Li Ping, Zeng Jun, Zulipiya Yunus, Gao Xiaoqi, Dong Xiuhuang, Xue Juan, Lou Kai
College of Life Science and Technology of Xinjiang University, Urumqi 830046, China.
Wei Sheng Wu Xue Bao. 2013 Mar 4;53(3):230-40.
We explored the composition and diversity of archaea in a cold sulfur spring water in Xinjiang earthquake fault zone.
Environmental total DNA was extracted directly with enzymatic lysis method from a cold sulfur spring water. We constructed clone library of 16S rRNA gene amplified with archaeal-specific primers. A total of 115 positive clones were selected randomly from the library and identified by restriction length polymorphism (RFLP) with enzyme Alu I and Afa I. The unique RFLP patterns corresponded clones were selected for sequencing, BLAS alignment and constructing 16S rRNA gene phylogenetic tree.
In total, 44 operational taxonomic units (OTUs) were determined from the library. BLAST and phylogenetic analysis indicated that these OTUs were affiliated with Euryarchaeota (94.78%) and Thaumarchaeota (4.35%). Only one Thaumarchaeotal clone was detected and most related to the genus Nitrosopumilus with 93% similarity. Euryarchaeotal clones were abundant and diverse. Of them, 42.61% of clones belonged to RC-V cluster; 13.91% of clones, 20.87% of clones were classified into LDS cluster and Methanomicrobiales respectively; 4.35% of clones had high similarity with ANME-1a-FW, which were involved in Anaerobic oxidation of methane (AOM). In addition, we also detected some (13.05%) unknown Euryarchaotal clones.
Euryarchaeota in the environment were diverse, and possibly with a large fraction of potential novel species.
我们探究了新疆地震断裂带一处冷硫泉水中古菌的组成与多样性。
采用酶解裂解法直接从冷硫泉水中提取环境总DNA。我们构建了用古菌特异性引物扩增的16S rRNA基因克隆文库。从文库中随机挑选了115个阳性克隆，并用Alu I和Afa I酶通过限制性片段长度多态性（RFLP）进行鉴定。选择具有独特RFLP模式的相应克隆进行测序、BLAST比对并构建16S rRNA基因系统发育树。
文库中共确定了44个操作分类单元（OTU）。BLAST和系统发育分析表明，这些OTU隶属于广古菌门（94.78%）和奇古菌门（4.35%）。仅检测到一个奇古菌门克隆，与亚硝化侏儒菌属的相似度最高，为93%。广古菌门克隆丰富多样。其中，42.61%的克隆属于RC-V簇；13.91%的克隆、20.87%的克隆分别被归类到LDS簇和甲烷微菌目；4.35%的克隆与ANME-1a-FW具有高度相似性，它们参与甲烷厌氧氧化（AOM）。此外，我们还检测到一些（13.05%）未知的广古菌门克隆。
环境中的广古菌门多样，可能存在很大一部分潜在新物种。
