Biologia plantarum 2016, 60:181-189 | DOI: 10.1007/s10535-015-0571-5

Overexpression of a harpin-encoding gene popW in tobacco enhances resistance against Ralstonia solanacearum

Chao Wang1, Cui Wang1, H. -W. Li1, T. Wei1, Y. -P. Wang2, H. -X. Liu1,*
1 College of Plant Protection and Key Laboratory of Integrated Management of Crop Diseases and Pests, Nanjing Agricultural University, Nanjing, P.R. China
2 College of Life Science and Chemical Engineering, Huaiyin Institute of Technology, Huai'an, P.R. China

PopW, a harpin protein identified from Ralstonia solanacearum, has multiple beneficial effects in plants, promoting plant growth and development, increasing crop yield, and inducing resistance to pathogens. Tobacco plants transformed with popW, the PopW-encoding gene, exhibited a promoted growth rate and enhanced resistance to Tobacco mosaic virus (TMV). Here, it is documented that the transgenic tobacco plants overexpressing popW exhibited a higher resistance to R. solanacearum YN10 infection compared with that of the wild-type plants. In the popW-expressing tobacco lines, an enhanced H2O2 accumulation and hypersensitive reaction (HR) were activated in the inoculated site. In addition, the resistance was accompanied with increased transcripts in numbers of genes related to defense (including HR), reactive oxygen species (ROS) scavenging, and salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) production. These results suggest that popW acted as positive regulator in tobacco resistance against R. solanacearum via modulation of SA-, JA-, and ET-mediated signaling pathways. We report for the first time that the expression of a harpin-encoding gene in vivo improved plant resistance to R. solanacearum.

Keywords: ethylene; jasmonic acid; hypersensitive reaction; Nicotiana tabacum; salicylic acid; transgenic plant
Subjects: harpin; disease resistance; ethylene; jasmonic acid; hypersensitive reaction; salicylic acid; transgenic plant; gene expression; tobacco
Species: Nicotiana tabacum

Received: September 5, 2014; Revised: June 7, 2015; Accepted: July 31, 2015; Published: January 1, 2016Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Wang, C., Wang, C., Li, H.-W., Wei, T., Wang, Y.-P., & Liu, H.-X. (2016). Overexpression of a harpin-encoding gene popW in tobacco enhances resistance against Ralstonia solanacearum. Biologia plantarum, 60(1), 181-189. doi: 10.1007/s10535-015-0571-5.
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)

