大庆油田聚合物驱后油藏微生物多样性研究

Microbial Diversity of the Daqing Oilfi eld after Polymer Flooding

从大庆油田4个聚合物驱后的采油井中提取总DNA,扩增其16S rDNA片段,克隆到大肠杆菌,然后用ARDRA法对所得的298个克隆中的插入片段进行分析,建立了16SrDNA克隆文库.采用文库库容值(C)、香农–威纳指数(H)和均匀度指数(E)对文库微生物多样性进行了评价.结果表明,4个样品中共得出22个操作分类单元(OTU).其中4个OTU在整个群落中占78.18%,9个OTU只有一个克隆.对22个OTU的代表克隆序列、GenBank数据库比对和系统发育分析表明,聚驱后油井中的细菌基本属于α变形菌纲(24.83%)、γ变形菌纲(23.49%)、δ变形菌纲(35.56%)和未培养微生物(16.78%).与石油烃降解和产生物表面活性剂相关的假单胞菌(Pseudomonas)和不动杆菌(Acinetobacter),在文库中占的比例分别是17.79%和6.02%.

DNA was extracted from four oil well samples after polymer flooding in the Daqing Oilfield, Heilongjiang, China. A 16S rDNA clone library was constructed by PCR amplification with universal primers, cloned in Escherichia coli and characterized subsequently by amplified rDNA restriction analysis （ARDRA）. 298 clones were obtained and classified into 22 different operational taxonomic units （OTUs）. Four OTUs occupied 78.18 % of the 16S rDNA clone library, while nine OTUs had only one clone. At the same time, the microbial diversity of the 16S rDNA clone library was evaluated by coverage value （C）, Shannon - Weiner index （H） and evenness index （E）. Phylogenetic analysis revealed that the dominant microbes of the library belonged to Alphaproteobacteria （24.83%）, Gammaproteobacteria （23.49%）, Deltaproteobacteria （34.56%） and the uncultured bacteria （16.78%）. Pseudomonas and Acinetobacter related with hydrocarbon biodegradation and surfactant production accounted for 17.79 % and 6.02 % in the clone library, respectively.