Keinänen-Toivola Minna M, Revetta Randy P, Santo Domingo Jorge W
US Environmental Protection Agency, NRMRL, WSWRD, MCCB, Cincinnati, OH, USA.
FEMS Microbiol Lett. 2006 Apr;257(2):182-8. doi: 10.1111/j.1574-6968.2006.00167.x.
Recent phylogenetic studies have used DNA as the target molecule for the development of environmental 16S rRNA gene clone libraries. As DNA may persist in the environment, DNA-based libraries cannot be used to identify metabolically active bacteria in water systems. In this study, an annular reactor was used to generate model drinking water biofilms grown on polycarbonate slides. High-quality RNA was extracted from 2-month-old biofilms and used to generate 16S rRNA-based clones. Sequencing analyses of 16S rRNA-based clones suggested that the active bacterial fraction consisted of a few dominant bacterial groups related to Nevskia ramosa and to uncultured bacteria. Several of these bacterial groups were closely related to clones characterized in a DNA-based clone library also generated in this study. Altogether, these results suggest that some of the predominant drinking water bacteria identified using DNA-based techniques are indeed active.
近期的系统发育研究已将DNA用作构建环境16S rRNA基因克隆文库的目标分子。由于DNA可能在环境中持续存在,基于DNA的文库无法用于识别水系统中具有代谢活性的细菌。在本研究中,使用环形反应器在聚碳酸酯载玻片上生成模型饮用水生物膜。从两个月大的生物膜中提取高质量RNA,并用于生成基于16S rRNA的克隆。对基于16S rRNA的克隆进行测序分析表明,活性细菌部分由一些与枝状涅斯捷连科氏菌和未培养细菌相关的优势细菌群组成。这些细菌群中的几个与本研究中生成的基于DNA的克隆文库中鉴定的克隆密切相关。总之,这些结果表明,使用基于DNA的技术鉴定出的一些主要饮用水细菌确实具有活性。