    • Open full article

    Supplementary files

    Download filebpl-201601-0020_S1.pdf

    File size: 223.43 kB

    References

    1. Alvarez, M.E., Pennell, R.I., Meijer, P.J., Ishikawa, A., Dixon, R.A., Lamb, C.: Reactive oxygen intermediates mediate a systemic signal network in the establishment of plant immunity. - Cell 92: 773-784, 1998. Go to original source...
    2. Bernoux, M., Ellis, J.G., Dodds, P.N.: New insights in plant immunity signaling activation. - Curr. Opin. Plant Biol. 14: 512-518, 2011. Go to original source...
    3. Chen, N., Goodwin, P.H., Hsiang, T.: The role of ethylene during the infection of Nicotiana tabacum by Colletotrichum destructivum. - J. exp. Bot. 54: 2449-2456, 2003. Go to original source...
    4. Choi, M.S., Heu, S., Paek, N.C., Koh, H.J., Lee, J.S., Oh, C.S.: Expression of hpa1 gene encoding a bacterial harpin protein in Xanthomonas oryzae pv. oryzae enhances disease resistance to both fungal and bacterial pathogens in rice and Arabidopsis. - Plant Pathol. J. 28: 364-372, 2012. Go to original source...
    5. Choi, M.S., Kim, W., Lee, C., Oh, C.S.: Harpins, multifunctional proteins secreted by gram-negative plantpathogenic bacteria. - Mol. Plant-Microb. Interact. 26: 1115-1122, 2013.
    6. Dang, F., Wang, Y., She, J., Lei, Y., Liu, Z., Eulgem, T., Lai, Y., Lin, J., Yu, L., Lei, D., Guan, D., Li, X., Yuan, Q., He, S.: Overexpression of CaWRKY27, a subgroup IIe WRKY transcription factors of Capsicum annuum, positively regulates tobacco resistance to Ralstonia solanacearum infection. - Physiol. Plant. 150: 397-411, 2014. Go to original source...
    7. Dodds, P.N., Rathjen, J.P.: Plant immunity: towards an integrated view of plant-pathogen interactions. - Natur. Rev. Genet. 11: 539-548, 2010. Go to original source...
    8. Dong, H., Delaney, T.P., Bauer, D.W., Beer, S.V.: 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, 1999. Go to original source...
    9. Dong, X.: SA, JA, ethylene, and disease resistance in plants. - Curr. Opin. Plant Biol. 1: 316-323, 1998. Go to original source...
    10. Farmer, E.E., Almeras, E., Krishnamurthy, V.: Jasmonates and related oxylipins in plant responses to pathogenesis and herbivory. - Curr. Opin. Plant Biol. 6: 372-378, 2003. Go to original source...
    11. Feys, B., Parker, J.: Interplay of signaling pathways in plant disease resistance. - Trends Genet. 16: 449-455, 2000. Go to original source...
    12. Flores-Cruz, Z., Allen, C.: Ralstonia solanacearum encounters an oxidative environment during tomato infection. - Mol. Plant-Microb. Interact. 22: 773-782, 2009.
    13. Fu, M., Xu, M., Zhou, T., Wang, D., Tian, S., Han, L., Dong, H., Zhang, C.: Transgenic expression of a functional fragment of harpin protein Hpa1 in wheat induces the phloem-based defence against English grain aphid. - J. exp. Bot. 65: 1439-1453, 2014. Go to original source...
    14. Fu, Z.Q., Dong, X.: Systemic acquired resistance: turning local infection into global defense. - Annu. Rev. Plant Biol. 64: 839-863, 2013. Go to original source...
    15. Genin, S., Boucher, C.: Ralstonia solanacearum: secrets of a major pathogen unveiled by analysis of its genome. - Mol. Plant Pathol. 3: 111-118, 2002. Go to original source...
    16. Greenberg, J.T., Yao, N.: The role and regulation of programmed cell death in plant-pathogen interactions. - Cell Microbiol. 6: 201-211, 2004. Go to original source...
    17. He, S.Y.: Elicitation of the plant hypersensitive response by bacteria. - Plant Physiol. 112: 865-869, 1996. Go to original source...
    18. Heath, M.C.: Hypersensitive response-related death. - Plant mol. Biol. 44: 321-334, 2000. Go to original source...
    19. Jenkins, S.F., Averre, C.W.: Root diseases of vegetables in hydroponic culture systems in North Carolina greenhouse. - Plant Dis. 9: 968-970, 1983. Go to original source...
    20. Jones, J.D., Dangl, J.L.: The plant immune system. - Nature 444: 323-329, 2006. Go to original source...
