Abstract
Calcium-dependent protein kinases (CDPK or CPK) are involved in protecting plants from abiotic stresses, but the properties and functions of CDPK are still poorly understood. Analysis of the Amur grape Vitis amurensis Rupr. is of great interest, since it demonstrates high tolerance to adverse environmental conditions. It was demonstrated that exposure of V. amurensis to salt and, to a lesser extent, osmotic and cold stress resulted in a considerable increase in the VaCPK1 and VaCPK26 transcription level. The present study was focused on the analysis of the effect of constitutive expression of the recombinant VaCPK1 and VaCPK26 genes on the resistance of V. amurensis cell cultures and Arabidopsis thaliana plants to salt, osmotic, and temperature stresses. It was demonstrated that expression of recombinant VaCPK26, as well as to a lesser extent VaCPK1, led to a 1.2–1.7 times increase in biomass accumulation by V. amurensis cell cultures and to a 1.2–2.1 times increase in survival of A. thaliana plants under salt stress. In addition, VaCPK26-expressing lines of A. thaliana lines demonstrated a slight degree of drought tolerance and were characterized by increased expression of some stress-induced protective genes involved in the formation of salt and dehydration tolerance. The data obtained indicate that the VaCPK1 and VaCPK26 genes can act as positive regulators of plant response to salt stress.
Similar content being viewed by others
REFERENCES
Medvedev, S.S., Calcium signaling system in plants, Russ. J. Plant Physiol., 2005, vol. 52, no. 2, pp. 249—270.
Hashimoto, K. and Kudla, J., Calcium decoding mechanisms in plants, Biochimie, 2011, vol. 93, pp. 2054–2059. https://doi.org/10.1016/j.biochi.2011.05.019
Liese, A. and Romeis, T. Biochemical regulation of in vivo function of plant calcium-dependent protein kinases (CDPK). Biochim. Biophys. Acta—Mol. Cell Res., 2013, vol. 7, pp. 1582—1589. https://doi.org/10.1016/j.bbamcr.2012.10.024
Harper, J.F. and Harmon, A., Plants, symbiosis and parasites: a calcium signalling connection, Nat. Rev. Mol. Cell Biol., 2005, vol. 6, pp. 555—566. https://doi.org/10.1038/nrm1679
Schulz, P., Herde, M., and Romeis, T., Calcium-dependent protein kinases: hubs in plant stress signaling and development, Plant Physiol., 2013, vol. 163, pp. 523—530. https://doi.org/10.1104/pp.113.222539
Boudsocq, M. and Sheen, J., CDPKs in immune and stress signaling, Trends Plant Sci., 2013, vol. 18, pp. 30—40. https://doi.org/10.1016/j.tplants.2012.08.008
Asano, T., Hayashi, N., Kobayashi, M., et al., A rice calcium-dependent protein kinase OsCPK12 oppositely modulates salt-stress tolerance and blast disease resistance, Plant J., 2012, vol. 69, pp. 26—36. https://doi.org/10.1111/j.1365-313X.2011.04766.x
Dubrovina, A.S., Kiselev, K.V., and Khristenko, V.S., Expression of calcium-dependent protein kinase (CDPK) genes under abiotic stress conditions in wild-growing grapevine Vitis amurensis, J. Plant Physiol., 2013, vol. 170, pp. 1491—1500. https://doi.org/10.1016/j.jplph.2013.06.014
Aleynova, O.A., Dubrovina, A.S., and Kiselev, K.V., Activation of stilbene synthesis in cell cultures of Vitis amurensis by calcium-dependent protein kinases VaCPK1 and VaCPK26, Plant Cell Tissue Organ Cult., 2017, vol. 130, pp. 141—152. https://doi.org/10.1007/s11240-017-1210-y
Tzfira, T., Tian, G.W., Lacroix, B., et al., PSAT vectors: a modular series of plasmids for autofluorescent protein tagging and expression of multiple genes in plants, Plant Mol. Biol., 2005, vol. 57, pp. 503—516. https://doi.org/10.1007/s11103-005-0340-5
Kiselev, K.V., Dubrovina, A.S., and Bulgakov, V.P., Phenylalanine ammonia-lyase and stilbene synthase gene expression in rolB transgenic cell cultures of Vitis amurensis, Appl. Microbiol. Biotechnol., 2009, vol. 82, pp. 647—655. https://doi.org/10.1007/s00253-008-1788-4
