Korean Journal of Environmental Agriculture (한국환경농학회지)

The Korean Society of Environmental Agriculture (한국환경농학회)
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Assessment of Korean Paddy Soil Microbial Community Structure by Use of Quantitative Real-time PCR Assays

한국의 논 토양 미생물 다양성 분석을 위한 Quantitative Real-time PCR의 응용

  • Choe, Myeong-Eun (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, In-Jung (School of Applied Biosciences, Kyungpook National University) ;
  • Shin, Jae-Ho (School of Applied Biosciences, Kyungpook National University)
  • 최명은 (경북대학교 농업생명과학대학 응용생명과학부) ;
  • 이인중 (경북대학교 농업생명과학대학 응용생명과학부) ;
  • 신재호 (경북대학교 농업생명과학대학 응용생명과학부)
  • Received : 2011.11.30
  • Accepted : 2011.12.12
  • Published : 2011.12.31
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Abstract

BACKGROUND: In order to develop effective assessment method for Korean paddy soil microbial community structure, reliable genomic DNA extraction method from paddy soil and quantitative real-time PCR (qRT-PCR) method are needed to establish METHODS AND RESULTS: Out of six conventional soil genomic DNA extraction methods, anion exchange resin purification method was turn to be the most reliable. Various PCR primers for distinguishing five bacterial phylum (${\alpha}$-Proteobacteria, ${\beta}$-Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes), all bacteria, and all fungi were tested. Various qRT-PCR temperature conditions were also tested by repeating experiment. Finally, both genomic DNA extraction and qRT-PCR methods for paddy soil were well established. CONCLUSION: Quantitative real-time PCR (qRT-PCR) method to assess paddy soil microbial community was established.

논 토양의 미생물 생태 다양성을 조사하기 위한 효과적인 방법으로 qRT-PCR을 적용하고자 본 연구를 수행하였다. 논 토양 미생물의 gDNA를 분리하기 위하여 Mo Bio kit를 사용한 효과적이고 안정적인 gDNA 분리 방법을 확립하였다. 논 토양 미생물 다양성을 qRT-PCR로 검출하기 위하여 bacteria를 세분한 ${\alpha}$-Proteobacteria, ${\beta}$-Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes 다섯 가지 문과 전체 bacteria, 전체 fungi를 구분할 수 있는 특이 primer set을 선정하여 다양한 조건의 시험을 통하여 최종 조건을 확립하였으며 재현성 실험을 통하여 방법의 유의성을 검증하였다.

Keywords

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