Takai K, Moser D P, DeFlaun M, Onstott T C, Fredrickson J K
Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Appl Environ Microbiol. 2001 Dec;67(12):5750-60. doi: 10.1128/AEM.67.21.5750-5760.2001.
A culture-independent molecular analysis of archaeal communities in waters collected from deep South African gold mines was performed by performing a PCR-mediated terminal restriction fragment length polymorphism (T-RFLP) analysis of rRNA genes (rDNA) in conjunction with a sequencing analysis of archaeal rDNA clone libraries. The water samples used represented various environments, including deep fissure water, mine service water, and water from an overlying dolomite aquifer. T-RFLP analysis revealed that the ribotype distribution of archaea varied with the source of water. The archaeal communities in the deep gold mine environments exhibited great phylogenetic diversity; the majority of the members were most closely related to uncultivated species. Some archaeal rDNA clones obtained from mine service water and dolomite aquifer water samples were most closely related to environmental rDNA clones from surface soil (soil clones) and marine environments (marine group I [MGI]). Other clones exhibited intermediate phylogenetic affiliation between soil clones and MGI in the Crenarchaeota. Fissure water samples, derived from active or dormant geothermal environments, yielded archaeal sequences that exhibited novel phylogeny, including a novel lineage of Euryarchaeota. These results suggest that deep South African gold mines harbor novel archaeal communities distinct from those observed in other environments. Based on the phylogenetic analysis of archaeal strains and rDNA clones, including the newly discovered archaeal rDNA clones, the evolutionary relationship and the phylogenetic organization of the domain Archaea are reevaluated.
通过对rRNA基因(rDNA)进行PCR介导的末端限制性片段长度多态性(T-RFLP)分析,并结合古菌rDNA克隆文库的测序分析,对从南非深部金矿采集的水样中的古菌群落进行了非培养分子分析。所用水样代表了各种环境,包括深部裂隙水、矿井服务用水以及来自上覆白云岩含水层的水。T-RFLP分析表明,古菌的核糖体类型分布随水源不同而变化。深部金矿环境中的古菌群落表现出极大的系统发育多样性;大多数成员与未培养物种关系最为密切。从矿井服务用水和白云岩含水层水样中获得的一些古菌rDNA克隆与来自表层土壤的环境rDNA克隆(土壤克隆)和海洋环境(海洋第一类群[MGI])关系最为密切。其他克隆在泉古菌门中表现出介于土壤克隆和MGI之间的中间系统发育归属。来自活跃或休眠地热环境的裂隙水样产生了具有新系统发育的古菌序列,包括一个新的广古菌门谱系。这些结果表明,南非深部金矿中存在与其他环境中观察到的不同的新型古菌群落。基于对古菌菌株和rDNA克隆(包括新发现的古菌rDNA克隆)的系统发育分析,对古菌域的进化关系和系统发育组织进行了重新评估。