# A drought stress-responsive metabolite malate modulates stomatal responses through G-protein-dependent pathway in grapevine and Arabidopsis

## TL;DR

The study shows that the TCA cycle metabolite malate helps plants respond to drought by triggering stomatal closure through a G-protein-dependent signaling pathway in grapevine and Arabidopsis.

## Contribution

The novel finding is that malate specifically activates stomatal closure via G-protein signaling, linking metabolic changes to drought tolerance in plants.

## Key findings

- Malate induces stomatal closure by increasing cytosolic Ca2+ and activating the SLAC1 anion channel in guard cells.
- G-proteins are essential for malate signaling by regulating second messenger production in drought responses.
- TCA cycle metabolites are sensed individually by guard cells, with malate playing a central role in drought adaptation.

## 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. Ca2+ imaging and ion channel measurements indicated that fumarate, malate, and α-ketoglutarate induced cytosolic free Ca2+ concentration ([Ca2+]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 [Ca2+]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.

The online version contains supplementary material available at 10.1186/s43897-025-00181-z.