Cytology and Genetics

, Volume 53, Issue 2, pp 99–105 | Cite as

Calcium and Components of Lipid Signaling in Implementation of Hydrogen Sulfide Influence on the State of Stomata in Arabidopsis thaliana

  • T. O. YastrebEmail author
  • Yu. E. KolupaevEmail author
  • E. N. HavvaEmail author
  • M. A. ShkliarevskyiEmail author
  • A. P. DmitrievEmail author


A time and concentration dependence of the hydrogen sulfide donor sodium hydrosulfide (NaHS) influence on the state of stomata of Arabidopsis thaliana (Col-0) leaves, as well as the role of calcium and phospholipases in the implementation of its effects, was studied. Treatment of leaves with NaHS in the concentration range of 5–250 μM caused a decrease in the size of the stomatal aperture. The maximal effect of the stomatal closure was observed 90 min after the beginning of the treatment with H2S donor, and the stomatal aperture in NaHS variants was, on the contrary, much wider than in the control after 180 min of exposure. The effect of treatment with NaHS solutions on the stomata state was completely eliminated by hydroxylamine, the hydrogen sulfide scavenger, which indicates the specificity of NaHS effects as an H2S donor. The decrease in stomatal aperture and relative number of open stomata caused by the donor of hydrogen sulfide was almost completely leveled off by the pretreatment of leaves with the calcium channel blocker lanthanum chloride, the extracellular calcium chelator EGTA, the phospholipase C inhibitor neomycin, and the antagonist of the formation of cyclic adenosine-5'-diphosphate ribose—nicotinamide. In addition, the stomatal effect of theH2S donor was partially eliminated by the calmodulin antagonist chlorpromazine. The leveling of the hydrogen sulfide donor action on the state of stomatal apparatus of Arabidopsis leaves was also noted at the pretreatment of leaves with butanol-1, an inhibitor of phospholipase D-dependent formation of phosphatidic acid. A conclusion about the importance of calcium intake into the cytosol from various compartments, as well as lipid signaling mediators formed with the involvement of phospholipases C and D, in the implementation of hydrogen sulfide action on the state of stomata was made.



The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.


