[논문]16S rRNA 분석을 통한 인도네시아의 Cisolok, Kamojang, Likupang 지열지대 내 미생물 다양성 분석

서명지; 김정녀; 변유량

doi:10.4014/kjmb.1205.05001

16S rRNA 분석을 통한 인도네시아의 Cisolok, Kamojang, Likupang 지열지대 내 미생물 다양성 분석
16S rRNA Gene Sequence-based Microbial Diversity Analyses of the Geothermal Areas of Cisolok, Kamojang, and Likupang in Indonesia 원문보기

한국미생물·생명공학회지 = Korean journal of microbiology and biotechnology, v.40 no.3, 2012년, pp.268 - 273

서명지 (한국식품연구원 발효기능연구단) , 김정녀 (연세대학교 생명공학과) , 변유량 (연세대학교 생명공학과)

초록
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인도네시아 지열지대의 미생물 다양성을 16S rRNA 염기서열 분석을 통해 조사하였다. 전체적으로 어떠한 phylum 계통군에도 포함되지 않는 unclassified bacteria가 20.0-26.5% 존재하였으며 sampling 지역에 상관없이 Proteobacteria가 우점 phylum 계통군으로 나타났다. Cisolok 주변의 지열 지역을 조사한 결과 ${\beta}$-Proteobacteria (27.1%)와 Cyanobacteria (11.0%)가 높은 비율을 차지한 반면 Kamojang의 화산 주변 지역의 경우에는 ${\gamma}$-Proteobacteria (51.5%) 그리고 Aquificales (12.9%)가 우점 계통군으로 나타났다. 또한 Likupang 열수구의 경우에는 ${\gamma}$-Proteobacteria (33.3%)와 Bacteroidetes (27.6%)가 높은 비율로 나타났다. 본 연구를 통해 인도네시아 각 지열지대에 분포하는 미생물 군집은 각 지역의 환경적인 특징 (극한 지열 및 해양 서식지)과 밀접한 연관성이 있음을 알 수 있었다.

Abstract ▼ AI-Helper

Microbial diversity analyses were performed in several geothermal areas in Indonesia using a culture-independent approach with 16S rRNA gene sequencing. All areas and the majority of samples were noted as being affiliated with Proteobacteria. In addition, unclassified bacteria with no phylum affiliation were detected at an incidence rate of 20.0-26.5% in every location. The majority groupings in the geothermal hot stream in Cisolok belonged to ${\beta}$-Proteobacteria (27.1%) and Cyanobacteria (11.0%), whereas the majority from the volcanic area in Kamojang was ${\gamma}$-Proteobacteria (51.5%) followed by Aquificales (12.9%). The predominant groups around an underwater thermal vent in the sea at Likupang were ${\gamma}$-Proteobacteria (33.3%) and then Bacteroidetes (27.6%). This detailed microbial community analyses of each area strongly support a possible association with plausible community groups and environmental habitats, such as extremely geothermal or marine habitats. This study has significantly contributed to the expansion of scientific knowledge of the microbial community in Indonesia.

주제어

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제안 방법

Despite these specific geographical characteristics of Indonesia, there have been few studies on microbial diversity from Indonesian geothermal areas [1, 3]. In this study, we therefore analyzed the microbial diversity of geothermal areas in Indonesia. The analysis was based on culture-independent approach using 16S rRNA gene sequencing after direct extraction of environmental DNA.
In this study, we therefore analyzed the microbial diversity of geothermal areas in Indonesia. The analysis was based on culture-independent approach using 16S rRNA gene sequencing after direct extraction of environmental DNA.
After sequencing each 16S rRNA gene from the libraries, too short sequences of less than 350 bp in length were excluded for the further analysis. The resulting sequences were compared to 16S rRNA gene sequence from the nonredundant sequence database by using BLASTN program at National Centre of Biotechnological Information (NCBI) to determine their approximate taxonomic identifications and investigate the microbial diversity.

대상 데이터

Environmental samples were collected from geothermal areas (Cisolok, Kamojang, and Likupang) in Indonesia (Fig. 1). The environmental temperature and pH of each sample was 70℃ and pH 7.
Environmental samples were collected from geothermal hot stream at Cisolok (6º57’23”S, 106º28’46”E), volcano area at Kamojang (7º07’30”S, 107º47’60”E), West Java, and thermal vent under the sea water at Likupang (1º41’35”N, 125º00’47”E), north Sulawesi, Indonesia.

성능/효과

After sequencing 171 representatives, the two most abundant microbial orders were Acidithiobacillales (46.2%) belonged to γ-Proteobacteria, followed by Aquificales (12.9%), whereas unclassified bacteria also accounted for a high portion (21.6%) (Table 1 and Fig. 2).
Based on 155 representative 16S rRNA gene sequences, two most major abundant phyla were shown in Cisolok; Proteobacteria (38.7%) with major class, β-Proteobacteria (27.1%) and Cyanobacteria (11.0%) with major genus, Thermosynechococcus (10.3%) (Table 1 and Fig. 2).
The explanation of extremely acidic condition based on the distribution of Acidithiobacillales could be strongly supported by the previous study on microbial diversity of acidic hot spring (Kawah Hujan B) in Kamojang area [1]. The study described that the majority of the sequences was closely related to Crenarchaeota frequently found in extremely acidic environments [19] and capable to utilize elemental sulfur as energy source [5].
Total of 105 representative clones from thermal vent under the sea water at Likupang, north Sulawesi, Indonesia were sequenced and analyzed, resulting that the highest proportion was γ-Proteobacteria (33.3%), followed by Bacteroidetes (27.6%) and unclassified bacteria group (20%).

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