1887

Abstract

Previously, 12 protease-deficient mutants of the pv. () RS105 strain were recovered from a Tn-tagged mutant library. In the current study, the Tn insertion site in each mutant was mapped. Mutations in genes encoding components of the type II secretion apparatus, cAMP regulatory protein, integral membrane protease subunit, -adenosylmethionine decarboxylase proenzyme and extracellular protease () either partially or completely abolished extracellular protease activity (ECPA) and reduced virulence in rice. Transcription of was induced in all the mutants except RΔ Complementation of RΔ with restored ECPA, virulence and bacterial growth . Purified EcpA induced chlorosis- and necrosis-like symptoms similar to those induced by the pathogen when injected into rice leaves. Heterologous expression of conferred ECPA upon the vascular bacterium pv. () and upon non-pathogenic . Genetic analysis demonstrated that the C-terminal residues of EcpA in PXO99 and RS105 are different, and a frame shift in may explain the absence of EcpA activity in . Collectively, these results suggest that EcpA is a tissue-specific virulence factor for but not , although the two pathovars are closely related bacterial pathogens of rice.

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2012-09-01
2019-10-18
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References

  1. Bogdanove A. J. , Koebnik R. , Lu H. , Furutani A. , Angiuoli S. V. , Patil P. B. , Van Sluys M. A. , Ryan R. P. , Meyer D. F. . & other authors ( 2011; ). Two new complete genome sequences offer insight into host and tissue specificity of plant pathogenic Xanthomonas spp. . J Bacteriol 193:, 5450–5464. [CrossRef] [PubMed]
    [Google Scholar]
  2. Büttner D. , Bonas U. . ( 2010; ). Regulation and secretion of Xanthomonas virulence factors. . FEMS Microbiol Rev 34:, 107–133. [CrossRef] [PubMed]
    [Google Scholar]
  3. Cho H. J. , Park Y. J. , Noh T. H. , Kim Y. T. , Kim J. G. , Song E. S. , Lee D. H. , Lee B. M. . ( 2008; ). Molecular analysis of the hrp gene cluster in Xanthomonas oryzae pathovar oryzae KACC10859. . Microb Pathog 44:, 473–483. [CrossRef] [PubMed]
    [Google Scholar]
  4. Cianciotto N. P. . ( 2005; ). Type II secretion: a protein secretion system for all seasons. . Trends Microbiol 13:, 581–588. [CrossRef] [PubMed]
    [Google Scholar]
  5. da Silva A. C. R. , Ferro J. A. , Reinach F. C. , Farah C. S. , Furlan L. R. , Quaggio R. B. , Monteiro-Vitorello C. B. , Van Sluys M. A. , Almeida N. F. . & other authors ( 2002; ). Comparison of the genomes of two Xanthomonas pathogens with differing host specificities. . Nature 417:, 459–463. [CrossRef] [PubMed]
    [Google Scholar]
  6. de Groot A. , Filloux A. , Tommassen J. . ( 1991; ). Conservation of xcp genes, involved in the two-step protein secretion process, in different Pseudomonas species and other Gram-negative bacteria. . Mol Gen Genet 229:, 278–284. [CrossRef] [PubMed]
    [Google Scholar]
  7. DebRoy S. , Dao J. , Söderberg M. , Rossier O. , Cianciotto N. P. . ( 2006; ). Legionella pneumophila type II secretome reveals unique exoproteins and a chitinase that promotes bacterial persistence in the lung. . Proc Natl Acad Sci U S A 103:, 19146–19151. [CrossRef] [PubMed]
    [Google Scholar]
  8. Dow J. M. , Daniels M. J. , Dums F. , Turner P. C. , Gough C. . ( 1989; ). Genetic and biochemical analysis of protein export from Xanthomonas campestris . . J Cell Sci Suppl 11:, 59–72.[PubMed] [CrossRef]
    [Google Scholar]
  9. Dow J. M. , Clarke B. R. , Milligan D. E. , Tang J. L. , Daniels M. J. . ( 1990; ). Extracellular proteases from Xanthomonas campestris pv. campestris, the black rot pathogen. . Appl Environ Microbiol 56:, 2994–2998.[PubMed]
    [Google Scholar]
