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Identification of two-component system elements downstream of AHK5 in the stomatal closure response of Arabidopsis thaliana.

ABSTRACT: To optimize water use efficiency, plants regulate stomatal closure through a complex signaling process. Hydrogen peroxide (H?O?) 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 H?O? and other stimuli that depend on H?O?. 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 H?O?- 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 H?O? response pathways of guard cells. Furthermore, we demonstrate that aspartate phosphorylation of ARR4 is only required for the H?O?- 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 H?O? and ethylene.

SUBMITTER: Mira-Rodado V

PROVIDER: S-EPMC3548872 | biostudies-other | 2012 Nov

REPOSITORIES: biostudies-other

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Identification of two-component system elements downstream of AHK5 in the stomatal closure response of Arabidopsis thaliana.

Mira-Rodado Virtudes V Veerabagu Manikandan M Witthöft Janika J Teply Julia J Harter Klaus K Desikan Radhika R

Plant signaling & behavior 20120905 11

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. Th ...[more]

PMID: 22951399

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Project description:The Mg-chelatase H subunit (CHLH) has been shown to mediate chlorophyll biosynthesis, as well as plastid-to-nucleus and abscisic acid (ABA)-mediated signaling. A recent study using a novel CHLH mutant, rtl1, indicated that CHLH specifically affects ABA-induced stomatal closure, but also that CHLH did not serve as an ABA receptor in Arabidopsis thaliana. However, the molecular mechanism by which CHLH engages in ABA-mediated signaling in guard cells remains largely unknown. In the present study, we examined CHLH function in guard cells and explored whether CHLH expression might influence stomatal aperture. Incubation of rtl1 guard cell protoplasts with ABA induced expression of the ABA-responsive genes RAB18 and RD29B, as also observed in wild-type (WT) cells, indicating that CHLH did not affect the expression of ABA-responsive genes. Earlier, ABA was reported to inhibit blue light (BL)-mediated stomatal opening, at least in part through dephosphorylating/inhibiting guard cell H(+)-ATPase (which drives opening). Therefore, we immunohistochemically examined the phosphorylation status of guard cell H(+)-ATPase. Notably, ABA inhibition of BL-induced phosphorylation of H(+)-ATPase was impaired in rtl1 cells, suggesting that CHLH influences not only ABA-induced stomatal closure but also inhibition of BL-mediated stomatal opening by ABA. Next, we generated CHLH-GFP-overexpressing plants using CER6 promoter, which induces gene expression in the epidermis including guard cells. CHLH-transgenic plants exhibited a closed stomata phenotype even when brightly illuminated. Moreover, plant growth experiments conducted under water-deficient conditions showed that CHLH transgenic plants were more tolerant of drought than WT plants. In summary, we show that CHLH is involved in the regulation of stomatal aperture in response to ABA, but not in ABA-induced gene expression, and that manipulation of stomatal aperture via overexpression of CHLH in guard cells improves plant drought tolerance.

Dataset Information

Identification of two-component system elements downstream of AHK5 in the stomatal closure response of Arabidopsis thaliana.

Publications

Identification of two-component system elements downstream of AHK5 in the stomatal closure response of Arabidopsis thaliana.

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