Abstract
Apepper zinc-finger protein gene, CAZFP1, encoding the Cys2/His2-type zinc-finger transcription factor was isolated from pepper leaves inoculated with an avirulent strain Bv5-4a of Xanthomonas campestris pv. vesicatoria. The CAZFP1 protein is a nuclear targeting protein, which functions as a transcriptional regulator. The full-length CAZFP1 had no transcriptional activation activity, whereas the C-terminal region of CAZFP1 had transactivation activity. The CAZFP1 transcripts were constitutively expressed in the pepper stem, root, flower and red fruit, but were not detectable in the leaf and green fruit. The CAZFP1 transcripts accumulated earlier than the CABPR1 (PR-1) gene in the incompatible interaction of the pepper leaves with X. campestris pv. vesicatoria. The CAZFP1 transcripts were significantly induced in the systemic, uninoculated leaf tissues early after inoculation with bacterial pathogens, but gradually declined thereafter. The CAZFP1 transcripts were localized, and con.ned to the phloem cells of the vascular bundle in the pepper leaf midrib in response to Colletotrichum. coccodes infection, ethylene and abscisic acid. The CAZFP1 gene was also induced much earlier by abiotic elicitors and environmental stresses, compared with the CABPR1 gene. Overexpression of the CAZFP1 gene in the transgenic Arabidopsis plants enhanced not only the resistance against infection by Pseudomonas syringae pv. tomato, but also the drought tolerance. These results suggest that the CAZFP1 gene functions as an early-defense gene to enhance disease resistance and drought tolerance.
Similar content being viewed by others
References
Alexander, D., Goodman, R.M., Gutrella, M., Glascock, C., Weymann, K., Friedrich, L., Maddox, D., Ahlgoy, P., Luntz, T., Ward, E. and Ryals, J. 1993. Increased tolerance to 2 oomycete pathogens in transgenic tobacco expressing pathogenesis-related protein-1a. Proc. Natl. Acad. Sci. USA 90: 7327–7331.
Altschul, S.F., Madden, T.L., Schaäffer, A.A., Zhang, J., Zhang, Z, Miller, W. and Lipman, D.J. 1997. “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs“. Nucleic Acids Res. 25: 3389–3402.
Alvarez, M.E., Penell, R.I., Meijer, P.J., Ishikawa, A., Dixon, R.A. and Lamb, C. 1998. Reactive oxygen intermediates mediate a systemic signal network in the establishment of plant immunity. Cell 92: 773–784.
Barrieu, F., Chaumont, F. and Chrispeels, M.J. 1998. High expression of the tonoplast aquaporin ZmTIP1 in epidermal and conducting tissues of maize. Plant Physiol. 117: 1153–1163.
Berg, J.M. 1990. Zinc fingers and other metal binding domains. J. Biol. Chem. 265: 6513–6516.
Chen, W. and Singh, K.B. 1999. The auxin, hydrogen peroxide and salicylic acid induced expression of the Arabidopsis GST6 promoter is mediated in part by an ocs element. Plant J. 19: 667–677.
Chomczynski, P. and Sacchi, N. 1987. Single step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162: 156–159.
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
Cox, K.H. and Goldberg, R.B. 1988. Analysis of plant gene expression. In: C. H. Shaw (Ed), Plant Molecular Biology: A Practical Approach, IRL Press Oxford, UK, pp. 1–34.
Daniel, X., Lacomme, C., Morel, J.B. and Roby, D. 1999. A novel myb oncogene homologue in Arabidopsis thaliana related to hypersensitive cell death. Plant J. 20: 57–66.
Desjarlais, J.R. and Berg, J.M. 1992. Toward rules relating zinc finger protein sequences and DNA binding site preferences. Proc. Natl. Acad. Sci. USA 89: 7345–7349.
Do, H.M., Hong, J.K., Jung, H.W., Kim, S.H., Ham, J.H. and Hwang, B.K. 2003. Expression of peroxidase-like genes, H2O2 production, and peroxidase activity during the hypersensitive response to Xanthomonas campestris pv. vesicatoria in Capsicum annuum. Mol. Plant Microbe Interact. 16: 196–205.
Durrant, W.E., Rowland, O., Piedras, P., Hammond-Kosack, K.E. and Jones, J.D.G. 2000. cDNA-AFLP reveals a striking overlap in race specific resistance and wound response gene expression profiles. Plant Cell 12: 963–977.