Zhang, X., Henriques, R., Lin, S.S., et al., Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method, Nat. Protoc., 2006, vol. 1, pp. 641—646. https://doi.org/10.1038/nprot.2006.97
Ma, L. and Chung, W.K., Quantitative analysis of copy number variants based on real-time LightCycler PCR, Curr. Protoc. Hum. Genet., 2014, vol. 80, pp. 1—10. https://doi.org/10.1002/0471142905.hg0721s80
Dubrovina, A.S., Kiselev, K.V., Khristenko, V.S., and Aleynova, O.A., VaCPK20, a calcium-dependent protein kinase gene of wild grapevine Vitis amurensis Rupr., mediates cold and drought stress tolerance, J. Plant Physiol., 2015, vol. 185, pp. 1—12. https://doi.org/10.1016/j.jplph.2015.05.020
Dubrovina, A.S., Kiselev, K.V., Khristenko, V.S., and Aleynova, O.A., VaCPK21, a calcium-dependent protein kinase gene of wild grapevine Vitis amurensis Rupr., is involved in grape response to salt stress, Plant Cell Tissue Organ Cult., 2016, vol. 124, pp. 137—150. https://doi.org/10.1007/s11240-015-0882-4
Dubrovina, A.S., Kiselev, K.V., Khristenko, V.S., and Aleynova, O.A., The calcium-dependent protein kinase gene VaCPK29 is involved in grapevine responses to heat and osmotic stresses, Plant Growth Regul., 2017, vol. 82, pp. 79—89. https://doi.org/10.1007/s10725-016-0240-5
Kiselev, K.V., Dubrovina, A.S., Shumakova, O.A., et al., Structure and expression profiling of a novel calcium-dependent protein kinase gene, CDPK3a, in leaves, stems, grapes, and cell cultures of wild-growing grapevine Vitis amurensis Rupr., Plant Cell Rep., 2013, vol. 32, pp. 431—442. https://doi.org/10.1007/s00299-012-1375-0
Kiselev, K.V., Aleinova, O.A., and Tyunin, A.P., Expression of the R2R3 MYB transcription factors in Vitis amurensis Rupr. plants and cell cultures with different resveratrol content, Russ. J. Genet., 2017, vol. 53, no. 5, pp. 465—471. https://doi.org/10.1134/S1022795417040093
Tyunin, A.P., Kiselev, K.V., and Zhuravlev, Y.N., Effects of 5-azacytidine induced DNA demethylation on methyltransferase gene expression and resveratrol production in cell cultures of Vitis amurensis, Plant Cell Tissue Organ Cult., 2012, vol. 111, pp. 91—100. https://doi.org/10.1007/s11240-012-0175-0
Livak, K.J. and Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method, Methods, 2001, vol. 25, pp. 402—408. https://doi.org/10.1006/meth.2001.1262
Zhang, H., Liu, W.Z., Zhang, Y., et al., Identification, expression and interaction analyses of calcium-dependent protein kinase (CDPK) genes in canola (Brassica napus L.), BMC Genomics, 2014, vol. 15, p. 211. https://doi.org/10.1186/1471-2164-15-211
Ingram, J. and Bartels, D., The molecular basis of dehydration tolerance in plants, Annu. Rev. Plant Physiol. Plant Mol. Biol., 1996, vol. 47, pp. 377—403. https://doi.org/10.1146/annurev.arplant.47.1.377
Fujita, M., Fujita, Y., Maruyama, K., et al., A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway, Plant J., 2004, vol. 39, pp. 863—876. https://doi.org/10.1111/j.1365-313X.2004.02171.x
Xiong, L., Schumaker, K.S., and Zhu, J.K., Cell signaling during cold, drought, and salt stress, Plant Cell, 2002, vol. 14, pp. S165—S183. https://doi.org/10.1105/tpc.000596
Kasuga, M., Liu, Q., Miura, S., et al., Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor, Nat. Biotechnol., 1999, vol. 17, pp. 287—291. https://doi.org/10.1038/7036
Akhtar, M., Jaiswal, A., Taj, G., et al., DREB1/CBF transcription factors: their structure, function and role in abiotic stress tolerance in plants, J. Genet., 2012, vol. 91, pp. 385—395. https://doi.org/10.1007/s12041-012-0201-3
Liu, L. and Li, H., Review: research progress in Amur grape, Vitis amurensis Rupr., Can. J. Plant Sci., 2013, vol. 93, pp. 565—575. https://doi.org/10.4141/CJPS2012-202
Khristenko, V.S., Dubrovina, A.S., Aleinova, O.A., and Kiselev, K.V., The effect of overexpression of the Ca2+-dependent protein kinase gene VaCDPK13 on the resistance of the Amur Vitis amurensis grapes cell cultures to abiotic stress, Vestn. Krasnodar. Gos. Agrar. Univ., 2015, vol. 2, pp. 132—138.
