Abstract
Perception of biotic and abiotic stresses often leads to stomatal closure in plants1,2. Rapid influx of calcium ions (Ca2+) across the plasma membrane has an important role in this response, but the identity of the Ca2+ channels involved has remained elusive3,4. Here we report that the Arabidopsis thaliana Ca2+-permeable channel OSCA1.3 controls stomatal closure during immune signalling. OSCA1.3 is rapidly phosphorylated upon perception of pathogen-associated molecular patterns (PAMPs). Biochemical and quantitative phosphoproteomics analyses reveal that the immune receptor-associated cytosolic kinase BIK1 interacts with and phosphorylates the N-terminal cytosolic loop of OSCA1.3 within minutes of treatment with the peptidic PAMP flg22, which is derived from bacterial flagellin. Genetic and electrophysiological data reveal that OSCA1.3 is permeable to Ca2+, and that BIK1-mediated phosphorylation on its N terminus increases this channel activity. Notably, OSCA1.3 and its phosphorylation by BIK1 are critical for stomatal closure during immune signalling, and OSCA1.3 does not regulate stomatal closure upon perception of abscisic acid—a plant hormone associated with abiotic stresses. This study thus identifies a plant Ca2+ channel and its activation mechanisms underlying stomatal closure during immune signalling, and suggests specificity in Ca2+ influx mechanisms in response to different stresses.
This is a preview of subscription content, access via your institution
Access options
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
Blot source images are presented in Supplementary Fig. 1. Identifiers for publicly available Arabidopsis lines are provided in Methods. Raw data and a detailed description of the analysis presented in Fig. 4a have been deposited on GitHub: https://github.com/TeamMacLean/peak_analysis. All SRM assay information and raw data have been deposited to the Panorama Skyline server and can be accessed via https://panoramaweb.org/Vzao3P.url. Source data are provided with this paper.
Code availability
All codes used for the wavelet analysis are available at https://github.com/TeamMacLean/peak_analysis.
Change history
17 November 2020
A Correction to this paper has been published: https://doi.org/10.1038/s41586-020-2954-9
References
Melotto, M., Zhang, L., Oblessuc, P. R. & He, S. Y. Stomatal defense a decade later. Plant Physiol. 174, 561–571 (2017).
Sussmilch, F. C., Schultz, J., Hedrich, R. & Roelfsema, M. R. G. Acquiring control: the evolution of stomatal signalling pathways. Trends Plant Sci. 24, 342–351 (2019).
Hedrich, R. Ion channels in plants. Physiol. Rev. 92, 1777–1811 (2012).
Jezek, M. & Blatt, M. R. The membrane transport system of the guard cell and its integration for stomatal dynamics. Plant Physiol. 174, 487–519 (2017).
Berridge, M. J., Lipp, P. & Bootman, M. D. The versatility and universality of calcium signalling. Nat. Rev. Mol. Cell Biol. 1, 11–21 (2000).
Dodd, A. N., Kudla, J. & Sanders, D. The language of calcium signaling. Annu. Rev. Plant Biol. 61, 593–620 (2010).
Kudla, J. et al. Advances and current challenges in calcium signaling. New Phytol. 218, 414–431 (2018).
Liang, X. & Zhou, J.-M. Receptor-like cytoplasmic kinases: central players in plant receptor kinase–mediated signaling. Annu. Rev. Plant Biol. 69, 267–299 (2018).
Yu, X., Feng, B., He, P. & Shan, L. From chaos to harmony: responses and signaling upon microbial pattern recognition. Annu. Rev. Phytopathol. 55, 109–137 (2017).
Kadota, Y. et al. Direct regulation of the NADPH oxidase RBOHD by the PRR-associated kinase BIK1 during plant immunity. Mol. Cell 54, 43–55 (2014).
Li, L. et al. The FLS2-associated kinase BIK1 directly phosphorylates the NADPH oxidase RbohD to control plant immunity. Cell Host Microbe 15, 329–338 (2014).
Ranf, S. et al. Microbe-associated molecular pattern-induced calcium signaling requires the receptor-like cytoplasmic kinases, PBL1 and BIK1. BMC Plant Biol. 14, 374 (2014).
Monaghan, J., Matschi, S., Romeis, T. & Zipfel, C. The calcium-dependent protein kinase CPK28 negatively regulates the BIK1-mediated PAMP-induced calcium burst. Plant Signal. Behav. 10, e1018497 (2015).
