Dijkstra / Jager Practical Plant Virology

Protocols and Exercises
Erscheinungsjahr 2012
ISBN: 978-3-642-72030-7
Verlag: Springer
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 459 Seiten, Web PDF

Reihe: Springer Lab Manuals

The idea for this book arose from what we perceived as the need for an up-to-date guide to class exercises in plant virology. We were encouraged to proceed after receiving 29 positive responses (out of 30 replies to our enquiries) from colleagues worldwide. To the best of our knowledge, no such publications have appeared since D. Noordam's book containing practical exercises (Noordam 1973) and the latest (1988) edition of the American Phytopathological Society's Laboratory Exercises in Plant Pathology, in which 4 out of its 31 chapters discuss plant viruses. Our original plan was to aim this publication at students and teachers of plant virology, plant pathology, plant breeding and microbiology. How ever, both colleagues and our publisher suggested widening the scope of the book by making it useful also for research workers and laboratory technicians. Therefore, we decided to prepare a laboratory manual of interest to all groups. We have tried to cover all relevant branches of plant virology, including the molecular aspects, in as far as they pertain to the detection and basic characterisation of plant viruses. We have not included protocols for the molecular biology of plant viruses (sequencing, construction of recombi nants, transgenic plants, etc.), as they are presented adequately in many other recent publications. The protocols in this book are described in a manner which should be understandable to those with a basic knowledge of biology and chemistry.

Zielgruppe

Research

Autoren/Hrsg.

Dijkstra, Jeanne

Jager, Cees de

Inhaltsverzeichnis

I Virus Inoculation.- Protocol 1 Mechanical Inoculation of Plants.- Protocol 2 Inoculation of Protoplasts Isolated from Leaves.- Protocol 3 Differential Temperature Treatment of Infected Plants.- Exercise 1 Infectivity of Tobacco Mosaic Tobamovirus.- II Symptomatology.- Protocol 4 Description of External Symptoms.- Protocol 5 Determination of Internal Symptoms.- Exercise 2 Systemic Acquired Resistance (SAR).- Exercise 3 Demonstration of Phloem Necrosis in Plants Affected by Potato Leafroll.- Exercise 4 Inclusions in Hair Cells of Leaves of Tobacco Infected with Tobacco Mosaic Tobamovirus.- III Infectivity Assays.- Protocol 6 Infectivity Assay on Local-Lesion Hosts.- Protocol 7 Infectivity Assay on Hosts Producing Starch Lesions.- Protocol 8 Infectivity Assay on Hosts Producing Systemic Symptoms Only.- Exercise 5 Effect of Temperature on the Susceptibility of Plants.- Exercise 6 Effect of Abrasives on the Infection.- Exercise 7 Effect of Inhibitors on the Infection.- Exercise 8 Effect of Light on the Susceptibility of Plants.- Exercise 9 Effect of Time of the Day on the Susceptibility of Plants.- IV Infectivity of Virus in Crude Sap.- Protocol 9 Determination of the Dilution End-Point.- Protocol 10 Determination of the Thermal Inactivation Point.- Protocol 11 Determination of the Longevity In Vitro.- Exercise 10 Determination of the Dilution End-Point.- Exercise 11 Determination of the Thermal Inactivation Point.- Exercise 12 Determination of the Longevity In Vitro.- V Transmission of Viruses.- Protocol 12 Virus Transmission by Grafting.- Protocol 13 Virus Transmission Through Soil.- Protocol 14 Virus Transmission by Nematodes.- Protocol 15 Virus Transmission by Fungi.- Protocol 16 Virus Transmission by Aphids.- Protocol 17 Transmission by Leafhoppers (Cicadellidae).- Protocol 18 Transmission by Whiteflies (Aleyrodidae).- Protocol 19 Transmission by Thrips (Thripidae).- Exercise 13 Wedge-Grafting of Tobacco.- Exercise 14 Separation of a Nonpersistently and a Persistently Transmitted Virus by Differential Transmission.- Exercise 15 Visualisation of Salivary Sheaths Formed by Aphids.- Exercise 16 Separation of a Semipersistently Transmitted and a Circulative-Propagative Virus by Differential Transmission.- VI Maintenance of Virus Isolates.- Protocol 20 Freezing.- Protocol 21 Dehydration.- Protocol 22 Freeze-Drying.- Protocol 23 Short-Term Storage.- VII Production of Virus-Free Material from Infected Plants.- Protocol 24 Heat Treatment.- Protocol 25 Meristem-Tip Culture.- Protocol 26 Heat Treatment Combined with Meristem-Tip Culture.- VIII Virus Isolation and Purification.- Protocol 27 Density-Gradient Centrifugation.- Protocol 28 Virus Purification Protocols.- IX Physicochemical Properties of Virus Particle.- Protocol 29 UV Light Absorption.- Protocol 30 Buoyant Density in Caesium Chloride.- Protocol 31 Particle Morphology and Size as Determined with the Electron Microscope.- X Properties of Viral Components.- Protocol 32 Determination of the Molecular Mass of Coat Proteins by Gel Electrophoresis.- Protocol 33 RNA Extraction from Purified Virus Particles.- Protocol 34 Determination of the Molecular Mass of Ribonucleic Acids by Gel Electrophoresis.- XI Serological Techniques for Detection and Identification of Viruses and Viral Products.- Protocol 35 Production of Antiserum.- Protocol 36 Absorption of Antiserum.- Protocol 37 Chloroplast Agglutination Test.- Protocol 38 Microprecipitin Test.- Protocol 39 Gel Double-Diffusion Test in Plates.- Protocol 40 Enzyme-Linked Immunosorbent Assay.- Protocol 41 Immunoblotting Assays.- Exercise 17 Standard DAS-ELISA.- XII Electron Microscope and Light Microscope Serology.- Protocol 42 Electron Microscope Serology.- Protocol 43 Light Microscope Serology.- XIII Detection of Viral RNA.- Protocol 44 Northern Blotting.- Protocol 45 Northern Hybridisation.- Protocol 46 Dot-Blot Hybridisation (Nucleic Acid Spot Hybridisation).- Protocol 47 Polymerase Chain Reaction.- XIV Diagnosis of a Disease and Identification of the Causal Virus(es).- XIV Diagnosis of a Disease and Identification of the Causal Virus(es).