Li Hui, Zeng Jun, Gao Xiaoqi, Yang Hongmei, Zhang Tao, Yang Xiaofang, Sun Jian, Lou Kai
Institute of Microbiology, Xinjiang Academy of Agriculture Science, Urumqi 830091, China.
Wei Sheng Wu Xue Bao. 2011 May;51(5):595-602.
In order to investigated composition and diversity of bacterial in a cold sulfur spring in Xinjiang faulting zone.
Environmental total DNA was directly extracted from the water of the No. 10 cold sulfur spring. The 16S rRNA genes were amplified from the total DNA by PCR with bacteria-specific primers and construction a clone library. Positive clones were randomly selected from the library and identified by restriction fragment length polymorphism (RFLP). The unique RFLP pattern corresponded sequences were sequenced, BLAST and then constructed phylogenetic tree.
In total, 228 positive clones were screened and grouped into 33 Operational Taxonomic Units (OTUs). The clone coverage C value was 92%. 33 Operational Taxonomic Units were divided into 3 phyla with Blast analysis and RDP classifer: Proteobacteria, Bacteroidetes, and Firmicutes. Proteobacteria (98%) was the absolutely dominant group, of which 20% of the clones were highly related to the known photoautotrophic and chemoautotrophic bacteria ( > 97% sequence similarity). Besides, 64% of the clones showed less than 96% of sequence similarity with sequence deposited in GenBank database, of them 54% sequences were affiliated to genus Legionella spp.
Bacterial diversity in No. 10 cold sulfur spring was low, but maybe have a diversity of novel species and lineages. In addition, large number of novel species of Legionella were detected in the spring water may suggest the water potentially a source of Legionnaires disease and may constitute a menace to the health of human and livestock that lived down the spring.
为研究新疆断裂带一处冷硫泉中细菌的组成与多样性。
直接从10号冷硫泉水中提取环境总DNA。使用细菌特异性引物通过PCR从总DNA中扩增16S rRNA基因，并构建克隆文库。从文库中随机挑选阳性克隆，通过限制性片段长度多态性（RFLP）进行鉴定。对具有独特RFLP模式的相应序列进行测序、BLAST分析，然后构建系统发育树。
共筛选出228个阳性克隆，分为33个操作分类单元（OTU）。克隆覆盖率C值为92%。通过Blast分析和RDP分类器将33个操作分类单元分为3个门：变形菌门、拟杆菌门和厚壁菌门。变形菌门（98%）是绝对优势菌群，其中20%的克隆与已知的光合自养和化能自养细菌高度相关（序列相似性>97%）。此外，64%的克隆与GenBank数据库中存储的序列的序列相似性低于96%，其中54%的序列隶属于军团菌属。
10号冷硫泉中的细菌多样性较低，但可能存在新物种和新谱系的多样性。此外，在泉水中检测到大量军团菌新物种可能表明该水体可能是军团病的一个来源，并可能对生活在泉水下游的人类和牲畜健康构成威胁。