Tian, W. et al. A calmodulin-gated calcium channel links pathogen patterns to plant immunity. Nature 572, 131–135 (2019).
Yuan, F. et al. OSCA1 mediates osmotic-stress-evoked Ca2+ increases vital for osmosensing in Arabidopsis. Nature 514, 367–371 (2014).
Hou, C. et al. DUF221 proteins are a family of osmosensitive calcium-permeable cation channels conserved across eukaryotes. Cell Res. 24, 632–635 (2014).
Li, Y. et al. Genome-wide survey and expression analysis of the OSCA gene family in rice. BMC Plant Biol. 15, 261 (2015).
Ganie, S. A., Pani, D. R. & Mondal, T. K. Genome-wide analysis of DUF221 domain-containing gene family in Oryza species and identification of its salinity stress-responsive members in rice. PLoS ONE 12, e0182469 (2017).
Ding, S., Feng, X., Du, H. & Wang, H. Genome-wide analysis of maize OSCA family members and their involvement in drought stress. PeerJ 7, e6765 (2019).
Benschop, J. J. et al. Quantitative phosphoproteomics of early elicitor signaling in Arabidopsis. Mol. Cell. Proteomics 6, 1198–1214 (2007).
Gómez-Gómez, L. & Boller, T. FLS2: an LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Mol. Cell 5, 1003–1011 (2000).
Zhang, J. et al. Receptor-like cytoplasmic kinases integrate signaling from multiple plant immune receptors and are targeted by a Pseudomonas syringae effector. Cell Host Microbe 7, 290–301 (2010).
Lu, D. et al. A receptor-like cytoplasmic kinase, BIK1, associates with a flagellin receptor complex to initiate plant innate immunity. Proc. Natl Acad. Sci. USA 107, 496–501 (2010).
Murthy, S. E. et al. OSCA/TMEM63 are an evolutionarily conserved family of mechanically activated ion channels. eLife 7, e41844 (2018).
Zhang, M. et al. Structure of the mechanosensitive OSCA channels. Nat. Struct. Mol. Biol. 25, 850–858 (2018).
Jojoa-Cruz, S. et al. Cryo-EM structure of the mechanically activated ion channel OSCA1.2. eLife 7, e41845 (2018).
Liu, X., Wang, J. & Sun, L. Structure of the hyperosmolality-gated calcium-permeable channel OSCA1.2. Nat. Commun. 9, 5060 (2018).
Maity, K. et al. Cryo-EM structure of OSCA1.2 from Oryza sativa elucidates the mechanical basis of potential membrane hyperosmolality gating. Proc. Natl Acad. Sci. USA 116, 14309–14318 (2019).
Iida, H., Nakamura, H., Ono, T., Okumura, M. S. & Anraku, Y. MID1, a novel Saccharomyces cerevisiae gene encoding a plasma membrane protein, is required for Ca2+ influx and mating. Mol. Cell. Biol. 14, 8259–8271 (1994).
Knight, M. R., Campbell, A. K., Smith, S. M. & Trewavas, A. J. Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. Nature 352, 524–526 (1991).
Thor, K. & Peiter, E. Cytosolic calcium signals elicited by the pathogen-associated molecular pattern flg22 in stomatal guard cells are of an oscillatory nature. New Phytol. 204, 873–881 (2014).
Montillet, J.-L. et al. An abscisic acid-independent oxylipin pathway controls stomatal closure and immune defense in Arabidopsis. PLoS Biol. 11, e1001513 (2013).
Liu, Y. et al. Anion channel SLAH3 is a regulatory target of chitin receptor-associated kinase PBL27 in microbial stomatal closure. eLife 8, e44474 (2019).
Wang, J. et al. A cyclic nucleotide-gated channel mediates cytoplasmic calcium elevation and disease resistance in rice. Cell Res. 29, 820–831 (2019).
Kwaaitaal, M., Huisman, R., Maintz, J., Reinstädler, A. & Panstruga, R. Ionotropic glutamate receptor (iGluR)-like channels mediate MAMP-induced calcium influx in Arabidopsis thaliana. Biochem. J. 440, 355–373 (2011).
