Skip to main content
Log in

Ntlim1, a PAL-box binding factor, controls promoter activity of the horseradish wound-inducible peroxidase gene

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

To understand molecular mechanisms underlying wound-induced expression of plant peroxidase genes, the promoter of a horseradish C2 peroxidase (prxC2) gene was analyzed. We had previously isolated a tobacco nuclear protein, Ntlim1, as a trans factor binding to a PAL-box motif of the prxC2 promoter; however, the function of the Ntlim1 trans factor and the PAL-box motif in wound-responsive expression of the prxC2 gene remains unclear. Here, we found that the prxC2 promoter without the intact PAL-box motif failed to direct a normal level of both the basal and the wound-induced expression of β-glucuronidase (GUS) reporter gene in transgenic tobacco plants, indicating that the PAL-box motif functions as an essential cis element of the prxC2 promoter. We also found that antisense expression of Ntlim1 in transgenic plants carrying the prxC2 promoter::GUS chimeric construct decreased not only the level of the basal and the wound-induced expression of the GUSreporter gene but also the extent of wound inducibility of the prxC2 promoter itself. This result indicates that Ntlim1 is required for the basal level of prxC2 promoter activity as well as its up-regulation under wound stress. Moreover, consistent with the results obtained in planta, result from super-shift assay indicates that the Ntlim1 binds to the PAL-box motif independently of wound stress.

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

  • Arias, J.A., Dixon, R.A. and Lamb, C.J. 1993. Dissection of the functional architecture of a plant defense gene promoter using a homologous in vitro transcription initiation system. Plant Cell 5: 485–496.

    Google Scholar 

  • Baker, C.J., Deahl, K., Domek, J. and Orlandi, E.W. 2000. Scavenging of H2O2 and production of oxygen by horseradish peroxidase. [In process citation.] Arch. Biochem. Biophys. 382: 232–237.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Church, G.M. and Gilbert, W. 1984. Genomic sequencing. Proc. Natl. Acad. Sci. USA 81: 1991–1995.

    Google Scholar 

  • Clarke, H.R., Davis, J.M., Wilbert, S.M., Bradshaw, H.D. Jr. and Gordon, M.P. 1994. Wound-induced and developmental activation of a poplar tree chitinase gene promoter in transgenic tobacco. Plant Mol. Biol. 25: 799–815.

    Google Scholar 

  • Cramer, C.L., Edwards, K., Dron, M., Liang, X., Dildine, S.L., Bolwell, G.P., Dixon, R.A., Lamb, J. and Schuch, W. 1989 Phenylalanine ammonia-lyase gene organization and structure. Plant Mol. Biol. 12: 367–383.

    Google Scholar 

  • Curtis, M.D., Rae, A.L., Rusu, A.G., Harrison, S.J. and Manners, J.M. 1997. A peroxidase gene promoter induced by phytopathogens and methyl jasmonate in transgenic plants. Mol. Plant-Microbe Interact. 10: 326–338.

    Google Scholar 

  • da Costa e Silva, O., Klein, L., Schmelzer, E., Trezzini, G.F. and Hahlbrock, K. 1993. BPF-1, a pathogen-induced DNA-binding protein involved in the plant defense response. Plant J. 4: 125–135.

    Google Scholar 

  • Dawid, I.B., Toyama, R. and Taira, M. 1995. LIM domain proteins. C.R. Acad. Sci. III 318: 295–306.

    Google Scholar 

  • De Leo, F., Ceci, L.R., Jouanin, L. and Gallerani, R. 2001. Analysis of mustard trypsin inhibitor-2 gene expression in response to developmental or environmental induction. Planta 212: 710–717.

    Google Scholar 

  • Douglas, C., Hoffman, H., Schulz, W. and Hahlbrock, K. 1987. Structure and elicitor or UV-light stimulated expression of two 4-coumarate:CoA ligase genes in parsley. EMBO J. 6: 1189–1195.

    Google Scholar 

  • Droge-Laser, W., Kaiser, A., Lindsay, W.P., Halkier, B.A., Loake, G.J., Doerner, P., Dixon, R.A. and Lamb, C. 1997. Rapid stimulation of a soybean protein-serine kinase that phosphorylates a novel bZIP DNA-binding protein, G/HBF-1, during the induction of early transcription-dependent defenses. EMBO J. 16: 726–738.

    Google Scholar 

  • Dron, M., Clouse, S.D., Dixon, R.A., Lawton, M.A. and Lamb, C.J. 1988 Glutathione and fungal elicitor regulation of a plant defense gene promoter in electroporated protoplasts. Proc. Natl. Acad. Sci. USA 85: 6738–6742.

