Techniques in Plant Virology at CIP - Capacity Strengthening Department

Materials

Techniques in Plant Virology at CIP

Cover
Foreword
Introducction

PART 1: Transmission

1.1	Transmission of Plant Viruses
1.2	Aphid Transmission of Potato Viruses

PART 2: Detection

2.1 Symptomatology

2.1.1	Symptoms of Plant Virus Diseases
2.1.2	Diseases Caused by Phytoplasmas Organisms in Potato

2.2 Indicator Plants

2.2.1	Production and Maintenance of Indicator Plants
2.2.2	Disinfecting Soil
2.2.3	Inoculation of Indicator Plants for detection of Plant Viruses

2.3 Serology

2.3.1	An Introduction to Serology
2.3.2	Antiserum Procuction
2.3.3	Purification and Conjugation of Antiserum
2.3.4	Double Diffusion Test in Gels (Ouchterlony)
2.3.5	Latex Test
2.3.6	Enzyme-Linked Immunosorbent Assays: Double Antibody "Sandwich" (DAS-ELISA)
2.3.7	Enzyme-Linked Immunosorbent Assays on Nitrocellulose Membranes (NCM-ELISA)
2.3.8	Serological Assays with Enzymatic Conjugates: Penicillinase Enzymatic Conjugate (PNC-DAS-ELISA)

2.4 Potato Spindle Tuber Viroid

2.4.1	Potato Spindle Tuber Viroid (PSTVd)
2.4.2	Detection of PSTVd by Polyacrylamide Gel Electrophoresis (PAGE)

2.5 Molecular Methods

2.5.1	Nucleic Acid Hybridization (NASH)
2.5.2	Sample Preparation for PSTVd Detection by Nucleic Acid Hybridization
2.5.3	Preparation of ³²P-Labeled Probes by RNA Transcription
2.5.4	Preparation of ³²P-Labeled Probes by Nick Translation
2.5.5	Non-Radioactive PSTVd Detection Using Biotin-Labeled Probes
2.5.6	Non-Radioactive PSTVd Detection Using Fluorescein-Labeled-Probes

PART 3 : Microscopy

3.1

Electron Microscopy

PART 4 : Control

4.1	Potato Seedling Screening for Resistance to Potato Viruses X and Y by the Mass Inoculation Technique with Spray Gun
4.2	Virus Eradication: Tissue Culture of Meristems Thermotherapy, and Chemotherapy

PART 5 : Virus Purification

5.1	Fundamentals of Purification of Plant Viruses
5.2	Protocols for the Purification of Plant Viruses
5.3	Basic Principles Centrifugation

International Potato Center - Capacity Strengthening Department • Postal Address: Apartado 1558, Lima 12, Peru • Tel.: (511) 3496017 • Email: cip-csd@cgiar.org