Ma, Y., Walker, R. K., Zhao, Y. & Berkowitz, G. A. Linking ligand perception by PEPR pattern recognition receptors to cytosolic Ca2+ elevation and downstream immune signaling in plants. Proc. Natl Acad. Sci. USA 109, 19852–19857 (2012).
Espinoza, C., Liang, Y. & Stacey, G. Chitin receptor CERK1 links salt stress and chitin-triggered innate immunity in Arabidopsis. Plant J. 89, 984–995 (2017).
Meena, M. K. et al. The Ca2+ channel CNGC19 regulates Arabidopsis defense against Spodoptera herbivory. Plant Cell 31, 1539–1562 (2019).
Schwessinger, B. et al. Phosphorylation-dependent differential regulation of plant growth, cell death, and innate immunity by the regulatory receptor-like kinase BAK1. PLoS Genet. 7, e1002046 (2011).
Cutler, S. R., Ehrhardt, D. W., Griffitts, J. S. & Somerville, C. R. Random GFP:cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency. Proc. Natl Acad. Sci. USA 97, 3718–3723 (2000).
Waterhouse, A. et al. SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res. 46 (W1), W296–W303 (2018).
Söding, J., Biegert, A. & Lupas, A. N. The HHpred interactive server for protein homology detection and structure prediction. Nucleic Acids Res. 33, W244–W248 (2005).
Pettersen, E. F. et al. UCSF Chimera—a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).
Monaghan, J. et al. The calcium-dependent protein kinase CPK28 buffers plant immunity and regulates BIK1 turnover. Cell Host Microbe 16, 605–615 (2014).
Nakagawa, T. et al. Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. J. Biosci. Bioeng. 104, 34–41 (2007).
Bender, K. W. et al. Autophosphorylation-based calcium (Ca2+) sensitivity priming and Ca2+/calmodulin inhibition of Arabidopsis thaliana Ca2+-dependent protein kinase 28 (CPK28). J. Biol. Chem. 292, 3988–4002 (2017).
MacLean, B. et al. Skyline: an open source document editor for creating and analyzing targeted proteomics experiments. Bioinformatics 26, 966–968 (2010).
Charpentier, M. et al. Nuclear-localized cyclic nucleotide-gated channels mediate symbiotic calcium oscillations. Science 352, 1102–1105 (2016).
Fischer, M. et al. The Saccharomyces cerevisiae CCH1 gene is involved in calcium influx and mating. FEBS Lett. 419, 259–262 (1997).
Gietz, R. D. & Woods, R. A. Genetic transformation of yeast. Biotechniques 30, 816–831 (2001).
Reyes, R. et al. Cloning and expression of a novel pH-sensitive two pore domain K+ channel from human kidney. J. Biol. Chem. 273, 30863–30869 (1998).
Jurman, M. E., Boland, L. M., Liu, Y. & Yellen, G. Visual identification of individual transfected cells for electrophysiology using antibody-coated beads. Biotechniques 17, 876–881 (1994).
Hamill, O. P., Marty, A., Neher, E., Sakmann, B. & Sigworth, F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 391, 85–100 (1981).
Moyen, C., Hammond-Kosack, K. E., Jones, J., Knight, M. R. & Johannes, E. Systemin triggers an increase of cytoplasmic calcium in tomato mesophyll cells: Ca2+ mobilization from intra- and extracellular compartments. Plant Cell Environ. 21, 1101–1111 (1998).
Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
Miwa, H., Sun, J., Oldroyd, G. E. D. & Downie, J. A. Analysis of calcium spiking using a cameleon calcium sensor reveals that nodulation gene expression is regulated by calcium spike number and the developmental status of the cell. Plant J. 48, 883–894 (2006).
Böhm, J. et al. Understanding the molecular basis of salt sequestration in epidermal bladder cells of Chenopodium quinoa. Curr. Biol. 28, 3075–3085.e7 (2018).
Dindas, J. et al. AUX1-mediated root hair auxin influx governs SCFTIR1/AFB-type Ca2+ signaling. Nat. Commun. 9, 1174 (2018).
Newman, I. A. Ion transport in roots: measurement of fluxes using ion-selective microelectrodes to characterize transporter function. Plant Cell Environ. 24, 1–14 (2001).
Arif, I., Newman, I. A. & Keenlyside, N. Proton flux measurements from tissues in buffered solution. Plant Cell Environ. 18, 1319–1324 (1995).
