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Phosphorylation of WDR48 by phototropins drives starch degradation to promote stomatal opening

ABSTRACT: Plant stomata open in response to blue light under a background of red light. Red light induces starch accumulation through guard cell photosynthesis, whereas blue light causes rapid starch degradation via the activation of plasma membrane H+-ATPase through phosphorylation of BLUS1 by phototropins, thus promoting stomatal opening. However, how phototropins mediate starch degradation remains unclear. In this study, we identified WD-repeat protein 48 (WDR48), which is essential for starch degradation. Phosphoproteome analysis revealed that WDR48 was phosphorylated in response to blue light. Mutation of WDR48 impaired starch degradation and stomatal opening by blue light. Phototropins interacted with and phosphorylated WDR48. WDR48 and BLUS1 constituted separate signalling pathways required for starch degradation. We propose that the coordinated control of starch synthesis by red light and its degradation by blue light signalling through WDR48 and BLUS1 is a key mechanism for stomatal opening by red and blue light.

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

SUBMITTER: Atsushi Takemiya

PROVIDER: PXD073138 | JPOST Repository | Wed Feb 18 00:00:00 GMT 2026

REPOSITORIES: jPOST

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			Action	DRS
	240511NS01WT_RL.raw	Raw
	240511NS02WT_BL.raw	Raw
	240511NS03p1p2_RL.raw	Raw
	240511NS04p1p2_BL.raw	Raw
	240511NS07WT_RL.raw	Raw

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Phosphorylation of two Thr residues in the C-terminal auto-inhibitory domain of plasma membrane H+-ATPase is crucial for light-induced stomatal opening-2

Project description:	Plasma membrane H+-ATPase provides the driving force for light-induced stomatal opening. However, the mechanisms underlying the regulation of its activity remain unclear. Here, we showed that the phosphorylation of two Thr residues in the C-terminal autoinhibitory domain is crucial for H+-ATPase activation and stomatal opening. Our phosphoproteome analysis revealed that blue light induces the phosphorylation of Thr-881 within the C-terminal region I domain and penultimate Thr-948 in AUTOINHIBITED H+-ATPASE 1 (AHA1). The site-directed mutagenesis showed that both Thr phosphorylations are essential for H+ pumping and stomatal opening in response to blue light. The phosphorylation of Thr-948 is a prerequisite for the phosphorylation of Thr-881 by blue light. Furthermore, red-light-driven guard cell photosynthesis also caused Thr-881 phosphorylation, possibly contributing to red-light-dependent stomatal opening. Taken together, our findings provide mechanistic insights into H+-ATPase activation that exploits the ion transport across the plasma membrane and light signalling network in guard cells.

2023-12-25 | PXD044476 | JPOST Repository

Phosphorylation of two Thr residues in the C-terminal auto-inhibitory domain of plasma membrane H+-ATPase is crucial for light-induced stomatal opening

Project description:Plasma membrane H+-ATPase provides the driving force for light-induced stomatal opening. However, the mechanisms underlying the regulation of its activity remain unclear. Here, we showed that the phosphorylation of two Thr residues in the C-terminal autoinhibitory domain is crucial for H+-ATPase activation and stomatal opening. Our phosphoproteome analysis revealed that blue light induces the phosphorylation of Thr-881 within the C-terminal region I domain and penultimate Thr-948 in AUTOINHIBITED H+-ATPASE 1 (AHA1). The site-directed mutagenesis showed that both Thr phosphorylations are essential for H+ pumping and stomatal opening in response to blue light. The phosphorylation of Thr-948 is a prerequisite for the phosphorylation of Thr-881 by blue light. Furthermore, red-light-driven guard cell photosynthesis also caused Thr-881 phosphorylation, possibly contributing to red-light-dependent stomatal opening. Taken together, our findings provide mechanistic insights into H+-ATPase activation that exploits the ion transport across the plasma membrane and light signalling network in guard cells.

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Project description:Environmental stimuli-triggered stomatal movement is a key physiological process that regulates CO<sub>2</sub> uptake and water loss in plants. Stomata are defined by pairs of guard cells that perceive and transduce external signals, leading to cellular volume changes and consequent stomatal aperture change. Within the visible light spectrum, red light induces stomatal opening in intact leaves. However, there has been debate regarding the extent to which red-light-induced stomatal opening arises from direct guard cell sensing of red light versus indirect responses as a result of red light influences on mesophyll photosynthesis. Here we identify conditions that result in red-light-stimulated stomatal opening in isolated epidermal peels and enlargement of protoplasts, firmly establishing a direct guard cell response to red light. We then employ metabolomics workflows utilizing gas chromatography mass spectrometry and liquid chromatography mass spectrometry for metabolite profiling and identification of Arabidopsis guard cell metabolic signatures in response to red light in the absence of the mesophyll. We quantified 223 metabolites in Arabidopsis guard cells, with 104 found to be red light responsive. These red-light-modulated metabolites participate in the tricarboxylic acid cycle, carbon balance, phytohormone biosynthesis and redox homeostasis. We next analyzed selected Arabidopsis mutants, and discovered that stomatal opening response to red light is correlated with a decrease in guard cell abscisic acid content and an increase in jasmonic acid content. The red-light-modulated guard cell metabolome reported here provides fundamental information concerning autonomous red light signaling pathways in guard cells.

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