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Full Description
This book presents a set of modern protocols forming a solid background for who want to start or improve research programme on phytoplasmas. Chapters guide readers through detailed techniques for maintaining phytoplasma collections, border inspection, detection of different phytoplasma strains, new pipelines to produce phytoplasma genome draft, protocols for phytoplasma gene expression analyses, and methods for the investigation of the phloem tissue. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Phytoplasmas: Methods and Protocols aims to ensure successful results in the further study of this vital field.
Contents
Phytoplasmas: An Introduction.- Micro-tom Tomato Grafting for Stolbur-phytoplasma Transmission: Different Grafting Techniques.- Phytoplasma Transmission: Insect Rearing and Infection Protocols.- Sampling Methods for Leafhopper, Planthopper, and Psyllid Vectors.- Symptoms of Phytoplasma Diseases.- Comparison of Different Procedures for DNA Extraction for Routine Diagnosis of Phytoplasmas.- Standard Detection Protocol: PCR and RFLP Analyses Based on 16S rRNA Gene.- PCR-based Sequence Analysis on Multiple Genes other than 16S rRNA Gene for Differentiation of Phytoplasmas.- Real-time PCR Protocol for Phytoplasma Detection and Quantification.- Duplex TaqMan Real-Time PCR for Rapid Quantitative Analysis of a Phytoplasma in its Host Plant without External Standard Curves.- A Multiplex-PCR Method for Diagnosis of AY-Group Phytoplasmas.- One-step Multiplex Quantitative RT-PCR for the Simultaneous Detection of Viroids and Phytoplasmas.- A Rapid Protocol of Crude RNA/DNA Extraction for RT-qPCR Detection and Quantification.- Quantitative Analysis with Droplet Digital PCR.- Rapid Sample Preparation and LAMP for Phytoplasma Detection.- Assembly of Phytoplasma Genome Drafts from Illumina Reads using Phytoassembly.- Protocol for the Definition of a Multi-Spectral Sensor for Specific Foliar Disease Detection: Case of 'Flavescence Dorée'.- Transcriptomic Analyses of Phytoplasmas.- Transcriptomic Analyses of Phytoplasmas.- Laser Microdissection of Phytoplasma-infected Grapevine Leaf Phloem Tissue for Gene Expression Study.- Collection of Phloem Sap in Phytoplasma-infected Plants.- DAPI and Confocal Laser-Scanning Microscopy for in vivo Imaging of Phytoplasmas.- Immunofluorescence Assay to Study Early Events of Vector Salivary Gland Colonization by Phytoplasmas.- Characterization of Phytoplasmal Effector Protein Interaction with Proteinaceous Plant Host Targets using Bimolecular Fluorescence Complementation (BiFC).- Collection, Identification, and Statistical Analysis of Volatile Organic Compound Patterns Emitted by Phytoplasma Infected Plants.- Quantification of Phytohormones by HPLC-MS/MS Including Phytoplasma-infected Plants.