Müller, H. M. et al. The desert plant Phoenix dactylifera closes stomata via nitrate-regulated SLAC1 anion channel. New Phytol. 216, 150–162 (2017).
Acknowledgements
We thank H. Krutinová and S. Vanneste for assistance in early stages of this project; J. P. Kukkonen for assistance for setting up the HEK293T cell assays; J. Sun for assistance with Ca2+ measurements in guard cells; B. Brandt for help with structural modelling of OSCA1.3; P. He and E. Peiter for providing published materials; J. –M. Zhou for early strategic discussions on this project and for providing published materials; M. Smoker, J. Taylor and J. Lopez from the TSL Plant Transformation support group for plant transformation; the John Innes Centre Horticultural Services for plant care; and all past and current members of the Zipfel group for technical help and fruitful discussions. This work was supported by the European Research Council under the Grant Agreements No. 309858 and 773153 (grants ‘PHOSPHinnATE’ and ‘IMMUNO-PEPTALK’ to C.Z.), The Gatsby Charitable Foundation (to C.Z.), the University of Zürich (to C.Z.), and the Swiss National Science Foundation (grant 31003A_182625 to C.Z.). The Biotechnology and Biological Research Council supported C.Z. and G.E.D.O. with BB/P012574/1. S.J., J.D., T.A.D. and J. Gronnier were supported by post-doctoral fellowships from the European Molecular Biology Organization (EMBO-LTF no. 225-2015; EMBO-LTF no. 683-2018; EMBO-LTF no. 100-2017 and EMBO-LTF no. 438-2018, respectively). Y.K. was supported by JSPS KAKENHI Grant Numbers JP16H06186 and JP16KT0037. Work in the J.F. laboratory was supported by the National Institutes of Health (NIH R01 GM131043), the National Science Foundation (MCB1616437/2016 and MCB1930165/2019) and the University of Maryland. Work in the M.W. laboratory was supported by the Academy of Finland (grant numbers 275632, 283139 and 312498). R.H. and M.R.G.R. were supported by the German Research Foundation (DFG, HE 1640/34-1; HE 1640/40-1; RO2381/6-1 and RO2381/8-1).
Author information
Authors and Affiliations
Contributions
C.Z. designed and supervised the project, and obtained funding. K.T. and S.J. conceived, designed and performed the majority of the plant and biochemical experiments. E.M. and J.F. provided the patch-clamp data in COS-7 cells; J. George performed some of the genetic and phenotypic characterization of the osca1.3/1.7 mutant. P.D. and F.L.H.M. performed the SRM assays. N.L., M.C. and G.E.D.O. provided the yeast complementation assays. K.H. and M.W. provided the HEK cell data. T.A.D. and J.D. performed aequorin and YC3.6 measurements in leaf discs. P.K. and J. Gronnier generated expression constructs for OSCA1.7. L.S. assisted with the genetic characterization of the mutants. Y.K. provided initial data on the BIK1–OSCA1.3 interaction. C.A.B. provided OSCA1.3 localization data. S.S., S.H., M.R.G.R. and R.H. assisted with initial electrophysiological characterization, conducted ion flux measurements and carried out gas-exchange recordings. D.M. analysed the guard-cell Ca2+ measurements. K.T. and C.Z. wrote the manuscript. All authors commented and agreed on the manuscript before submission.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Peer review information Nature thanks Thorsten Nürnberger, Yumou Qiu, Keiko Yoshioka and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended data figures and tables
Extended Data Fig. 1 Predicted topology of OSCA1.3 with possible BIK1 phosphorylation sites and multiple alignment of loop 1 from Clade 1 OSCA proteins.
a, Topology was visualized using Protter (www.wlab.ethz.ch/protter) version 1.0 on the basis of information from ref. 26. Blue numbers indicate transmembrane regions. Possible BIK1 phosphorylation sites are highlighted in red. b, Protein sequence alignment of OSCA1.1 to OSCA1.8 showing amino acids 30 to 95. Clustal Omega alignments were visualized with Jalview 2.10.5. Possible BIK1 phosphorylation motifs (SxxL/I) are highlighted in red. Blue colour denotes % identity. c, Structural model for OSCA1.3. Arrows indicate the position of S54 located in the cytosolic loop.
Extended Data Fig. 2 OSCA1.3 localizes to the plasma membrane.
