新疆乌鲁木齐10号冷泉古菌群落结构多样性  

Archaeal community structure and diversity in Urumqi No.10 cold sulfur Spring analyzed by culture-independent approach

作　　者：李萍[1,2] 曾军[2] 祖丽皮亚.玉努斯[1] 高小其[3] 董秀黄[1,2] 薛娟[2,4] 娄恺[2] 

机构地区：[1]新疆大学生命科学与技术学院,乌鲁木齐830046 [2]新疆农业科学院微生物研究所,乌鲁木齐830091 [3]新疆维吾尔自治区地震局,乌鲁木齐830011 [4]新疆石河子大学生命科学学院,石河子832003

出　　处：《微生物学报》2013年第3期230-240,共11页Acta Microbiologica Sinica

基　　金：新疆维吾尔自治区高技术研究发展项目(201216145)~~

摘　　要：【目的】探究新疆地震断裂带含硫冷泉泉水古菌群落组成及多样性。【方法】利用酶解法直接从冷泉泉水样品中提取环境总DNA,采用古菌通用引物对16S rRNA基因进行扩增,构建16S rRNA基因克隆文库,通过Alu I和Afa I两种限制性内切酶对随机挑选的115个阳性克隆子进行酶切分型,将不同酶切带型对应的克隆子送样测序,测序结果与GenBank序列进行比对并构建16S rRNA基因系统发育树。【结果】古菌克隆文库中共得到44个不同的酶切带型,BLAST序列比对和系统发育分析将它们划分于广古菌门(Euryarchaeota,94.78%)和奇古菌门(Thaumarchaeota,4.35%)。奇古菌门克隆与Nitrosopumilus.sp序列相似性达到了93%;而广古菌门类群较为多样,其中,42.61%的克隆子属于RC-V cluster,20.87%与13.91%的克隆子分别属于LDS cluster和Methanomicrobiales,4.35%的克隆子与甲烷厌氧氧化相关的类群(ANME-1a-FW)具有较高的的相似。另外,13.05%的克隆子属于广古菌门中的未知类群。【结论】乌鲁木齐10号泉水体中广古菌类群多样,可能蕴藏有大量潜在的古菌新类群。[ Objective] We explored the composition and diversity of archaea in a cold sulfur spring water in Xinjiang earthquake fault zone. [ Methods] 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 phylogenetie tree. [ Results] In total, 44 operational taxonomic units （OTUs） were determined from the library. BLAST and phylogenetie 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-la-FW, which were involved in Anaerobic oxidation of methane （AOM）. In addition, we also detected some （13.05%） unknown Euryarchaeotal clones. [ Conclusion] Euryarchaeota in the environment were diverse, and possibly with a large fraction of potential novel species.

关 键 词：含硫冷泉 古菌 16SrRNA基因克隆文库 断裂带 

分 类 号：Q938.8[生物学—微生物学]