PCR-RFLP 방법을 이용한 활성 슬러지의 세균군집 분석

Molecular Characterization of the Bacterial Community in Activated Sludges by PCR­RFLP

  • 이현경 (충북대학교 미생물학과 및 기초과학연구소) ;
  • 김준호 (충북대학교 미생물학과 및 기초과학연구소) ;
  • 김치경 (충북대학교 미생물학과 및 기초과학연구소) ;
  • 이동훈 (충북대학교 미생물학과 및 기초과학연구소)
  • Lee Hyun-Kyung (Department of Microbiology and Basic Science Research Institute, Chungbuk National University) ;
  • Kim Jun-Ho (Department of Microbiology and Basic Science Research Institute, Chungbuk National University) ;
  • Kim Chi-Kyung (Department of Microbiology and Basic Science Research Institute, Chungbuk National University) ;
  • Lee Dong-Hun (Department of Microbiology and Basic Science Research Institute, Chungbuk National University)
  • 발행 : 2004.12.01

초록

폐수처리에 중요한 역할을 담당하고 있는 세균 군집의 다양성과 폐수종류에 따른 군집차이를 알아보기 위해 분자생물학적 분석방법을 사용하였다. 국내 폐수처리장 슬러지 시료로부터 16S rDNA 클론 라이브러리를 구축하였고, HaeIII RFLP pattern과 염기서열을 분석하였다. 하수처리장 시료에서는 총 1,151개의 클론에서 699개의 서로 다른 RFLP pattern이, 화학산업 폐수처리장 시료에서는 총 1,228개의 클론에서 300개의 서로 다른 RFLP pattern이 관찰되었다. Shannon-Weiner diversity index의 계산결과 하수처리장 슬러지 시료는 8.7,화학산업 폐수처리장 슬러지 시료는 6.1로 하수처리장시료가 더 다양한 군집을 구성하고 있었다. 두 시료에서 우점하는 RFLP pattern에 해당되는 40개의 클론을 선정하여 염기서열 분석과 상동성 검색을 수행하였다. 분석된 서열의 $70\%$인 28개의 클론은 배양이 보고되지 않은 균주의 16S rRNA와 유사도가 높았고, 두 시료 모두 ${\beta}-Proteobacteria$가 우점하였다. 그러나, 하수처리장의 경우 활성슬러지에서 분리된 균주들과 유사한 군집이 많았던 반면, 화학산업 페수처리장의 경우 고온이며, 혐기성이고,탄화수소나 황이 많이 존재하는 환경에서 분리된 균주들과 유사한 군집이 많았다. 이러한 결과는 유입수의 조성에 따른 차이로 생각된다.

Diversity of the bacterial communities and the relation between community structure and components of waste­water were analyzed by 16S rRNA-based molecular techniques. Clone libraries of the 16S rDNAs from the sludges were constructed by PCR and cloning. The 1,151 clones from a sludge sample of sewage treatment plant were clustered into 699 RFLP phylotypes and the 1,228 clones from the wastewater disposal plant of chemical industry were clustered into 300 RFLP phylotypes. Shannon-Weiner diversity indices of two sampling sites were 8.7 and 6.1, indicating that the bacterial community structure of sewage treatment plant was more diverse than that of wastewater disposal plant of chemical industry. Forty clones belonging to predominant RFLP types were selected and sequenced. Seventy percent (28 clones) of the sequenced clones were related to the uncultured bacteria in public databases. The ${\beta}-Proteobacteria$ dominated in the bacterial communities of investigated two sludge samples. 16S rDNA sequences of the sewage treatment plant were similar to those of other activated sludges, while the bacterial community in wastewater disposal plant of chemical industry rep­resented the strains identified from high-temperature, anaerobic, hydrocarbon-rich, and sulfur-rich environ­ments. This result suggested that bacterial communities depended upon the components of wastewater.

키워드

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