Confocal microscopy of osca1.3 cotyledons expressing OSCA1.3-GFP under the control of the OSCA1.3 promoter. Right Panel: Plasmolysis with 2 M NaCl underlines plasma membrane localization. Green: GFP; magenta: chlorophyll autofluorescence. The experiment was performed once.
Extended Data Fig. 3 PBL1 also phosphorylates OSCA1.3.
Differences in PBL1-mediated incorporation of radioactive phosphate in OSCA1.3 and its mutation variants. In vitro kinase assay performed with the corresponding recombinant proteins. For blot source data, see Supplementary Fig. 1. The experiment was performed twice with similar results.
Extended Data Fig. 4 OSCA1.3 promotes calcium influx in HEK cells.
HEK293T cells loaded with the calcium indicator Fura-2 and transfected with OSCA1.3-myc show an increase in fluorescence intensity ratio at 340/380 nm excitation compared to non-transfected cells after addition of sorbitol and calcium to the culture medium, indicating an increase in calcium influx. Data show mean ± s.d. (n = 4 technical replicates). Similar results were obtained in 3 independent biological repeats.
Extended Data Fig. 5 OSCA1.3 and OSCA1.7 are BIK1-activated calcium-permeable channels.
a, Typical currents (left panel) and corresponding I/V curves (right panel) recorded in OSCA1.3 plus BIK1 expressing COS-7 cells increase with increasing calcium concentrations as indicated on the figure legend (n = 3 cells, mean ± s.e.m.). Currents were normalized with current intensities recorded at -100 mV in the standard bath solution (5 mM calcium), and consequently expressed in normalized arbitrary units for easier comparison of reverse potential changes. Note the inward currents increase and the reverse potentials shift to positive values when extracellular calcium concentration increases, indicating a calcium permeation of the channel. See methods for solutions composition. b, Typical traces (left panel) and corresponding statistical analysis (right panel) of currents recorded in whole-cell configuration in COS-7 cells co-transfected with pCI-OSCA1.7 plus pCI-BIK1 (n = 17 cells, mean ± s.e.m.) or plus pCI-BIK1(KD) (n = 9 cells, mean ± s.e.m.) as indicated on the figure legend. OSCA1.7 is a BIK1-activated channel. I/V curves recorded on cells. c, BIK1 kinase activity activates currents in cells expressing both OSCA1.3 and OSCA1.7. Typical currents (left panel) and corresponding I/V curves (right panel) recorded in cells co-transfected with both pCI-OSCA1.3 and pCI-OSCA1.7 plus pCI-BIK1 (n = 10 cells, mean ± s.e.m.) or plus pCI-BIK1(KD) (n = 9 cells, mean ± s.e.m.) as indicated on the figure legend. Note that current intensities are not higher than current intensities recorded in cells expressing either OSCA1.3+BIK1 (Fig. 3b, c) or OSCA1.7+BIK1 (a), giving no indication on functional heteromerization of OSCA1.3 and OSCA1.7. Whole-cell patch clamp protocols used in b and c were identical to the one used in Fig. 3b, c.
Extended Data Fig. 6 T-DNA insertion lines used in this study and transcript levels.
a, Gene structure of OSCA1.3 and OSCA1.7 showing exons (black boxes) and introns (lines) as well as location of T-DNA insertions. Line osca1.3/1.7 was obtained by crossing osca1.3 and osca1.7. Arrows denote location of primers used for genotyping. b, Transcript levels of OSCA1.3 and OSCA1.7 in Col-0, osca1.3, osca1.7 and osca1.3/1.7 as determined by quantitative real-time PCR with reverse transcription. Values are mean +/− s.d. (n = 6, representing 2 independent experiments with 3 biological repeats each). c, Transcript levels of OSCA1.3 in Col-0, osca1.3/1.7 and osca1.3/1.7 complemented with OSCA1.3(WT) or OSCA1.3(S54A), respectively. Expression levels for three independent T1 plants corresponding to Fig. 4f are shown separately, with two technical replicates (leaves). This experiment was repeated three times. Shown are quantitative real-time RT–PCR data relative to U-box (At5g15400). Primers used in b and c are listed in Supplementary Table 2.
Extended Data Fig. 7 Expression pattern of OSCA genes from Clade 1.
Tissue-specific expression patterns were obtained from Genevestigator (www.genevestigator.com). OSCA1.3 shows high expression levels in guard cells and guard cell protoplasts.