Frugier, F., Poirier, S., Satiat-Jeunemaitre, B., Kondorosi, A. and Crepsi, M. 2000. A Kruppel-like zinc-finger protein is involved in nitrogen-fixing root nodule organogenesis. Genes Dev. 14: 475–482.
Gorlach, J., Volrath, S., Knauf-Beiter, G., Hengy, G., Beckhove, U., Kogel, K.H., Oostendorp, M., Staub, T., Ward, E., Kessmann, H. and Ryals, J. 1996. Benzothiadiazole, a novel class of inducers of systemic acquired resistance, activates gene expression and disease resistance in wheat. Plant Cell 8: 629–643.
Heinekamp, T., Kuhlmann, M., Lenk, A., Strathmann, A. and Droge-Laser, W. 2002. The tobacco bZIP transcription factor BZI-1 binds to G-box elements in the promoters of phenylpropanoid pathway genes in vitro, but it is not involved in their regulation in vivo. Mol. Genet. Genomics 267: 16–26.
Hong, J.K., Hwang, B.K. and Kim, C.H. 1999. Induction of local and systemic resistance to Colletotrichum coccodes in pepper plants by DL-β-amino-n-butyric acid. J. Phytopathol. 147: 193–198.
Hong, J.K., Jung, H.W., Kim, Y.J. and Hwang, B.K. 2000. Pepper gene encoding a basic class II chitinase is inducible by pathogen and ethephon. Plant Sci. 159: 39–49.
Horvath, D.M., Huang, D.J. and Chua, N.H. 1998. Four classes of salicylate-induced genes. Mol. Plant-Microbe Interact. 11: 895–905.
Iwabuchi, K., Li, B., Bartel, P. and Fields, S. 1993. Use of the two-hybrid system to identify the domain of p53 involved in oligomerization. Oncogene 8: 1693–1696.
Jung, H.W. and Hwang, B.K. 2000a. Isolation, partial sequencing, and expression of pathogenesis-related cDNA genes from pepper leaves infected by Xanthomonas campestris pv. vesicatoria. Mol. Plant Microbe Interact. 13: 136–142.
Jung, H.W. and Hwang, B. K. 2000b. Pepper gene encoding a basic β-1,3-glucanase is differentially expressed in pepper tissues upon pathogen infection and ethephon or methyl jasmonate treatment. Plant Sci. 159: 97–106.
Kalderon, D., Richardson, W.D., Markham, A.F. and Smith, A.E. 1984. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature 311: 33–38.
Kang, S.G., Jin, J.B., Piao, H.L., Pih, K.T., Jang, H.J., Lim, J.H. and Hwang, I. 1998. Molecular cloning of an Arabidopsis cDNA encoding a dynamin-like protein that is localized to plastids. Plant Mol. Biol. 38: 437–447.
Karin, M. and Hunter, T. 1995. Transcriptional control by protein phosphorylation: signal transmission from the cell surface to the nucleus. Curr. Biol. 5: 747–757.
Kim, J.C., Lee, S.H., Cheong, Y.H., Yoo, C.M., Lee, S.I., Chun, H.J., Yun, D.J., Hong, J.C., Lee, S.Y., Lim, C.O. and Cho, M.J. 2001. A novel cold-inducible zinc finger protein from soybean, SCOF-1, enhances cold tolerance in transgenic plants. Plant J. 25: 247–259.
Kim, Y.J. and Hwang, B.K. 2000. Pepper gene encoding a basic pathogenesis-related 1 protein is pathogen and ethylene inducible. Physiol. Planta. 108: 51–60.
Kranz, H.D., Denekamp, M., Greco, R., Jin, H., Leyva, A., Meissner, R.C., Petroni, K., Urzainqui, A., Bevan, M., Martin, C., Smeekens, S., Tonelli, C., Paz-Ares, J. and Weisshaar, B. 1998. Towards functional characterization of the members of the R2R3-MYB gene family from Arabidopsis thaliana. Plant J. 16: 263–276.
Kubo, K., Sakamoto, A., Kobayashi, A., Rybka, Z., Kanno, Y., Nakagawa, H., Nishino, T. and Takatsuji, H. 1998. Cys2/His2 zinc-finger protein family of petunia: evolution and general mechanism of target sequence recognition. Nucleic Acids Res. 26: 608–615.
