Abstract
Pepper (Capsicum annuum) plants exhibit hypersensitive response (HR) against infection by many tobamoviruses. A clone encoding a putative nonspecific lipid transfer protein (CaLTP1) was isolated by differential screening of a cDNA library from resistant pepper leaves when inoculated with tobacco mosaic virus (TMV) pathotype P0. The predicted amino acid sequence of CaLTP1 is highly similar to that of the other plant LTPs. Southern blot analysis showed that a small gene family of LTP-related sequences was present in the pepper genome. Transcripts homologous to CaLTP1 accumulated abundantly in old leaves and flowers. CaLTP1 expression was induced in the incompatible interaction with TMV-P0 but was not induced in the compatible interaction with TMV-P1.2. In correlation with the temporal progression of HR in the inoculated leaves, CaLTP1 transcripts started to accumulate at 24 h after TMV-P0 inoculation, reaching a maximal level at 48 h. A strain of Xanthomonas campestris pv. vesicatoria (Xcv) that carries the bacterial avirulence gene, avrBs2, was infiltrated into leaves of a pepper cultivar containing the Bs2 resistance gene. A marked induction of CaLTP1 expression was observed in Xcv-infiltrated leaves. Effects of exogenously applied abiotic elicitors on CaLTP1 expression were also examined. Salicylic acid caused a rapid accumulation of CaLTP1 transcripts in pepper leaves and ethephon treatment also induced the expression of the CaLTP1 gene. Transient expression in the detached pepper leaves by biolistic gene bombardment indicated that CaLTP1 is localized mostly at the plant cell surface, possibly in the cell wall. These results suggest possible role(s) for LTPs in plant defense against pathogens including viruses.
Similar content being viewed by others
References
Altschul, S.F., Gish, W., Miller, W., Myers, E.W. and Lipman, D.J. 1990. Basic local alignment search tool. J. Mol. Biol. 215: 403–410.
Ashfield, M., Hammond-Kosack, K.E., Harrison, K. and Joned, J.D.G. 1994. Cf gene-dependent induction of a ?-1,2-glucanase promoter in tomato plants infected with Cladosporium fulvum. Mol. Plant-Microbe Interact. 7: 645–657.
Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 1995. Short Protocols in Molecular Biology. John Wiley, New York.
Bernhard, W.R. and Somerville, C.R. 1989. Coidentity of putative amylase inhibitors from barley and finger millet with phospholipid transfer proteins inferred from amino acid sequence homology. Arch. Biochem. Biophys. 269: 695–697.
Berzal-Herranz, A., de la Cruz, A., Tenllado, F., Diaz-Ruiz, J.R., Lopez, L., Sanz, A.I., Vaquero, C., Serra, M.T. and Garcia-Luque, I. 1995. The Capsicum L3 gene-mediated resistance against the tobamoviruses is elicited by the coat protein. Virology 209: 498–505.
Boller, T. and Metraux, J.P. 1988. Extracellular localization of chitinase in cucumber. Physiol. Mol. Plant Path. 33: 11–16.
Boukema, I.W. 1982. Resistance to TMV in Capsicum chacoense Hunz. is governed by an allele of the L-locus. Capsicum Newsl. 3: 47–48.
Brederode, F.T., Linthorst, H.J.M. and Bol, J.F. 1991. Differential induction of acquired resistance and PR gene expression in tobacco by virus infection, ethephon treatment, UV light and wounding. Plant Mol. Biol. 17: 1117–1125.
Creelman, R.A. and Mullet, J.E. 1997. Biosynthesis and action of jasmonates in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 355–381.
Davis, S.J. and Vierstra, R.D. 1996. Soluble derivatives of green fluorescent protein (GFP) for use in Arabidopsis thaliana. Weeds World 3: 43–48.
Davis, S.J. and Vierstra, R.D. 1998. Soluble, highly fluorescent variants of green fluorescent protein (GFP) for use in higher plants. Plant Mol. Biol. 36: 521–528.
Dong, X., Mindrinos, M., Davis, K.R. and Ausubel, F.M. 1991. Induction of Arabidopsis defense genes by virulent and avirulent Pseudomonas syringae strain and by a cloned avirulence gene. Plant Cell 8: 155–168.
Durner, J., Shah, J. and Klessig, D.F. 1997. Salicylic acid and disease resistance in plants. Trends Plant Sci. 2: 266–274.
