Dawn regulates guard cell proteins in Arabidopsis thaliana that function in ATP production from fatty acid beta-oxidation
Based on the nature of the proteins that are altered in abundance, we conclude that guard cells switch their energy source from fatty acid metabolism to chloroplast activity, at the onset of dawn.
During stomatal opening at dawn, evidence was recently presented for a breakdown and liquidation of stored triacylglycerols in guard cells to supply ATP for use in stomatal opening. However, proteome changes that happen in the guard cells during dawn were until now poorly understood. Bad accessibility to pure and intact guard cell samples can be considered as the primary reason behind this lack of knowledge. To overcome these technical constraints, epidermal guard cell samples with ruptured pavement cells were isolated at 1 h pre-dawn, 15 min post-dawn and 1 h post-dawn from Arabidopsis thaliana. Proteomic changes were analysed by ultra-performance-liquid–chromatography-mass-spectrometry. With 994 confidently identified proteins, we present the first analysis of the A. thaliana guard cell proteome that is not influenced by side effects of guard cell protoplasting. Data are available via ProteomeXchange with identifier PXD009918. By elucidating the identities of enzymes that change in abundance by the transition from dark to light, we corroborate the hypothesis that respiratory ATP production for stomatal opening results from fatty acid beta-oxidation. Moreover, we identified many proteins that were never reported in the context of guard cell biology. Among them are proteins that might play a role in signalling or circadian rhythm.
KeywordsGuard cell Stomatal opening ATPase Beta-oxidation Triacylglycerol Dawn
Principal component analysis
PRoteomics IDEntifications database
Singular enrichment analysis
Cross comparison of SEA
Partial least squares analysis
Chloroplast stem-loop binding proteins
Database for annotation, visualization and integrated discovery
Plastidial glyceraldehyde-3-phosphate dehydrogenases
The authors are grateful to Kusay Arat for excellent technical assistance and the Coopération européenne dans le domaine de la recherche scientifique et technique is acknowledged for granting a STSM scholarship to Christoph-Martin Geilfus (Action FA1306). Dr. Deirdre McLachlan, University Bristol, is acknowledged for giving critical comments. We are grateful to Bastian Franzisky, University Hohenheim, for helping with guard cell preparation. This work was supported by the DFG research Grant GE 3111/1–1 to C.-M. Geilfus.
JL designed the study, grew plants and collected epidermal peels; CMG collected proteomic data; CMG and SC contributed data analysis, SC calculated statistics; CMG, SC and JL wrote the paper.
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