Plant Molecular Biology

, Volume 47, Issue 5, pp 641–652 | Cite as

The promoter of a basic PR1-like gene, AtPRB1, from Arabidopsis establishes an organ-specific expression pattern and responsiveness to ethylene and methyl jasmonate

  • Marjorie Santamaria
  • Catherine J. Thomson
  • Nick D. Read
  • Gary J. Loake


Antimicrobial proteins are a key feature underlying the deployment of both pre-formed and inducible defence responses. Probably the most well characterised class are the pathogenesis-related (PR) proteins, which are found in both basic and acidic isoforms. Here we describe the isolation and characterisation of a gene, designated AtPRB1, encoding a basic PR1-like protein from Arabidopsis. This protein showed high amino acid sequence identity with basic and acidic PR1 proteins from other plant species, for example PRB1 from Nicotiana tabacum and PR1 from Brassica napus, at 64% and 78% identity respectively. A genomic DNA fragment containing 2345 bp upstream from the putative transcriptional start site was fused to the gene encoding the luciferase (LUC) gene from Photinus pyralis in order to test for promoter activity. The resulting construct was transformed into Arabidopsis accession Col-0 and analysis of LUC activity, using an ultra-low-light imaging camera system, revealed that the AtPRB1 promoter established an exquisite organ-specific expression pattern. LUC activity was observed in flowers, stems and roots but not in leaf tissue. Superimposed upon this organ-specific expression pattern was responsiveness, in root tissue, to ethylene and methyl jasmonate (MeJA), important cues during the establishment of plant disease resistance. In contrast, AtPRB1::LUC gene expression was repressed in response to salicylic acid treatment. Analysis of a limited series of AtPRB1 5′-promoter deletion mutants, identified a number of promoter regions important for both the establishment of organ-specific expression and responsiveness to ethylene and MeJA. While AtPRB1 gene expression was not induced in response to an avirulent isolate of Peronospora parasitica in leaf tissue, this gene may contribute to horizontal resistance in other tissues and/or to MeJA- and ethylene-dependent defence responses engaged against necrotrophic pathogens in root tissue. It is anticipated that transgenic plants containing AtPRB1-based promoter::reporter constructs will provide useful tools for the future dissection of the cognate signalling networks regulating the expression of this gene.

disease resistance ethylene methyl jasmonate PR proteins 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alexander, D., Goodman, R.M., Gut-Rela, M., Glascock, C., Weymann, K., Friedrich, L., Maddox, D., Ahl Goy, P., Luntz, T., Ward, E. and Ryals, J. 1993. Increased tolerance to two oomycete pathogens in transgenic tobacco expressing pathogenesis related protein 1a. Proc. Natl. Acad. Sci. USA 90: 7327–7331.Google Scholar
  2. Altschul, S.F., Boguski, M.S., Gish, W., Miller, Meyers, E.W. and Lipman, D.J. 1990. Basic local alignment search tool. J. Mol. Biol. 215: 403–410.Google Scholar
  3. Brederode, F.T., Linthorst, H. 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.Google Scholar
  4. Bowling, S.A., Clarke, J.D., Liu, Y., Klessig, D.F. and Dong, X. 1997. The cpr5 mutant of Arabidopsis expresses both NPR1-dependent and NPR1-independent resistance. Plant Cell 9: 1573–1584.Google Scholar
  5. 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: 284–254.Google Scholar
  6. Brady, J.D. and Fry, S. 1997. Formation of di-isodityrosine and loss of isodityrosine in the cell walls of tomato cell-suspension cultures treated with fungal elicitors or H2O2. Plant Physiol. 115: 87–92.Google Scholar
  7. Broekaert, W.F., Terras, F.R.G., Cammue, B. and Osborn, R.W. 1995. Plant defensins: novel antimicrobial peptides as components of the host defence system. Plant Physiol. 108: 1353–1358.Google Scholar
