Skip to main content
Log in

Sensitization of defense responses and activation of programmed cell death by a pathogen-induced receptor-like protein kinase in Arabidopsis

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

During the search for potential target genes of WRKY DNA-binding transcription factors, we have previously identified four pathogen-induced Arabidopsis genes (CRK5, CRK6, CRK10 and CRK11) encoding receptor-like protein kinases (RLKs) containing novel cysteine-rich repeats in their extracellular domains. In the present study, we transformed Arabidopsis plants with the RLK genes under control of the constitutive CaMV 35S promoter or a steroid-inducible Gal4 promoter. Expression of CRK5, but not the three other RLK genes, resulted in significant alterations in defense responses and leaf growth in transgenic plants. In transgenic plants harboring the 35S::CRK5 construct, significantly elevated and constitutive expression of CRK5 correlated with enhanced leaf growth and increased resistance to the bacterial pathogen Pseudomonas syringae. The enhanced disease resistance in the transgenic plants was associated with more rapidly induced expression of the PR1 gene after pathogen infection. In transgenic plants transformed with CRK5 under control of the steroid-inducible promoter, expression of the transgene was induced at relatively high levels after the steroid application and this induced expression of CRK5 triggered hypersensitive response-like cell death. Induced CRK5 expression also activated cell death in the npr1, ndr1 and eds1 mutants and in the transgenic nahG plants that fail to accumulate salicylic acid. Thus, the novel RLK is capable of activating multiple distinct defense responses depending on the manner and/or the levels of its over-expression in transgenic plants.

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.

Institutional subscriptions

Similar content being viewed by others

References

  • Aarts, N., Metz, M., Holub, E., Staskawicz, B.J., Daniels, M.J. and Parker, J.E. 1998. Different requirements for EDS1 and NDR1 by disease resistance genes define at least two R gene-mediated signaling pathways in Arabidopsis. Proc. Natl. Acad. Sci. US 95: 10306–10311.

    Google Scholar 

  • Arabidopsis Genome Initiative. 2000. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408: 796–815.

    Google Scholar 

  • Aoyama, T. and Chua, N.-H. 1997. A glucocorticoid-mediated transcriptional induction system in transgenic plants. Plant J. 11: 605–612.

    Google Scholar 

  • Bechtold, N. and Pelletier, G. 1998. In planta Agrobacterium mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Meth. Mol. Biol. 82: 259–266.

    Google Scholar 

  • Becraft, P.W., Stinard, P.S. and McCarty, D.R. 1996. CRINKLY4: a TNFR-like receptor kinase involved in maize epidermal differentiation [see comments]. Science 273: 1406–1409.

    Google Scholar 

  • 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 

  • Cao, H., Li, X. and Dong, X. 1998. Generation of broad-spectrum disease resistance by over-expression of an essential regulatory gene in systemic acquired resistance. Proc. Natl. Acad. Sci. USA 95: 6531–6536.

    Google Scholar 

  • Cao, H., Glazebrook, J., Clarke, J.D., Volko, S. and Dong, X. 1997. TheArabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. Cell 88: 57–63.

    Google Scholar 

  • Chen, C. and Chen, Z. 2000. Isolation and characterization of two pathogen-and salicylic acid-induced genes encoding WRKY DNA-binding proteins from tobacco. Plant Mol. Biol. 42: 387–396.

    Google Scholar 

  • Chen, C. and Chen, Z. 2002. Potentiation of developmentally regulated plant defense response by AtWRKY18, a pathogeninduced Arabidopsis transcription factor. Plant Physiol. 129: 706–716.

    Google Scholar 

  • Chen, Z. 2001. A superfamily of proteins with novel cysteine-rich repeats. Plant Physiol. 126: 473–476.

    Google Scholar 

  • Clark, S.E., Running, M.P. and Meyerowitz, E.M. 1993. CLAVATA1, a regulator of meristem and flower development in Arabidopsis. Development 119: 397–418.

    Google Scholar 

  • Clark, S.E., Williams, R.W. and Meyerowitz, E.M. 1997. The CLAVATA1 gene encodes a putative receptor kinase that controls shoot and floral meristem size in Arabidopsis. Cell 89: 575–585.

    Google Scholar 

  • Clay, N.K. and Nelson, T. 2002. VH1, a provascular cell-specific receptor kinase that influences leaf cell patterns in Arabidopsis. Plant Cell 14: 2707–2722.

