Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya.
Curr Microbiol. 2010 Jan;60(1):47-52. doi: 10.1007/s00284-009-9500-1. Epub 2009 Oct 6.
A non-culture approach was used to study the archaeal diversity in Lake Elmenteita, Kenya. Five different sampling points were selected randomly within the lake. Wet sediments and water samples were collected from each sampling point. In addition, dry mud cake was collected from three points where the lake had dried. DNA was extracted from these samples and the 16S rRNA genes were amplified using primers described to be Domain-specific for Archaea. Eleven clone libraries were constructed using PCR-amplified 16S rRNA genes. A total of 1,399 clones were picked and analysed via ARDRA. 170 ARDRA patterns were unique and the respective clones were selected for sequencing. 149 clones gave analysable sequences. BLAST analysis showed that 49 belong to the Domain Archaea while the others were either chimera or affiliated to eukaryotic taxa. Comparative sequence analysis of archaeal clones affiliated them to a wide range of genera. The order Halobacteriales was represented by members of the genera Natronococcus, Halovivax, Halobiforma, Halorubrum, and Halalkalicoccus. The highest percentage (46%) of the clones, however, belonged to uncultured members of the Domain Archaea in the order Halobacteriales. The results show that the archaeal diversity in the lake could be higher than previously reported.
本研究采用非培养方法对肯尼亚埃尔门泰塔湖的古菌多样性进行了研究。在湖中随机选择了五个不同的采样点。从每个采样点采集了湿沉积物和水样。此外，还从三个干涸的湖点采集了干泥饼。从这些样品中提取 DNA，并使用描述为专用于古菌的引物扩增 16S rRNA 基因。使用 PCR 扩增的 16S rRNA 基因构建了 11 个克隆文库。共挑选了 1399 个克隆，并通过 ARDRA 进行了分析。有 170 种 ARDRA 模式是独特的，相应的克隆被选择进行测序。有 149 个克隆给出了可分析的序列。BLAST 分析表明，其中 49 个属于古菌域，而其他的则是嵌合体或与真核生物分类群有关。古菌克隆的比较序列分析将它们归入了广泛的属。嗜盐菌目由纳塔尔球菌属、盐单胞菌属、盐形菌属、盐杆菌属和盐生球菌属的成员组成。然而，克隆的最高比例（46%）属于嗜盐菌目中未培养的古菌成员。结果表明，湖中古菌的多样性可能高于先前的报道。
