DOI QR코드

DOI QR Code

Changes in pathogenic characters of Pseudomonas tolaasii 6264 strain by storage period

저장 기간에 따른 Pseudomonas tolaasii 6264 균주의 병원 특성 변화

  • Yun, Yeong-Bae (Department of Environmental and Biological Chemistry, Chungbuk National University) ;
  • Huh, Jeong-Hun (Department of Environmental and Biological Chemistry, Chungbuk National University) ;
  • Kim, Young-Kee (Department of Environmental and Biological Chemistry, Chungbuk National University)
  • Received : 2018.10.30
  • Accepted : 2018.11.14
  • Published : 2018.12.31

Abstract

Pseudomonas tolaasii strain No. 6264 has been isolated from mushroom tissue and identified as one of the major pathogen causing brown blotch disease. It secretes peptide toxins, known as tolaasin and its analogue peptides. P. tolaasii 6264 has been used as a typical pathogenic strain to study the brown blotch disease for last 20 years after confirming its blotch-forming ability, hemolytic activity, and white line formation. In this study, the characteristics of P. tolaasii 6264 strain were analyzed and compared according to storage period. Strains of P. tolaasii 6264 stored annually since 2012 were cultured and their pathogenic characters were analyzed. When the 16S rRNA sequences were compared, all strains were divided into two groups. Pathogenic characters including hemolytic activity, blotch-forming ability, and white line test were also investigated. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, had all three activities; however, the rest of stored strains showed only blotch-forming ability losing other pathogenic characters. Tolaasin peptides were purified from the bacterial cultures and analyzed by mass spectrometry. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, secreted Tol I (1987 Da), Tol II (1943 Da), and its analogues (1973 Da, 2005 Da) while some of these peptides were not found in the media cultured other strains. These results indicate that the pathogenicity of P. tolaasii could be varied during the storage period.

Pseudomonas tolaasii 6264 균주는 버섯 조직에서 분리하였고, 갈반병을 일으키는 주요 병원균 중 하나로 확인되었다. 이 균주는 펩티드 독소인 tolaasin과 유사 펩티드를 분비한다. P. tolaasii 6264는 지난 20여년간 갈반병 연구를 위한 대표균주로 사용되었으며, 이 균주는 갈반형성능, 용혈활성, 흰색침강선형성능을 확인한 후 사용하였다. 본 연구에서는, 저장 기간에 따라 P. tolaasii 6264 균주의 특성을 비교분석하였다. 2012년 이후 저장된 P. tolaasii 6264 균주를 매년 배양하여 이들 균주의 병원성을 분석하였다. 16S rRNA 염기서열을 비교하였을 때, 모든 균주들은 두 그룹으로 나뉘어졌다. 용혈활성, 갈반형성능, 흰색침강선형성능을 포함한 균주의 병원 특성도 조사하였다. P. tolaasii 6264-15-2와 P. tolaasii 6264-17 균주는 세 가지 활성을 모두 가졌으나, 그 외 균주들은 갈반형성능만 보였으며 다른 병원 특성들은 잃었다. 각 균주 배양액으로부터 tolaasin 펩티드를 순수분리하였으며, 질량분석기를 이용하여 분석하였다. P. tolaasii 6264-15-2와 P. tolaasii 6264-17 균주는 Tol I (1987 Da), Tol II (1943 Da)을 비롯한 유사체(1973 Da, 2005 Da)를 분비하였으며, 다른 균주의 배양액에서는 이 펩티드들의 일부가 발견되지 않았다. 이러한 결과는 P. tolaasii의 병원성이 저장기간동안 변할 수 있음을 보여준다.

