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KMID : 0364820120480040275
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Korean Journal of Microbiology
2012 Volume.48 No. 4 p.275 ~ p.283
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Phylogenetic Analysis of Bacterial Diversity in the Marine Sponge, Asteropus simplex, Collected from Jeju Island
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Jeong In-Hye
Park Jin-Sook
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Abstract
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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 cultureindependent
method than in culture-dependent method
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KEYWORD
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Asteropus simplex, bacterial diversity, DGGE, RFLP, sponge
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