Browse > Article
http://dx.doi.org/10.5423/PPJ.2007.23.2.076

Expression and Promoter Analyses of Pepper CaCDPK4 (Capsicum annuum calcium dependent protein kinase 4) during Plant Defense Response to Incompatible Pathogen  

Chung, Eun-Sook (BK21 Center for Silver-Bio Industrialization, Dong-A University)
Oh, Sang-Keun (Plant Genome Research Center, Korea Research Institute of Bioscience & Biotechnology)
Park, Jeong-Mee (Plant Genome Research Center, Korea Research Institute of Bioscience & Biotechnology)
Choi, Do-Il (Department of Plant Sciences, Seoul National University)
Publication Information
The Plant Pathology Journal / v.23, no.2, 2007 , pp. 76-89 More about this Journal
Abstract
CaCDPK4, a full-length cDNA clone encoding Capsicum annuum calcium-dependent protein kinase 4, was isolated from chili pepper (Capsicum annuum L.). Deduced amino acid sequence of CaCDPK4 shares the highest homology with tobacco NpCDPK8 and chickpea CaCDPK2 with 79% identity. Genomic blot analyses revealed that CaCDPK4 is present as a single copy in pepper genome, but it belongs to a multigene family. CaCDPK4 was highly induced when pepper plants were inoculated with an incompatible bacterial pathogen. Induced levels of CaCDPK4 transcripts were also detected in pepper leaves by the treatment of ethephon, an ethylene-inducing agent, and high-salt stress condition. The bacterial-expressed GST-CaCDPK4 protein showed to retain the autophosphorylation activity in vitro. GUS expression driven by CaCDPK4 promoter was examined in transgenic Arabidopsis containing transcriptional fusion of CaCDPK4 promoter. GUS expression under CaCDPK4 promoter was strong in the root and veins of the seedlings. GW (-1965) and D3 (-1377) promoters conferred on GUS expression in response to inoculation of an incompatible bacterial pathogen, but D4-GUS (-913) and DS-GUS (-833) did not. Taken together, our results suggest that CaCDPK4 can be implicated on signal transduction pathway of defense response against an incompatible bacterial pathogen in pepper.
Keywords
CDPK; chili pepper; non-host resistance; promoter;
Citations & Related Records

Times Cited By Web Of Science : 16  (Related Records In Web of Science)
연도 인용수 순위
  • Reference
1 Choi, D., Kim, H. M., Yun, H. K., Park, J. A., Kim, W. T. and Bok, S. H. 1996. Molecular cloning of a metallothionein-like gene from Nicotiana glutinosa L. and its induction by wounding and tobacco moasaic virus infection. Plant Physiol. 112:353-359   DOI
2 Chung, E., Kim, S. Y, Yi, S. and Choi, D. 2003. Capsicum annuum dehydrin, an osmotic-stress gene in chili pepper plants. Mol. Cells 15:327-332
3 Chung, E., Park, J.M., Oh, S.-K., Joung, Y.H., Lee, S. and Choi, D. 2004. Molecular and biochemical characterization of the Capsicum annuum calcium-dependent protein kinase 3 (CaCDPK3) gene induced by abiotic and biotic stresses. Planta 220:286-295   DOI
4 Church, G. M. and Gilbert, W. 1984. Genomic sequencing. Proc. Natl. Aca. Sci. USA 81:1991-1995
5 Dammann, C., Ichida, A., Hong, B., Romanowsky, S. M., Hrabak, E. M., Harmon, A. C., Pickard, B. G. and Harper, J. F. 2003. Subcellular targeting of nine calcium-dependent protein kinase isoforms from Arabidopsis. Plant Physiol. 132:1840-1848   DOI
6 Dellaporta, S. L., Wood, J. and Hicks, J. B. 1983. A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1: 19-21   DOI
7 Harmon, A. C., Putnam-Evans, C. and Cormier, M. J. 1987. A calcium-dependent but calmodulin-independent protein kinase from soybean. Plant Physiol. 83:830-837   DOI   ScienceOn
8 Harper, J. F., Huang, J.-F. and Lloyd, S. J. 1994. Genetic identification of an autoinhibitor in CDPK, a protein kinase with a calmodulin-like domain. Biochem. 33:7267-7277   DOI   ScienceOn
9 Harper, J. F., Breton, G. and Harmon, A. 2004. Decoding $Ca^{2+}$ sigrials through plant protein kinase. Annu. Rev. Plant Biol. 55:263-288   DOI
10 Hrabak, E. M., Chan, C. W, Gribskov, M., Harper, J. F., Choi, J. H., Halford, N., Kudla, J., Luan, S., Nimmo, H. G, Sussman, M. R., Thomas, M., Walker-Simmons, K., Zhu, J. K. and Harmon, A. C. 2003. The Arabidopsis CDPK-SnRK superfamily of protein kinases. Plant Physiol. 132:666-680   DOI   ScienceOn
