Plant Immunity

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.

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.