    Google Scholar 

  • Czernic, P., Visser, B., Sun, W., Savoure, A., Deslandes, L., Marco, Y., Van Montagu, M. and Verbruggen, N. 1999. Characterization of an Arabidopsis thaliana receptor-like protein kinase gene activated by oxidative stress and pathogen attack. Plant J. 18: 321–327.

    Google Scholar 

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

    Google Scholar 

  • Dietrich, R.A., Delaney, T.P., Uknes, S.J., Ward, E.R., Ryals, J.A. and Dangl, J.L. 1994. Arabidopsis mutants simulating disease resistance response. Cell 77: 565–577.

    Google Scholar 

  • Dong, H., Delaney, T.P., Bauer, D.W. and Beer, S.V. 1999. Harpin induces disease resistance in Arabidopsis through the systemic acquired resistance pathway mediated by salicylic acid and the NIM1 gene. Plant J. 20: 207–215.

    Google Scholar 

  • Dong, J., Chen, C. and Chen, Z. 2003. Expression profile of the Arabidopsis WRKY gene superfamily during plant defense response. Plant Mol. Biol. 51: 21–37.

    Google Scholar 

  • Du, L. and Chen, Z. 2000. Identification of genes encoding novel receptor-like protein kinases as possible target genes of pathogen-induced WRKY DNA-binding proteins. Plant J. 24: 837–848.

    Google Scholar 

  • Falk, A., Feys, B.J., Frost, L.N., Jones, J.D., Daniels, M.J. and Parker, J.E. 1999. EDS1, an essential component of R genemediated disease resistance in Arabidopsis has homology to eukaryotic lipases. Proc. Natl. Acad. Sci. USA 96: 3292–3297.

    Google Scholar 

  • Frary, A., Nesbitt, T.C., Grandillo, S., Knaap, E., Cong, B., Liu, J., Meller, J., Elber, R., Alpert, K.B. and Tanksley, S.D. 2000.fw2.2: a quantitative trait locus key to the evolution of tomato fruit size. Science 289: 85–88.

    Google Scholar 

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

    Google Scholar 

  • Gomez-Gomez, L. and Boller, T. 2000. FLS2: an LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Mol. Cell 5: 1003–1011.

    Google Scholar 

  • He, Z., Wang, Z.Y., Li, J., Zhu, Q., Lamb, C., Ronald, P. and Chory, J. 2000. Perception of brassinosteroids by the extracellular domain of the receptor kinase BRI1. Science 288: 2360–2363.

    Google Scholar 

  • Heath, M.C. 2000. Hypersensitive response-related death. Plant Mol. Biol. 44: 321–334.

    Google Scholar 

  • Hunt, M.D., Delaney, T.P., Dietrich, R.A., Weymann, K.B., Dangl, J.L. and Ryals, J.A. 1997. Salicylate-independent lesion formation in Arabidopsis lsd mutants. Mol. Plant-Microbe Interact. 10: 531–536.

    Google Scholar 

  • Jabs, T. 1999. Reactive oxygen intermediates as mediators of programmed cell death in plants and animals. Biochem. Pharmacol. 57: 231–245.

    Google Scholar 

  • Jinn, T.L., Stone, J.M. and Walker, J.C. 2000. HAESA, an Arabidopsis leucine-rich repeat receptor kinase, controls floral organ abscission. Genes Dev. 14: 108–117.

    Google Scholar 

  • Knoester, M., Pieterse Pieterse, C.M., Bol, J.F. and van Loon, L.C. 1999. Systemic resistance in Arabidopsis induced by rhizobacteria requires ethylene-dependent signaling at the site of application. Mol. Plant-Microbe Interact. 12: 720–727.

    Google Scholar 

  • Krusell, L., Madsen, L.H., Sato, S., Aubert, G., Genua, A., Szczyglowski, K., Duc, G., Kaneko, T., Tabata, S., de Bruijn, F., Pajuelo, E., Sandal, N. and Stougaard, J. 2002. Shoot control of root development and nodulation is mediated by a receptor-like kinase. Nature 420: 422–426.

    Google Scholar 

  • Lally, D., Ingmire, P., Tong, H.Y. and He, Z.H. 2001. Antisense expression of a cell wall-associated protein kinase, WAK4, inhibits cell elongation and alters morphology. Plant Cell 13: 1317–1331.

    Google Scholar 

  • Lange, J., Xie, Z.-P., Broughton, W.J., Vogeli-Lange, R. and Boller, T. 1999. A gene encoding a receptor-like protein kinase in the roots of common bean is differentially regulated in response to pathogens, symbionts and nodulation factors. Plant Sci. 142: 133–145.

