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KMID : 0364820080440040352
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Korean Journal of Microbiology
2008 Volume.44 No. 4 p.352 ~ p.357
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Evaluation of Various Oligotrophic Media for Cultivation of Previously Uncultured Soil Bacteria
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Kim Do-Hyoung
Lee Sang-Hoon
Cho Jae-Chang
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Abstract
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We evaluated cultivation methods to obtain pure cultures of previously uncultivated bacteria from soil. Soil bacteria (suspensions) were inoculated onto various oligotrophic media with one of the following additives: 1) soil extract; 2) anthraquinone disulfonate (humic acid analogue); 3) acyl homoserine lactones (quorum-signaling compounds); 4) catalase (for the protection of bacteria from exogenous peroxides). After the relatively long period (60 days) of incubation with elevated concentrations of CO2 (5%, v/v), the media containing catalase showed the highest colony count. We purified 147 randomly selected colonies from the media and the isolates were subjected to the phylogenetic analyses of their 16S rRNA gene sequences. Phylogenetic analysis revealed that approximately 30% of the isolates might belong to novel species or novel family, suggesting that the media and incubation conditions used could be useful for the cultivation of as-yet-uncultured bacteria. Especially, bacteria belonging to the phylum Acidobacteria, ubiquitous bacterial taxon known as an uncultured bacterial group (at least difficult to culture from environmental samples), were successfully cultured in this study.
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KEYWORD
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oligotrophic media, phylogenetic analysis, uncultured bacteria
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