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Expression of the Hypersensitive Response-assisting Protein in Arabidopsis Results in Harpin-dependent Hypersensitive Cell Death in Response to Erwinia carotovora

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Abstract

Active defense mechanisms of plants against pathogens often include a rapid plant cell death known as the hypersensitive cell death (HCD). Hypersensitive response-assisting protein (HRAP) isolated from sweet pepper intensifies the harpinPss-mediated HCD. Here we demonstrate that constitutive expression of the hrap gene in Arabidopsis results in an enhanced disease resistance towards soft rot pathogen, E. carotovora subsp. carotovora. This resistance was due to the induction of HCD since different HCD markers viz. Athsr3, Athsr4, ion leakage, H2O2 and protein kinase were induced. One of the elicitor harpin proteins, HrpN, from Erwinia carotovora subsp. carotovora was able to induce a stronger HCD in hrap-Arabidopsis than non-transgenic controls. To elucidate the role of HrpN, we used E. carotovora subsp. carotovora defective in HrpN production. The hrpN− mutant did not induce disease resistance or HCD markers in hrap-Arabidopsis. These results imply that the disease resistance of hrap-Arabidopsis against a virulent pathogen is harpin dependent.

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Abbreviations

ATP:

adenosine tri-phosphate

DTT:

dithiothreitol

EGTA:

Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid

HRAP:

hypersensitive response-assisting protein

HCD:

hypersensitive cell death

MAPK:

mitogen activated protein kinase

PCR:

polymerase chain reaction

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

References

  • C.J. Baker E.W. Orlandi N.M. Mock (1993) ArticleTitleHarpin, an elicitor of the hypersensitive response in tobacco caused by Erwinia amylovora, elicits active oxygen production in suspension cells Plant Physiol. 102 1341–1344 Occurrence Handle12231911

    PubMed  Google Scholar 

  • F. Barras F. Gijsegem ParticleVan A. Chatterjee (1994) ArticleTitleExtracellular enzymes and pathogenesis of soft rot Erwinia Annu. Rev. Phytopathol. 32 201–234

    Google Scholar 

  • D.W. Bauer A.J. Bogdanove S.V. Beer A. Collmer (1994) ArticleTitleErwinia chrysanthemi hrp genes and their involvement in soft rot pathogenesis and elicitation of the hypersensitive response Mol. Plant–Microbe Interact. 7 573–581 Occurrence Handle7949326

    PubMed  Google Scholar 

  • D.W. Bauer Z.-M. Wei S.V. Beer A. Collmer (1995) ArticleTitleErwinia chrysanthemi Harpin Ech: an elicitor of the hypersensitive response that contributes to soft-rot pathogenesis Mol. Plant–Microbe Interact. 8 484–491 Occurrence Handle8589405

    PubMed  Google Scholar 

  • N. Bechtold J. Ellis G. Pellettier (1993) ArticleTitleIn planta Agrobacterium-mediated gene transfer by infiltration of adult Arabidopsis thaliana plants CR Acad. Sci. 316 1194–1199

    Google Scholar 

  • A.O. Charkowski J.R. Alfano G. Preston J. Yuan S.Y. He A. Collmer (1998) ArticleTitleThe Pseudomonas syringae pv. tomato HrpW protein has domains similar to harpins and pectate lyases and can elicit the plant hypersensitive response and bind to pectate J. Bacteriol. 180 5211–5217 Occurrence Handle9748456

    PubMed  Google Scholar 

  • C.H. Chen H.J. Lin T.Y. Feng (1998) ArticleTitleAn amphipathic protein from sweet pepper can dissociate harpinPss multimeric forms and intensify the harpinPss-mediated hypersensitive response Physiol. Mol. Plant Pathol. 52 139–149 Occurrence Handle10.1006/pmpp.1997.0120

    Article  Google Scholar 

  • C.H. Chen H.J. Lin M.J. Ger D. Chow T.Y. Feng (2000) ArticleTitlecDNA cloning and characterization of a plant protein that may be associated with the harpinPSS-mediated hypersensitive response Plant Mol. Biol. 43 429–438 Occurrence Handle10.1023/A:1006448611432 Occurrence Handle11052195

