【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。更多还原

【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.更多还原