . . . . . . . . . . . . . . . . . . 145
Chapter 11 Specific Cytological Alterations in Virus-Infected Plant Cells
D.-E. LESEMANN (With 13 Figures) . . . . . . . . . . . . . . . .. 147
1 Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . .. 147
2 Viral Genome Products as Infection-Specific Constituents
of Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . 148
3 Virus-Specific Alterations of the Cellular Membrane
System . . . 152
4 Conclusions 157
References . 157
Chapter 12 Structure, Cellular Location, and Cytopathology of Viroids
D. RIESNER, 1. HARDERS, N. LUKACS, R. GRUNER, U. 1. SANTORE,
G. KLOTZ, M. ROBERT-NICOUD, T. M. lOVIN, and H. L. SANGER
(With 9 Figures) . . . . . . . . . . . . . . . . . . . . . . . . . . .. 161
1 Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . .. 161