    Article  PubMed  Google Scholar 

  • G.R. Cornelis F. Gijsegem ParticleVan (2000) ArticleTitleAssembly and function of type III secretory systems Annu. Rev. Microbiol. 54 735–774 Occurrence Handle10.1146/annurev.micro.54.1.735 Occurrence Handle11018143

    Article  PubMed  Google Scholar 

  • Y. Cui L. Madi A. Mukherjee C.K. Dumenyo A. Chatterjee (1996) ArticleTitleThe RsmA-mutants of Erwinia carotovora subsp. carotovora strain Ecc71 overexpress hrpN Ecc and elicit a hypersensitive reaction-like response in tobacco leaves Mol. Plant–Microbe Interact. 9 565–573 Occurrence Handle8810071

    PubMed  Google Scholar 

  • J.L. Dangl J.D. Jones (2001) ArticleTitlePlant pathogens and integrated defence response to infection Nature 411 826–833 Occurrence Handle11459065

    PubMed  Google Scholar 

  • J.L. Dangl R.A. Diretrich M.H. Richberg (1996) ArticleTitleDeath don’t have no mercy: cell death programs in plant–microbe interactions Plant Cell 8 1793–1807 Occurrence Handle10.1105/tpc.8.10.1793 Occurrence Handle12239362

    Article  PubMed  Google Scholar 

  • R. Desikan A. Clarke P. Atherfold J.T. Hancock S.J. Neill (1999) ArticleTitleHarpin induces mitogen-activated protein kinase activity during defence responses in Arabidopsis thaliana suspension cultures Planta 210 97–103 Occurrence Handle10.1007/s004250050658 Occurrence Handle10592037

    Article  PubMed  Google Scholar 

  • R. Desikan J.T. Hancock K. Ichimura K. Shinozaki S.J. Neill (2001) ArticleTitleHarpin induces activation of the Arabidopsis mitogen-activated protein kinases AtMPK4 and AtMPK6 Plant Physiol. 126 1579–1587 Occurrence Handle10.1104/pp.126.4.1579 Occurrence Handle11500556

    Article  PubMed  Google Scholar 

  • R.A. Dixon M.J. Harrison (1990) ArticleTitleActivation, structure and organization of genes involved in microbial defense in plants Adv. Genet. 28 165–234 Occurrence Handle2239449

    PubMed  Google Scholar 

  • J.E. Galan A. Collmer (1999) ArticleTitleType III secretion machines: bacterial devices for protein delivery into host cells Science 21 1322–1328

    Google Scholar 

  • S. Gaudriault M.N. Brisset M.A. Barny (1998) ArticleTitleHrpW of Erwinia amylovora, a new Hrp-secreted protein FEBS Lett. 428 224–228 Occurrence Handle10.1016/S0014-5793(98)00534-1 Occurrence Handle9654138

    Article  PubMed  Google Scholar 

  • M.J. Ger C.H. Chen S.Y. Hwang H.E. Huang A.R. Podile B.V. Dayakar T.Y. Feng (2002) ArticleTitleConstitutive expression of hrap gene in transgenic tobacco plant enhances resistance against virulent bacterial pathogens by induction of a hypersensitive response Mol. Plant–Microbe Interact. 15 764–773 Occurrence Handle12182333

    PubMed  Google Scholar 

  • S. Gopalan D.W. Bauer J.R. Alfano A.O. Loniello S.Y. He A. Collmer (1996) ArticleTitleExpression of the Pseudomonas syringae avirulence protein AvrB in plant cells alleviates its dependence on the hypersensitive response and pathogenicity (Hrp) secretion system in eliciting genotype-specific hypersensitive cell death Plant Cell 8 1095–1105 Occurrence Handle10.1105/tpc.8.7.1095 Occurrence Handle8768370