  1. 1.
    Lisjak, M., Teklic, T., Wilson, I.D., Whiteman, M., and Hancock, J.T., Hydrogen sulfide: environmental factor or signalling molecule?, Plant Cell Environ., 2013, vol. 36, no. 9, pp. 1607–1616. CrossRefPubMedGoogle Scholar
  2. 2.
    Zhang, H., Hydrogen sulfide in plant biology, in Signaling and Communication in Plants. Gasotransmitters in Plants, Lamattina, L. and Garcia-Mata, C., Eds., Switzerland: Springer Int. Publ., 2016, pp. 23–51.
  3. 3.
    Jin, Z.P., Shen, J.J., Qiao, Z.J., Yang, G.D., Wang, R., and Pei, Y.X., Hydrogen sulfide improves drought resistance in Arabidopsis thaliana, Biochem. Biophys. Res. Commun., 2011, vol. 414, no. 3, pp. 481–486. CrossRefPubMedGoogle Scholar
  4. 4.
    Lai, D.W., Mao, Y., Zhou, H., Li, F., Wu, M., Zhang, J., He, Z., Cui, W., and Xie, Y., Endogenous hydrogen sulfide enhances salt tolerance by coupling the reestablishment of redox homeostasis and preventing salt-induced K+ loss in seedlings of Medicago sativa, Plant Sci., 2014, vol. 225, pp. 117–129. CrossRefPubMedGoogle Scholar
  5. 5.
    Fang, H.H., Pei, Y.X., Tian, B.H., Zhang, L.P., Qiao, Z.J., and Liu, Z.Q., Ca2+ participates in H2S induced Cr6+ tolerance in Setaria italica, Chin. J. Cell Biol., 2014, vol. 36, no. 6, pp. 758–765.Google Scholar
  6. 6.
    Shi, H., Ye, T., and Chan, Z., Nitric oxide-activated hydrogen sulfide is essential for cadmium stress response in bermudagrass (Cynodon dactylon (L). Pers.), Plant Physiol. Biochem., 2014, vol. 74, pp. 99–107. CrossRefPubMedGoogle Scholar
  7. 7.
    Li, Z.G. and Zhu, L.P., Hydrogen sulfide donor sodium hydrosulfide-induced accumulation of betaine is involved in the acquisition of heat tolerance in maize seedlings. Braz. J. Bot., 2014, vol. 38, no. 1, pp. 31–38.
  8. 8.
    Yang, M., Qin, B.P., Ma, X.L., Wang, P., Li, M.L., Chen, L.L., Chen, L.T., Sun, A.Q., Wang, Z.L., and Yin, Y.P., Foliar application of sodium hydrosulfide (NaHS), a hydrogen sulfide (H2S) donor, can protect seedlings against heat stress in wheat (Triticum aestivum L.), J. Integr. Agricult., 2015. V. 15. no. 12. P. 2745–58.
  9. 9.
    Kolupaev, Yu.E., Firsova, E.N., Yastreb, T.O., and Lugovaya, A.A., The participation of calcium ions and reactive oxygen species in the induction of antioxidant enzymes and heat resistance in plant cells by hydrogen sulfide donor. Appl. Biochem. Microbiol., 2017, vol. 53, no. 5, pp. 573–579.
  10. 10.
    Wang, Y., Li, L., Cui, W., Xu, S., Shen, W., and Wang, R., Hydrogen sulfide enhances alfalfa (Medicago sativa) tolerance against salinity during seed germination by nitric oxide pathway, Plant Soil., 2012, vol. 351, nos. 1–2, pp. 107–119.
  11. 11.
    Lai, D.W., Mao, Y., Zhou, H., Li, F., Wu, M., Zhang, J., He, Z., Cui, W., and Xie, Y., Endogenous hydrogen sulfide enhances salt tolerance by coupling the reestablishment of redox homeostasis and preventing salt-induced K+ loss in seedlings of Medicago sativa, Plant Sci., 2014, vol. 225, pp. 117–129. CrossRefPubMedGoogle Scholar
  12. 12.
    Shi, H., Ye, T., and Chan, Z., Exogenous application of hydrogen sulfide donor sodium hydrosulfide enhanced multiple abiotic stress tolerance in bermudagrass (Cynodon dactylon (L.). Pers.), Plant Physiol. Biochem., 2013, vol. 71, pp. 226–234. CrossRefPubMedGoogle Scholar
  13. 13.
    Li, Z.G., Min, X., and Zhou, Z.H., Hydrogen sulfide: a signal molecule in plant cross-adaptation, Front. Plant Sci., 2016, vol. 7, p. 1621. PubMedGoogle Scholar
  14. 14.
    Zhang, H., Ye, Y.K., Wang, S.H., Luo, J.P., Tang, J., and Ma, D.F., Hydrogen sulfide counteracts chlorophyll loss in sweet potato seedling leaves and alleviates oxidative damage against osmotic stress, Plant Growth Regul., 2009, vol. 58, no. 3, pp. 243–250.
  15. 15.
    Kolupaev, Yu.E., Firsova, E.N., Yastreb, T.O., Kirichenko, V.V., and Ryabchun, N.I., Influence of hydrogen sulfide donor on state of antioxidant system and resistance of wheat plants to soil drought. Russ. J. Plant Physiol., 2019, vol. 66, no. 1, pp. 59–66. doi 10.1134/S1021443719010084Google Scholar
  16. 16.
    Jin, Z., Wang, Z., Ma, Q., Sun, L., Zhang, L., Liu, Z., Liu, D., Hao, X., and Pei, Y., Hydrogen sulfide mediates ion fluxes inducing stomatal closure in response to drought stress in Arabidopsis thaliana, Plant Soil, 2017, vol. 419, nos. 1–2, pp. 141–152.
  17. 17.
    Scuffi, D., Nietzel, T., Di Fino, L.M., Meyer, A.J., Lamattina, L., Schwarzländer, M., Laxalt, A.M., and García-Mata, C., Hydrogen sulfide increases production of NADPH oxidase-dependent hydrogen peroxide and phospholipase D-derived phosphatidic acid in guard cell signaling, Plant Physiol., 2018, vol. 176, no. 3, pp. 2532–2542. CrossRefPubMedGoogle Scholar
  18. 18.
    Hu, K.D., Tang, J., Zhao, D.L., Hu, L.Y., Li, Y.H., Liu, Y.S., Jones, R., and Zhang, H., Stomatal closure in sweet potato leaves induced by sulfur dioxide involves H2S and NO signaling pathways, Biol. Plant., 2014, vol. 58, no. 4, pp. 676–680.