  10. Dow J. M. , Fan M. J. , Newman M. A. , Daniels M. J. . ( 1993; ). Differential expression of conserved protease genes in crucifer-attacking pathovars of Xanthomonas campestris . . Appl Environ Microbiol 59:, 3996–4003.[PubMed]
    [Google Scholar]
  11. Dow J. M. , Davies H. A. , Daniels M. J. . ( 1998; ). A metalloprotease from Xanthomonas campestris that specifically degrades proline/hydroxyproline-rich glycoproteins of the plant extracellular matrix. . Mol Plant Microbe Interact 11:, 1085–1093. [CrossRef] [PubMed]
    [Google Scholar]
  12. Furutani A. , Tsuge S. , Ohnishi K. , Hikichi Y. , Oku T. , Tsuno K. , Inoue Y. , Ochiai H. , Kaku H. , Kubo Y. . ( 2004; ). Evidence for HrpXo-dependent expression of type II secretory proteins in Xanthomonas oryzae pv. oryzae . . J Bacteriol 186:, 1374–1380. [CrossRef] [PubMed]
    [Google Scholar]
  13. Furutani A. , Takaoka M. , Sanada H. , Noguchi Y. , Oku T. , Tsuno K. , Ochiai H. , Tsuge S. . ( 2009; ). Identification of novel type III secretion effectors in Xanthomonas oryzae pv. oryzae . . Mol Plant Microbe Interact 22:, 96–106. [CrossRef] [PubMed]
    [Google Scholar]
  14. Gnanamanickam S. S. , Pryiyadarasani V. , Narayanan N. N. , Vasudevan P. , Kavitha S. . ( 1999; ). An overview of bacterial blight disease of rice and strategies for management. . Curr Sci 77:, 1435–1444.
    [Google Scholar]
  15. Hajri A. , Brin C. , Hunault G. , Lardeux F. , Lemaire C. , Manceau C. , Boureau T. , Poussier S. . ( 2009; ). A “repertoire for repertoire” hypothesis: repertoires of type three effectors are candidate determinants of host specificity in Xanthomonas . . PLoS ONE 4:, e6632. [CrossRef] [PubMed]
    [Google Scholar]
  16. Hu J. , Qian W. , He C. Z. . ( 2007; ). The Xanthomonas oryzae pv. oryzae eglXoB endoglucanase gene is required for virulence to rice. . FEMS Microbiol Lett 269:, 273–279. [CrossRef] [PubMed]
    [Google Scholar]
  17. Jha G. , Rajeshwari R. , Sonti R. V. . ( 2007; ). Functional interplay between two Xanthomonas oryzae pv. oryzae secretion systems in modulating virulence on rice. . Mol Plant Microbe Interact 20:, 31–40. [CrossRef] [PubMed]
    [Google Scholar]
  18. Jiang J. , Zou H. S. , Li Y. R. , Chen G. Y. . ( 2009; ). [Expression of the hrcC, hrpE and hpa3 genes is not regulated by the hrpG and hrpX genes in a rice pathogen Xanthomonas oryzae pv. oryzicola]. . Wei Sheng Wu Xue Bao 49:, 1018–1025 (in Chinese).[PubMed]
    [Google Scholar]
  19. Johnson T. L. , Abendroth J. , Hol W. G. J. , Sandkvist M. . ( 2006; ). Type II secretion: from structure to function. . FEMS Microbiol Lett 255:, 175–186. [CrossRef] [PubMed]
    [Google Scholar]
  20. Lee B. M. , Park Y. J. , Park D. S. , Kang H. W. , Kim J. G. , Song E. S. , Park I. C. , Yoon U. H. , Hahn J. H. . & other authors ( 2005; ). The genome sequence of Xanthomonas oryzae pathovar oryzae KACC10331, the bacterial blight pathogen of rice. . Nucleic Acids Res 33:, 577–586. [CrossRef] [PubMed]
    [Google Scholar]
  21. Li Y. R. , Zou H. S. , Che Y. Z. , Cui Y. P. , Guo W. , Zou L. F. , Chatterjee S. , Biddle E. M. , Yang C. H. , Chen G. Y. . ( 2011; ). A novel regulatory role of HrpD6 in regulating hrp-hrc-hpa genes in Xanthomonas oryzae pv. oryzicola . . Mol Plant Microbe Interact 24:, 1086–1101. [CrossRef] [PubMed]
    [Google Scholar]
  22. Li Y. R. , Xiao Y. L. , Zou L. , Zou H. , Chen G. . ( 2012; ). Identification of HrpX regulon genes in Xanthomonas oryzae pv. oryzicola using a GFP visualization technique. . Arch Microbiol 194:, 281–291. [CrossRef] [PubMed]
    [Google Scholar]
  23. Liang C. Y. , Li D. C. , Yi J. P. , Xu Z. G. . ( 1994; ). Studies on virulence types of Xanthomonas oryzae pv. oryzicola collected from diseased area in China. . Journal of Nanjing Agricultural University 17:, 48–52.
