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Calcium-dependent protein kinase CPK9 negatively functions in stomatal abscisic acid signaling by regulating ion channel activity in Arabidopsis

  • Dong-Hua Chen
  • Hui-Ping Liu
  • Chun-Long Li
Article
  • 28 Downloads

Abstract

Key message

In this manuscript, we demonstrated the negative role of CPK9 in stomatal ABA signaling, and both CPK9 and CPK33 for accurate guard cell function was explored via cpk9/cpk33 double mutants’ phenotype.

Abstract

Abscisic acid (ABA) can inhibit stomatal opening and promote stomatal closure by regulating ion channel activity in guard cell membranes. As an important second messenger, calcium (Ca2+) is essentially needed in ABA regulation of stomatal movement. Calcium-dependent protein kinases (CDPKs) have been proposed to contribute to central Ca2+ signal transduction in plants. Here, we report the functional characterization of CPK9 in Arabidopsis stomatal ABA signaling. CPK9 had high expression in guard cells and the protein was subcellularly located in the cell membrane. A loss-of-function mutant cpk9 showed a much more sensitive phenotype to ABA regulation of stomatal movement and ion channel activity, while CPK9 overexpression lines had opposite phonotypes. These findings demonstrated the negative role of CPK9 in stomatal ABA signaling. As the closest homolog of CPK33, we also proved that stomatal movement of the cpk9/cpk33 double mutants was more sensitive to ABA than either single mutants. These results revealed the role of CPK9 in guard cells, and the need of both CPK9 and CPK33 for accurate guard cell function.

Keywords

CPK9 Stomatal movement Guard cell Abscisic acid Ion channel 

Notes

Acknowledgements

This work was supported by National Natural Science Foundation of China (31500211), the START-UP Fund from Jiangsu Normal University (17XLR039) and the Science Fund for Distinguished Young Scholars from Shandong University.

Author contributions

CL designed the experiments; CL, DC, HL performed the experiments, analyzed and discussed the results; CL and DC wrote the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11103_2018_805_MOESM1_ESM.docx (415 kb)
Supplementary material 1 (DOCX 414 KB)
11103_2018_805_MOESM2_ESM.docx (24 kb)
Supplementary Table 1 (DOCX 23 KB)

