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作 者 ：徐宏翔,吴敏,王小谷,杨俊毅,王春生

期 刊 ：生态学报 2008年 28卷 2期 页码：479~485

关键词：多金属结核合同区；深海沉积物；细菌多样性；16S rRNA；

Keywords:polymetallic nodule province, deep-sea sediment, bacterial diversity, 16S rRNA gene analysis,

摘 要 ：采用两种方法提取中国结核合同区东区沉积物不同层次总DNA，通过克隆测序构建了含有79个克隆子的细菌16S rRNA基因文库，分析了该海域沉积物中细菌的多样性。79个克隆在系统发育树中形成了11个大分支，包括Gamma proteobacteria(22.8%)，Alpha proteobacteria(16.5%)，Planctomycetacia(7.6%)，Delta proteobacteria(6.3%)， Nitrospira(6.3%)，Actinobacteria(6.3%)，Beta proteobacteria(5%)，Acidobacteria(5.1%)，Sphingobacteria(3.8%)，Firmicutes(2.5%)，Other bacteria(177%)，其中Gamma proteobacteria在总文库中所占比例最高，该分支细菌在0～2cm、4～6cm层也是优势菌种。Gamma proteobacteria中假单胞菌（Pseudomonas）为优势属（22.2%）。各个层次中所含细菌类群有所不同，Alpha proteobacteria、Gamma proteobacteria、Delta proteobacteria 、Planctomycetacia、Nitrospira 、Actinobacteria和Acidobacteria为三层样品共有类群。

Abstract:The development of culture-independent technique using nucleic acids has led to many new findings in studies of microbial ecology. The 16S rDNA sequencing method is one of the effectively used culture-independent techniques in recent years. In this study, 16S rDNA sequencing method was used to investigate the bacterial diversity in deep-sea sediment from Northeast Pacific polymetallic nodule province (145.3968 W, 8.3751 N, water depth of 5307m). Sediment samples were collected by a TV-multicorer and total DNA was extracted using two different methods (chemical method and DNA extracting kit method). After purification genomic DNA was amplified using the universal primers (27F and 1492R). PCR products (1.5kb) were recovered and cloned into pMD18-T vector (TaKaRa). Clones were randomly selected and sequenced. After the sequences were checked using the Chimera Check program of the RDP database, a bacterial 16S rDNA gene library of 79 clones was established. Phylogenetic analysis using Mega3.1 indicated that 79 clones can be divided into 11 phylotypes. Gamma proteobacteria (22.8%) and Alpha proteobacteria (16.5%) were the dominant components of the sediment bacterial community, followed by Planctomycetacia (7.6%), Delta proteobacteria (6.3%), Nitrospira (6.3%), Actinobacteria (6.3%), Beta proteobacteria (5%), Acidobacteria (5.1%), Sphingobacteria (38%), Firmicutes (2.5%) and Other bacteria (17.7%). Gamma proteobacteria also dominated at layer 0-2cm and 4-6cm. Different layers had different types of bacteria, but Alpha proteobacteria, Gamma proteobacteria, Delta proteobacteria, Planctomycetacia, Nitrospira, Actinobacteria and Acidobacteria appeared in all layers. Pseudomonas is common in many different deep-sea environments. In this study, it accounted for 22.2% of total Gamma proteobacteria.

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