  19. 19.
    Lisjak, M., Srivastava, N., Teklic, T., Civale, L., Lewandowski, K., Wilson, I., Wood, M.E., Whiteman, M., and Hancock, J.T., A novel hydrogen sulfide donor causes stomatal opening and reduces nitric oxide accumulation, Plant Physiol. Biochem., 2010, vol. 48, no. 12, pp. 931–935. CrossRefPubMedGoogle Scholar
  20. 20.
    Lisjak, M., Teklić, T., Wilson, I.D., Wood, M.E., Whiteman, M., and Hancock, J.T., Hydrogen sulfide effects on stomatal apertures, Plant Signal. Behav., 2011, vol. 6, no. 10, pp. 1444–1446. CrossRefPubMedGoogle Scholar
  21. 21.
    Duan, B., Ma, Y., Jiang, M., Yang, F., Ni, L., and Lu, W., Improvement of photosynthesis in rice (Oryza sativa L.) as a result of an increase in stomatal aperture and density by exogenous hydrogen sulfide treatment, Plant Growth Regul., 2015, vol. 75, no. 1, pp. 33–44.
  22. 22.
    Honda, K., Yamada, N., Yoshida, R., Ihara, H., Sawa, T., Akaike, T., and Iwai, S., 8-Mercapto-Cyclic GMP mediates hydrogen sulfide-induced stomatal closure in Arabidopsis, Plant Cell Physiol., 2015, vol. 56, no. 8, pp. 1481–1489. CrossRefPubMedGoogle Scholar
  23. 23.
    Papanatsiou, M., Scuffi, D., Blatt, M.R., and Garcia-Mata, C., Hydrogen sulfide regulates inward-rectifying K+ channels in conjunction with stomatal closure, Plant Physiol., 2015, vol. 168, no. 1, pp. 29–35.
  24. 24.
    Wang, L., Ma, X., Che, Y., Hou, L., Liu, X., and Zhang, W., Extracellular ATP mediates H2S-regulated stomatal movements and guard cell K+ current in a H2O2-dependent manner in Arabidopsis, Sci. Bull., 2015, vol. 60, no. 4, pp. 419–27.
  25. 25.
    Suhita, D., Kolla, V.A., Vavasseur, A., and Raghavendra, A.S., Different signaling pathways involved during the suppression of stomatal opening by methyl jasmonate or abscisic acid, Plant Sci., 2003, vol. 164, no. 4, pp. 481–488.
  26. 26.
    Marino, D., Dunand, C., Puppo, A., and Pauly, N., A burst of plant NADPH oxidases, Trends Plant Sci., 2012, vol. 17, no. 1, pp. 9–15. CrossRefPubMedGoogle Scholar
  27. 27.
    Yastreb, T.O., Kolupaev, Yu.E., Shvidenko, N.V., Lugovaya, A.A., and Dmitriev, A.P., Salt stress response in Arabidopsis thaliana plants with defective jasmonate signaling, Appl. Biochem. Microbiol., 2015, vol. 51, no. 4, pp. 451–454.
  28. 28.
    Yastreb, T.O., Kolupaev, Yu.E., Lugovaya, A.A., and Dmitriev, A.P., Formation of adaptive reactions in Arabidopsis thaliana wild-type and mutant jin1 plants under action of abscisic acid and salt stress, Cytol. Genet., 2017, vol. 51, no. 5, pp. 325–330.
  29. 29.
    Neill, S., Bright, J., Desikan, R., Hancock, J., Harrison, J., and Wilson, I., Nitric oxide, evolution and perception, J. Exp. Bot., 2008, vol. 59, no. 1, pp. 25–35.
  30. 30.
    Lanteri, M.L., Laxalt, A.M., and Lamattina, L., Nitric oxide triggers phosphatidic acid accumulation via phospholipase D during auxin-induced adventitious root formation in cucumber, Plant Physiol., 2008, vol. 147, no. 1, pp. 188–198. CrossRefPubMedGoogle Scholar
  31. 31.
    Liu, H.T., Huang, W.D., Pan Q.H., Weng F.H., Zhan J.C., Liu Y., Wan S.B., Liu Y.Y. Contributions of PIP2-specific-phospholipase C and free salicylic acid to heat acclimation-induced thermotolerance in pea leaves, J. Plant Physiol., 2006, vol. 163, no. 4, pp. 405–416.
  32. 32.
    Lee, Y. and Lee, Y., Roles of phosphoinositides in regulation of stomatal movements, Plant Signal. Behav., 2008, vol. 3, no. 4, pp. 211–213.CrossRefPubMedGoogle Scholar
  33. 33.
    Lecourieux, D., Mazars, C., Pauly, N., Ranjeva, R., and Pugin, A., Analysis and effects of cytosolic free calcium increases in response to elicitors in Nicotiana plumbaginifolia cells, Plant Cell, 2002, vol. 14, no. 10, pp. 2627–2641. CrossRefPubMedGoogle Scholar
  34. 34.
    Iakovenko, O.M., Kretynin, S.V., Kabachevskaya, E.M., Lyakhnovich, G.V., Volotovski, D.I., and Kravets, V.S., Role of phospholipase C in ABA regulation of stomata function, Ukr. Bot. J., 2008, vol. 65, no. 4, pp. 605–613.Google Scholar
  35. 35.
    Arisz, S.A., Wijk, R., Roels, W., Zhu, J.K., Haring, M.A., and Munnik, T., Rapid phosphatidic acid accumulation in response to low temperature stress in Arabidopsis is generated through diacylglycerol kinase, Front. Plant Sci., 2013, vol. 4, p. 1. CrossRefPubMedGoogle Scholar
  36. 36.
    Pappan, K., Zheng, S., and Wang, X., Identification and characterization of a novel plant phospholipase D that requires polyphosphoinositides and submicromolar calcium for activity in Arabidopsis, J. Biol. Chem., 1997, vol. 272, no. 11, pp. 7048–7054. CrossRefPubMedGoogle Scholar

Copyright information

© Allerton Press, Inc. 2019

Authors and Affiliations

  1. 1.Dokuchaev Kharkiv National Agrarian UniversityKharkivUkraine
  2. 2.Karazin Kharkiv National UniversityKharkivUkraine
  3. 3.Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of UkraineKyivUkraine

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