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Phylogenetic Analysis of Bacterial Diversity in the Marine Sponge, Asteropus simplex, Collected from Jeju Island

제주도에서 채집한 해양 해면, Asteropus simplex의 공생세균에 관한 계통학적 분석

  • Jeong, In-Hye (Department of Biological Science and Biotechnology, Hannam University) ;
  • Park, Jin-Sook (Department of Biological Science and Biotechnology, Hannam University)
  • 정인혜 (한남대학교 생명시스템과학과) ;
  • 박진숙 (한남대학교 생명시스템과학과)
  • Received : 2012.12.15
  • Accepted : 2012.12.26
  • Published : 2012.12.31

Abstract

Culture-dependent RFLP and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Asteropus simplex collected from Jeju Island. A total of 120 bacterial strains associated with the sponge were cultivated using modified Zobell and MA media. PCR amplicons of the 16S rDNA from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rDNA sequences derived from RFLP patterns showed more than 94% similarities compared with known bacterial species, and the isolates belonged to five phyla, Alphaproteobacteria, Gammaproteobacteria Actinobacteria, Bacteroidetes, and Firmicutes, of which Gammaproteobacteria was dominant. DGGE fingerprinting of 16S rDNAs amplified from the sponge-derived total gDNA showed 12 DGGE bands, and their sequences showed more than 90% similarities compared with available sequences. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of seven phyla, including Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteira, Chloroflexi, and Nitrospira. Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria were commonly found in bacteria associated with A. simplex by both RFLP and DGGE methods, however, overall bacterial community in the sponge differed depending on the analysis methods. Sponge showed more various bacterial community structures in culture-independent method than in culture-dependent method.

Keywords

Asteropus simplex;bacterial diversity;DGGE;RFLP;sponge

Acknowledgement

Supported by : 한남대학교

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