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Full Description
This volume provides a comprehensive understanding of the methods used in studying phytoplasma diseases and transmission. Chapters cover a wide range of topics, from basic detection methods to advanced genomics and emerging technologies, vectors, plant interactions, and disease management. 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.
Cutting-edge and comprehensive, Phytoplasma: Methods and Protocols is a valuable resource for all researchers and practitioners with a keen interest in practical methodologies for exploring various facets of phytoplasma disease and its transmission.
Contents
Phytoplasmas: An Introduction.- Phytoplasma Detection and Identification: Tools for their Classification.- End Point PCR for Universal Detection of Phytoplasmas Based on 16S rRNA Gene.- Imp Gene Analysis for Diagnosis and Genotyping of Flavescence Dorée Related Phytoplasmas.- Triplex qPCR for Grapevine Yellows Phytoplasmas Combined with a Universal Endogenous Control.- Rapid and Sensitive Detection of Phytoplasma Diseases Using a CRISPR/Cas12a DETECTR Assay Combined with Isothermal Recombinase Polymerase Amplification.- On-site Universal Phytoplasma Detection by 23S rRNA-gene-targeted Loop-mediated Isothermal Amplification (LAMP).- Phytoplasma Transmission Trials via Insect Vectors Under Controlled Conditionn.- A Workflow for Phytoplasma Discovery and Characterization in Potential Insect Vectors: From Non-destructive DNA Extraction and Vouchering to Next-generation Sequencing.- Insect Cell Cultures as Platforms for the Study of Insect Vectors' Physiology and Vector-phytoplasma Interactionn.- Protocols for Grapevine Transformation: Tissue Preparation, Protoplast Isolation, and Cultivation.- Expression of Recombinant Phytoplasma Proteins in E. coli.- Transient Expression of Phytoplasma Proteins in Planta Using an Agro-infiltration Method.- An In Vitro Ubiquitination Assay to Study Phytoplasmal Effector Function.- Post-embedding Immunolabeling of Thin Sections of Phytoplasma-infected Plant Tissues for Transmission Electron Microscopy.- The use of Spiroplasma melliferum as a Model Organism to Study Potential Antimicrobials Against Phytoplasma.- High-resolution Identification of Phytoplasmas With Multiplexed Multilocus Sequence Typing Using Hybridization Probe-based Enrichment.- Phytoplasma Genome Reconstruction: From DNA Enrichment to Sequencing and Genome Assembly. Complete Genome Sequence of Phytoplasma Obtained by Nanopore Sequencing Using High Molecular Weight DNA.
