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This volume explores plant immunity through protocols for recently established genetic and biochemical techniques to elucidate immune mechanisms mediated by pattern-recognition receptors (PRRs) and nucleotide-binding leucine-rich repeat receptors (NLRs). The book covers the identification of PRRs, their associated downstream immune regulators, PRR-ligand interaction-based immune responses, identification of NLR complex components, pan-NLRome analysis, resistosome assays, and cell death evaluation. Additionally, it presents protocols for co-transcriptome analysis of plant-pathogen interactions, single-nucleus multi-omics, metabolite imaging, and bioluminescence-based quantification of pathogen growth, as well as genome editing tools that contribute to the development of disease-resistant plants. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Plant Immunity: Methods and Protocols serves as an ideal guide for researchers aiming to accelerate ongoing research and advances in the field of plant immunity.
Sommario
Lumi-Map, a Luciferase Bioluminescence Screen of Mutants Combined with MutMap Allows Rapid Dissection of Plant Immune Signaling Pathways.- Identification of Phosphorylation and Protein Interactions of Plant Pattern Recognition Receptors Through Immunoprecipitation.- Plant-Based Ligand-Depletion Binding Assay for Assessing Pattern Recognition Receptor-Ligand Interactions.- Plate Reader-Based Quantification of Cellular Calcium Dynamics in Immune Responses Using the Genetically-Encoded Calcium Indicator GCaMP.- Visualization of Calcium Decoding via a FRET-Based Reporter of Calcium-Decoder Conformation.- Visualization and Quantification of Apoplastic ROS by Plant Immune NADPH Oxidase Utilizing Chemiluminescence.- Detection of Phosphorylated NADPH Oxidase by Antibody During Plant Immune Responses.- The Application of RenSeq for Pan-NLRome Analysis.- Blue Native-PAGE Assay of NLR Protein Complexes Combined with Genetic Complementation in Nicotiana benthamiana.- Quantification of NLR-Induced Cell Death Using Rice Protoplasts.- Split-TurboID-Based Proximity Labeling for Probing the Composition of Spatiotemporally Defined Protein Complexes In Planta.- Virus-Induced Gene Silencing for Identifying Autoimmune Regulators in Arabidopsis.- Bioinformatics Workflow for Co-Transcriptome Analysis of Plant-Bacterial Interactions.- Nuclei Isolation from Mature Leaves of Arabidopsis thaliana Infected by Bacterial Pathogens for Single-Nucleus Multiomics.- Multimodal Analysis of Phytoalexin Synthesis in Arabidopsis by Mass Spectrometry Imaging and Fluorescent Microscopy.- Bioluminescence-Based Growth Quantification of the Phytopathogenic Bacterium Pseudomonas syringae pv. tomato DC3000.- Genome Editing for Developing Disease-Resistant Plants.