    Article  PubMed  Google Scholar 

  • S.Y. He D.W. Bauer A. Collmer S.V. Beer (1994) ArticleTitleHypersensitive response elicited by Erwinia amylovora harpin requires active plant metabolism Mol. Plant–Microbe Interact. 7 289–292

    Google Scholar 

  • M. Holsters D. Waele ParticleDe A. Depicker E. Messens M. Montagu ParticleVan J. Schell (1978) ArticleTitleTransfection and transformation of Agrobacterium tumefaciens Mol. Gen. Genet. 163 182–187

    Google Scholar 

  • S. Jana M.A. Choudhuri (1982) ArticleTitleGlycolate metabolism of three submerged aquatic angiosperm during aging Aquat. Bot. 12 345–354 Occurrence Handle10.1016/0304-3770(82)90026-2

    Article  Google Scholar 

  • T. Kawano (2003) ArticleTitleRoles of reactive oxygen species-generating peroxidase reaction in plant defense and growth induction Plant Cell Rep. 21 829–837 Occurrence Handle12789499

    PubMed  Google Scholar 

  • H. Keller N. Pamboukdjian M. Ponchet A. Poupet R. Delon J.L. Verrier D. Roby P. Ricci (1999) ArticleTitlePathogen-induced elicitin production in transgenic tobacco generates a hypersensitivity response and nonspecific disease resistance Plant Cell 11 223–235 Occurrence Handle10.1105/tpc.11.2.223 Occurrence Handle9927640

    Article  PubMed  Google Scholar 

  • J.F. Kim S.V. Beer (1998) ArticleTitleHrpW of Erwinia amylovora, a new harpin that contains a domain homologous to pectate lyases of a distinct class J. Bacteriol. 180 5203–5210 Occurrence Handle9748455

    PubMed  Google Scholar 

  • A. Kotoujansky (1987) ArticleTitleMolecular genetics of pathogenesis by soft-rot Erwinias Annu. Rev. Phytopathol. 25 405–430 Occurrence Handle10.1146/annurev.py.25.090187.002201

    Article  Google Scholar 

  • C. Lacomme D. Roby (1999) ArticleTitleIdentification of new early markers of the hypersensitive response in Arabidopsis thaliana FEBS Lett. 459 149–153 Occurrence Handle10.1016/S0014-5793(99)01233-8 Occurrence Handle10518009

    Article  PubMed  Google Scholar 

  • C. Lamb R.A. Dixon (1997) ArticleTitleThe oxidative burst in plant disease resistance Annu. Rev. Plant Physiol. Plant Mol. Biol. 48 223–250 Occurrence Handle10.1146/annurev.arplant.48.1.251 Occurrence Handle15012263

    Article  PubMed  Google Scholar 

  • F.L. Menke J.A. Pelt Particlevan C.M. Pieterse D.F. Klessig (2004) ArticleTitleSilencing of the mitogen-activated protein kinase MPK6 compromises disease resistance in Arabidopsis Plant Cell 16 897–907 Occurrence Handle10.1105/tpc.015552 Occurrence Handle15020743

    Article  PubMed  Google Scholar 

  • A. Mukherjee Y. Cui Y. Liu A.K. Chatterjee (1997) ArticleTitleMolecular characterization and expression of the Erwinia carotovora hrpN Ecc gene, which encodes an elicitor of the hypersensitive reaction Mol. Plant–Microbe Interact. 10 462–471 Occurrence Handle9150595

    PubMed  Google Scholar 

  • D. Pontier L. Godiard Y. Marco D. Roby (1994) ArticleTitlehsr203J, a tobacco gene whose activation is rapid, highly localized and specific for incompatible plant/pathogen interactions Plant J. 5 507–521 Occurrence Handle8012404

    PubMed  Google Scholar 

  • D. Pontier M. Tronchet P. Rogowsky E. Lam D. Roby (1998) ArticleTitleActivation of hsr203, a plant gene expressed during incompatible plant pathogen interaction, is correlated with program cell death Mol. Plant–Microbe Interact. 11 544–554 Occurrence Handle9612953