  8. 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.Google Scholar
  9. Cornelissen, B.J.C. Horowitz, J., van Kan, J.A.L., Goldberg, R.B. and Bol, J.F. 1997. Structure of tobacco genes encoding pathogenesis-related proteins from the PR-1 group. Nucl. Acids Res. 15: 6799–6811.Google Scholar
  10. Cutt, J.R. and Klessig, D.F. 1992. Pathogenesis related proteins. In: T. Boller and F. Meins Jr. (Eds.) Genes Involved in Plant Defence. Springer-Verlag, Vienna, Austria, pp. 209–243.Google Scholar
  11. de Jong, A.J., Cordewener, J., Lo Schiavo, F., Terzi, M., Van-dekerckhove, J., van Kammen, A. and de Vries, S.C. 1992. A carrot somatic embryo mutant is rescued by chitinase. Plant Cell 4: 425–433.Google Scholar
  12. 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
  13. 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
  14. Eyal, Y., Meller, Y., Lev-Yadun, S. and Fluhr, R. 1993. A basic-type PR-1 promoter directs ethylene responsiveness, vascular and abcission zone-specific expression. Plant J. 4: 225–234.Google Scholar
  15. Faktor, O., Loake, G., Dixon, R.A. and Lamb, C.J. 1997. The G-box and H-box in a 39 bp region of a French bean chalcone synthase promoter constitute a tissue-specific regulatory element. Plant J. 11: 1105–1113.Google Scholar
  16. Felton, G.W., Korth, K.L., Bi, J.L., Wesley, S.V., Huhman, D.V., Mathews, M.C., Murphy, J.B., Lamb, C. and Dixon, R.A. 1999. Inverse relationship between systemic resistance of plants to microorganisms and to insect herbivory. Curr. Biol. 9: 317–321.Google Scholar
  17. Foster, R., Izawa, T. and Chua, N.H. 1994. Plant bZip proteins gather at ACGT elements. FASEB J. 8: 192–200.Google Scholar
  18. Gleave, A.P. 1992. A versatile binary vector system with a T-DNA organizational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol. Biol. 20: 1203–1207.Google Scholar
  19. Grand, R.J.A. 1989. Acylation of viral and eukaryotic proteins. Biochem. J. 258: 625–638.Google Scholar
  20. Grant, J.J., Yun, B. W. and Loake, G.J. 2000. Oxidative burst and cognate redox signalling reported by luciferase imaging: identification of a signal network that functions independently of ethylene, SA and Me-JA but is dependent on MAPKK activity. Plant J. 24: 569–582.Google Scholar
  21. Hammond-Kosack, K.E. and Jones, J.D.G. 1994. Resistance gene-dependent plant defense responses. Plant Cell 8: 1773–1791.Google Scholar
  22. Harrison, M.J., Lawton, M.A., Lamb, C.J. and Dixon, R.A. 1991. Characterisation of a nuclear protein that binds to three elements within the silencer region of a bean chalcone synthase gene promoter. Proc. Natl. Acad. Sci. USA 88: 2515–2519.Google Scholar
  23. Henikoff, S. 1984. Unidirectional digestion with exonuclease III creates targetted breakpoints for DNA sequencing. Gene 28: 351–359.Google Scholar
  24. Hertig, C., Rebmann, G., Bull, J., Mauch, F. and Dudler, R. 1991. Sequence and tissue-specific expression of a putative peroxidase gene from wheat (Triticum aestivum L.). Plant Mol. Biol. 16: 171–174.Google Scholar
  25. Holub, E.B., Beynon, J.L. and Crute, I.R. 1994. Phenotypic and genotypic characterisation of interactions between isolates of Peronospora parasitica and ecotypes of Arabidopsis thaliana. Mol. Plant-Microbe Interact. 8: 916–928.Google Scholar
  26. Joshi, C.P. 1987. An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucl. Acids Res. 15: 6643–6653.Google Scholar
  27. Lamb C.J., Lawton M.A., Dron M. and Dixon R.A. 1989. Signals and transduction mechanisms for activation of plant defenses against microbial attack. Cell 56: 215–224.Google Scholar
  28. Lebel, E., Heifetz, P., Thorne, 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.Google Scholar
  29. Legrand, M., Kaufmann, S., Geoffroy, P. and Fritig, B. 1987. Biological function of ‘pathogenesis-related’ proteins: four tobacco PR proteins are chitinases. Proc. Natl. Acad. Sci. USA 84: 6750–6754.Google Scholar
