Targeted Metabolomics of Plant Hormones and Redox Metabolites in Stomatal Immunity

Phytohormones and redox metabolites are important molecules in a number of biological processes related to plant growth, development, and stress responses. Understanding how these metabolites are involved in abiotic and biotic stress is a frequent topic of plant biology research. However, many factors, such as low physiological concentrations and the inherent complexity of plant samples, make identification and quantification of these important metabolites difficult. Here, we describe a method for metabolite extraction from whole leaves and guard cell–enriched samples and a targeted metabolomics strategy for the identification and quantification of specific hormone- and redox-related metabolites. In our experiment, we used the reference plant Arabidopsis thaliana infected with the biotrophic pathogen Pseudomonas syringe pv. tomato (Pst) DC3000, and examined the changes in hormone and redox metabolites in systemic leaves, using the targeted metabolomics strategy in order to investigate potential functions of these metabolites in systemic acquired resistance (SAR) during a plant’s immune responses. The methods reported here can be expanded to other metabolites and other biological systems beyond plants and bacterial pathogens.