Skip to main content
Log in

Molecular cloning of a novel pathogen-inducible cDNA encoding a putative acyl-CoA synthetase from Capsicum annuum L.

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

By means of differential display, a pool of salicylic acid (SA)-induced mRNAs were identified and subsequently their cDNAs were isolated from a cDNA library prepared from SA-induced leaf tissues of hot pepper. One of these cDNA clones, designated CaSIG4, was 1900 bp and contained an open reading frame encoding 523 amino acids with a calculated molecular mass of 56.3 kDa. The predicted amino acid sequence of CaSIG4 showed high sequence similarity to the AMP-binding protein family of both prokaryotic and eukaryotic acyl-CoA synthetases. CaSIG4 transcripts accumulated rapidly after SA treatment and in response to both incompatible and compatible interactions with Xanthomonas campestris pv. vesicatoria race 1. To investigate the cis-acting elements mediating CaSIG4 expression, the CaSIG4 5′-flanking region was isolated by inverse PCR. Database searches indicated that a potential cis-regulatory element is almost identical to the consensus core sequences ACC(A/T)ACC(A/C) which are conserved among promoters of other phenylpropanoid biosynthetic genes. The subcellular localization of the CaSIG4 protein was studied by using a soluble modified GFP gene fusion delivered into epidermal cells of onion by biolistic bombardment. The CaSIG4-smGFP fusion protein was localized to the plasma membrane. Taken together, CaSIG4 encoding a putative acyl-CoA synthetase could function as a plasma membrane-bound protein with a role in signaling in plant defense.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Blobel, F. and Erdmann, R. 1996. Identification of a yeast peroxisomal member of the family of AMP-binding proteins. Eur. J. Biochem. 240: 468–476.

    PubMed  Google Scholar 

  • Casey, P.J. 1995. Protein lipidation in cell signaling. Science 268: 221–225.

    Google Scholar 

  • 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 mosaic virus infection. Plant Physiol. 112: 353–359.

    PubMed  Google Scholar 

  • 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.

    Google Scholar 

  • David, S.J. and Vierstra, R.D. 1996. Soluble derivatives of green fluorescent protein (GFP) for use in Arabidopsis thaliana. Weeds World 3: 43–48.

    Google Scholar 

  • Delaney, T.P., Uknes, S.J., Vernooij, B., Friedrich, L., Weymann, K., Negrotto, D., Gaffney, T., Gut-Rella, M., Kessmann, H., Ward, E.R. and Ryals, J.A. 1994. A central role of salicylic acid in plant disease resistance. Science 266: 1247–1250.

    Google Scholar 

  • Douglas, C.J. 1996. Phenylpropanoid metabolism and lignin biosynthesis: from weeds to trees. Trends Plant Sci. 1: 171–178.

    Google Scholar 

  • Durner, J., Shah, J. and Klessig, D.F. 1997. Salicylic acid and disease resistance in plants. Trends Plant Sci. 2: 266–274.

    Google Scholar 

  • Ecker, J.R. 1995. The ethylene signal transduction pathway in plants. Science 268: 667–675.

    PubMed  Google Scholar 

  • Fang, G., Hammar, S. and Grumet, R. 1992. A quick and inexpensive method for removing polysaccharides from plant genomic DNA. BioTechniques 13: 52–54.

    PubMed  Google Scholar 

  • Feinberg, A.P. and Vogelstein, B. 1983. A technique for radiolabeling a DNA restriction endonuclease fragment to high specific activity. Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Feuillet, C. 1995. Tissue-and cell-specific expression of a cinnamyl alcohol dehydrogenase promoter in transgenic poplar trees. Plant Mol. Biol. 27: 651–667.

    PubMed  Google Scholar 

  • Fulda, M., Heinz, E. and Wolter, F.P. 1997. Brassica napus cDNAs encoding fatty acyl-CoA synthetase. Plant Mol. Biol. 33: 911–922.

    Google Scholar 

  • Gaffney, T., Friedrich, L., Vernooij, B., Negrotto, D., Nye, G., Uknes, S.J., Ward, E.R., Kessmann, H. and Ryals, J.A. 1993. Requirement of salicylic acid for the induction of systemic acquired resistance. Science 261: 754-756.

    Google Scholar 

  • Gould, S.J., Keller, J.A., Schneider, M., Howell, S.H., Garrard, L.J., Goodman, J., Distel, B., Tabak, H. and Subramani, S. 1990. Peroxisomal protein import is conserved between yeast, plants, insects and mammals. EMBO J. 9: 85–90.

    PubMed  Google Scholar 

  • Grotewald, E., Drummond, B.J., Bowen, B. and Peterson, T. 1994. The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell76: 543–553.

    Article  PubMed  Google Scholar 

  • Hauffe, K.D., Paszkowski, U., Schulze-Lefert, P., Hahlbrock, K., Dangl, J.L. and Douglas, C.J. 1991. A parsley 4CL-1 promoter fragment specifies complex expression patterns in transgenic tobacco. Plant Cell 3: 435–443.

    Article  PubMed  Google Scholar 

  • Lamb, C. and Dixon, R.A. 1997. The oxidative burst in plant disease resistance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 251–275.

    PubMed  Google Scholar 

  • Liang, P. and Pardee, A.B. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257: 967–971.

    PubMed  Google Scholar 

  • Logemann, E., Parniske, M. and Hahlbrock, K. 1995. Modes of expression and common structural features of the complete phenylalanine ammonia-lyase gene family in parsley. Proc. Natl. Acad. Sci. USA 92: 5905–5909.

    PubMed  Google Scholar 

  • Reymond, P. and Farmer, E.E. 1998. Jasmonate and salicylate as global signals for defense gene expression. Curr. Opin. Plant Biol. 1: 404–411.

    PubMed  Google Scholar 

  • Rogers, S., Wells, R. and Rechsteiner, M. 1986. Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234: 364–368.

    PubMed  Google Scholar 

  • Rushton, P.J. and Somssich, I.E. 1998. Transcriptional control of plant genes responsive to pathogens. Curr. Opin. Plant Biol. 1: 311–315.

    PubMed  Google Scholar 

  • Sablowski, R.W.M., Moyano, E., Culianez-Macia, F.A., Schuch, W., Martin, C. and Bevan, M. 1994. A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. EMBO J. 13: 128–137.

    PubMed  Google Scholar 

  • Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • 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.

    Google Scholar 

  • van den Ackerveken, G., Marois, E. and Bonas, U. 1996. Recognition of the bacterial avirulence protein AvrBs3 occurs inside the host plant cell. Cell 87: 1307–1316.

    PubMed  Google Scholar 

  • Ward, E.R., Uknes, S.J., Williams, S.C., Dincher, S.S., Wiederhold, D.L., Alexander, D.C., Ahl-Goy, P., Métraux, J.P. and Ryals, J.A. 1991. Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell 3: 1085–1094.

    Article  PubMed  Google Scholar 

  • White, R.F. 1979. Acetylsalicylic acid (aspirin) induces resistance to TMV in tobacco. Virology 99: 410–412.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lee, S.J., Suh, MC., Kim, S. et al. Molecular cloning of a novel pathogen-inducible cDNA encoding a putative acyl-CoA synthetase from Capsicum annuum L.. Plant Mol Biol 46, 661–671 (2001). https://doi.org/10.1023/A:1011677028605

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1011677028605

Navigation