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

摘要:

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。

关键词:关键词:含硫冷泉，古菌，16S rRNA基因克隆文库，断裂带

Abstract:

Abstract:［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 phylogenetic tree．［Results］ 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 Euryarchaeotal clones．Conclusion］ Euryarchaeota in the environment were diverse，and possibly with a large fraction of potential novel species．

Key words:Keywords: sulfur cold spring，archaea，16S rRNA genes clones library，Earthquake fault zone