Laskey, R.A. and Dingwall, C. 1993. Nucleus shuttling: the default pathway for nucleus proteins? Cell 74: 585–586
Lebel, E., Heifetz, P., Throne, L., Uknes, S., Ryals, J. and Ward, E. 1998. Functional analysis of regulatory sequences controlling PR-1 gene expression in Arabidopsis. Plant J. 16: 223–233.
Lee, S.C., Hong, J.K., Kim, Y.J. and Hwang, B.K. 2000. Pepper gene encoding thionin is differentially induced by pathogens, ethylene and methyl jasmonate. Physiol. Mol. Plant Pathol. 56: 207–216.
Lee, S.C., Kim, Y.J. and Hwang, B.K. 2001. A pathogeninduced chitin-binding protein gene from pepper: its isolation and differential expression in pepper tissues treated with pathogens, ethephon, methyl jasmonate or wounding. Plant Cell Physiol. 42: 1321–1330.
Lichtenthaler, H.K. 1987. Cholrophylls and carotenoids: pigments or photosynthetic biomembranes. Methods Enzymol. 148: 350–382.
Maldonado, A.M., Doerner, P., Dixon, R.A., Lamb, C.J. and Cameron, R.K. 2002. A putative lipid transfer protein involved in systemic resistance signaling in Arabidopsis. Nature 419: 399–403.
Norman-Setterblad, C., Vidal, S. and Palva, E.T. 2000. Interacting signal pathways control defense gene expression in Arabidopsis in response to cell wall-degrading enzymes from Erwinia. Mol. Plant-Microbe Interact. 13: 430–438.
Ohta, M., Matsui, K., Hiratsu, K., Shinshi, H. and Ohme-Takagi, M. 2001. Repression domains of class 11 ERF transcriptional repressor share an essential motif for active repression. Plant Cell 13: 1959–1968.
Okubara, P.A. and Berry, A.M. 1999. An mRNA (Accession no. AF119050) from symbiotic root nodules of Datisca glomerata with homology to zinc finger transcription factor genes. Plant Physiol. 119: 1149.
Pavletich, N. and Pabo, C. 1991. Zinc finger-DNA recognition: crystal structure of a Zif268-DNA complex at 2.1 Å. Science 252: 809–817.
Penninckx, I.A., Thomma B.P., Buchala, A., Metraux, J.P. and Broekaert, W.F. 1998. Concomitant activation of jasmonate and ethylene response pathways is required for induction of a plant defensin gene in Arabidopsis. Plant cell 10: 2103–2113.
Raikhel, N. 1992. Nucleus targeting in plants. Plant Physiol. 100: 1627–1632.
Reymond, P. and Farmer, E.E. 1998. Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant Biol. 1: 404–411.
Reymond, P., Weber, H., Damond, M. and Farmer, E.E. 2000. Differential gene expression in response to mechanical wounding and insect feeding in Arabidopsis. Plant Cell 12: 707–719.
Rickauer, M., Brodschelm, W., Bottin, A., Veronesi, C., Grimal, H. and Esquerre-Tugaye, M.T. 1997. The jasmonate pathway is involved differentially in the regulation of different defense responses in tobacco cells. Planta 202: 155–162.
Riechmann, J.L., Heard, J., Martin, G., Reuber, L., Jiang, C.Z., Keddie, J., Adam, L., Pineda, O., Ratcliffe, O.J., Samaha, R.R., Creelman, R., Pilgrim, M., Broun, P., Zhang, J.Z., Ghandeharl, D., Sherman, B.K. and Yu, G.L. 2000. Arabidopsis transcription factor: genome-wide comparative analysis among eukaryotes. Science 290: 2105–2110.
Rogers, E.E. and Ausubel, E.M. 1997. Arabidopsis enhanced disease susceptibility mutant exhibit enhanced susceptibility to several bacterial pathogens and alterations in PR-1 gene expression. Plant Cell 9: 305–316.
Rushton, P.J. and Somssich, I.E. 1998. Transcriptional control of plant genes responsive to pathogens. Curr. Opin. Plant Biol. 1: 311–315.
Ryals, J.A., Neuenschwander, U.H., Willits, M.G., Molina, A., Steiner, H.Y. and Hunt, M.D. 1996. Systemic acquired resistance. Plant Cell 8: 1809–1819.
Ryan, C.A. 2000. The systemin signaling pathway: differential activation of plant defensive genes. Biochem. Biophys. Acta 1477: 112–121.
