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Updated: May 18, 2026

Direct Observation and Automated Measurement of Stomatal Responses to Pseudomonas syringae pv. tomato DC3000 in Arabidopsis thaliana
Published on: February 9, 2024
Identification of two-component system elements downstream of AHK5 in the stomatal closure response of Arabidopsis
Virtudes Mira-Rodado1, Manikandan Veerabagu, Janika Witthöft
1Center for Plant Molecular Biology (ZMBP), Department of Plant Physiology, University of Tübingen, Tübingen, Germany.
Abstract:
To optimize water use efficiency, plants regulate stomatal closure through a complex signaling process. Hydrogen peroxide (H₂O₂) is produced in response to several environmental stimuli, and has been identified as a key second messenger involved in the regulation of stomatal aperture. The Arabidopsis histidine kinase 5 (AHK5) has been shown to regulate stomatal closure in response to H₂O₂ and other stimuli that depend on H₂O₂. AHK5 is a member of the two-component system (TCS) in Arabidopsis. The plant TCS comprises three different protein types: the hybrid histidine kinases (HKs), the phosphotransfer proteins (HPs) and the response regulators (RRs). Here we determined TCS elements involved in H₂O₂- and ethylene-dependent stomatal closure downstream of AHK5. By yeast and in planta interaction assays and functional studies, AHP1, 2 and 5 as well as the response regulators ARR4 and ARR7 were identified acting downstream of AHK5 in the ethylene and H₂O₂ response pathways of guard cells. Furthermore, we demonstrate that aspartate phosphorylation of ARR4 is only required for the H₂O₂- but not for the ethylene-induced stomatal closure response. Our data suggest the presence of a complex TCS signaling network comprising of at least AHK5, several AHPs and response regulators, which modulate stomatal closure in response to H₂O₂ and ethylene.
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