Phytophthora Methods and Protocols

This comprehensive volume examines techniques for studying Phytophthora species, a devastating group of plant pathogens that cause diseases in crops and natural ecosystems worldwide. The book compiles essential molecular techniques and methods, such as CRISPR-Cas genome editing for investigating pathogenicity and resistance, as well as tools for detecting, visualizing, and quantifying disease. Written for the highly successful Methods in Molecular Biology series, each chapter includes an introduction to its respective topic, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Phytophthora: Methods and Protocols equips researchers with the tools needed to address the challenges posed by these destructive microorganisms.

Promoter Analysis and Dissection Using Reporter Genes, Comparative Genomics,

and Gel Shift Assays in Phytophthora.- RNA Silencing Strategies in Phytophthora: Experimental Guidelines and Insights.- Protoplast Transformation of Phytophthora spp.- Gene Editing and Protein Tagging in the Oomycete Phytophthora infestans Using CRISPR-Cas12a.- CRISPR/Cas9-Mediated Gene-Knockout and In Situ Complementation System for Phytophthora sojae.- Fluorescence In Situ Hybridization Protocol for Visualization of Oomycetes In Vitro and In Planta.- Rapid and Robust Monitoring of Phytophthora Infectivity through Detached Leaf Assays with Automated Image Analysis.- Quantification of Phytophthora palmivora Infection in Barley and Related Monocot Roots.- The Use of eDNA Metabarcoding to Detect and Identify Phytophthora in Water Samples.- A Method for the Separation of Phytophthora Oospores from Soil for DNA-Based Detection.- Annotation of RxLR Effectors in Oomycete Genomes.- Pinus radiata cDNA Library for the Screening of Phytophthora Effector Protein Targets in Yeast.- Immunoisolation of Plant Endosomal Vesicles to Explore Uptake of Pathogen Effector Proteins during Infection of Nicotiana benthamiana.- Fabrication of Lab-on-a-Chip Devices to Study the Forces Imparted by Growing Phytophthora Hyphae.- Application of Proteomic Methods in Oomycete Biology.- Heterologous Expression of Secreted Proteins from Phytophthora in Pichia pastoris Followed by Protein Purification by Immobilized Metal Ion Affinity (IMAC).