    21. Kempe, J., Sequeira, L.: Biological control of baterial wilt of potatoes: attempts to induce resistance by treating tubes with bacteria. - Plant Dis. 67: 499-501, 1983. Go to original source...
    22. Koornneef, A., Pieterse, C.M.J.: Cross talk in defense signaling. - Plant Physiol. 146: 839-844, 2008. Go to original source...
    23. Leon-Reyes, A., Du, Y., Koorneef, A., Proietti, S., Körbes, A.P., Memelink, J.: Ethylene signaling renders the jasmonate response of Arabidopsis insensitive to future suppression by salicylic acid. - Mol. Plant-Microbe Interact. 23:187-197, 2010. Go to original source...
    24. Li, J.G., Cao, J., Sun, F.F., Niu, D.D., Yan, F., Liu, H.X., Guo, J.H.: Control of Tobacco mosaic virus by PopW as a result of induced resistance in tobacco under greenhouse and field conditions. - Phytopathology 101: 1202-1208, 2011. Go to original source...
    25. Li, J.G., Liu, H.X., Cao, J., Chen, L.F., Gu, C., Allen, C., Guo, J.H.: PopW of Ralstonia solanacearum, a new two-domain harpin targeting the plant cell wall. - Mol. Plant Pathol. 11: 371-381, 2010. Go to original source...
    26. Li, L., Feng, X., Tang, M., Hao, W., Han, Y., Zhang, G., Wan, S.: Antibacterial activity of lansiumamide B to tobacco bacterial wilt (Ralstonia solanacearum). - Microbiol. Res. 169: 522-526, 2014. Go to original source...
    27. Liang, Z., Xu, J.P., Meng, X.L., Lu, W., Wang, J., Xia, H.: Improve bioavailability of Harpin protein on plant use PLGA based nanoparticle. - J. Biotechnol. 143: 296-301, 2009. Go to original source...
    28. Lumbreras, V., Vilela, B., Irar, S., Sole, M., Capellades, M., Valls, M., Coca, M., Pages, M.: MAPK phosphatase MKP2 mediates disease responses in Arabidopsis and functionally interacts with MPK3 and MPK6. - Plant J. 63: 1017-1030, 2010. Go to original source...
    29. Miao, W., Wang, X., Song, C., Wang, Y., Ren, Y., Wang, J.: Transcriptome analysis of Hpa1Xoo transformed cotton revealed constitutive expression of genes in multiple signalling pathways related to disease resistance. - J. exp. Bot. 61: 4263-4275, 2010. Go to original source...
    30. Pavli, O.I., Kelaidi, G.I., Tampakaki, A.P., Skaracis, G.N.: The hrpZ gene of Pseudomonas syringae pv. phaseolicola enhances resistance to rhizomania disease in transgenic Nicotiana benthamiana and sugar beet. - PLoS ONE 6: e17306, 2011. Go to original source...
    31. Peng, J.L., Bao, Z.L., Ren, H.Y., Wang, J.S., Dong, H.S.: Expression of HarpinXoo in transgenic tobacco induces pathogen defense in the absence of hypersensitive cell death. - Phytopathology 94: 1048-1055, 2004. Go to original source...
    32. Pontier, D., Tronchet, M., Rogowsky, P., Lam, E., Roby, D.: Activation of hsr203, a plant gene expressed during incompatible plant-pathogen interactions, is correlated with programmed cell death. - Mol. Plant-Microbe Interact. 11: 544-554, 1998. Go to original source...
    33. Schell, M.A.: Control of virulence and pathogenicity genes of Ralstonia solanacearum by an elaborate sensory network. - Annu. Rev. Phytopathol. 38: 263-292, 2000. Go to original source...
    34. Shah, J.: Plants under attack: systemic signals in defence. - Curr. Opin. Plant Biol. 12: 459-464, 2009. Go to original source...
    35. Shao, M., Wang, J.S., Dean, R.A., Lin, Y.G., Gao, X.W., Hu, S.J.: Expression of a harpin-encoding gene in rice confers durable nonspecific resistance to Magnaporthe grisea. - Plant Biotechnol. J. 6: 73-81, 2008.
    36. Shi, W.N., Hao, L.L., Li, J., Liu, D.D., Guo, X.Q., Li, H.: The Gossypium hirsutum WRKY gene GhWRKY39-1 promotes pathogen infection defense responses and mediates salt stress tolerance in transgenic Nicotiana benthamiana. - Plant Cell Rep. 33: 483-498, 2014. Go to original source...
    37. Sohn, S., Kim, Y., Kim, B., Lee, S., Lim, C.K., Hur, J.H., Lee, J.: Transgenic tobacco expressing the hrpNEP gene from Erwinia pyrifoliae triggers defense responses against Botrytis cinerea. - Mol. Cells 24: 232-239, 2007.