Lang, V. and Palva, E.T., The expression of a rab-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.) Heynh., Plant Mol. Biol., 1992, vol. 20, pp. 951—962. https://doi.org/10.1007/BF00027165
Fujita, M., Fujita, Y., Maruyama, K., et al., A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway, Plant J., 2004, vol. 39, pp. 863—876. https://doi.org/10.1111/j.1365-313X.2004.02171.x
Msanne, J., Lin, J., Stone, J.M., and Awada, T., Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes, Planta, 2011, vol. 234, pp. 97—107. https://doi.org/10.1007/s00425-011-1387-y
Liu, D., Li, W., Cheng, J., and Hou, L., Expression analysis and functional characterization of a cold-responsive gene COR15A from Arabidopsis thaliana, Acta Physiol. Plant., 2014, vol. 36, pp. 2421—2432. https://doi.org/10.1007/s11738-014-1615-8
Finkelstein, R.R. and Rock, C.D., Abscisic acid biosynthesis and response, Arabidopsis Book, 2002, vol. 1. e0058. https://doi.org/10.1199/tab.0058
Coca, M. and San Segundo, B., AtCPK1 calcium-dependent protein kinase mediates pathogen resistance in Arabidopsis, Plant J., 2010, vol. 63, pp. 526—540. https://doi.org/10.1111/j.1365-313X.2010.04255.x
Xu, J., Tian, Y.S., Peng, R.H., et al., AtCPK6, a functionally redundant and positive regulator involved in salt/drought stress tolerance in Arabidopsis, Planta, 2010, vol. 231, pp. 1251—1260. https://doi.org/10.1007/s00425-010-1122-0
Zou, J.J., Wei, F.J., Wang, C., et al., Arabidopsis calcium-dependent protein kinase CPK10 functions in abscisic acid- and Ca2+-mediated stomatal regulation in response to drought stress, Plant Physiol., 2010, vol. 154, pp. 1232—1243. https://doi.org/10.1104/pp.110.157545
Chen, J., Xue, B., Xia, X., and Yin, W., A novel calcium-dependent protein kinase gene from Populus euphratica, confers both drought and cold stress tolerance, Biochem. Biophys. Res. Commun., 2013, vol. 441, pp. 630—636. https://doi.org/10.1016/j.bbrc.2013.10.103
Wei, S., Hu, W., Deng, X., et al., A rice calcium-dependent protein kinase OsCPK9 positively regulates drought stress tolerance and spikelet fertility, BMC Plant Biol., 2014, vol. 14, p. 133. https://doi.org/10.1186/1471-2229-14-133
Ma, S.Y. and Wu, W.H., AtCPK23 functions in Arabidopsis responses to drought and salt stresses, Plant Mol. Biol., 2007, vol. 65, pp. 511—518. https://doi.org/10.1007/s11103-007-9187-2
Franz, S., Ehlert, B., Liese, A., et al., Calcium-dependent protein kinase CPK21 functions in abiotic stress response in Arabidopsis thaliana, Mol. Plant, 2011, vol. 4, pp. 83—96. https://doi.org/10.1093/mp/ssq064
Weckwerth, P., Ehlert, B., and Romeis, T., ZmCPK1, a calcium-independent kinase member of the Zea mays CDPK gene family, functions as a negative regulator in cold stress signalling, Plant Cell Environ., 2015, vol. 38, pp. 544—558. https://doi.org/10.1111/pce.12414
ACKNOWLEDGMENTS
We thank O.A. Aleynova and V.S. Khristenko for assistance in producing transgenic grapevine cell cultures and Arabidopsis plants.
This study was supported by the Russian Foundation for Basic Research (grant no. 18-04-00284).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
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.
Additional information
Translated by N. Maleeva
Rights and permissions
About this article
Cite this article
Dubrovina, A.S., Kiselev, K.V. The Role of Calcium-Dependent Protein Kinase Genes VaCPK1 and VaCPK26 in the Response of Vitis amurensis (in vitro) and Arabidopsis thaliana (in vivo) to Abiotic Stresses. Russ J Genet 55, 319–329 (2019). https://doi.org/10.1134/S1022795419030049
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1022795419030049