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
李萍 1曾军 2祖丽皮亚·玉努斯 3高小其 4董秀黄 1薛娟 5娄恺2
作者信息
- 1. 新疆大学生命科学与技术学院,乌鲁木齐830046;新疆农业科学院微生物研究所,乌鲁木齐830091
- 2. 新疆农业科学院微生物研究所,乌鲁木齐830091
- 3. 新疆大学生命科学与技术学院,乌鲁木齐830046
- 4. 新疆维吾尔自治区地震局,乌鲁木齐830011
- 5. 新疆农业科学院微生物研究所,乌鲁木齐830091;新疆石河子大学生命科学学院,石河子832003
- 折叠
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AfaⅠ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.
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-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/16S/rRNA基因克隆文库/断裂带Key words
sulfur cold spring/archaea/16S rRNA genes clones library/Earthquake fault zone引用本文复制引用
基金项目
新疆维吾尔自治区高技术研究发展项目(201216145)
出版年
2013
NETL
NSTL
维普
万方数据