Ecker, J.R. 1995. The ethylene signal transduction pathway in plants. Science 268: 667–675.
Fleming, A.J., Mandel, T., Hofmann, S., Sterk, P., de Vries, S.C. and Kuhlemeier, C.P. 1992. Expression pattern of a tobacco lipid transfer protein gene within the shoot apex. Plant J. 2: 855–862.
García-Olmedo, F., Molina, A., Segura, A. and Moreno, M. 1995. The defensive role of nonspecific lipid-transfer proteins in plants. Trends Microbiol. 3: 72–74.
Gausing, K. 1994. Lipid transfer protein genes specifically expressed in barley leaves and coleoptiles. Planta 192: 574–580.
Green, T.R. and Ryan, C.A. 1972. Wound-induced proteinase inhibitors in plant leaves. Science 175: 776–777.
Hughes, M.A., Dunn, M.A., Pearce, RS., White, A.J. and Zhang, L. 1992. An abscisic acid-responsive, low temperature barley gene has homology with a maize phospholipid transfer protein. Plant Cell Environ. 15: 861–865.
Itaya, A., Hickman, H., Bao, Y., Nelson, R. and Ding, B. 1997. Cell-to-cell trafficking of cucumber mosaic virus movement pro254 tein: green fluorescent protein fusion produced by biolistic gene bombardment in tobacco. Plant J. 12: 1223–1230.
Jung, H.W. and Hwang, B.K. 2000. 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.
Kader, J.C. 1996. Lipid-transfer proteins in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 627–654.
Kim, Y.J. and Hwang, B.K. 2000. Pepper gene encoding a basic pathogenesis-related 1 protein is pathogen and ethylene inducible. Physiol. Plant. 108: 51–60.
Kinlaw, C.S., Gerttulla, S.M. and Carter, M.C. 1994. Lipid transfer protein genes of loblolly pine are members of a complex gene family. Plant Mol. Biol. 26: 1213–1216.
Lamb, C. and Dixon, R.A. 1997. The oxidative burst in plant disease resistance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 251–275.
Lawton, K.A., Potter, S.L., Uknes, S. and Ryals, J. 1994. Acquired resistance signal transduction in Arabidopsis is ethylene independent. Plant Cell 6: 581–588.
Leon, J., Yalpani, N., Raskin, I. and Lawton, M.A. 1993. Induction of benzoic acid 2-hydroxylase in virus-inoculated tobacco. Plant Physiol. 103: 323–328.
Memelink, J., Linthorst, H.J.M., Schilperoort, R.A. and Hoge, J.H.C. 1990. Tobacco genes encoding acidic and basic pathogenesis-related proteins display different expression patterns. Plant Mol. Biol. 14: 119–126.
Molina, A. and Garcia-Olmedo, F. 1993. Developmental and pathogen induced expression of three barley genes encoding lipid transfer proteins. Plant J. 4: 983–991.
Molina, A. and Garcia-Olmedo, F. 1997. Enhanced tolerance to bacterial pathogens caused by the transgenic expression of barley lipid transfer protein LTP2. Plant J. 12: 669–675.
Molina, A., Segura, A. and Garcia-Olmedo F. 1993. Lipid transfer proteins (nsLTPs) from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens. FEBS Lett. 316: 119–122.
Molina, A., Diaz, I., Vasil, I.K., Carbonero, P. and Garcia-Olmedo, F. 1996. Two cold-inducible genes encoding lipid transfer protein LTP4 from barley show differential responses to bacterial pathogens. Mol. Gen. Genet. 252: 162–168.
Mundy, J. and Rogers, J.C. 1986. Selective expression of a probable amylase/protease inhibitor in barley aleurone cells. Planta 169: 51–63.
Park, C.-J., Shin, R., Park, J.M., Lee, G.-J., Yoo, T.H. and Paek, K.-H. 2001. A hot pepper cDNA encoding a pathogenesis-related protein 4 is induced during the resistance response to Tobacco Mosaic Virus. Mol. Cells 11: 122–127.
Penninckx, I.A., Eggermont, K., Terras, F.R., Thomma, B.P., De Samblanx, G.W., Buchala, A., Métraux, J.P., Manners, J.M. and Broekaert, W.F. 1996. Pathogen-induced systemic activation of a plant defensin gene in Arabidopsis follows a salicylic acid-independent pathway. Plant Cell 8: 2309–2323.
