Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Diversity of Uncultured Microorganisms Associated with the Anaerobic Pentachlorophenol Degradation Estimated by Comparative RELP Analysis of PCR-Amplified 16S rDNA Clones

16S rDNA 클론들의 RFLP 비교분석에서 얻어진 Pentachlorophenol의 혐기성 분해에 따른 미생물군집의 변화

  • 성창수 (인제대학교 자연과학대학 미생물학과) ;
  • 권오섭 (인제대학교 자연과학대학 환경학과) ;
  • 박영식 (인제대학교 자연과학대학 미생물학과)
  • Published : 1997.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

We have employed comparative RFL,P(Restriction Fragment Ixngth Pol~iniorphism) analysis and molecular phylogenetic techniques to investigate the diversity of uncultured microorganisms associated with the anaerobic PCP degradation in PCP-adapted enrichment cultures inoculated by samples from anaerobic cewage sludgc(Jangrim, Pusan) and leachate of landfill site(Kimhae). 16s rDNA cloncs were obtairted by PCR amplification of mixed population DNAs extracted directly from the nonactive and active stage ol each PCP-adapted culture. After three rounds of comparative RFLP analyses. two RFLP types. designated as Ala and Hld, were found prevalent and common in both active stage samples. Thc analysis of phylogenctic diversity bawd on the 5'-terminal 180 nt of sequences from whole clones of the Ala and Bld RFLP types showed close similarity among themselves. In case of Bld clones, 7XQ of them shared identical sequences. Thcse resuliq suggest that the clones of both RFLP types wcre originated from highly affiliated microorganisms which are e~iriched as a result of metabolic activity to PCP. The full-length 16s rRNA sequence of each representative clone from both RFLP types was determined. and an Ala clone w i n found to he related to Clo.strrdiurn ulfutzac~(Genk~ank No. Z69203) and a Bld clone to Thermobacteroides proteolyticus(Genbank No. X09335), with sequence similarities of 89%' and 97%. respectively.

Pentachlorophenol(PCP)가 첨가된 혐기성 무기배지에 혐기성 소화조슬럿지와 쓰레기 매립장의 침출수를 각각 접종한 후, 활성을 보이기 전과 후의 각 시료 내에 존재하는 총유전물질로부터 16S rRNA 유전자를 PCR 증폭하여 이들에 대한 restriction fragment length polymorphism(RFLP) 비교분석과 계통수분석을 실시하였다. 3차에 걸친 RFLP 분석 결과 Ala와 Bld로 명명된 두 가지의 FRLP 유형이 두 가지의 활성시료 모두에서 높은 빈도로 발견되었다. 이들 유형에 속하는 모든 클론들의 5'쪽에 해당되는 180개씩의 염기서열분석을 실시하여 계통수 분석을 실시한 결과, 각각의 유형에 속하는 클론들간에는 높은 상동성을 가지는 것으로 확인되었다. 특히 Bld 유형에서는 78%에 해당되는 클론들이 동일한 염기서열을 가지는 것으로 확인되었다. 이들 Ala와 Bld 유형에 속하는 클론들은 PCP에 대한 분해활성의 결과로서 증식된 유사종의 미생물 군집에서 비롯된 것으로 추정된다. 이 두 가지 유형에 속하는 클론들 중 하나씩의 16S rDNA 전체으 염기서열을 분석한 결과 Ala의 클론은 Clostridium ultunae (Genbank No. Z69293)의 염기서열과 89%의 상동성을 보였으며, Bld의 클론은 Thermobacteroides proteolyticus (Genbank No. X69335)의 것과 97%의 상동성을 가지는 것으로 나타났다.

