1887

Abstract

is an emerging plant-pathogenic bacterium that causes disease in rice in several of the major rice-producing areas throughout the world. In the southern United States, is the major causal agent of bacterial panicle blight of rice and has caused severe yield losses in recent decades. Despite its importance, few management options are available for diseases caused by , and knowledge of how this pathogen causes disease is limited. In an effort to identify novel factors that contribute to the pathogenicity of , random mutagenesis using the miniTn transposon was performed on two strains of . Resultant mutants were screened in the laboratory for altered phenotypes in various known or putative virulence factors, including toxoflavin, lipase and extracellular polysaccharides. Mutants that exhibited altered phenotypes compared to their parent strain were selected and subsequently characterized using a PCR-based method to identify the approximate location of the transposon insertion. Altogether, approximately 20 000 random mutants were screened and 51 different genes were identified as having potential involvement in the production of toxoflavin, lipase and/or extracellular polysaccharide. Especially, two regulatory genes, and , encoding a LysR-type transcriptional regulator and a σ-dependent response regulator, respectively, were discovered in this study as new negative regulatory factors for the production of toxoflavin, the major phytotoxin synthesized by and involved in bacterial pathogenesis.

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2017-02-01
2024-03-28
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References

  1. Ham JH, Melanson RA, Rush MC. Burkholderia glumae: next major pathogen of rice?. Mol Plant Pathol 2011; 12:329–339 [View Article][PubMed]
    [Google Scholar]
  2. Nandakumar R, Shahjahan AKM, Yuan XL, Dickstein ER, Groth DE et al. Burkholderia glumae and B. gladioli cause bacterial panicle blight in rice in the southern United States. Plant Dis 2009; 93:896–905 [CrossRef]
    [Google Scholar]
  3. Saichuk J. Louisiana Rice Production Handbook Baton Rouge, LA, USA: LSU AgCenter; 2009 p. 128
    [Google Scholar]
  4. Saichuk J, Blanche B, Courville B, Harrell D, Groth D et al. 2010 Rice Varieties and Management Tips Baton Rouge, LA, USA: LSU AgCenter; 2009 p. 24
    [Google Scholar]
  5. Hikichi Y, Egami H, Oguri Y, Okuno T. Fitness for survival of Burkholderia glumae resistant to oxolinic acid in rice plants. Jpn J Phytopath 1998; 64:147–152 [CrossRef]
    [Google Scholar]
  6. Maeda Y, Kiba A, Ohnish K, Hikichi Y. New method to detect oxolinic acid-resistant Burkholderia glumae infecting rice seeds using a mismatch amplification mutation assay polymerase chain reaction. J Gen Plant Pathol 2004; 70:215–217 [CrossRef]
    [Google Scholar]
  7. Kim J, Kim JG, Kang Y, Jang JY, Jog GJ et al. Quorum sensing and the LysR-type transcriptional activator ToxR regulate toxoflavin biosynthesis and transport in Burkholderia glumae. Mol Microbiol 2004; 54:921–934 [View Article][PubMed]
    [Google Scholar]
  8. Suzuki F, Sawada HA, Zegami K, Tsuchiya K. Molecular characterization of the tox operon involved in toxoflavin biosynthesis of Burkholderia glumae. J Gen Plant Pathol 2004; 70:97–107 [CrossRef]
    [Google Scholar]
  9. Cottyn B, Cerez MT, Van Outryve MF, Barroga J. Bacterial diseases of rice I. Pathogenic bacteria associated with sheath rot complex and grain discoloration of rice in the Philippines. Plant Dis 1996; 80:429–437 [CrossRef]
    [Google Scholar]