    38. Sonnewald, S., Priller, J.P.R., Schuster, J., Glickmann, E., Hajirezaei, M.R., Siebig, S., Mudgett, M.B., Sonnewald, U.: Regulation of cell wall-bound invertase in pepper leaves by Xanthomonas campestris pv. vesicatoria type three effectors. - PLoS ONE 7: e51763, 2012. Go to original source...
    39. Takahashi, H., Chen, Z., Du, H., Liu, Y., Klessig, D.F.: Development of necrosis and activation of disease resistance in transgenic tobacco plants with severely reduced catalase levels. - Plant J. 11: 993-1005, 1997. Go to original source...
    40. Takahashi, Y., Nagata, T.: parB: an auxin-regulated gene encoding glutathione S-transferase. - Proc. nat. Acad. Sci. USA 89: 56-59, 1992. Go to original source...
    41. Ton, J., De Vos, M., Robben, C., Buchala, A., Métraux, J.P., Van Loon, L.C., Pieterse, C.M.: Characterization of Arabidopsis enhanced disease susceptibility mutants that are affected in systemically induced resistance. - Plant J. 29: 11-21, 2002. Go to original source...
    42. Torres, M.A., Jones, J.D.G., Dangl, J.L.: Reactive oxygen species signalling in response to pathogens. - Plant Physiol. 141: 373-378, 2006. Go to original source...
    43. Vlot, A.C., Dempsey, D.M.A., Klessig, D.F.: Salicylic acid, a multifaceted hormone to combat disease. - Annu. Rev. Phytopathol. 47: 177-206, 2009. Go to original source...
    44. Wang, C., Cao, J., Wang, T.T., Zheng, L., Wang, C., Liu, H.X.: [Effect of PopW from Ralstonia solanacearum on inducing plants disease resistance.] - Chin. J. biol. Control 30: 79-85, 2014a. [In Chin.]
    45. Wang, C., Liu, H.X., Cao, J., Wang, C., Guo, J.H.: [Construction of transgenic tobacco expressing popW and analysis of its biological phenotype.] - Chin. J. Biotechnol. 30: 569-580, 2014b. [In Chin.]
    46. Wang, D., Wang, Y., Fu, M., Mu, S., Han, B., Ji, H., Cai, H., Dong, H., Zhang, C.: Transgenic expression of the functional fragment Hpa1(10-42) of the harpin protein Hpa1 imparts enhanced resistance to powdery mildew in wheat. - Plant Dis. 98: 448-455, 2014c. Go to original source...
    47. Ward, E.R., Payne, G.B., Moyer, M.B., Williams, S.C., Dincher, S.S., Sharkey, K.C., Beck, J.J., Taylor, H.T., Ahl-Goy, P., Meins, F., Ryals, J.A.: Differential regulation of beta-1,3-glucanase messenger RNAs in response to pathogen infection. - Plant Physiol. 96: 390-397, 1991. Go to original source...
    48. Wei, Z.M., Laby, R.J., Zumoff, C.H., Bauer, D.W., He, S.Y., Collmer, A., Beer, S.V.: Harpin, elicitor of the hypersensitive response produced by the plant pathogen Ervinia amylovora. - Science 257: 85-88, 1992. Go to original source...
    49. Xiao, S., Chye, M.L.: Overexpression of Arabidopsis ACBP3 enhances NPR1-dependent plant resistance to Pseudomonas syringe pv. tomato DC3000. - Plant Physiol. 156: 2069-2081, 2011. Go to original source...
    50. Yao, J., Allen, C.: Chemotaxis is required for virulence and competitive fitness of the bacterial wilt pathogen Ralstonia solanacearum. - J. Bacteriol. 188: 3697-3708, 2006. Go to original source...
    51. Zhang, L., Xiao, S., Li, W., Feng, W., Li, J., Wu, Z., Gao, X., Liu, F., Shao, M.: Overexpression of a Harpin-encoding gene hrf1 in rice enhances drought tolerance. - J. exp. Bot. 62: 4229-4238, 2011a. Go to original source...
    52. Zhang, W., Yang, X., Qiu, D., Guo, L., Zeng, H., Mao, J., Gao, Q.: PeaT1-induced systemic acquired resistance in tobacco follows salicylic acid-dependent pathway. - Mol. Biol. Rep. 38: 2549-2556, 2011b. Go to original source...
    53. Zheng, L., Luo, Y.M., Xue, Q.Y., Li, S.M., Liu, H.X., Guo, J.H.: [Control and growth promotion of PopW to cucumber downy mildew under greenhouse and field conditions.] - Acat. phytopathol. sin. 43: 179-186, 2013. [In Chinese]
    54. Zipfel, C.: Early molecular events in PAMP-triggered immunity. - Curr. Opin. Plant Biol. 12: 414-420, 2009. Go to original source...

    Return to the content