References

  1. Acharya BR, Jeon BW, Zhang W, Assmann SM (2013) Open stomata 1 (OST1) is limiting in abscisic acid responses of Arabidopsis guard cells. New Phytol 200:1049–1063CrossRefPubMedGoogle Scholar
  2. Almadanim MC, Alexandre BM, Rosa MT, Sapeta H, Leitão AE, Ramalho JC, Lam TT, Negrão S, Abreu IA, Oliveira MM (2017) Rice calcium-dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose-phosphate synthase and is required for a proper cold stress response. Plant Cell Environ 40:1197–1213CrossRefPubMedGoogle Scholar
  3. Asano T, Kunieda N, Omura Y, Ibe H, Kawasaki T, Takano M, Sato M, Furuhashi H, Mujin T, Takaiwa F, Wu Cy CY, Tada Y, Satozawa T, Sakamoto M, Shimada H (2002) Rice SPK, a calmodulin-like domain protein kinase, is required for storage product accumulation during seed development: phosphorylation of sucrose synthase is a possible factor. Plant Cell 14:619–628CrossRefPubMedPubMedCentralGoogle Scholar
  4. Asano T, Tanaka N, Yang G, Hayashi N, Komatsu S (2005) Genome-wide identification of the rice calcium-dependent protein kinase and its closely related kinase gene families: comprehensive analysis of the CDPKs gene family in rice. Plant Cell Physiol 46:356–366CrossRefPubMedGoogle Scholar
  5. Asano T, Hayashi N, Kobayashi M, Aoki N, Miyao A, Mitsuhara I, Ichikawa H, Komatsu S, Hirochika H, Kikuchi S (2012) A rice calcium-dependent protein kinase OsCPK12 oppositely modulates salt-stress tolerance and blast disease resistance. Plant J 69:26–36CrossRefPubMedGoogle Scholar
  6. Benetka W, Mehlmer N, Maurer-Stroh S, Sammer M, Koranda M, Neumüller R, Betschinger J, Knoblich JA, Teige M, Eisenhaber F (2008) Experimental testing of predicted myristoylation targets involved in asymmetric cell division and calcium-dependent signalling. Cell Cycle 7:3709–3719CrossRefPubMedGoogle Scholar
  7. Blatt MR, Armstrong F (1993) K+ channels of stomatal guard cells: abscisic acid-evoked control of the outward rectifier mediated by cytoplasmic pH. Planta 191:330–341CrossRefGoogle Scholar
  8. Boudsocq M, Sheen J (2013) CDPKs in immune and stress signaling. Trends Plant Sci 18:30–40CrossRefPubMedGoogle Scholar
  9. Cheng SH, Willmann MR, Chen HC, Sheen J (2002) Calcium signaling through protein kinases. The Arabidopsis calcium-dependent protein kinase gene family. Plant Physiol 129:469–485CrossRefPubMedPubMedCentralGoogle Scholar
  10. Choi HI, Park HJ, Park JH, Kim S, Im MY, Seo HH, Kim YW, Hwang I, Kim SY (2005) Arabidopsis calcium-dependent protein kinase AtCPK32 interacts with ABF4, a transcriptional regulator of abscisic acid-responsive gene expression, and modulates its activity. Plant Physiol 139:1750–1761CrossRefPubMedPubMedCentralGoogle Scholar
  11. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefGoogle Scholar
  12. Corratgé-Faillie C, Ronzier E, Sanchez F, Prado K, Kim JH, Lanciano S, Leonhardt N, Lacombe B, Xiong TC (2017) The Arabidopsis guard cell outward potassium channel GORK is regulated by CPK33. FEBS Lett 591:1982–1992CrossRefPubMedGoogle Scholar
  13. Cousson A, Vavasseur A (1998) Putative involvement of cytosolic Ca2+ and GTP-binding proteins in cyclic-GMP-mediated induction of stomatal opening by auxin in Commelina communis L. Planta 206:308–314CrossRefGoogle Scholar
  14. Dammann C, Ichida A, Hong B, Romanowsky SM, Hrabak EM, Harmon AC, Pickard BG, Harper JF (2003) Subcellular targeting of nine calcium-dependent protein kinase isoforms from Arabidopsis. Plant Physiol 132:1840–1848CrossRefPubMedPubMedCentralGoogle Scholar
  15. Dubiella U, Seybold H, Durian G, Komander E, Lassig R, Witte C, Schulze WX, Romeis T (2013) Calcium-dependent protein kinase/NADPH oxidase activation circuit is required for rapid defense signal propagation. Proc Natl Acad Sci USA 110:8744–8749CrossRefPubMedGoogle Scholar