  30. Lund, S.T., Stall, R.E. and Klee, H.J. 1998. Ethylene regulates the susceptible response to pathogen infection in tomato. Plant Cell 10: 371–382.Google Scholar
  31. MacDonald, M.H., Mogen, B.D. and Hunt, A.G. 1991. Characterisation of the polyadenylation signal from the T-DNA encoded octopine synthase gene. Nucl. Acids. Res. 19: 5575–5581.Google Scholar
  32. Malamy, J., Carr, J.P., Klessig, D.F. and Raskin, I. 1990. Salicylic acid: a likely endogenous signal in the resistance response of tobacco to viral infection. Science 250: 1002–1004.Google Scholar
  33. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratories, Cold Spring Harbor, NY.Google Scholar
  34. Manners, J.M., Penninckx, I.A., Vermaere, K., Kazan, K., Brown, R.L., Morgan, A., Maclean, D.J., Curtis, M.D., Cammue, B.P. and Broekaert, W.F. 1998. The promoter of the plant defensin gene PDF1.2 from Arabidopsis is systemically activated by fungal pathogens and responds to methyl jasmonate but not to salicylic acid. Plant Mol. Biol. 38: 1071–1080Google Scholar
  35. Mason, H.S., DeWald, D.B. and Mullet, J.E. 1993. Identification of a methyl-jasmonate responsive domain in the soybean vspB promoter. Plant Cell 5: 241–251.Google Scholar
  36. Memelink, J., Hoge, J.H.C. and Schilperoort, R.A. 1987. Cytokinin stress changes the developmental regulation of several defence-related genes in tobacco. EMBO 6: 3579–3583.Google Scholar
  37. Memelink, J., Linthorst, H., Schilperoort, R.A. and Hoge, J.H. 1990. Tobacco genes encoding acidic and basic isoforms of pathogenesis-related proteins display different expression patterns. Plant Mol. Biol. 14: 119–126.Google Scholar
  38. Metzler, M.C., Cutt, J.R. and Klessig, D.F. 1991. Isolation and characterisation of a gene encoding a PR-1 like protein from Arabidopsis thaliana. Plant Physiol. 96: 346–348.Google Scholar
  39. Niderman, T., Genetet, J., Bruyere, R., Gees, R., Stintzi, A., Legrand, M., Fritig, B. and Moesinger, E. 1995. Pathogenesis related PR-1 proteins are anti-fungal. Plant Physiol. 108: 17–27.Google Scholar
  40. Niki, T., Mitsuhara, I., Seo, S., Ohtsubo, N. and Ohashi, Y. 1998. Antagonistic effect of salicylic acid and jasmonic acid on the expression of pathogenesis-related (PR) protein genes in wounded mature tobacco leaves. Plant Cell Physiol. 39: 500–507.Google Scholar
  41. Ohshima, M., Harada, N., Matsuoka, M. and Ohashi, Y. 1990a. The nucleotide sequence of pathogenesis-related (PR) 1b protein gene of tobacco. Nucl. Acids Res. 18: 181.Google Scholar
  42. Ohshima, M., Harada, N., Matsuoka, M. and Ohashi, Y. 1990b. The nucleotide sequence of pathogenesis-related (PR) 1c protein gene of tobacco. Nucl. Acids Res. 18: 182.Google Scholar
  43. Ohta, M., Ohme-Takagi, M. and Shinshi, H. 2000. Three ethylene responsive transcription factors in tobacco with distinct transactivation functions. Plant J. 22: 29–38.Google Scholar
  44. Ohtsubo, N., Mitsuhara, I., Koga, M., Seo, S. and Ohashi, Y. 1999. Ethylene promotes the necrotic lesion formation and basic PR gene expression in TMV-infected tobacco. Plant Cell Physiol. 40: 808–817.Google Scholar
  45. Osbourn, A.E. 1996. Preformed antimicrobial compounds and plant defense against fungal attack. Plant Cell 8: 1821–1831.Google Scholar
  46. Parent, J.G. and Asselin, A. 1984. Detection of pathogenesis related proteins and other proteins in the intercellular fluid of hypersensitive plants infected with tobacco mosaic virus. Can J. Bot. 62: 564–569.Google Scholar
  47. Parker, J.E., Szabo, V., Staskawicz, B.J., Lister, C., Dean, C., Daniels, M.J. and Jones, J.D.G. 1993. Phenotypic characterisation and molecular mapping of the Arabidopsis thaliana locus, RPP5, determining disease resistance to Peronospora parasitica. Plant J. 4: 821–831.Google Scholar