    PubMed  Google Scholar 

  • J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular Cloning: A Laboratory Manual EditionNumber2nd Cold Spring Harbor Laboratory Press Cold Spring Harbor NY, USA 931–957

    Google Scholar 

  • S. Shen Q. Li S.Y. He K.R. Barker D. Li A.G. Hunt (2000) ArticleTitleConversion of compatible plant–pathogen interactions into incompatible interactions by expression of the Pseudomonas syringae pv. syringae 61 hrmA gene in transgenic tobacco plants Plant J. 23 205–213 Occurrence Handle10.1046/j.1365-313x.2000.00772.x Occurrence Handle10929114

    Article  PubMed  Google Scholar 

  • B.J. Staskawicz (2001) ArticleTitleGenetics of plant–pathogen interactions specifying plant disease resistance Plant Physiol. 125 73–76 Occurrence Handle10.1104/pp.125.1.73 Occurrence Handle11154300

    Article  PubMed  Google Scholar 

  • R.N. Strobel J.S. Gopalan J.A. Kuc S.Y. He (1996) ArticleTitleInduction of systemic acquired resistance in cucumber by Pseudomonas syringae pv. syringae 61 HrpZPss protein Plant J. 9 431–439 Occurrence Handle10.1046/j.1365-313X.1996.09040431.x

    Article  Google Scholar 

  • F. Vailleau X. Daniel M. Tronchet J.L. Montillet C. Triantaphylides D. Roby (2002) ArticleTitleA R2R3-MYB gene, AtMYB30, acts as a positive regulator of the hypersensitive cell death program in plants in response to pathogen attack Proc. Natl. Acad. Sci. USA 99 10179–10184 Occurrence Handle10.1073/pnas.152047199 Occurrence Handle12119395

    Article  PubMed  Google Scholar 

  • M.C. Verberne R. Verpoorte J.F. Bol J. Mercado-Blanco H.J.M. Linthorst (2000) ArticleTitleOverproduction of salicylic acid in plants by bacterial transgenes enhances pathogen resistance Nat. Biotechnol. 18 779–783 Occurrence Handle10.1038/77347 Occurrence Handle10888849

    Article  PubMed  Google Scholar 

  • Z.-M. Wei S.V. Beer (1996) ArticleTitleHarpin from Erwinia amylovora induces plant resistance Acta Hortic. 411 223–225

    Google Scholar 

  • S. Zhang C.D. Jin S.J. Roux (1993) ArticleTitleCasein kinase II-type protein kinase from pea cytoplasm and its inactivation by alkaline phosphatase in vitro Plant Physiol. 103 955–962 Occurrence Handle10.1104/pp.103.3.1029 Occurrence Handle12231993

    Article  PubMed  Google Scholar 

  • J. Zhou Y.T. Loh R.A. Bressan G.B. Martin (1995) ArticleTitleThe tomato gene Pti1 encodes a serine/threonine kinase that is phosphorylated by Pto and is involved in the hypersensitive response Cell 15 925–935 Occurrence Handle10.1016/0092-8674(95)90208-2

    Article  Google Scholar 

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Authors and Affiliations

  1. Institute of Plant and Microbial Biology, Academia Sinica 115, Taipei, Taiwan

    Ajay-Kumar Pandey, Hsiang-En Huang, Mei-Kuen Yip, Jiqing Zeng & Teng-Yung Feng

  2. Department of Life Science, National University of Kaohsiung, 811, Kaohsiung, Taiwan

    Mang-Jye Ger

Authors
  1. Ajay-Kumar Pandey
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  2. Mang-Jye Ger
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  3. Hsiang-En Huang
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  4. Mei-Kuen Yip
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  5. Jiqing Zeng
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  6. Teng-Yung Feng
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Correspondence to Teng-Yung Feng.

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Pandey, AK., Ger, MJ., Huang, HE. et al. Expression of the Hypersensitive Response-assisting Protein in Arabidopsis Results in Harpin-dependent Hypersensitive Cell Death in Response to Erwinia carotovora. Plant Mol Biol 59, 771–780 (2005). https://doi.org/10.1007/s11103-005-1002-3

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