  16. Franz S, Ehlert B, Liese A, Kurth J, Cazale A, Romeis T (2011) Calcium-Dependent protein kinase CPK21 functions in abiotic stress response in Arabidopsis thaliana. Mol Plant 4:83–96CrossRefGoogle Scholar
  17. Gambale F, Uozumi N (2006) Properties of shaker-type potassium channels in higher plants. J Membr Biol 210:1–19CrossRefPubMedGoogle Scholar
  18. Hrabak EM, Chan CWM, Gribskov M, Harper JF, Choi JH, Halford NG, Kudla J, Luan S, Nimmo HG, Sussman MR (2003) The Arabidopsis CDPK-SnRK superfamily of protein kinases. Plant Physiol 132:666–680CrossRefPubMedPubMedCentralGoogle Scholar
  19. Jiang S, Zhang D, Wang L, Pan J, Liu Y, Kong X, Zhou Y, Li D (2013) A maize calcium-dependent protein kinase gene, ZmCPK4, positively regulated abscisic acid signaling and enhanced drought stress tolerance in transgenic Arabidopsis. Plant Physiol Biochem 71:112–120CrossRefPubMedPubMedCentralGoogle Scholar
  20. Johnson DR, Bhatnagar RS, Knoll LJ, Gordon JI (1994) Genetic and biochemical studies of protein N-myristoylation. Annu Rev Biochem 63:869–914CrossRefPubMedGoogle Scholar
  21. Lee SC, Lim CW, Lan W, He K, Luan S (2013) ABA signaling in guard cells entails a dynamic protein–protein interaction relay from the PYL-RCAR family receptors to ion channels. Mol Plant 6:528–538CrossRefPubMedGoogle Scholar
  22. Li J, Wang X, Watson MB, Assmann SM (2000) Regulation of abscisic acid-induced stomatal closure and anion channels by guard cell AAPK Kinase. Science 287:300–303CrossRefPubMedGoogle Scholar
  23. Li CL, Wang M, Ma XY, Zhang W (2014) NRGA1, a putative mitochondrial pyruvate carrier, mediates ABA regulation of guard cell ion channels and drought stress responses in Arabidopsis. Mol Plant 7:1508–1521CrossRefPubMedGoogle Scholar
  24. Li CL, Wang M, Wu XM, Chen DH, Lv HJ, Shen JL, Qiao Z, Zhang W (2016) THI1, a thiamine thiazole synthase, interacts with Ca2+-dependent protein kinase CPK33 and modulates the S-type anion channels and stomatal closure in Arabidopsis. Plant Physiol 170:1090–1104CrossRefPubMedGoogle Scholar
  25. Liu K-h, Niu Y, Konishi M, Wu Y, Du H, Chung HS, Li L, Boudsocq M, McCormack M, Maekawa S (2017) Discovery of nitrate–CPK–NLP signalling in central nutrient–growth networks. Nature 545:311CrossRefPubMedPubMedCentralGoogle Scholar
  26. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2– ∆∆Ct method. Methods 25:402–408CrossRefGoogle Scholar
  27. Ma S, Wu W (2007) AtCPK23 functions in Arabidopsis responses to drought and salt stresses. Plant Mol Biol 65:511–518CrossRefPubMedGoogle Scholar
  28. McAinsh MR, Webb AAR, Taylor JE, Hetherington AM (1995) Stimulus-induced oscillations in guard cell cytosolic free calcium. Plant Cell 7:1207–1219CrossRefPubMedPubMedCentralGoogle Scholar
  29. Mehlmer N, Wurzinger B, Stael S, Hofmann-Rodrigues D, Csaszar E, Pfister B, Bayer R, Teige M (2010) The Ca2+-dependent protein kinase CPK3 is required for MAPK-independent salt-stress acclimation in Arabidopsis. Plant J 63:484–498CrossRefPubMedPubMedCentralGoogle Scholar
  30. Mori IC, Murata Y, Yang Y, Munemasa S, Wang Y, Andreoli S, Tiriac H, Alonso JM, Harper JF, Ecker JR (2006) CDPKs CPK6 and CPK3 function in ABA regulation of guard cell S-type anion- and Ca2+-permeable channels and stomatal closure. PLoS Biol 4:1749–1762CrossRefGoogle Scholar
  31. Oliver B, Jorg K (2012) Analysis of calcium signaling pathways in plants. Biochim Biophys Acta 1820:1283–1293CrossRefGoogle Scholar
  32. Pandey S, Zhang W, Assmann SM (2007) Roles of ion channels and transporters in guard cell signal transduction. FEBS Lett 581:2325–2336CrossRefPubMedGoogle Scholar
