Abstract
Drought stress is a significant environmental threat to global agricultural production and distribution. Plant adaptation to dehydration stress involves intricate biological processes with substantial changes in metabolite composition. In this study, we investigated the role of tricarboxylic acid (TCA) cycle metabolites in drought tolerance in grapevine and Arabidopsis by metabolome, live cell imaging, electrophysiological and pharmacological approaches. Metabolome analysis revealed that amount of malate, citrate, and isocitrate increased over time in detached grapevine leaves. Ca(2+) imaging and ion channel measurements indicated that fumarate, malate, and α-ketoglutarate induced cytosolic free Ca(2+) concentration ([Ca(2+)](cyt)) elevation in guard cells and directly activated a guard-cell anion channel SLOW ANION CHANNEL-ASSOCIATED 1 (SLAC1). However, only malate induced stomatal closure, which required increases in [Ca(2+)](cyt) in guard cells and activation of SLAC1. Through pharmacological experiments and reverse genetics analyses, G-proteins were identified as essential components of malate signaling by regulating second messenger production. These results indicate that TCA cycle metabolites are sensed individually by guard cells and that malate plays a key role in connecting metabolic regulation and drought tolerance through G-protein-dependent signal cascades.