Molecular analysis of deep-sea hydrothermal vent aerobic methanotrophs by targeting genes of 16S rRNA and particulate methane monooxygenase
Molecular analysis of deep-sea hydrothermal vent aerobic methanotrophs by targeting genes of 16S rRNA and particulate methane monooxygenase
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DOI：
10.1007/s10126-004-3042-0
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发表时间：
2004-09-01
影响因子：
3
通讯作者：
Naganuma, T
中科院分区：
文献类型：
--
作者：
Elsaied, HE;Hayashi, T;Naganuma, T
使用 16S 核糖体 DNA 和编码颗粒甲烷单加氧酶 (pMOA) 亚基 A 的 pmoA 研究了深海热液喷口需氧甲烷氧化菌的分子多样性。热液喷口羽流和烟囱样本分别从日本冲绳中部海槽 (MOT) 的弧后喷口和沿大西洋中脊的跨大西洋地理穿越 (TAG) 站点采集。使用特定引物通过聚合酶链式反应从大量 DNA 中扩增靶基因并进行克隆。对来自每个克隆文库的五十个克隆进行直接测序。 16S rDNA 序列分为 3 个操作分类单元 (OTU)，其中 2 个来自 MOT，I 来自 TAG。两个 OTU（I MOT 和 I TAG）位于 I 型甲烷氧化伽玛变形菌的分支内。另一个 MOT OTU 形成了与 I 型甲烷氧化菌相关的独特系统发育谱系。对来自 MOT 和 TAG 样本的 50 个克隆进行直接测序，分别产生 17 个和 4 个可操作的 pmoA 单位 (OPU)。基于OPU推导的pMOA氨基酸序列的系统发育树在I型甲烷氧化菌分支内形成了不同的系统发育谱系，除了与X型甲烷氧化菌相关的OPU MOT-pmoA-8之外。推断该基因的 pMOA 拓扑结构与所有已知的 pMOA 相似，这可能表明 pmoA g 在进化过程中是保守的。 16 rDNA 和 pmoA 分子分析均无法检测到 II 型甲烷氧化菌，这表明收集的喷口样品中不存在 II 型甲烷氧化菌。
Molecular diversity of deep-sea hydrothermal vent aerobic methanotrophs was studied using both 16S ribosomalDNA and pmoA encoding the subunit A of particulate methane monooxygenase (pMOA). Hydrothermal vent plume and chimney samples were collected from back-arc vent at Mid-Okinawa Trough (MOT), Japan, and the Trans-Atlantic Geotraverse (TAG) site along Mid-Atlantic Ridge, respectively. The target genes were amplified by polymerase chain reaction from the bulk DNA using specific primers and cloned. Fifty clones from each clone library were directly sequenced. The 16S rDNA sequences were grouped into 3 operational taxonomic units (OTUs), 2 from MOT and I from TAG. Two OTUs (I MOT and I TAG) were located within the branch of type I methanotrophic gamma-Proteobacteria. Another MOT OTU formed a unique phylogenetic lineage related to type I methanotrophs. Direct sequencing of 50 clones each from the MOT and TAG samples yielded 17 and 4 operational pmoA units (OPUs), respectively. The phylogenetic tree based on the pMOA amino acid sequences deduced from OPUs formed diverse phyogenetic lineages within the branch of type I methanotrophs, except for the OPU MOT-pmoA-8 related to type X methanotrophs. The gene is deduced pMOA topologies were similar to those of all known pMOA, which may suggest that the pmoA g conserved through evolution. Neither the 16 rDNA nor pmoA molecular analysis could detect type II methanotrophs, which suggests the absence of type II methanotrophs in the collected vent samples.