  10. Cottyn B, Van Outryve MF, Cerez MT, De Cleene M, Swings J. Bacterial diseases of rice II. Characterization of pathogenic bacteria associated with sheath rot complex and grain discoloration of rice in the Philippines. Plant Dis 1996; 80:438–445 [CrossRef]
    [Google Scholar]
  11. Zeigler RS, Alvarez E. Grain discoloration of rice caused by Pseudomonas glumae in Latin America. Plant Dis 1989; 73:368 [CrossRef]
    [Google Scholar]
  12. Zhou XG. First report of bacterial panicle blight of rice caused by Burkholderia glumae in South Africa. Plant Dis 2014; 98:566 [CrossRef]
    [Google Scholar]
  13. Schaad NW. Emerging plant pathogenic bacteria and global warming. In Fatmi M, Collmer A, Iacobellis NS, Mansfield JW, Murillo J. et al (editors) Pseudomonas syringae Pathovars and Related Pathogens – Identification, Epidemiology and Genomics Netherlands: Springer; 2008 pp. 369–379 [CrossRef]
    [Google Scholar]
  14. Karki HS, Shrestha BK, Han JW, Groth DE, Barphagha IK et al. Diversities in virulence, antifungal activity, pigmentation and DNA fingerprint among strains of Burkholderia glumae. PLoS One 2012; 7:e45376 [View Article][PubMed]
    [Google Scholar]
  15. Devescovi G, Bigirimana J, Degrassi G, Cabrio L, LiPuma JJ et al. Involvement of a quorum-sensing-regulated lipase secreted by a clinical isolate of Burkholderia glumae in severe disease symptoms in rice. Appl Env Microbiol 2007; 73:4950–4958 [CrossRef]
    [Google Scholar]
  16. Kim J, Kang Y, Choi O, Jeong Y, Jeong JE et al. Regulation of polar flagellum genes is mediated by quorum sensing and FlhDC in Burkholderia glumae. Mol Microbiol 2007; 64:165–179 [View Article][PubMed]
    [Google Scholar]
  17. Chen R, Barphagha IK, Karki HS, Ham JH. Dissection of quorum-sensing genes in Burkholderia glumae reveals non-canonical regulation and the new regulatory gene tofM for toxoflavin production. PLoS One 2012; 7:e52150 [View Article][PubMed]
    [Google Scholar]
  18. Bernier SP, Nguyen DT, Sokol PA. A LysR-type transcriptional regulator in Burkholderia cenocepacia influences colony morphology and virulence. Infect and Immun 2008; 76:38–47 [CrossRef]
    [Google Scholar]
  19. Rott P, Fleites L, Marlow G, Royer M, Gabriel DW. Identification of new candidate pathogenicity factors in the xylem-invading pathogen Xanthomonas albilineans by transposon mutagenesis. Mol Plant Microbe Interact 2011; 24:594–605 [View Article][PubMed]
    [Google Scholar]
  20. Sambrook J. Molecular Cloning: a Laboratory Manual, 3rd ed. Cold Spring Harbor Press; 2001
    [Google Scholar]
  21. Correa VR, Majerczak DR, Ammar el D, Merighi M, Pratt RC et al. The bacterium Pantoea stewartii uses two different type III secretion systems to colonize its plant host and insect vector. Appl Environ Microbiol 2012; 78:6327–6336 [CrossRef]
    [Google Scholar]
  22. Smibert RM, Krieg NR. Phenotypic characterization. In Gerhardt P, Murray RGE, Wood WA, Krieg NR. (editors) Methods for General and Molecular Bacteriology Washington, DC, USA: American Society for Microbiology; 1994
    [Google Scholar]
  23. Kwon YM, Ricke SC. Efficient amplification of multiple transposon-flanking sequences. J Microbiol Meth 2000; 41:195–199 [CrossRef]
    [Google Scholar]
  24. Iiyama K, Furuya N, Takanami Y, Matsuyama N. A role of phytotoxin in virulence of Pseudomonas glumae Kurita et Tabei. JpnJ Phytopath 1995; 61:470–476 [CrossRef]
    [Google Scholar]
  25. Jung WS, Lee J, Kim MI, Ma J, Nagamatsu T et al. Structural and functional analysis of phytotoxin toxoflavin-degrading enzyme. PLoS One 2011; 6:e22443 [View Article][PubMed]
