Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Characterization of Phage-Resistant Strains Derived from Pseudomonas tolaasii 6264, which Causes Brown Blotch Disease

  • Yun, Yeong-Bae (Department of Environmental and Biological Chemistry, Chungbuk National University) ;
  • Han, Ji-Hye (Department of Environmental and Biological Chemistry, Chungbuk National University) ;
  • Kim, Young-Kee (Department of Environmental and Biological Chemistry, Chungbuk National University)
  • Received : 2018.09.14
  • Accepted : 2018.10.01
  • Published : 2018.12.28
Download PDF
⟨ Previous Next ⟩

Abstract

Pseudomonas tolaasii 6264 is a representative strain that causes bacterial blotch disease on the cultivated oyster mushroom, Pleurotus ostreatus. Bacteriophages are able to sterilize the pathogenic P. tolaasii strains, and therefore, they can be applied in creating disease-free mushroom cultivation farms, through a method known as "phage therapy". For successful phage therapy, the characterization of phage-resistant strains is necessary, since they are frequently induced from the original pathogenic bacteria in the presence of phages. When 10 different phages were incubated with P. tolaasii 6264, their corresponding phage-resistant strains were obtained. In this study, changes in pathogenic, genetic, and biochemical characteristics as well as the acquired phage resistance of these strains were investigated. In the phylogenetic analyses, all phage-resistant strains were identical to the original parent strain based on the sequence comparison of 16S rRNA genes. When various phage-resistant strains were examined by three different methods, pitting test, white line test, and hemolytic activity, they were divided into three groups: strains showing all positive results in three tests, two positive in the first two tests, and all negative. Nevertheless, all phage-resistant strains showed that their pathogenic activities were reduced or completely lost.