Plant, A.L., Cohen, A., Moses, M.S. and Bray, E.A. 1991. Nucleotide sequence and spatial expression pattern of a droughtand abscisic acid-induced gene of tomato. Plant Physiol. 97: 900–906.
Poirier, G.M.-C., Pyati, J., Wan J.S. and Erlander, M.G. 1997 Screening differentially expressed cDNA clones obtained by differential display using amplified RNA. Nucl. Acids Res. 25: 913–914.
Reuber, T.L. and Ausubel, F.M. 1996. Isolation of Arabidopsis genes that differentiate between resistance genes. Plant Cell 8: 241–249.
Reymond, P. and Farmer, E.E. 1998. Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant Biol. 1: 404–411.
Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.
Sanger, F., Nicklen, S. and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.
Sohal, A.K., Love, A.J., Cecchini, E., Covey, S.N., Jenkins, G.I. and Milner, J.J. 1999a. Cauliflower mosaic virus infection stimulates lipid transfer protein gene expression in Arabidopsis. J. Exp. Bot. 50: 1727–1733.
Sohal, A.K., Pallas, J.A. and Jenkins, G.I. 1999b. The promoter of a Brassica napus lipid transfer protein gene is active in a range of tissues and stimulated by light and viral infection in transgenic Arabidopsis. Plant Mol. Biol. 41: 75–87.
Sterk, P., Booij, H., Schellekens, G.A., van Kammen, A. and de Vries, S.C. 1991. Cell-specific expression of the carrot EP2 lipid transfer protein gene. Plant Cell 3: 907–921.
Sticher, L., Mauch-Mani, B. and Metraux, J.P. 1997. Systemic acquired resistance. Annu. Rev. Phytopath. 35: 235–270.
Tai, T.H., Dahlbeck, D., Clark. E.T., Gajiwala, P., Pasion, R., Whalen, M.C., Stall, R.E. and Staskawicz, B.J. 1999. Expression of the Bs2 pepper gene confers resistance to bacterial spot disease in tomato. Proc. Natl. Acad. Sci. USA 96: 14153–14158.
Takeuchi, Y., Dotson, M. and Keen, N.T. 1992. Plant transformation: a simple particle bombardment device based on flowing helium. Plant Mol. Biol. 18: 835–839.
Thoma, S., Kaneto, Y. and Somerville, C.R. 1993. A nonspecific lipid transfer protein from Arabidopsis is a cell wall protein. Plant J. 3: 427–436.
Thoma, S., Hecht, U., Kippers, A., Botella, J., de Vries, S. and Somerville, S. 1994. Tissue-specific expression of a gene encoding a cell wall-localised lipid transfer protein from Arabidopsis. Plant Physiol. 105: 35–45.
Torres-Schumann, S., Godoy, J.A. and Pintor-Toro, J.A. 1992. A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants. Plant Mol. Biol. 18: 749–757.
Tsuboi, S., Osafune, T., Tsugeki, R., Nishimura, M. and Yamada, M. 1992. Nonspecific lipid transfer protein in castor bean cotyledon cells: subcellular localization and a possible role in lipid metabolism. J. Biochem. 111: 500–508.
Vignols, F., Wigger, M., Garcia-Garrido, J.M., Grellet, F., Kader, J.C. and Delseny, M. 1997. Rice lipid transfer protein (LTP) genes belong to a complex multigene family and are differentially regulated. Gene 195: 177–186.
White, A., Dunn, M.A., Brown, K. and Hughes, M.A. 1994. Comparative analysis of genomic sequence and expression of a lipid transfer protein gene family in winter barley. J. Exp. Bot. 45: 1885–1892.
Wirtz, K.W.A. 1991. Phospholipid transfer proteins. Annu. Rev. Biochem. 60: 73–99.
Zhang, H., Zhang, R. and Liang, P. 1996. Differential screening of gene expression difference enriched by differential display. Nucl. Acids Res. 24: 2454–2455.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Park, CJ., Shin, R., Park, J.M. et al. Induction of pepper cDNA encoding a lipid transfer protein during the resistance response to tobacco mosaic virus. Plant Mol Biol 48, 243–254 (2002). https://doi.org/10.1023/A:1013383329361
Issue Date:
DOI: https://doi.org/10.1023/A:1013383329361