    Google Scholar 

  • Eyal, Y., Sagee, O. and Fluhr, R. 1992. Dark-induced accumulation of a basic pathogenesis-related (PR-1) transcript and a light requirement for its induction by ethylene. Plant Mol. Biol. 19: 589–599.

    Google Scholar 

  • Feuillet, C., Lauvergeat, V., Deswarte, C., Pilate, G., Boudet, A. and Grima-Pettenati, J. 1995. Tissue-and cell-specific expression of a cinnamyl alcohol dehydrogenase promoter in transgenic poplar plants. Plant Mol. Biol. 27: 651–667.

    Google Scholar 

  • Guevara-Garcia, A., Lopez-Ochoa, L., Lopez-Bucio, J., Simpson, J. and Herrera-Estrella, L. 1998. A 42 bp fragment of the pmas1' promoter containing an ocs-like element confers a developmental, wound-and chemically inducible expression pattern. Plant Mol. Biol. 38: 743–753.

    Google Scholar 

  • Hara, K., Yagi, M., Kusano, T. and Sano, H. 2000. Rapid systemic accumulation of transcripts encoding a tobacco WRKY transcription factor upon wounding. Mol. Gen. Genet. 263: 30–37.

    Google Scholar 

  • Hatton, D., Sablowski, R., Yung, M.H., Smith, C., Schuch, W. and Bevan, M. 1995. Two classes of cis sequences contribute to tissue-specific expression of a PAL2 promoter in transgenic tobacco. Plant J. 7: 859–876.

    Google Scholar 

  • Hiraga, S., Ito, H., Sasaki, K., Yamakawa, H., Mitsuhara, I., Toshima, H., Matsui, H., Honma, M. and Ohashi, Y. 2000. Wound-induced expression of a tobacco peroxidase is not enhanced by ethephon and suppressed by methyl jasmonate and coronatine. fication of a basic glycoprotein induced by ethylene in primary leaves of azuki bean as a cationic peroxidase. Plant Physiol. 101: 193–199.

    Google Scholar 

  • Ito, H., Kimizuka, F., Ohbayashi, A., Matsui, H., Honma, M., Shinmyo, A., Ohashi, Y., Caplan, A.B. and Rodrigues, R.L. 1994. Molecular cloning and characterization of two complementary DNAs encoding putative peroxidases from rice (Oryza sativa L.) shoots. Plant Cell Rep. 13: 361–366.

    Google Scholar 

  • Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: ?-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.

    Google Scholar 

  • Kanofsky, J.R. 1988. Singlet oxygen production from the peroxidase-catalyzed oxidation of indole-3-acetic acid. J. Biol. Chem. 263: 14171–14175.

    Google Scholar 

  • Kaothien, P., Shimokawatoko, Y., Kawaoka, A., Yoshida, K. and Shinmyo, A. 2000. A cis-element containing PAL-box functions in the expression of the wound-inducible peroxidase gene of horseradish. Plant Cell Rep. 19: 558–562.

    Google Scholar 

  • Kawaoka, A., Shinichi, S., Nakahara, K. Matsushima, N., Okada, N., Sekine, M., Shinmyo, A. and Takano, M. 1992. Expression and promoter activity of genes for isozymes of horseradish peroxidase. Plant Cell Physiol. 33: 1143–1150.

    Google Scholar 

  • Kawaoka, A., Kawamoto, T., Ohta, H., Sekine, M., Takano, M. and Shinmyo, A. 1994a. Wound-induced expression of horseradish peroxidase. Plant Cell Rep. 13: 149–154.

    Google Scholar 

  • Kawaoka, A., Kawamoto, T., Sekine, M., Yoshida, K., Takano, M., and Shinmyo, A. 1994b. A cis-acting element and a trans-acting factor involved in the wound-induced expression of a horseradish peroxidase gene. Plant J. 6: 87–97.

    Google Scholar 

  • Kawaoka, A., Kaothien, P., Yoshida, K., Endo, S., Yamada, K. and Ebinuma, H. 2000. Functional analysis of tobacco LIM protein Ntlim1 involved in lignin biosynthesis. Plant J. 22: 289–301.

    Google Scholar 

  • Lois, R., Dietrich, A., Hahlbrock, K. and Schulz, W. 1989. A phenylalanine ammonia-lyase gene from parsley: structure, regulation and identification of elicitor and light responsive cis-acting elements. EMBO J. 8: 1641–1648.

    Google Scholar 

  • Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • Mohan, R., Bajar, A.M. and Kolattukudy, P. E. 1993. Induction of a tomato anionic peroxidase gene (tap1) by wounding in transgenic tobacco and activation of tap1/GUS and tap2/GUS chimeric gene fusions in transgenic tobacco by wounding and pathogen attack. Plant Mol. Biol. 21: 341–354.

    Google Scholar 

  • Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 18: 473–479.

