The role of Arabidopsis ABA receptors from the PYR/PYL/RCAR family in stomatal acclimation and closure signal integration

Abstract

Stomata are microscopic pores found on the surfaces of leaves that act to control CO2 uptake and water loss. By integrating information derived from endogenous signals with cues from the surrounding environment, the guard cells, which surround the pore, ‘set’ the stomatal aperture to suit the prevailing conditions. Much research has concentrated on understanding the rapid intracellular changes that result in immediate changes to the stomatal aperture. In this study, we look instead at how stomata acclimate to longer timescale variations in their environment. We show that the closure-inducing signals abscisic acid (ABA), increased CO2, decreased relative air humidity and darkness each access a unique gene network made up of clusters (or modules) of common cellular processes. However, within these networks some gene clusters are shared amongst all four stimuli. All stimuli modulate the expression of members of the PYR/PYL/RCAR family of ABA receptors. However, they are modulated differentially in a stimulus-specific manner. Of the six members of the PYR/PYL/RCAR family expressed in guard cells, PYL2 is sufficient for guard cell ABA-induced responses, whereas in the responses to CO2, PYL4 and PYL5 are essential. Overall, our work shows the importance of ABA as a central regulator and integrator of long-term changes in stomatal behaviour, including sensitivity, elicited by external signals. Understanding this architecture may aid in breeding crops with improved water and nutrient efficiency.

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Fig. 1: Guard cell signalling pathway analysis.
Fig. 2: Integrative networks analysis identifies distinct stomatal closing signal modules.
Fig. 3: PYL2 is sufficient for stomatal ABA sensitivity.
Fig. 4: ABA-induced movements of individual stoma.
Fig. 5: PYL4 and PYL5 render guard cells CO2 sensitive.

Data availability

The data that support the findings of this study are available from the corresponding authors upon reasonable request. Microarray data of the ABA and low air humidity treatments that were taken from ref. 6 were deposited in the Gene Expression Omnibus (GEO) database http://www.ncbi.nlm.nih.gov/geo with accession no. GSE41054. The microarray data from CO2 and darkness experiments were deposited in the same database under GSE118520.

Code availability

Algorithms and statistics used in the analyses are based on published approaches available in R packages (mainly Bioconductor framework) and other cited publicly available repositories.

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Acknowledgements

Work in the Hedrich laboratory was supported by a grant from King Saud University Deanship for Scientific Research, International Research Group Programme (IRG14-22), Riyadh, Saudi Arabia, and that of the Rodriguez laboratory was supported by the Ministerio de Ciencia e Innovacion, Fondo Europeo de Desarrollo Regional and Consejo Superior de Investigaciones Cientificas (grants BIO2014-52537-R and BIO2017-82503-R, to P.L.R.). M.D. was supported by the CRC/Transregio 124 – FungiNet funded by the Deutsche Forschungsgemeinschaft (project B2). A.M.H. acknowledges support from the UK BBSRC (grant no. BB/N001168/1). E.M. was supported by the Estonian Research Council (grant no. PUT1133).

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M.D. and T.M. conceived and conducted bioinformatics. H.M.M., H.B. and P.A. conceived, performed and analysed the expression studies. H.M.M. and E.M. conducted and analysed gas exchange measurements. M.P.-L. and P.L.R. conceived and conducted the generation of transgenic plants. C.-M.G. and S.C.C. conceived and conducted proteomic analyses. J.H. conceived, conducted and analysed electro-infusion experiments. P.A., P.L.R., H.K., K.A.S.A.-R., T.M., A.M.H. and R.H. designed and conceived the study. M.D., T.M., P.A., A.M.H. and R.H. wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Tobias Müller or Alistair M. Hetherington.

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Supplementary Information

Supplementary Notes, Supplementary References, legends for Supplementary Videos and Supplementary Figs. 1–9.

Reporting Summary

Supplementary Video 1

Stomatal movement following ABA electro-infusion of the 11,458 and 12,458 mutants.

Supplementary Video 2

Stomatal movement following ABA electro-infusion of wild type and the PYL2::12458 complementation line.

Supplementary Table 1

Table with three sheets.

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Dittrich, M., Mueller, H.M., Bauer, H. et al. The role of Arabidopsis ABA receptors from the PYR/PYL/RCAR family in stomatal acclimation and closure signal integration. Nat. Plants 5, 1002–1011 (2019). https://doi.org/10.1038/s41477-019-0490-0

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