11 Lee, S., Kim, S. Y., Chung, E., Joung, Y. H., Pai, H. S., Hur, C. G. and Choi, D. 2004. EST and microarray analyses of pathogen responsive genes in hot pepper (Capsicum annuum L.) nonhost resistance against soybean pustule pathogen (Xanthomonas axonopodis pv. glycines). Funct. Integrative Genomics 4:196-205
12 Notredame, C., Higgins, D. and Heringa, J. 2000. A novel method for multiple sequence alignments. J. Mol. Biol. 302:205-217   DOI   ScienceOn
13 Oh, S.-K., Park, J. M., Joung, Y. H., Lee, S., Chung, E., Kim, S.-Y., Yu, S. H. and Choi, D. 2005. A plant EPF-type zinc-finger protein, CaPIF1, involved in defence against pathogens. Mol. Plant Pathol. 6:269-285   DOI   ScienceOn
14 Yoon, G. M., Cho, H. S., Ha, H. J., Liu, J. R. and Pai, H.-S. 1999. Characterization of NtCDPK1, a calcium-dependent protein kinase gene in Nicotiana tabacum, and the activity of its encoded protein. Plant Mol. Biol. 39:991-1001   DOI   ScienceOn
15 Roberts, D. M. and Harmon, A C. 1992. Calcium-modulated proteins: targets of intracellular calcium signals in higher plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43:375-414   DOI
16 Saijo, Y., Hata, S., Kyozuka, J., Shimamoto, K. and Izui, K. 2000. Over-expression of a single $Ca^{2+}$ -dependent kinase confers both cold and salt/drought tolerance. Plant J. 23:319-327   DOI   ScienceOn
17 Sambrook, J., Fritsch, E. F. and Maniatis, T. 1989. Molecular cloning: a laboratory manual, Cold Spring Harbor, Cold Spring Harbor Laboratory Press, USA
18 Botella, J. R., Arteca, J. M., Somodevilla, M. and Arteca, R. N. 1996. Calcium-dependent protein kinase gene expression in response to physical and chemical stimuli in mungbean (Vigna radiata). Plant Mol. Biol. 30:1129-1137   DOI
19 Cheng, S. H., Willman, M. R., Chen, H. C. and Sheen, J. 2002. Calcium signaling through protein kinases. The Arabidopsis Calcium-dependent protein kinase gene family. Plant Physiol. 129:49-55
20 Marcotte, W. R. Jr., Russell, S. H. and Qatrano, R. S. 1989. Abscisic acid-responsive sequences from the Em gene of wheat. Plant Cell 1:969-976   DOI   ScienceOn
21 Clough, S. J. and Bent, A. F. 1998. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16:735-743   DOI   ScienceOn
22 Heath, M. C. 2000. Hypersensitive response-related death. Plant Mol. Biol. 44:321-334   DOI   ScienceOn
23 Putnam-Evans, C., Harmon, A C., Palevitz, B. A, Fechheimer, M. and Cormier, M. J. 1989. Calcium-dependent protein kinase is localized with F-actin in plant cells. Cell Motil. Cytoskeleton 12: 12-22   DOI
24 Jefferson, R. A., Kavanagh, T. A. and Bevan, M. W 1987. GUS fusion: ${\beta}$ as a sensitive and versatile gene fusion marker in higher plants. EMBO. J. 6:3901-3907
25 Laemmli, U. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685   DOI   ScienceOn
26 Ecker, J. R. 1995. The ethylene signal transduction pathway in plants. Science 268:667-675   DOI   ScienceOn
27 Yi, S. Y., Kim, J. H., Joung, Y. H., Lee, S., Kim, W. T., Yu, S. H. and Choi, D. 2004. The pepper (Capsicum annuum L.) transcription factor, CaPF1, confers pathogen and freezing tolerance in Arabidopsis. Plant Physiol. 136:2862-2874   DOI   ScienceOn
28 Yu, D., Chen, C. and Chen, Z. 2001. Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant Cell 13:1527-1539   DOI
29 Asano, T., Tanaka, N., Yang, G, Hayashi, N. and 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. 46:356-366   DOI   ScienceOn
30 Evans, N. H., McAinsh, M. R. and Hetherington, A. M. 2001. Calcium oscillations in higher plants. Curr. Opin. Plant Biol. 4:415-420   DOI   ScienceOn
31 Putnam-Evans, C. L., Harmon, A C. and Cormier, M. J. 1990. Purification and characterization of a novel calcium-dependent protein kinase from soybean. Biochem. 29:2488-2495   DOI   ScienceOn
32 Whitelaw, C. A., Le Huquet, J. A., Thurman, D. A. and Tomsett, A. B. 1997. The isolation and characterization of type II metal-lothionein-like genes from tomato (Lycopersicon esculentum L.). Plant Mol. Biol. 33:503-511   DOI   ScienceOn
33 Murillo, I., Cordero, E. J. M. and Segundo, B. S. 2001. Transcriptional activation of a maize calcium-dependent protein kinase gene in response to fungal elicitors and infection. Plant Mol. Biol. 45:145-158   DOI   ScienceOn