Keywords

References

  1. Paine SG. 1919. Studies in bacteriosis. II. A brown blotch disease of cultivated mushrooms. Ann. Appl. Biol. 5: 206-219. https://doi.org/10.1111/j.1744-7348.1919.tb05291.x
  2. Tolaas AG. 1915. A bacterial disease of cultivated mushrooms. Phytopathol. 5: 51-54.
  3. Housby JN, Mann NH. 2009. Phage therapy. Drug Discov. Today 14: 536-540. https://doi.org/10.1016/j.drudis.2009.03.006
  4. Lu TK, Koeris MS. 2011. The next generation of bacteriophage therapy. Curr. Opin. Micorbiol. 14: 524-531. https://doi.org/10.1016/j.mib.2011.07.028
  5. Wright A, Hawkins CH, Anggard EE, Harper DR. 2009. A controlled clinical trial of a therapeutic bacteriophage preparation in chronic otitis due to antibiotic-resistant Pseudomonas aeruginosa; a preliminary report of efficacy. Clin. Otolaryngol. 34: 349-357. https://doi.org/10.1111/j.1749-4486.2009.01973.x
  6. Fujiwara A, Fujisawa M, Hamasaki R, Kawasaki T, Fujie M, Yamada T. 2011. Biocontrol of Ralstonia solanacearum by treatment with lytic bacteriophages. Appl. Environ. Microbiol. 77: 4155-4162. https://doi.org/10.1128/AEM.02847-10
  7. Yosef I, Kiro R, Molshanski-Mor S, Edgar R, Qimron U. 2014. Different approaches for using bacteriophages against antibiotic-resistant bacteria. Bacteriophage 4: e28491. https://doi.org/10.4161/bact.28491
  8. Parracho HM, Burrowes BH, Enright MC, McConville ML, Harper DR. 2012. The role of regulated clinical trials in the development of bacteriophage therapeutics. Mol. Genet. Med. 6: 279-286.
  9. Kelly D, McAuliffe O, Ross RP, O'Mahony J, Coffey A. 2011. Development of a broad-host-range phage cocktail for biocontrol. Bioeng. Bugs 2: 31-37. https://doi.org/10.4161/bbug.2.1.13657
  10. Kim MH, Park SW, Kim YK. 2011. Bacteriophages of Pseudomonas tolaasii for the biological control of brown blotch disease. J. Korean Soc. Appl. Biol. Chem. 54: 99-104.
  11. Park SJ, Han JH, Kim YK. 2016. Isolation of bacteriophageresistant Pseudomonas tolaasii strains and their pathogenic characters. J. Appl. Biol. Chem. 59: 351-356. https://doi.org/10.3839/jabc.2016.059
  12. Khan A, Jett J. 2004. Cycle sequencing using bigdye v3.1: Performed on fosmid DNA template. http://www.jgi.doe.gov/sequencing/protocols/BigDyev3.1FosmidCycleSequencingSOP.doc.
  13. Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, et al. 2012. Introducing EzTaxon-e: a prokaryotic 16s rRNA gene sequence database with phylotypes that represent uncultured species. Int. J. Syst. Evol. Microbiol. 62: 716-721. https://doi.org/10.1099/ijs.0.038075-0
  14. Thompson JD, Higgins DG, Gibson TJ. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673-4680. https://doi.org/10.1093/nar/22.22.4673
  15. Gandy DG. 1968. pp. 150-154. A Technique for Screening Bacteria Causing Brown Blotch of Cultivated Mushrooms. Annual Report Glasshouse Crops Research Institute 1967.
  16. Rainey PB, Brodey CL, Johnstone K. 1991. Biological properties and spectrum of activity of tolaasin, a lipodepsipeptide toxin produced by the mushroom pathogen Pseudomonas tolaasii. Physiol. Mol. Plant. Pathol. 39: 57-70. https://doi.org/10.1016/0885-5765(91)90031-C
  17. Lo Cantore P, Giorgio A, Iacobellis NS. 2015. Bioactivity of volatile organic compounds produced by Pseudomonas tolaasii. Front. Microbiol. 6: 1802.
  18. Simon JL, Julie ES, Sylvain M. 2010. Bacteriophage resistance mechanisms. Nat. Rev. Microbiol. 8: 317-327. https://doi.org/10.1038/nrmicro2315
  19. Koskella B, Brockhurst MA. 2014. Bacteria-phage coevolution as a driver of ecological and evolutionary processes in microbial communities. FEMS Microbiol. Rev. 38: 916-931. https://doi.org/10.1111/1574-6976.12072
  20. Brüssow H, Canchaya C, Hardt W, Bru H. 2004. Phages and the evolution of bacterial pathogens: from genomic rearrangements to lysogenic conversion. Microbiol. Mol. Biol. Rev. 68: 560-602. https://doi.org/10.1128/MMBR.68.3.560-602.2004
  21. Filippov AA, Sergueev KV, He Y, Huang XZ, Gnade BT, Mueller AJ, et al. 2011. Bacteriophage-resistant mutants in Yersinia pestis: identification of phage receptors and attenuation for mice. PLoS One 6: e25486. https://doi.org/10.1371/journal.pone.0025486
  22. Capparelli R, Nocerino N, Iannaccone M, Ercolini D, Parlato M, Chiara M, et al. 2010. Bacteriophage therapy of Salmonella enterica: a fresh appraisal of bacteriophage therapy. J. Infect. Dis. 201: 52-61. https://doi.org/10.1086/648478
  23. Leon M, Bastias R. 2015. Virulence reduction in bacteriophage resistant bacteria. Front. Microbiol. 6: 343.
  24. Yun YB, Park SW, Cha JS, Kim YK. 2013. Biological characterization of various strains of Pseudomonas tolaasii that causes brown blotch disease. J. Korean Soc. Appl. Biol. Chem. 56: 41-45. https://doi.org/10.1007/s13765-012-2242-y
  25. Nutkins JC, Mortishire-Smith RJ, Williams DH, Packman LC, Brodey CL, Rainey PB, et al. 1991. Structure determination of tolaasin, an extracellular lipodepsipeptide produced by the mushroom pathogen Pseudomonas tolaasii paine. J. Am. Chem. Soc. 113: 2621-2627. https://doi.org/10.1021/ja00007a040
  26. Fett WF, Wells JM, Cescutti P, Wijey C. 1995. Identification of exopolysaccharides produced by fluorescent Pseudomonads associated with commercial mushroom (Agaricus bisporus) production. Appl. Environ. Microbiol. 61: 513-517.
  27. Labrie SJ, Samson JE, Moineau S. 2010. Bacteriophage resistance mechanisms. Nat. Rev. Microbiol. 8: 317-327. https://doi.org/10.1038/nrmicro2315