Journal of Mushroom (한국버섯학회지)

The Korean Society of Mushroom Science (한국버섯학회)
권호 표지
DOI QR코드

DOI QR Code

Optimum cultivation conditions for mass production of antagonistic bacterium Pseudomonas azotoformans HC5 effective in antagonistic of brown blotch disease caused by Pseudomonas tolaasii

버섯 세균갈색무늬병균(Pseudomonas tolaasii)에 대한 길항세균 Pseudomonas azotoformans HC5의 대량배양을 위한 최적 배양조건

  • 이찬중 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 문지원 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 유영미 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 한주연 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 정종천 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 공원식 (농촌진흥청 국립원예특작과학원 버섯과)
  • Received : 2015.06.15
  • Accepted : 2015.06.29
  • Published : 2015.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

This study was conducted to investigate optimum conditions for mass production of antagonistic microbes Pseudomonas azotoformans HC5. P. azotoformans HC5 is a potent biological control agent to control brown blotch disease caused by Pseudomonas tolaasii. This markedly showed the antagonistic activity against P. tolaasii, the most destructive pathogen of cultivated mushrooms. To define the optimum conditions for the mass production of the P. azotoformans HC5, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 6.0 and $15^{\circ}C$, respectively. The optimal concentration of medium elements for the growth of pathogen inhibitor bacterium was determined as follows: 0.6% adonitol, 1.5% yeast extract, 0.8% $NH_4H_2PO_4$, 5mM $MgSO_4$, and 0.2% asparagine.

버섯 세균갈성색무늬병원균인 Pseudomonas tolaasii에 대한 길항미생물로 보고된 Pseudomonas azotoformans HC5 균주의 배양적 특성과 대량 배양을 위한 최적배양 조건을 설정하였다. HC5 균주의 생육온도는 $10{\sim}20^{\circ}C$, pH는 6.0~9.0의 범위에서 왕성한 생육을 보였다. 대량배양을 위한 효율적인 영양원 선발을 위하여 기본배지에 탄소원 fructose 등 18종, 무기질소원 $NH_4Cl$ 등 6종, 유기질소원 peptone 등 6종 그리고 아미노산 asparagine 등 11종을 각각 1%씩 첨가하였고, 무기염류 13종을 1 mM 농도로 첨가하여 각각에 대한 생육에 미치는 영향을 조사하였다. 또한 선발된 각각의 영양성분들에 대한 최적 농도를 조사하기 위하여 각각의 성분을 최소 0.1%에서 최대 4.0%까지 배지에 첨가하여 배양 후 생육정도를 조사하였다. 그 결과, 대량배양을 위한 생육최적조건은 온도 $15^{\circ}C$, pH 6, 탄소원 0.6% adonitol, 유기질소원 1.5% yeast extract, 무기질소원 0.8% $NH_4H_2PO_4$, 아미노산 0.2% asparagine 그리고 무기염류는 5 mM $MgSO_4$에서 왕성한 생육을 보였다.

