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Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells

Nature Cell Biology volume 12, pages 87–93 (2010)Cite this article

An Erratum to this article was published on 01 June 2011

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Abstract

The continuing rise in atmospheric CO2 causes stomatal pores in leaves to close and thus globally affects CO2 influx into plants, water use efficiency and leaf heat stress1,2,3,4. However, the CO2-binding proteins that control this response remain unknown. Moreover, which cell type responds to CO2, mesophyll or guard cells, and whether photosynthesis mediates this response are matters of debate5,6,7,8. We demonstrate that Arabidopsis thaliana double-mutant plants in the β-carbonic anhydrases βCA1 and βCA4 show impaired CO2-regulation of stomatal movements and increased stomatal density, but retain functional abscisic-acid and blue-light responses. βCA-mediated CO2-triggered stomatal movements are not, in first-order, linked to whole leaf photosynthesis and can function in guard cells. Furthermore, guard cell βca-overexpressing plants exhibit instantaneous enhanced water use efficiency. Guard cell expression of mammalian αCAII complements the reduced sensitivity of ca1 ca4 plants, showing that carbonic anhydrase-mediated catalysis is an important mechanism for βCA-mediated CO2-induced stomatal closure and patch clamp analyses indicate that CO2/HCO3− transfers the signal to anion channel regulation. These findings, together with ht1-2 (ref. 9) epistasis analysis demonstrate that carbonic anhydrases function early in the CO2 signalling pathway, which controls gas-exchange between plants and the atmosphere.

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Figure 1: Disruption of the carbonic anhydrases βCA1 and βCA4 greatly impairs CO2-induced stomatal movements, but not responses to blue light and abscisic acid.
Figure 2: Introduction of wild-type genomic βca complements the reduced CO2 sensitivity of ca1 ca4 plants.
Figure 3: Leaf photosynthesis is not directly linked to βCA-mediated CO2-triggered stomatal responses.
Figure 4: βca expression in ca1 ca4 guard cells restores CO2 responses and βca overexpressing plants show improved instantaneous water use efficiency.
Figure 5: HT1 epistasis analysis, human αCAII expression in guard cells restores CO2 responsiveness and HCO3− regulation of anion channels.

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  • 16 May 2011

    In the version of this letter initially published online and in print, the competing financial interests were incomplete.

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Acknowledgements

We thank M. Maktabi, J. Young and C. Engineer for preliminary analyses of βca mutants, R. Xu for assistance, S. Zeeman for suggestions and K. Iba (Kyushu University, Japan) for providing ht1-2 seeds. This research was supported by NSF (MCB0918220), NIH (GM060396) and in part by DOE (DE-FG02-03ER15449) grants (to J.I.S.) and by fellowships from the Swedish Research Council Formas (to M.I.-N.), the Deutsche Forschungsgemeinschaft (to M.B.), EMBO (to J.M.K.) and in part from the King Abdullah University of Science and Technology (KAUST; No. KUS-F1-021-31 to H.H.).

Author information

Author notes

  1. Shaowu Xue

    Present address: Permanent address: Institute of Molecular Science, Shanxi University, Taiyuan, 030006, China.,

  2. Aurélien Boisson-Dernier

    Present address: Current address: Department of Developmental Genetics, University of Zürich, CH-8008 Zürich, Switzerland.,

  3. Maria Israelsson-Nordström

    Present address: Current address: Department of Botany, Stockholm University, SE-10691 Stockholm, Sweden.,

  4. Honghong Hu, Aurélien Boisson-Dernier and Maria Israelsson-Nordström: These authors contributed equally to this work.

  5. Maik Böhmer and Shaowu Xue: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Biological Sciences, Cell and Developmental Biology Section, University of California San Diego, La Jolla, 92093-0116, CA, USA

    Honghong Hu, Aurélien Boisson-Dernier, Maria Israelsson-Nordström, Maik Böhmer, Shaowu Xue, Amber Ries, Jan Godoski, Josef M. Kuhn & Julian I. Schroeder

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Contributions

J.I.S. conceived the project and proposed the experimental design. H.H., A. B.-D. and M.I.-N. performed most of the experiments and contributed equally to the work. M.B. performed CA activity analyses. S.X. performed patch clamp experiments. A.R. contributed to stomatal movement and stomatal index measurements. J.G. performed norflurazon experiments. J.M.K. analysed CO2/HCO3−-binding protein-encoding gene expression patterns and isolated the initial CA, PEPC and Rubisco T-DNA insertion lines. J.I.S., H.H., A.B.-D. and M.I.-N. wrote the paper.

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Correspondence to Julian I. Schroeder.

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The University of California, San Diego, has submitted a patent form on behalf of J.I.S., M.I-N., J.M.K., H.H. and A.B.D. on aspects of the findings.

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Hu, H., Boisson-Dernier, A., Israelsson-Nordström, M. et al. Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells. Nat Cell Biol 12, 87–93 (2010). https://doi.org/10.1038/ncb2009

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