    Google Scholar 

  • Nakayama, H., Yoshida, K., Ono, H., Murooka, Y. and Shinmyo, A. 2000. Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells. Plant Physiol. 122: 1239–1247.

    Google Scholar 

  • Nishiuchi, T., Kodama, H., Yanagisawa, S. and Iba, K. 1999. Wound-induced expression of the FAD7 gene is mediated by different regulatory domains of its promoter in leaves/stems and roots. Plant Physiol. 121: 1239–1246.

    Google Scholar 

  • Ohl, S., Hedrick, S.A., Chory, J. and Lamb, C.J. 1990. Functional properties of a phenylalanine ammonia-lyase promoter from Arabidopsis. Plant Cell 2: 837–848.

    Google Scholar 

  • Palm, C.J., Costa, M.A., An, G. and Ryan, C.A. 1990. Woundinducible nuclear protein binds DNA fragments that regulate a proteinase inhibitor II gene from potato. Proc. Natl. Acad. Sci. USA 87: 603–607.

    Google Scholar 

  • Quiroga, M., Guerrero, C., Botella, M.A., Barcelo, A., Amaya, I., Medina, M.I., Alonso, F.J., de Forchetti, S.M., Tigier, H. and Valpuesta, V. 2000. A tomato peroxidase involved in the synthesis of lignin and suberin. Plant Physiol. 122: 1119–1127.

    Google Scholar 

  • Roberts, E. and Kolattukudy, P.E. 1989. Molecular cloning, nucleotide sequence, and abscisic acid induction of a suberizationassociated highly anionic peroxidase. Mol. Gen. Genet. 217: 223–232.

    Google Scholar 

  • Rogers, S.G., Horsch, R.B. and Fraley, R.T. 1986. Gene transfer in plant: production of transformed plant using Ti plasmid vectors. Meth. Enzymol. 118: 627–640.

    Google Scholar 

  • Ruiz-Rivero, O.J. and Prat, S. 1998. A-308 deletion of the tomato LAP promoters is able to direct flower-specific and MeJa-induced expression in transgenic plants. Plant Mol. Biol. 36: 639–648.

    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.

    Google Scholar 

  • Sablowski, R.W.M., Baulcombe, D.C. and Bevan, M. 1995. Expression of a flower-specific MYB protein in leaf cells using a viral vector causes ectopic activation of a target promoter. Proc. Natl. Acad. Sci. USA 92: 6901–6905.

    Google Scholar 

  • Siebertz, B., Logemann, J., Willmitzer, L. and Schell, J. 1989. cis-analysis of the wound-inducible promoter wun1 in transgenic tobacco plants and histochemical localization of its expression. Plant Cell 1: 961–968.

    Google Scholar 

  • Staiger, D., Kaulen, H. and Schell, J. 1989. A CACGTG motif of the Antirrhinum majus chalcone synthase promoter is recognized by an evolutionarily conserved nuclear protein. Proc. Natl. Acad. Sci. USA 86: 6930–6934.

    Google Scholar 

  • Stankovic, B., Vian, A., Henry-Vian, C. and Davies, E., 2000. Molecular cloning and characterization of a tomato cDNA encoding a systemically wound-inducible bZIP DNA-binding protein. Planta 212: 60–66.

    Google Scholar 

  • Sugimoto, K., Takeda, S. and Hirochika, H. 2000. MYB-related transcription factor NtMYB2 induced by wounding and elicitors is a regulator of the tobacco retrotransposon Tto1 and defenserelated genes. Plant Cell 12: 2511–2528.

    Google Scholar 

  • Wadman, I.A., Osada, H., Grutz, G.G., Agulnick, A.D., Westphal, H., Foster, A. and Rabbitts, T.H. 1997. The LIM-only protein Lmo2 is a bridging molecule assembling an erythroid, DNAbinding complex which includes the TAL1, E47, GATA-1 and Ldb1/NL1 proteins. EMBO J. 16: 3145–3157.

    Google Scholar 

  • Yamada, T., Sriprasertsak, P., Kato, H., Hashimoto, T., Shimizu, H. and Shiraishi, T. 1994. Functional analysis of the promoters of phenylalanine ammonia-lyase genes in pea. Plant Cell Physiol. 35: 917–926.

    Google Scholar 

  • Yu, L.M., Lamb, C.J. and Dixon, R.A. 1993. Purification and biochemical characterization of proteins which bind to H-box cis-element implicated in transcriptional activation of plant defence gene. Plant J. 3: 805–816.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kaothien, P., Kawaoka, A., Ebinuma, H. et al. Ntlim1, a PAL-box binding factor, controls promoter activity of the horseradish wound-inducible peroxidase gene. Plant Mol Biol 49, 591–599 (2002). https://doi.org/10.1023/A:1015504515492

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015504515492

Navigation