Keywords

References

  1. Microbial Rev. v.43 Methanogens: Reevaluation of a uniquc biological group Balch, W. E.;G. E. Fox;L. J. Magrum;C. R. Woese;R. S. Wolfe
  2. Int. J. Syst. Bacteriol. v.46 Isolation and characterization of Desulfitobacterium frappieri sp. an anaerobic bacterium which reductively dechlorinates pentachlorophenol to 3-chlorophenol Bounchard, B.;R. Beaudet;R. Villemur;G. McSween;F. Leqine;J. G. Bisaillon
  3. Computer Applications in the Biociences v.12 TREEVIEW: An application to display phylogenetic trees on personal computers Rage, R. D. M.
  4. Wat. Res. v.28 Effects of electron doners and inhibitors on reductive dechlorination of 2,4,6-trichlorophenol Ferguson, J. F.;S. J. Komisar;P. S. Perkins
  5. Chemosphere v.26 Determination of chlorophenols and chlorobenzenes in leacheate by headspacw analysis Forst, C.;H. Simon;L. Stiefhiz
  6. Soil. Sci. Soc. Amer. and Amer. Soc. Agro. Dechlorination of pestcide by anaerobic microorganism in soils groundwater a review. In: Reactions and Movement of Organic chemical in Soils Kuhn, E. P.;J. M. Suflita;B. L. Sawhney(ed.);K. Brown(ed.)
  7. Chemosphete v.24 Chlorophenols in lake botton sediments: a retorspective study of drinking water contamination Lampi, P.;K. Tolonen;T. Vartiainen;J. Tuomisto
  8. Appl. Environ. Microbial. v.62 Initial characterzation of a reductive dehalogenase from Desulfidobacterium chlororespirans Co23 Loffler, F. E.;R. A. Sanford;J. M. Tiedje
  9. Appl. Environ. Microbiol. v.58 Isolation and characterization of an aerobic chlorophenol-transforming bacterium Madsen, T.;D. Licht
  10. Appl. Environ. Microbiol. v.52 Complete reductive dechlorination and mineralization of pentachlorophenol by anaerobic microorganisms Mikesell, M. D.;S. A. Licht
  11. Appl. Environ. Microbiol. v.58 Limited degradation of chlorophenols by anaerobic studge granules Mohn, W. W.;K. J. Kennedy
  12. Microbiol. Res. v.56 Microbial reductive dehalogenation MOhn, W. W.;J. M. Tiedje
  13. Appl. Env. Microbiol. v.60 Estimation of diversity and community structure through RFLP distribution analysis of bacterial 16S rRNA genes from a microbial mal at an active, hydrothermal Vent System. Loihi Seamount, Hawaii Moyer, C. L.;F. C. Dobbs;D. M. Karl
  14. Appl. Env. Microbiol. v.57 Biodegadation of creosote and pentachlorophenol in contaminated groundwater: Chemical and biological assessment Muller, J. G.;D. P. Middaugh;S. E. Lantz;P. J. Chapman
  15. Apl. Environ. Microbiol. v.58 Reuctive dechlorination of chlorophenols by a pentachlorophenol-acclimated methanogenic consortium Nicholson, D. K.;S. L. Woods;J. D. Istok;D. C. Peek
  16. Lett. Appl. Microbiol. v.16 Phylogenetic analysis of the bacterial genus Thermobacteroides indicates an ancient origin of Thermobacteroides proteolyticus. Rainey, F.;E. Stackebrandt
  17. Molecular cloning, A laboratory Manual. (2nd ed.) Sambrook, J.;E. F. Fritsch;T. Maniatis
  18. Int. J. Syst. Bacteriol. v.46 Chlostridium ultunae sp. nov., a mesophilic bacterium oxidizing acetate in syntrophic association with a hydrogenotrophuc methanogenic bacterium Schunrer, A.;B. Schink;B. H. Svensson
  19. J. Gen. Microbiol. v.136 Partial 16S rRNA primary structure of five Actinomyces species: phylogenetic implications and development of a Actinomyces israelii-specific oligonucleotide probe Stackebrandt, E.;O. Charfreitag
  20. Notice Federal Register v.51 Creosote, pentachlorophenol and inorganic arsenicals: Amendment of notice intent to cancel registration USEPA Federal Register
  21. Appl. Environ. Microbiol. v.61 Specificity of reductive dehalogenation of substituted ortho-chlorophenols by Desulfidobacterium dehalofenana JW/IU-DC-1 Utkin, I.;D. D. Dalton;J. Wiegel
  22. Appl. Microbiol. Biotechnol. v.21 Studies on the physiology of microbial degradation of pentachlorophenol Valo, R.;J. A. Apajajalahti;M. S. Salkinoja-Salonen
  23. Appl. Env. Microbiol. v.61 Ditect DNA Extraction for PCR-Mediated Assays of Siol Organisms Volossiouk, T.;E. J. Robb;N. N. Rpss
  24. Chemosphere v.24 Pentachlorophenol in the UK enviroment: (I) A budgel and source inventory Wild, S. R.;S. J. Harrad;K. C. Jones