    Google Scholar 

  • Li, J. and Chory, J. 1997. A putative leucine-rich repeat receptor kinase involved in brassinosteroid signal transduction. Cell 90: 929–938.

    Google Scholar 

  • Maleck, K., Levine, A., Eulgem, T., Morgan, A., Schmid Schmid, J., Lawton, K.A., Dangl, J.L. and Dietrich, R.A. 2000. The transcriptome of Arabidopsis thaliana during systemic acquired resistance. Nature Genet. 26: 403–410.

    Google Scholar 

  • Montesano, M., Koiv, V., Maee, A. and Palva, T. 2002 Novel receptor-like kinases induced by Erwinia carotovora and short oligogalacturonides in potato. Mol. Plant Path. 2: 339–346.

    Google Scholar 

  • Montoya, T., Nomura, T., Farrar, K., Kaneta Kaneta, T., Yokota, T. and Bishop, G.J. 2002. Cloning the tomato Curl3 gene highlights the putative dual role of the leucine-rich repeat receptor kinase tBRI1/SR160 in plant steroid hormone and peptide hormone signaling. Plant Cell 14: 3163–3176.

    Google Scholar 

  • Nautlyal, C.S., Johri, J.K. and Singh, H.B. 2002. Survival of the rhizosphere-competent biocontrol strain Pseudomonas fluorescens NBRI2650 in the soil and phytosphere. Can. J. Microbiol. 48: 588–601.

    Google Scholar 

  • Nishimura, R., Hayashi, M., Wu, G.J., Kouchi, H., Imaizumi-Anraku, H., Murakami, Y., Kawasaki, S., Akao, S., Ohmori, M., Nagasawa, M., Harada, K. and Kawaguchi, M. 2002. HAR1 mediates systemic regulation of symbiotic organ development. Nature 420: 426–429.

    Google Scholar 

  • Ohtake, Y., Takahashi, T. and Komeda, Y. 2000. Salicylic acid induces the expression of a number of receptor-like kinase genes in Arabidopsis thaliana. [In process citation.] Plant Cell Physiol. 41: 1038–1044.

    Google Scholar 

  • Pal, K.K., Tilak, K.V., Saxena, A.K., Dey Dey, R. and Singh, C.S. 2001. Suppression of maize root diseases caused by Macrophomina phaseolina, Fusarium moniliforme and Fusarium graminearum by plant growth promoting rhizobacteria. Microbiol. Res. 156: 209–223.

    Google Scholar 

  • Pandey, A., Palni, L.M. and Hebbar, K.P. 2001. Suppression of damping-off in maize seedlings by Pseudomonas corrugata. Microbiol. Res. 156: 191–194.

    Google Scholar 

  • Pieterse, C.M., van Wees, S.C., Hoffland, E., van Pelt, J.A. and van Loon, L.C. 1996. Systemic resistance in Arabidopsis induced by biocontrol bacteria is independent of salicylic acid accumulation and pathogenesis-related gene expression. Plant Cell 8: 1225–1237.

    Google Scholar 

  • Rate, D.N. and Greenberg, J.T. 2001. The Arabidopsis aberrant growth and death2 mutant shows resistance to Pseudomonas syringae and reveals a role for NPR1 in suppressing hypersensitive cell death. Plant J. 27: 203–211.

    Google Scholar 

  • Robson, C.A. and Vanlerberghe, G.C. 2002. Transgenic plant cells lacking mitochondrial alternative oxidase have increased susceptibility to mitochondria-dependent and-independent pathways of programmed cell death. Plant Physiol. 129: 1908–1920.

    Google Scholar 

  • Scheer, J.M. and Ryan, C.A., Jr. 2002. The systemin receptor SR160 from Lycopersicon peruvianum is a member of the LRR receptor kinase family. Proc. Natl. Acad. Sci. USA 99: 9585–9590.

    Google Scholar 

  • Shah, J., Kachroo Kachroo, P., and Klessig, D.F. 1999. The Arabidopsis ssi1 mutation restores pathogenesis-related gene expression in npr1 plants and renders defensin gene expression salicylic acid dependent. Plant Cell 11: 191–206.

    Google Scholar 

  • Shah, K., Vervoort, J. and de Vries, S.C. 2001. Role of threonines in the Arabidopsis thaliana somatic embryogenesis receptor kinase 1 activation loop in phosphorylation. J. Biol. Chem. 276: 41263–41269.

    Google Scholar 

  • Shiu, S.H. and Bleecker, A.B. 2001. Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proc. Natl. Acad. Sci. USA 98: 10763–10768.

