Mitogen-activated protein kinases (MAPKs) are versatile phosphorylating enzymes which regulate multiple proteins involved in gene expression, cell architecture, plant development and reaction to diverse abiotic and biotic factors. The main aim of Plant MAP Kinases: Methods and Protocols is to provide established and new MAPK protocols adapted to the challenges posed by working with plants. The book contains 19 chapters which encompass a wide array of methods progressively scaling from the single gene, protein or cell level to large-scale arrays of proteomic, phosphoproteomic and interactomic data in order to uncover previously unidentified plant MAPK signaling pathways and to tackle with the challenging task of substrate identification. Techniques for MAPK sequence analysis and subcellular localization helping to identify their substrates and subcellular compartmentalization are also provided. 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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Plant MAP Kinases: Methods and Protocols represents a collection of useful plant MAPK protocols written by experts in the field for researchers and students.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt
Transient Expression in Arabidopsis Leaf Mesophyll Protoplast System for Cell-based Functional Analysis of MAPK Cascades Signaling.- Quantification of Stress Induced Mitogen-activated Protein Kinase Expressional Dynamic using Quantitative Real-time PCR.- Analysis of MAPK Activities using MAPK-specific Antibodies.- Detection of Protein Phosphorylation and Charge Isoforms using Vertical one-Dimensional Isoelectric Focusing Gels.- Affinity-based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag™.- Identification of Constitutively Active AtMPK6 Mutants using a Functional Screen in Saccharomyces Cerevisiae.- Virus-induced Gene Silencing in Plant MAPK Research.- RNA Interference of Plant MAPK Cascades for Functional Studies.- Immunofluorescent Localization of MAPKs and Colocalization with Microtubules in Arabidopsis Seedling Whole Mount Probes.- Immunofluorescent Localization of MAPKs in Steedman's Wax Sections.- Fluorescent Protein Tagging of Arabidopsis MAPKs for In Vivo Localization Studies.- Bimolecular Fluorescent Complementation (BiFC) by MAP Kinases and MAPK Phosphatases.- Determination of Phosphorylation Sites in Microtubule Associated Protein MAP65-1.- In vivo Phosphorylation of WRKY Transcription Factor by MAPK.- Rapid Mutagenesis-based Analysis of Phosphorylation Sites in Mitogen-activated Protein Kinase Substrates.- Yeast Two-hybrid System for Dissecting the Rice MAPK Interactome.- Experimental and Analytical Approaches to Characterize Plant Kinases using Protein Microarrays.