This detailed volume explores techniques to study reactive oxygen species (ROS) in plants and to characterize their roles in development and stress responses. Beginning with a section on strategies to induce ROS production, the book continues with methods to visualize ROS and detect changes in redox homeostasis, small-scale and targeted analyses for investigating the effects of ROS accumulation during stress on plant physiology and metabolism, as well as systems biology approaches to understand ROS functions. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Reactive Oxygen Species in Plants: Methods and Protocols serves as a vital resource that any researcher, and in particular young researchers, can use and adapt to further our knowledge of this dynamic area of plant science.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

Modification of Chloroplast Antioxidant Capacity by Plastid Transformation.- Analysis of Ascorbate Metabolism in Arabidopsis under High-Light-Stress.- Genetic Manipulation of Reactive Oxygen Species (ROS) Homeostasis Utilizing CRISPR/Cas9-Based Gene Editing in Rice.- Studying Plant Stress Reactions In Vivo by PAM Chlorophyll Fluorescence Imaging.- Live Monitoring of ROS-Induced Cytosolic Redox Changes with roGFP2-Based Sensors ** in Plants.- Quantitative Measurement of Ascorbate and Glutathione by Spectrophotometry.- Measurement of NAD(P)H and NADPH-Generating Enzymes.- Quantitative Analysis for ROS-Producing Activity and Regulation of Plant NADPH Oxidases in HEK293T Cells.- Estimation of the Level of Abasic Sites in Plant mRNA Using Aldehyde Reactive Probe.- A Simplified Method to Assay Protein Carbonylation by Spectrophotometry.- In Vitro Biochemical Analysis of Recombinant Plant Proteins under Oxidation.- Methods to Analyze the Redox Reactivity of Plant Proteins.- Determination of ROS-Induced Lipid Peroxidation by HPLC-Based Quantification of Hydroxy Polyunsaturated Fatty Acids.- Detection of Lipid Peroxidation-Derived Free Azelaic Acid, a Biotic Stress Marker and Other Dicarboxylic Acids in Tobacco by Reversed Phase HPLC-MS under Non-Derivatized Conditions.- Determination of Reactive Carbonyl Species, which Mediate Reactive Oxygen Species Signals in Plant Cells.- Measuring Stress-Induced Changes in Defense Phytohormones and Related Compounds.- Targeted Mass Spectrometry Analysis of Protein Phosphorylation by Selected Ion Monitoring Coupled to Parallel Reaction Monitoring (tSIM/PRM).- Quantitative Analysis of Post-Translational Modifications of Plant Histones.- Characterization of RBPome in Oxidative Stress Conditions.- Analysis of ROS-Triggered Changes in the Transcriptome.