Extended Data Fig. 8 Flg22-induced calcium influx measured in leaf discs is comparable between wild-type and osca1.3/1.7 plants.
a, Calcium influx in leaf discs taken of Col-0 and osca1.3/1.7 plants expressing the calcium reporter aequorin. flg22 was added at time point 10 min. Error bars represent mean ± s.d. (n = 12 leaf discs from 6 independent plants). The experiment was performed twice with similar results. b, Average values of FRET ratio changes in leaf discs of Col-0 and osca1.3/1.7 expressing the ratiometric calcium reporter YC3.6 obtained in plate reader-based assays. Error bars show s.e.m., n = 90 leaf discs (Col-0) and 47 leaf discs (osca1.3/1.7), with 6 leaf discs taken per individual plant. The experiment was performed twice with similar results.
Extended Data Fig. 9 Flg22-induced calcium fluxes in osca1.3/1.7 guard cells are reduced compared to wild-type guard cells.
a, Typical flg22-induced spiking patterns and their distribution in Col-0 and osca1.3/1.7 guard cells. Legends show ratio changes of the Yellow Cameleon 3.6 calcium reporter observed over time (flg22 added at time point 10 min, indicated by an arrow). The pattern of every cell (n = 64 for wild-type and n = 61 for osca1.3/1.7) was assigned to one of the categories based on visual assessment. b, Left panel, net calcium fluxes of a representative Col-0 and osca1.3/1.7 guard cell, respectively, measured using Scanning Ion Selective Electrodes (SISE). Right panel, integrated calcium fluxes over 7 min after addition of flg22 are reduced in osca1.3/1.7 compared to Col-0 (n = 29 cells for Col-0, n = 23 cells for osca1.3/1.7; error bars represent mean ± s.e.m. bootstrapped Welch two sample t-test, two-sided P = 0.0464.). c, Left panel, flg22-induced calcium fluxes are blocked by lanthanum. Representative calcium fluxes measured using Scanning Ion Selective Electrodes (SISE) of Col-0 guard cells with or without lanthanum pre-treatment (1 mM lanthanum applied 10 min before flg22 treatment). One micromolar flg22 was added at time point 0 to epidermal strips. Right panel, integrated calcium fluxes over 8 min after addition of flg22 are significantly blocked by lanthanum in Col-0 (n = 8 cells without lanthanum and n = 5 cells with lanthanum; error bars represent mean ± s.e.m.; bootstrapped Welch two sample t-test, two-sided P = 0.0026).
Extended Data Fig. 10 AtPep1-induced decrease in stomatal conductance is impaired in osca1.3/1.7.
Leaf transpiration was recorded in excised intact leaves. AtPep1 was added to the solution at the petioles to a concentration of 3 μM, water was used as control. Data show mean ± s.e.m. (Col-0 mock, Col-0 AtPep1, osca1.3/1.7 AtPep1: n = 8; osca1.3/1.7 mock: n = 11 leaves).
Supplementary information
Supplementary Information
This file contains Supplementary Figure 1: Source data for images for gels and blots. Original source images for all data obtained by SDS-PAGE, western blots, autoradiography scans and Coomassie Blue stained blots and gels; Supplementary Table 1: Specific transitions used for selected reaction monitoring (SRM) with OSCA1.3 and control peptide; Supplementary Table 2: Primers used in this study.
Rights and permissions
About this article
Cite this article
Thor, K., Jiang, S., Michard, E. et al. The calcium-permeable channel OSCA1.3 regulates plant stomatal immunity. Nature 585, 569–573 (2020). https://doi.org/10.1038/s41586-020-2702-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41586-020-2702-1
This article is cited by
-
Mechanisms of calcium homeostasis orchestrate plant growth and immunity
Nature (2024)
-
Genome-wide identification and expression analysis of the cyclic nucleotide-gated ion channel (CNGC) gene family in Saccharum spontaneum
BMC Genomics (2023)
-
Large-scale phosphoproteome analysis in wheat seedling leaves provides evidence for extensive phosphorylation of regulatory proteins during CWMV infection
BMC Plant Biology (2023)
-
RALF signaling pathway activates MLO calcium channels to maintain pollen tube integrity
Cell Research (2023)
-
BIK1 protein homeostasis is maintained by the interplay of different ubiquitin ligases in immune signaling
Nature Communications (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
