Phylogenetic Characterization of a Mixed Microbial Community Capable of Degrading Carbon Tetrachloride

Two bacterial communities (NO92 and GBS) capable of degrading carbon tetrachloride (CT) were enriched from in-house CT-contaminated water. These communities are able to degrade CT in the presence of toluene. To characterize the community structure and diversity, one enrichment (NO92) was subjected to 16S ribosomal RNA (rRNA) gene-based molecular analysis. The 16S rRNA genes were amplified from the bulk genomic community DNA and cloned into plasmid vectors. Unique 16S rRNA gene clones, i.e., phylotypes, were detected by four tetrameric restriction enzymes. Together, 123 16S rRNA gene clones were obtained; thirty-one showed different restriction fragment length polymorphism (RFLP) patterns. About 73% of the clones belong to two dominant RFLP patterns. Phylogenetic analysis based on the partial 16S rRNA gene sequences of 10 major phylotypes showed that all the phylotypes that were sequenced were affiliated with the high G+C Gram-positive bacteria. Whereas seven of the phylotypes (~80% of the clones) were closely related to Rhodococcus, the other three (~5% of the clones) were related to Curtobacterium. These results suggest that this CT-degrading community is diverse but is predominated by closely related bacterial groups. Index Entries: Carbon tetrachloride; bioremediation; phylogeny; microbial community.