    [Google Scholar]
  26. Karki HS, Barphagha IK, Ham JH. A conserved two-component regulatory system, PidS/PidR, globally regulates pigmentation and virulence-related phenotypes of Burkholderia glumae. Molecul Plant Pathol 2012; 13:785–794 [CrossRef]
    [Google Scholar]
  27. Saxton AM. A macro for converting mean separation output to letter groundings in proc mixed. In Proceedings of the 23rd Annual Users Group International Conference SAS Institute, Inc; 1998
    [Google Scholar]
  28. Francis F, Kim J, Ramaraj T, Farmer A, Rush MC et al. Comparative genomic analysis of two Burkholderia glumae strains from different geographic origins reveals a high degree of plasticity in genome structure associated with genomic islands. Mol Genet Genomics 2013; 288:195–203 [View Article][PubMed]
    [Google Scholar]
  29. Sandkvist M. Biology of type II secretion. Mol Microbiol 2001; 40:271–283[PubMed] [CrossRef]
    [Google Scholar]
  30. Von Bodman SB, Bauer WD, Coplin DL. Quorum sensing in plant-pathogenic bacteria. Annu Rev Phytopathol 2003; 41:455–482 [View Article][PubMed]
    [Google Scholar]
  31. Bernhard F, Coplin DL, Geider K. A gene cluster for amylovoran synthesis in Erwinia amylovora: characterization and relationship to cps genes in Erwinia stewartii. Mol Genet Genomics 1993; 239:158–168
    [Google Scholar]
  32. McWilliams R, Chapman M, Kowalczuk KM, Hersberger D, Sun J et al. Complementation analyses of Pseudomonas solanacearum extracellular polysaccharide mutants and identification of genes responsive to EpsR. Mol Plant Microbe Interact 1995; 8:837–844[PubMed] [CrossRef]
    [Google Scholar]
  33. Lu GT, Xie JR, Chen L, Hu JR, An SQ et al. Glyceraldehyde-3-phosphate dehydrogenase of Xanthomonas campestris pv. campestris is required for extracellular polysaccharide production and full virulence. Microbiology 2009; 155:1602–1612 [View Article][PubMed]
    [Google Scholar]
  34. Barreto M, Jedlicki E, Holmes DS. Identification of a gene cluster for the formation of extracellular polysaccharide precursors in the chemolithoautotroph Acidithiobacillus ferrooxidans. Appl Env Microbiol 2005; 71:2902–2909 [CrossRef]
    [Google Scholar]
  35. Chai Y, Beauregard PB, Vlamakis H, Losick R, Kolter R. Galactose metabolism plays a crucial role in biofilm formation by Bacillus subtilis. MBio 2012; 3:e00184-12 [View Article][PubMed]
    [Google Scholar]
  36. Li CT, Liao CT, Du SC, Hsiao YP, Lo HH et al. Functional characterization and transcriptional analysis of galE gene encoding a UDP-galactose 4-epimerase in Xanthomonas campestris pv. campestris. Microbiol Res 2014; 169:441–452 [View Article][PubMed]
    [Google Scholar]
  37. Pugsley AP. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev 1993; 57:50–108[PubMed]
    [Google Scholar]
  38. Ditta G, Stanfield S, Corbin D, Helinski DR. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 1980; 77:7347–7351[PubMed] [CrossRef]
    [Google Scholar]
  39. de Lorenzo V, Eltis L, Kessler B, Timmis KN. Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes. Gene 1993; 123:17–24 [View Article][PubMed]
    [Google Scholar]
  40. Fouts DE, Abramovitch RB, Alfano JR, Baldo AM, Buell CR et al. Genomewide identification of Pseudomonas syringae pv. tomato DC3000 promoters controlled by the HrpL alternative sigma factor. Proc Natl Acad Sci USA 2002; 99:2275–2280 [View Article][PubMed]
    [Google Scholar]
  41. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 1995; 166:175–176 [View Article][PubMed]
    [Google Scholar]
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