    [Google Scholar]
  24. Lu H. , Patil P. , Van Sluys M. A. , White F. F. , Ryan R. P. , Dow J. M. , Rabinowicz P. , Salzberg S. L. , Leach J. E. . & other authors ( 2008; ). Acquisition and evolution of plant pathogenesis-associated gene clusters and candidate determinants of tissue-specificity in Xanthomonas . . PLoS ONE 3:, e3828. [CrossRef] [PubMed]
    [Google Scholar]
  25. McNeil M. , Darvill A. G. , Fry S. C. , Albersheim P. . ( 1984; ). Structure and function of the primary cell walls of plants. . Annu Rev Biochem 53:, 625–663. [CrossRef] [PubMed]
    [Google Scholar]
  26. Niño-Liu D. O. , Ronald P. C. , Bogdanove A. J. . ( 2006; ). Xanthomonas oryzae pathovars: model pathogens of a model crop. . Mol Plant Pathol 7:, 303–324. [CrossRef] [PubMed]
    [Google Scholar]
  27. Noël L. , Thieme F. , Nennstiel D. , Bonas U. . ( 2001; ). cDNA-AFLP analysis unravels a genome-wide hrpG-regulon in the plant pathogen Xanthomonas campestris pv. vesicatoria . . Mol Microbiol 41:, 1271–1281. [CrossRef] [PubMed]
    [Google Scholar]
  28. Ochiai H. , Inoue Y. , Takeya M. , Sasaki A. , Kaku H. . ( 2005; ). Genome sequence of Xanthomonas oryzae pv. oryzae suggests contribution of large numbers of effector genes and insertion sequences to its race diversity. . Jpn Agric Res Q 39:, 275–287.[CrossRef]
    [Google Scholar]
  29. Ou S. H. . ( 1985; ). Bacterial leaf blight. , In Rice Diseases, , 2nd edn., pp. 61–93. Kew, UK:: Commonwealth Mycological Institute;.
    [Google Scholar]
  30. Qian W. , Jia Y. T. , Ren S. X. , He Y. Q. , Feng J. X. , Lu L. F. , Sun Q. H. , Ying G. , Tang D. J. . & other authors ( 2005; ). Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris . . Genome Res 15:, 757–767. [CrossRef] [PubMed]
    [Google Scholar]
  31. Rajeshwari R. , Jha G. , Sonti R. V. . ( 2005; ). Role of an in planta-expressed xylanase of Xanthomonas oryzae pv. oryzae in promoting virulence on rice. . Mol Plant Microbe Interact 18:, 830–837. [CrossRef] [PubMed]
    [Google Scholar]
  32. Ray S. K. , Rajeshwari R. , Sonti R. V. . ( 2000; ). Mutants of Xanthomonas oryzae pv. oryzae deficient in general secretory pathway are virulence deficient and unable to secrete xylanase. . Mol Plant Microbe Interact 13:, 394–401. [CrossRef] [PubMed]
    [Google Scholar]
  33. Salzberg S. L. , Sommer D. D. , Schatz M. C. , Phillippy A. M. , Rabinowicz P. D. , Tsuge S. , Furutani A. , Ochiai H. , Delcher A. L. . & other authors ( 2008; ). Genome sequence and rapid evolution of the rice pathogen Xanthomonas oryzae pv. oryzae PXO99A. . BMC Genomics 9:, 204–221. [CrossRef] [PubMed]
    [Google Scholar]
  34. Sambrook J. , Russell D. W. . ( 2001; ). Molecular Cloning: a Laboratory Manual, , 3rd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;.