  33. Patharkar OR, Cushman JC (2000) A stress-induced calcium-dependent protein kinase from Mesembryanthemum crystallinum phosphorylates a two-component pseudo-response regulator. Plant J 24:679–691CrossRefPubMedGoogle Scholar
  34. Pei ZM, Kuchitsu K, Ward JM, Schwarz M, Schroeder JI (1997) Differential abscisic acid regulation of guard cell slow anion channels in Arabidopsis wild-type and abi1 and abi2 mutants. Plant Cell 9:409–423CrossRefPubMedPubMedCentralGoogle Scholar
  35. Schroeder JI, Raschke K, Neher E (1987) Voltage dependence of K+ channels in guard-cell protoplasts. Proc Natl Acad Sci USA 84:4108–4112CrossRefPubMedGoogle Scholar
  36. Schroeder JI, Allen GJ, Hugouvieux V, Kwak JM, Waner D (2001) Guard cell signal transduction. Annu Rev Plant Physiol Plant Mol Biol 52:627–658CrossRefPubMedGoogle Scholar
  37. Schwartz A, Wu WH, Tucker EB, Assmann SM (1994) Inhibition of inward K+ channels and stomatal response by abscisic acid: an intracellular locus of phytohormone action. Proc Natl Acad Sci USA 91:4019–4023CrossRefPubMedGoogle Scholar
  38. Sheen J (1996) Ca2+-dependent protein kinases and stress signal transduction in plants. Science 274:1900–1902CrossRefPubMedGoogle Scholar
  39. Shi S, Li S, Asim M, Mao J, Xu D, Ullah Z, Liu G, Wang Q, Liu H (2018) The Arabidopsis calcium-dependent protein kinases (CDPKs) and their roles in plant growth regulation and abiotic stress responses. Int J Mol Sci 19:1900–1918CrossRefPubMedCentralGoogle Scholar
  40. Very AA, Sentenac H (2003) Molecular mechanisms and regulation of K+ transport in higher plants. Annu Rev Plant Biol 54:575–603CrossRefPubMedGoogle Scholar
  41. Wang XQ, Ullah H, Jones AM, Assmann SM (2001) G protein regulation of ion channels and abscisic acid signaling in Arabidopsis guard cells. Science 292:2070–2072CrossRefPubMedGoogle Scholar
  42. Wang L, Yu C, Xu S, Zhu Y, Huang W (2016) OsDi19-4 acts downstream of OsCDPK14 to positively regulate ABA response in rice. Plant Cell Environ 39:2740–2753CrossRefPubMedGoogle Scholar
  43. Wei S, Hu W, Deng X, Zhang Y, Liu X, Zhao X, Luo Q, Jin Z, Li Y, Zhou S (2014) A rice calcium-dependent protein kinase OsCPK9 positively regulates drought stress tolerance and spikelet fertility. BMC Plant Biol 14:133CrossRefPubMedPubMedCentralGoogle Scholar
  44. Zhang W, Nilson SE, Assmann SM (2008) Isolation and whole-cell patch clamping of Arabidopsis guard cell protoplasts. Cold Spring Harbor Protoc 2008:pdb prot5014CrossRefGoogle Scholar
  45. Zhu SY, Yu XC, Wang XJ, Zhao R, Li Y, Fan RC, Shang Y, Du SY, Wang XF, Wu FQ, Xu YH, Zhang XY, Zhang DP (2007) Two calcium-dependent protein kinases, CPK4 and CPK11, regulate abscisic acid signal transduction in Arabidopsis. Plant Cell 19:3019–3036CrossRefPubMedPubMedCentralGoogle Scholar
  46. Zou J, Wei F, Wang C, Wu J, Ratnasekera D, Liu W, Wu W (2010) Arabidopsis calcium-dependent protein kinase CPK10 functions in abscisic acid- and Ca2+-mediated stomatal regulation in response to drought stress. Plant Physiol 154:1232–1243CrossRefPubMedPubMedCentralGoogle Scholar
  47. Zou J, Li XD, Ratnasekera D, Wang C, Liu WX, Song LF, Zhang WZ, Wu WH (2015) Arabidopsis CALCIUM-DEPENDENT PROTEIN KINASE8 and CATALASE3 function in abscisic acid-mediated signaling and H2O2 homeostasis in stomatal guard cells under drought stress. Plant Cell 27:1445–1460CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education; School of Life ScienceShandong UniversityQingdaoChina
  2. 2.College of Life ScienceJiangsu Normal UniversityXuzhouChina

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