Sakamoto, A., Minami, M., Huh, G.H. and Iwabuchi, M., 1993. The putative zinc-finger protein WZF1 interacts with a cis-acting element of wheat histone genes. Eur. J. Biochem. 217: 1049–1056.
Sambrook, J. and Russel, D.W. 2001. Molecular Cloning: A Laboratory Manual, 3edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Somssich, I.E. and Hahlbrock, K. 1998. Pathogen defense in plants: a paradigm of biological complexity. Trends Plant Sci. 3: 86–90.
Sprenger-Haussels, M. and Weisshaar, B. 2000. Transactivation properties of parsley proline-rich bZIP transcription factors. Plant J. 22: 1–8.
Stanojevic, D., Hoey, T. and Levine, M. 1989. Sequence-specific DNA binding activities of the gap proteins encoded by hunchback and Kruppel in Drosophila. Nature 341: 331–335.
Sugano, S., Kaminaka, H., Rybka, Z., Catala, R., Salinas, J., Matsui, K., Ohme-Takagi, M. and Takatsuji, H. 2003. Stress-responsive zinc finger gene ZPT2-3 plays a role in drought tolerance in petunia. Plant J. 36: 830–841.
Takatsuji, H. 1998. Zinc-finger transcription factors in plants. Cell. Mol. Life Sci. 54: 582–596.
Takatsuji, H. and Matsumoto, T. 1996. Target-sequence recognition by separate-type Cys2/His2 zinc-finger proteins in plants. J. Biol. Chem. 271: 23368–23373.
Takatsuji, H., Mori, M., Benfey, P.N., Ren, L. and Chua, N.-H. 1992. Characterization of a zinc finger DNA-binding protein expressed specifically in Petunia petals and seedlings. EMBO J. 11: 241–249.
Takatsuji, H., Nakamura, N. and Katsumoto, Y. 1994. A new family of zinc finger proteins in petunia: structure, DNA sequence recognition and floral organ-specific expression. Plant Cell 6: 947–958.
Titarenko, E., Rojo, E., Leó, J. and Sánchez-Serrano, J.J. 1997. Jasmonic acid-dependent and -independent signaling pathways control wound-induced gene activation in Arabidopsis thaliana. Plant Physiol. 115: 817–826.
Ukness, S., Mauch-Mani, B., Moyer, M., Potter, S., Williams, S., Dincher, S., Chandler, D., Slusarenko, A., Ward, E. and Ryals, J. 1992 Acquired resistance in Arabidopsis. Plant Cell 4: 645–656.
van Der Krol, A.R., van Poecke, R.M., Vorst, O.F., Voogt, C., van Leeuwen, W., Borst-Vrensen, T.W., Takatsuji, H. and van Der Plas, L.H. 1999. Developmental and wound-, cold-, desiccation-, ultraviolet-B-stress-induced modulations in the expression of the petunia zinc finger transcription factor gene ZPT2-2. Plant Physiol. 121: 1153–1162.
Varet, A., Hause, B., Hause, G., Scheel, D. and Lee, J. 2003. The Arabidopsis NHL3 gene encodes a plasma membrane protein and its overexpression correlates with increased resistance to Pseudomonas syringae pv. tomato DC3000. Plant Physiol. 132: 2023–2033.
Ward, E.R., Uknes, S.J., Williams, S.C., Dincher, S.S., Wiederhold, D.L., Alexander, D.C., Ahl-Goy, P., Metraux, J. and Ryals, J. A. 1991. Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell 3: 1085–1094.
Yang, Y., Shah, J. and Klessig, D.F. 1997. Signal perception and transduction in plant defense responses. Genes. Dev. 11: 1621–1639.
Yun, J.Y., Weigel, D. and Lee, I.H. 2002. Ectopic expression of SUPERMAN suppresses development of petal and stamens. Plant Cell Physiol. 43: 52–57.
Zhou, J., Tang, X. and Martin, G.B. 1997. The Pto kinase conferring resistance to tomato bacterial speck disease interacts with proteins that bind a cis-element of pathogenesis-related genes. EMBO J. 16: 3207–3218.
Nucleotide sequence data are available in the EMBL database under Accession number AF539746 (CAZFP1).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, S.H., Hong, J.K., Lee, S.C. et al. CAZFP1, Cys2/His2-type zinc-finger transcription factor gene functions as a pathogen-induced early-defense gene in Capsicum annuum . Plant Mol Biol 55, 883–904 (2004). https://doi.org/10.1007/s11103-005-2151-0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11103-005-2151-0