    Google Scholar 

  • Song, W.Y., Wang, G.L., Chen, L.L., Kim, H.S., Pi, L.Y., Holsten, T., Gardner, J., Wang, B., Zhai Zhai,W.X., Zhu, L.H. et al. 1995. A receptor kinase-like 34 protein encoded by the rice disease resistance gene, Xa21. Science 270: 1804–1806.

    Google Scholar 

  • Stein, J.C. and Nasrallah, J.B. 1993. A plant receptor-like gene, the S-locus receptor kinase of Brassica oleracea L., encodes a functional serine/ threonine kinase. Plant Physiol 101: 1103–1106.

    Google Scholar 

  • Stein, J.C., Howlett, B., Boyes, D.C., Nasrallah, M.E. and Nasrallah, J.B. 1991. Molecular cloning of a putative receptor protein kinase gene encoded at the self-incompatibility locus of Brassica oleracea. Proc. Natl. Acad. Sci. USA 88: 8816–8820.

    Google Scholar 

  • Stone, J.M., Trotochaud, A.E., Walker, J.C. and Clark, S.E. 1998. Control of meristem development by CLAVATA1 receptor kinase and kinase-associated protein phosphatase interactions. Plant Physiol. 117: 1217–1225.

    Google Scholar 

  • Thordal-Christensen, H., Zhang, Z.G., Wei, Y.D. and Collinge, D.B. 1997. Subcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J. 11: 1187–1194.

    Google Scholar 

  • Timmermans, M.C., Maliga, P., Vieira, J. and Messing, J. 1990. The pFF plasmids: cassettes utilising CaMV sequences for expression of foreign genes in plants. J. Biotechnol. 14: 333–344.

    Google Scholar 

  • Torii, K.U., Mitsukawa, N., Oosumi, T., Matsuura, Y., Yokoyama, R., Whittier, R.F. and Komeda, Y. 1996. The Arabidopsis ERECTA gene encodes a putative 35 receptor protein kinase with extracellular leucine-rich repeats. Plant Cell 8: 735–746.

    Google Scholar 

  • Uknes, 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.

    Google Scholar 

  • van Wees, S.C., Pieterse, C.M., Trijssenaar, A., van 't Westende, Y.A., Hartog, F. and van Loon, L.C. 1997. Differential induction of systemic resistance in Arabidopsis by biocontrol bacteria. Mol. Plant-Microbe Interact. 10: 716–724.

    Google Scholar 

  • Vanacker, H., Lu, H., Rate, D.N. and Breenberg, J.T. 2001. A role for salicylic acid and NPR1 in regulating cell growth in Arabidopsis. Plant J. 28: 209–216.

    Google Scholar 

  • Wagner, T.A. and Kohorn, B.D. 2001. Wall-associated kinases are expressed throughout plant development and are required for cell expansion. Plant Cell 13: 303–318.

    Google Scholar 

  • Walker, J.C. 1994. Structure and function of the receptor-like protein kinase of higher plants. Plant Mol. Biol. 261: 1599–1609.

    Google Scholar 

  • Wang, Z.Y., Seto, H., Fujioka, S., Yoshida, S. and Chory, J. 2001. BRI1 is a critical component of a plasma-membrane receptor for plant steroids. Nature 410: 380–383.

    Google Scholar 

  • Wei, Z.M., Laby, R.J., Zumoff, C.H., Bauer, D.W., He, S.Y., Collmer, A. and Beer, S.V. 1992. Harpin, elicitor of the hypersensitive response produced by the plant pathogen Erwinia amylovora. Science 257: 85–88.

    Google Scholar 

  • Yang, P., Wang, Z., Fan, B., Chen, C. and Chen, Z. 1999. A pathogen-and salicylic acid-induced WRKY DNA-binding activity recognizes the elicitor response element of the tobacco class I chitinase gene promoter. Plant J. 18: 141–149.

    Google Scholar 

  • Yang, Y., Shah, J. and Klessig, D.F. 1997. Signal perception and transduction in plant defense responses. Genes Dev. 11: 1621–1639.

    Google Scholar 

  • Yu, D., Chen, C. and Chen, Z. 2001. Evidence for an important role Of WRKY DNA binding proteins in the regulation of NPR1 gene expression. Plant Cell 13: 1527–1540.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Zhixiang Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, K., Du, L. & Chen, Z. Sensitization of defense responses and activation of programmed cell death by a pathogen-induced receptor-like protein kinase in Arabidopsis . Plant Mol Biol 53, 61–74 (2003). https://doi.org/10.1023/B:PLAN.0000009265.72567.58

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:PLAN.0000009265.72567.58

Navigation