    [Google Scholar]
  35. Sandkvist M. . ( 2001; ). Type II secretion and pathogenesis. . Infect Immun 69:, 3523–3535. [CrossRef] [PubMed]
    [Google Scholar]
  36. Shao F. , Golstein C. , Ade J. , Stoutemyer M. , Dixon J. E. , Innes R. W. . ( 2003; ). Cleavage of Arabidopsis PBS1 by a bacterial type III effector. . Science 301:, 1230–1233. [CrossRef] [PubMed]
    [Google Scholar]
  37. Sun Q. H. , Hu J. , Huang G. X. , Ge C. , Fang R. X. , He C. Z. . ( 2005; ). Type II secretion pathway structural gene xpsE, xylanase- and cellulase secretion and virulence in Xanthomonas oryzae pv. oryzae . . Plant Pathol 54:, 15–21. [CrossRef]
    [Google Scholar]
  38. Takeuchi Y. , Tohbaru M. , Sato A. . ( 1994; ). Polysaccharide in primary cell walls of rice cells in suspension culture. . Phytochemistry 35:, 361–363. [CrossRef]
    [Google Scholar]
  39. Thanassi D. G. , Hultgren S. J. . ( 2000; ). Multiple pathways allow protein secretion across the bacterial outer membrane. . Curr Opin Cell Biol 12:, 420–430. [CrossRef] [PubMed]
    [Google Scholar]
  40. Tsuge S. , Terashima S. , Furutani A. , Ochiai H. , Oku T. , Tsuno K. , Kaku H. , Kubo Y. . ( 2005; ). Effects on promoter activity of base substitutions in the cis-acting regulatory element of HrpXo regulons in Xanthomonas oryzae pv. oryzae . . J Bacteriol 187:, 2308–2314. [CrossRef] [PubMed]
    [Google Scholar]
  41. Voulhoux R. , Ball G. , Ize B. , Vasil M. L. , Lazdunski A. , Wu L. F. , Filloux A. . ( 2001; ). Involvement of the twin-arginine translocation system in protein secretion via the type II pathway. . EMBO J 20:, 6735–6741. [CrossRef] [PubMed]
    [Google Scholar]
  42. Wang L. , Makino S. , Subedee A. , Bogdanove A. J. . ( 2007; ). Novel candidate virulence factors in rice pathogen Xanthomonas oryzae pv. oryzicola as revealed by mutational analysis. . Appl Environ Microbiol 73:, 8023–8027. [CrossRef] [PubMed]
    [Google Scholar]
  43. Wengelnik K. , Van den Ackerveken G. , Bonas U. . ( 1996; ). HrpG, a key hrp regulatory protein of Xanthomonas campestris pv. vesicatoria is homologous to two-component response regulators. . Mol Plant Microbe Interact 9:, 704–712. [CrossRef] [PubMed]
    [Google Scholar]
  44. Xiang T. , Zong N. , Zou Y. , Wu Y. , Zhang J. , Xing W. , Li Y. , Tang X. , Zhu L. . & other authors ( 2008; ). Pseudomonas syringae effector AvrPto blocks innate immunity by targeting receptor kinases. . Curr Biol 18:, 74–80. [CrossRef] [PubMed]
    [Google Scholar]
  45. Zou L. F. , Wang X. P. , Xiang Y. , Zhang B. , Li Y. R. , Xiao Y. L. , Wang J. S. , Walmsley A. R. , Chen G. Y. . ( 2006; ). Elucidation of the hrp clusters of Xanthomonas oryzae pv. oryzicola that control the hypersensitive response in nonhost tobacco and pathogenicity in susceptible host rice. . Appl Environ Microbiol 72:, 6212–6224. [CrossRef] [PubMed]
    [Google Scholar]
  46. Zou L. F. , Li Y. R. , Chen G. Y. . ( 2011a; ). A non-marker mutagenesis strategy to generate poly-hrp gene mutants in the rice pathogen Xanthomonas oryzae pv. oryzicola . . Agric Sci China 10:, 1139–1150. [CrossRef]
    [Google Scholar]
  47. Zou H. S. , Yuan L. , Guo W. , Li Y. R. , Che Y. Z. , Zou L. F. , Chen G. Y. . ( 2011b; ). Construction of a Tn5-tagged mutant library of Xanthomonas oryzae pv. oryzicola as an invaluable resource for functional genomics. . Curr Microbiol 62:, 908–916. [CrossRef] [PubMed]
    [Google Scholar]
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