Textbook in PDF format

This book provides detailed information on methodologies used in biological, serological and nucleic acid based assays for the detection, diagnosis and management of plant viruses. The content is divided into six main parts, the first of which presents techniques used in the biological characterization and transmission of viruses, while Part II covers purification and techniques concerning the physico-chemical properties of viruses. In turn, Part III focuses on in vitro expression, production of polyclonal and monoclonal antibodies; and on various serological assays including precipitin tests, ELISA, blot immunoassays, immunosorbent electron microscopy and lateral flow immunoassays. Part IV addresses the isolation of DNA and RNA from plants and nucleic acid based assays such as dot-blot, polymerase chain reaction, real-time PCR, loop-mediated isothermal amplification, rolling circle amplification, recombinase polymerase amplification, and next-generation sequencing approaches. Part V discusses cloning, sequencing, sequence analysis and the production of infectious clones, while the last part (Part VI) provides biotechnological approaches for the management of viruses. Given its scope, the book will be a valuable resource for every laboratory, student and teacher, and for everyone interested in plant virology, plant pathology, plant biology and molecular biology, offering them a practical manual on various aspects of plant viruses.
Foreword
Preface
About the Book
About the Authors
Abbreviations
Glasshouse for Maintenance of Virus and Insect Culture
Symptoms of Virus-Infected Plants
Isolation and Diagnosis of Virus Through Indicator Hosts
Host Range of Viruses
Physico-chemical Properties of Virus in Crude Sap
Mechanical Sap Transmission
Transmission Through Grafting and Budding
Transmission Through Dodder
Virus Transmission Through Pollen
Transmission Through Seeds
Transmission of Viruses by Aphids
Transmission of Viruses by Leafhoppers
Transmission of Viruses by Whiteflies
Transmission of Viruses by Thrips
Transmission of Viruses Through Mealybugs
Transmission of Viruses Through Beetles
Transmission of Viruses Through Mites
Transmission of Viruses Through Fungi
Transmission of Viruses Through Nematodes
Storage and Preservation of Plant Virus Cultures
Purification of Plant Viruses
Ultraviolet Absorption Spectra of Purified Virus Preparation
Electron Microscopy and Utramicrotomy
Determination of Coat Protein Molecular Weight of Viruses
Isolation of Nucleic Acid from Purified Virus and Determination of Its Nature
Agarose Gel Electrophoresis for Nucleic Acids
In Vitro Expression of Viral Coat Protein in Prokaryotic System and Its Purification
Production of Polyclonal Antiserum
Production of Monoclonal Antibody
Serological Tests
Isolation of Total DNA from Plants
Isolation of Total RNA from Plants
Isolation of Double-Stranded (ds) RNA from Virus-Infected Plants
Dot-Blot Hybridization Technique
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
DNA Microarray for Detection of Plant Viruses
Loop-Mediated Isothermal Amplification (LAMP)
Rolling Circle Amplification (RCA)
Recombinase Polymerase Amplification
Next-Generation Sequencing for Diagnosis of Viruses
Cloning of PCR Product
cDNA Synthesis and Cloning
DNA Sequencing
Sequence Analysis and Phylogenetic Studies
Development of Infectious Clone of Virus
Virus Elimination by Meristem-Tip Culture
Virus Elimination Through Somatic Embryogenesis
Production of Virus-Resistant Plants Through Transgenic Approaches
Production of Virus-Resistant Plants Through CRISPR-Cas Technology
Appendix: Common Conversions, Information Sources and Software of Nucleic Acids and ProteinsWeight conversion1 μg = 10-6 g1 ng...
DNA Data
Important DNA and Protein Information Sources and Software
Software for Sequence Analysis
Glossary