34 Niggeweg, R., Thurow, C., Weigel, R., Pfitzner, U. and Gatz, C. 2000. Tobacco TGA factors differ with respect to interaction with NPRl, activation potential and DNA-binding properties. Plant Mol. Biol. 42:775-788   DOI   ScienceOn
35 Patharkar, O. R. and Cushman, J. C. 2000. A stress-induced calcium-dependent protein kinase from Mesembryanthemum crystallinum phosphorylates a two-component pseudo-response regulator. Plant J. 24:679-691   DOI   ScienceOn
36 Prescott, A and Martin, C. 1987. A rapid method for the quantitative assessment of levels of specific mRNAs in plants. Plant Mol. Biol. Rep. 4:219-224   DOI
37 Satterlee, J. S. and Sussman, M. R. 1998. Unusual membrane-associated protein kinases in higher plants. J. Membrane Biol. 164:205-213   DOI
38 Xing, T., Wang, X.-J., Malik, K. and Miki, B. L. 2001. Ectopic expression of an Arabidopsis calmodulin-like domain protein kinase-enhanced NADPH oxidase activity and oxidative burst in tomato protoplasts. Mol. Plant-Microbe Interact. 14:1261-1264   DOI   ScienceOn
39 Arabidopsis Genome Initiative. 2000. Analysis of the genome of the flowering plant Arabidopsis thaliana. Nature 408:820-826   DOI   ScienceOn
40 King, E. O., Ward, M. K. and Raney, D. E. 1954. Two simple media for the demonstration of phycocyanin and fluorescin. J. Lab. Clin. Med. 44:301-307
41 Ludwig, A. A., Romeis, T. and Jones, J. D. G. 2004. CDPK-mediated signalling pathways: specificity and cross-talk. J. Exp. Botany 55:181-188   DOI   ScienceOn
42 Whalen, M. C., Innes, R. W., Bent, A F. and Staskawicz, B. 1991. Identification of Pseudomonas syringae pathogens of Arabido psis and a bacterial locus determining avirulence on both Arabidopsis and soybean. Plant Cell 3:49-59   DOI   ScienceOn
43 Chico, J. M., Raices, M., Tellez-Inon, M. T. and Ulloa, R. M. 2002. A calcium-dependent protein kinase is systemically induced upon wounding in tomato plants. Plant Physiol. 128:256-270   DOI
44 Bray, E. A. 1993. Molecular responses to water deficit. Plant Physiol. 103:1035-1040   DOI
45 Knight, H., Trewavas, A. J. and Knight, M. R. 1997. Calcium signaling in Arabidopsis thaliana responding to drought and salinity. Plant J. 12:1067-1078   DOI   ScienceOn
46 Reymond, P. and Farmer, E. E. 1998. Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant Biol. 1 :404-411   DOI   ScienceOn
47 Lee, S. S., Cho, H. S., Yoon, G. M., Ahn, J.-W., Kim, H.-H., Pai, H.-S. 2003. Interaction of NtCDPK1 calcium-dependent protein kinase with NtRpn3 regulatory subunit of the 26S proteasome in Nicotiana tabacum. Plant J. 33:825-840
48 Chehab, E. W., Patharkar, O. R., Hegeman, A. D., Taybi, T. and Cushman, J. C. 2004. Autophosphorylation and subcellular localization dynamics of a salt- and water deficit-induced calcium dependent protein kinase from ice plant. Plant Physiol. 135:1430-1446   DOI   ScienceOn
49 Park, B. K. and Hwang, I. 1999. Identification of hrcC, hrpF, and miaA genes of Xanthomonas campestris pv. glycines 8ra: roles in pathogenicity and inducing hypersensitive response on nonhost plants. Plant Pathol. 15:21-2
50 Sanders, D., Pelloux, J., Brownlee, C. and Harper, J. F. 2002. Calcium at the crossroads of signaling. Plant Cell 14:S401-S417
51 Lu, S. X. and Hrabak, E. M. 2002. An Arabidopsis calciumdependent protein kinase is associated with the endoplasmic reticulum. Plant Physiol. 128:1008-1021   DOI   ScienceOn
52 Romeis, T., Ludwig, A A, Martin, R. and Jones, J. D. G. 2001. Calcium-dependent protein kinases play an essential role in a plant defence response. EMBO. J. 20:5556-5567   DOI   ScienceOn
53 An, G, Ebert, P. R., Mitra, A. and Ha, S. B. 1988. Binary vector. In: Plant Molecular Biology Manual. A3. ed. by S. B. Gelvin, R. A. Schilperoot and D. P. S. Verma, pp. 1-19. Kluwer Academic Publishers
54 Kang, J. Y., Choi, H. I., Im, M. Y. and Kim, S. Y. 2002. Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling. Plant Cell 14:343-357   DOI
55 Urao, T., Katagiri, T., Mizoguchi, T., Yamaguchi-Shinozaki, K., Hayashida, N. and Shinozaki, K. 1994. Two genes that encode $Ca^{2+}$ -dependent protein kinases are induced by drought and high-salt stresses in Arabidopsis thaliana. Mol. Gen. Genet. 244:331-340