  48. Payne, G., Parks, T.D., Burkhart, W., Dincher, S., Ahl, P., Métreux, J.P. and Ryals, J. 1988. Isolation of the genomic clone for pathogenesis-related protein 1a from Nicotiana tabacum cv. Xanthi-nc. Plant Mol. Biol. 11: 89–94.Google Scholar
  49. Pieterse, C.M.J., van Wees, S.C.M., van Pelt J.A., Knoester, M., Laan, R., Gerrits, H., Weisbeek, P.J. and van Loon, L.C. 1998. A novel signaling pathway controlling induced systemic resistance in Arabidopsis. Plant Cell 10: 1517–1580.Google Scholar
  50. Reymond, P. and Farmer, E.E. 1998. Jasmonate and salicylate as global signals for defence gene expression. Curr. Opin. Plant Biol. 1: 404–411.Google Scholar
  51. Rouster, J., Leah, R., Mundy, J. and Cameron-Mills, V. 1997. Identification of a methyl-jasmonate responsive region in the promoter of a lipoxygenase I gene expressed in barley grain. Plant J. 11: 513–523.Google Scholar
  52. Samac, D.A., Hironaka, C.M., Yallaly, P.E. and Shah, D.M. 1990. Isolation and characterization of the genes encoding basic and acidic chitinase in Arabidopsis thaliana. Plant Physiol. 93: 907–914.Google Scholar
  53. Sato, F., Kitajima, S., Koyamu, T. and Yamada, Y. 1996. Ethylene-induced gene expression of osmotin-like protein, a neutral isoform of tobacco PR-5, is mediated by the AGCCGCC cis sequence. Plant Cell Physiol. 37: 249–255.Google Scholar
  54. Schulz, B., Bennett, M.J., Dilkes, B.P. and Feldmann, K.A. 1994. T-DNA tagging in Arabidopsis thaliana: cloning by gene disruption. In: S.B. Gelvin and R.A. Schilperoort (Eds.) Plant Molecular Biology Manual, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 209–243.Google Scholar
  55. Thomma, B.P.H.J., Eggermont, K., Penninckx, I.A.M.A., Mauch-Mani, B., Vogelsang, R., Cammue B.P.A. and Broekaert, W.F. 1998. Separate jasmonate-dependent and salicylate-dependent defense-response pathways in Arabidopsis are essential for resistance to distinct microbial pathogens. Proc. Natl. Acad. Sci. USA 95: 15107–15111.Google Scholar
  56. Tornero, P., Gadea, J., Conejero, V. and Vera, P. 1997. Two PR-1 genes from tomato are differentially regulated and reveal a novel mode of expression for a pathogenesis-related gene during the hypersensitive response and development. Mol. Plant-Microbe Interact. 10: 624–634.Google Scholar
  57. Uknes, S., Mauch-Mani, B., Moyr, 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.Google Scholar
  58. Uknes, S., Dincher, S., Friedrich, L., Negrotto, D., Williams, S., Thompson-Taylor, H., Potter, S., Ward, E. and Ryals J. 1993. Regulation of pathogenesis-related protein-1a gene expression in tobacco. Plant Cell5: 159–169.Google Scholar
  59. van Loon, L.C. 1985. Pathogenesis-related proteins. Plant Mol. Biol. 4: 111–116.Google Scholar
  60. van Loon, L.C. and van Strien, E.A. 1999. The families of pathogenesis-related proteins, their activities, and comparative analysis of PR1 type proteins. Physiol. Mol. Plant Path. 55: 85–97.Google Scholar
  61. van' t Klooster, J.W., Kamoun, S., Vleeshouwers, V.G.A.A. and Govers, F. 1999. Characterisation of a cDNA encoding a pathogenesis-related protein PR-1 from potato (Solanum tubero-sum). Plant Physiol. 121: 1383.Google Scholar
  62. Vögeli-Lange, R., Fründt, C., Hart, C.M., Beffa, R., Nagy, F. and Meins, F. Jr. 1994. Evidence for a role of β-1,3-glucanase in dicot seed germination. Plant J. 5: 273–278.Google Scholar
  63. Woodgate, J.R. Gould, K.L. and Hunter, T. 1986. Substrate specificity of protein kinase C. Eur. J. Biochem. 161: 177–184.Google Scholar
  64. Xu, Y., Zhu, Q., Panbangred, W., Shirasu, K. and Lamb, C. 1996. Regulation, expression and function of a new basic chitinase gene in rice (Oryza sativa L.). Plant Mol. Biol. 30: 387–401.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Marjorie Santamaria
    • 1
  • Catherine J. Thomson
    • 1
  • Nick D. Read
    • 1
  • Gary J. Loake
    • 1
  1. 1.Institute of Cell & Molecular BiologyUniversity of EdinburghEdinburghUK

Personalised recommendations