Keywords

References

  1. Lee GS, Byun HS, Yoon KH, Lee JS, Choi KC, Jeung EB (2009) Dietary calcium and vitamin D2 supplementation with enhanced Lentinula edodes improves osteoporosis-like symptoms and induces duodenal and renal active calcium transport gene expression in mice. Eur J Nutr 48: 75-83 https://doi.org/10.1007/s00394-008-0763-2
  2. Shi YL, James AE, Benzie IFF, Buswell JA (2002) Mushroom-derived preparations in the prevention of H2O2-induced oxidative damage to cellular DNA. Teretog Carcinog Mutagen 22: 103-111 https://doi.org/10.1002/tcm.10008
  3. Chen S, Oh SR, Phung S, Hur G, Ye JJ, Kwok SL, Shrode GE, Melury M, Adams LS, Williams D (2006) Anti-aromatase activity of phytochemicals in white button mushrooms (Agaricus bisporus). Cancer Res 66: 12026-12034 https://doi.org/10.1158/0008-5472.CAN-06-2206
  4. Zhang M, Huang J, Xie X, Holman J (2009) Dietary intakes of mushrooms and green tea combine to reduce the risk of breast cancer in Chinese women. Int J Cancer 124: 1404-1408 https://doi.org/10.1002/ijc.24047
  5. Annual Report for Production of Mushroom (2016) Agricultural information and statistics. Ministry of Agricultural and Forestry, Sejong
  6. Jourdan F, Lazzaroni S, Mendez BL, Lo Cantore P, de Julio M, Amodeo P, Iacobellis NS, Evidente A, Motta A (2003) A left-handed ${\alpha}$-helix containing both L- and D-amino acids: the solution structure of the antimicrobial lipodepsipeptide tolaasin. Proteins 52: 534-543 https://doi.org/10.1002/prot.10418
  7. Hiep HM, Endo T, Saito M, Chikae M, Kim DK, Yamamura S, Takamura Y, Tamiya E (2008) Label-free detection of melittin binding to a membrane using electrochemical-localized surface plasmon resonance. Anal Chem 80: 1859-1864 https://doi.org/10.1021/ac800087u
  8. Bak M, Bywater RP, Hohwy M, Thomsen JK, Adelhorst K, Jakobsen HJ, Sorensen OW, Nielsen NC (2001) Conformation of alamethicin in oriented phospholipid bilayers determined by 15N solid-state nuclear magnetic resonance. Biophys J 81: 1684-1698 https://doi.org/10.1016/S0006-3495(01)75822-5
  9. Silva T, Claro B, Silva BFB, Vale N, Gomes P, Gomes MS, Funari SS, Teixeira J, Uhrikova D, Bastos M (2018) Unravelling a mechanism of action for a cecropin A-melittin hybrid antimicrobial peptide: the induced formation of multilamellar lipid stacks. Langmuir 34: 2158-2170 https://doi.org/10.1021/acs.langmuir.7b03639
  10. Yamaguchi S, Huster D, Waring A, Lehrer RI, Kearney W, Tack BF, Hong M (2001) Orientation and dynamic of an antimicrobial peptide in the lipid bilayer by solid-state NMR spectroscopy. Biophys J 81: 2203-2214 https://doi.org/10.1016/S0006-3495(01)75868-7
  11. Nutkins JC, Mortishire-Smith RJ, Packman LC, Brodey CL, Rainey PB, Johnstone K, Williams DH (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
  12. Cho KH, Kim ST, Kim YK (2007) Purification of a pore-forming peptide toxin, tolaasin, produced by Pseudomonas tolaasii 6264. J Biochem Mol Biol 40: 113-118
  13. 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
  14. Mu LL, Yun YB, Park SJ, Cha JS, Kim YK (2015) Various pathogenic Pseudomonas strains that cause brown blotch disease in cultivated mushrooms. J Appl Biol Chem 58: 349-354 https://doi.org/10.3839/jabc.2015.055
  15. Kim MC, Chun JS (2014) 16S rRNA gene-based identification of bacteria and archaea using the EzTaxon server. In: Goodfellow M (ed) New approaches to prokaryotic systematics, 1st edn. Elsevier, Oxford, pp 61-74
  16. Lo Cantore P, Lazzaroni S, Coraiola M, Dalla Serra M, Cafarchia C, Evidente A, Iacobellis NS (2006) Biological characterization of white line-inducing principle (WLIP) produced by Pseudomonas reactans NCPPB1311. MPMI 19: 1113-1120 https://doi.org/10.1094/MPMI-19-1113
  17. Burlinson P, Studholme D, Cambray-Young J, Heavens D, Rathjen J, Hodgkin J, Preston GM (2013) Pseudomonas fluorescens NZ17 repels grazing by C. elegans, a natural predator. ISME J 7: 1126-1138 https://doi.org/10.1038/ismej.2013.9
  18. Shirata A, Sugaya K, Takasugi M, Monde K (1995) Isolation and biological activity of toxins produced by a Japanese strain of Pseudomonas tolaasii, the pathogen of bacterial rot of cultivated oyster mushroom. Ann Phytopathol Soc Jpn 61: 492-502
  19. Bassarello C, Lazzaroni S, Bifulco G, Lo Cantore P, Iacobellis NS, Riccio R, Gomez-Paloma L, Evidente A (2004) Tolaasins A-E, five new lipodepsipeptides produced by Pseudomonas tolaasii. J Nat Prod 67: 811-816 https://doi.org/10.1021/np0303557
  20. Somerville GA, Proctor RA (2009) At the crossroads of bacterial metabolism and virulence factor synthesis in staphylococci. Microbiol Mol Biol Rev 73: 233-248 https://doi.org/10.1128/MMBR.00005-09
  21. Rutherford ST, Bassler BL (2012) Bacterial quorum sensing: its role in virulence and possibilities for its control. Cold Spring Harb Perspect Med 2: a012427
  22. Wang X, Yue J, Ding C, Wang S, Liu B, Tian M, Yu S (2016) Deletion of AS87_03730 gene changed the bacterial virulence and gene expression of Riemerella anatipestifer. Sci Rep 6: 22438. doi:10.1038/srep22438
  23. Kim JK (2009) Disruption of genes related tolaasin biosynthesis by transposon mutagenesis of Pseudomonas tolaasii. Dissertation, Gyeongsang National University
  24. Fakruddin M (2012) Biosurfactant: production and application. J Pet Environ Biotechnol 3: 124. doi: 10.4172/2157-7463.1000124
  25. Wu YS, Ngai SC, Goh BH, Chan KG, Lee LH, Chuah LH (2017) Anticancer activities of surfactic and potential application of nanotechnology assisted surfactic delivery. Front Pharmocol 8: 761. doi: 10.3389/fphar2017.00761