Keywords

References

  1. Brodey CL, Rainey PB, Teste M, Johnstone K, 1991. Bacterial blotch disease of the cultivated mushroom is caused by an ion channel forming lipodepsip eptide toxin. Mol. Plant-Microbe Interact. 4:407-411. https://doi.org/10.1094/MPMI-4-407
  2. Cho KH, Kim YK. 2003. Two types of ion channel formation of tolaasin, a Pseudomonas peptide toxin. FEMS Microbiol. Lett. 221:221-226. https://doi.org/10.1016/S0378-1097(03)00182-4
  3. Cutri SS, Macauley BJ, Roberts WP. 1984. Characteristics of pathogenic non-fluorescent (smooth) and nonpathogenic fluorescent (rough) forms of Pseudomonas tolaasii and Pseudomonas 'gingeri'. J. Appl. Bacteriol. 57:91-298.
  4. Goor M, Vantomme R, Swings J, Gillis M, Kersters K, de Ley J. 1986. Phenotypic and genotypic diversity of Pseudomonas tolaasii and white line reacting organisms isolated from cultivated mushrooms. J. Gen. Microbiol. 132:2249-2264.
  5. Kim JW, Kim KH and Kang HJ. 1994. Studies on the pathogenic Pseudomonas causing bacterial disease of cultivated mushroom in Korea. 1. On the causal organisms of the rots of Agaricus bisporus, Pleurotus ostreatus and Lentinus edodes. Kor. J. Plant Pathol. 10:197-210 (in Korean).
  6. Lee CJ, Yoo YM, Han JY, Jhune CS, Cheong JC, Moon JW, Suh JS, Han HS, Cha JS. 2014a. Isolation of the bacterium Pseudomonas azotoformans HC5 effective in antagonistic of brown blotch disease caused by Pseudomonas tolaasii. Kor. J. of Mycology 42:219-224. https://doi.org/10.4489/KJM.2014.42.3.219
  7. Lee CJ, Yoo YM, Han JY, Jhune CS, Cheong JC, Moon JW, Suh JS, Han HS, Cha JS. 2013. Isolation of the bacterium Pseudomonas sp. HC1 effective in inactivation of tolaasin produced by Pseudomonas tolaasii. Kor. J. of Mycology 41: 248-254. https://doi.org/10.4489/KJM.2013.41.4.248
  8. Lee CJ, Yoo YM, Han JY, Jhune CS, Cheong JC, Moon JW, Kong WS, Suh JS, Han HS, Cha JS. 2014b. Cultivation conditions for mass production of detoxifying bacterium Pseudomonas sp. HC1 of tolaasin produced by Pseudomonas tolaasii. J. of Mushrooms. 12:35-40. https://doi.org/10.14480/JM.2014.12.1.35
  9. Liao YM, Tu CC, Jeng JJ. 1980. Control of bacterial blotch of mushroom. Taiwan Mushrooms 4: 34-41.
  10. Nair NG, Fahy PC. 1972. Bacteria antagonistic to Pseudomonas tolaasii and their control of brown blotch of the cultivated mushroom Agaricus bisporus. J. of Applied Bacteriology. 35: 439442.
  11. Nair NG, Fahy PC. 1973. Toxin production by Pseudomonas tolaasii Paine. Aust. J. biol. Sci. 26:509-512 https://doi.org/10.1071/BI9730509
  12. 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
  13. Olivier JM, Guillaumes J, Martin G. 1978. Study of a bacterial disease of mushroom caps. Proceedings of 4th International Conference Plant Pathogenic Bateria, Angers, France. 903-916.
  14. 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
  15. Park BS, Cho NC, Chun UH. 1992. Identification of Pseudomonas fluorescens antagonistic to Pseudomonas tolaasii and its cultivation. Kor. J. Biotechnol. Bioeng. 7:296-301.
  16. Rainey PB, Brodey CL, Johnstone K. 1992. Biology of Pseudomonas tolaasii, cause of brown blotch disease of cultivated mushroom. Pages 95-118 in: Advances in Plant Pathology, Vol. 8. J. H. Andrews and I. Tommerup, eds. Academic Press, Inc., New York.
  17. Reasoner DJ, Geldreich EE. 1985. "A new medium for the enumeration and subculture of bacteria from potable water," Appl. Environ. Microbiol., 49:1-7.
  18. Scherwinski K, Grosch R, Berg G. 2008. Effect of bacterial antagonists on lettuce: active biocontrol of Rhizoctonia solani and negligible, short-term effects on nontarget microorganisms. FEMS Microbiol Ecol. 64: 106116.
  19. Stainer RY, Palleroni NJ, Doudoroff M. 1966. The aerobic pseudomonads: a taxonomic study. J. of general Microbiol. 43:159-271. https://doi.org/10.1099/00221287-43-2-159
  20. Tolaas AG. 1915. A Bacterial disease of cultivated mushrooms. Phytopathology 5: 51-54
  21. Tsuneda A, Suyama K, Muradami S, Ohira I. 1995. Occurrence of Pseudomonas tolaasii on fruiting bodies of Lentinula edodes formed on Quercus logs. Mycoscience 36: 283-288. https://doi.org/10.1007/BF02268603
  22. Wells JM, Sapers GM, Fett WF, Butterfield JE, Jones JB, Bouzar H, Miller FC. 1996. Postharvest discolorization of the cultivated mushroom Agaricus bisporus caused by Pseudomonas tolaasii, P. 'reactans', and P. 'gingeri'. Phytopathology 86: 1098-1104. https://doi.org/10.1094/Phyto-86-1098
  23. Wong WC, Fletche, JT, Unsworth BA, Preece TF. 1982. A note on ginger blotch, a new bacterial disease of the cultivated mushroom, Agaricus bisporus. J. Appl. Bacteriol. 52:43-48. https://doi.org/10.1111/j.1365-2672.1982.tb04371.x
  24. Young JM. 1970. Drippy gill: a bacterial disease of cultivated mushrooms caused by Pseudomonas agarici n. sp. N. Z. J. Agr. Res. 13:977-990. https://doi.org/10.1080/00288233.1970.10430530