微生物学报Acta Microbiologica Sinica
48(8)：1082~1087; 4 August 2008
基金项目: 国家十五攻关课题(2003BA613A-08-1); 中科院知识创新项目(KSCX2-YW-G-011)
*通讯作者。Tel: +86-10-64807618; Fax: +86-10-64807616; E-mail:xueyf@sun.im.ac.cn
作者简介: 袁三青(1981. ), 男, 湖北荆门人, 硕士研究生, 主要从事油藏微生物生态的研究。E-mail: yuansanq@126.com
袁三青1，薛燕芬1*，汪卫东2，李希明2，马延和1
(1 中国科学院微生物研究所，北京 100101)
(2 胜利油田采油工艺研究院，东营 257000)
摘要：【目的】通过比较分析油藏样品的微生物群落结构特点，认识油藏微生物的生态功能。【方法】利用3 种油藏微生物研究中常用的富集培养方法，对胜利油田单12 区块S12-4 油井产出水样品进行了选择性富集培养，运用构建16S rRNA 基因文库的方法分析了富集样品和非培养样品的细菌多样性。【结果】通过16S rRNA 基因序列比对发现，非培养样品、异养菌富集样品、烃降解菌富集样品和硫酸盐还原菌富集样品中的优势菌分别为Pseudomonas 属，Thermotoga 属，Thermaerobacter 属和Thermotoga 属的成员。多样性分析结果表明，非培养样品的微生物多样性最丰富，同时非培养样品和富集样品的微生物群落结构存在很大的差异，富集样品中的微生物包括优势菌在油藏原位环境中含量很低。【结论】细菌组成差异的比较结果，对油藏微生物的生态功能研究和微生物驱油潜力评估具有重要意义。
关键词：油藏；非培养；富集培养；16S rRNA 基因文库
中图分类号：X172 文献标识码：A 文章编号：0001-6209 (2008) 08-1082-06
Characterization of bacterial diversity in the Shengli-S12 oil reservoir by
culture-dependent and culture-independent methods
Sangqing Yuan1, Yanfen Xue1*, Weidong Wang2, Ximing Li2, Yanhe Ma1
(1Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China)
(2Academy of Oil Extraction Engineering, ShengLi Oil Field Company, Dongying 257000, China)
Abstract: [Objective] Examining bacterial diversity in an oil reservoir of Shengli oil field by both culture-independent molecular technique and enrichment method. [Methods] The heterotrophic bacteria, hydrocarbon-degrading bacteria and sulfate-reducing bacteria were enriched from S12-4 oil-well samples by the corresponding media. Then the genomic DNAs of the enrichments were extracted and the 16S rRNA gene clone libraries were constructed. [Results] The phylogenetic analysis revealed that the bacterial 16S rRNA gene clone libraries of the 3 enrichments were dominated by clones of Thermotoga, Thermaerobacter and Thermotoga, respectively. Sequences of the other co-dominant clones observed only in the enrichments of hydrocarbon-degrading bacteria and sulfate-reducing bacteria were, respectively, associated with Marinobacter and Moorella. The uncultured 16S rRNA gene library was also generated directly from total DNA of S12-4 oil-well samples by bacterial primer set. Sequence analysis of this bacterial library indicated that a large percentage of clones were highly related to the genus Pseudomonas and the dominant species emerging in the enrichment
samples had a very low content in the tested oil reservoir. The significant difference of the bacterial composition between the samples obtained from independent-culture method and enrichment method implies that the specialized nutrient may lead to a distinctive selection of dominant organisms. [Conclusion] Through culture-dependent and culture-independent
methods, we acquired important information on the bacterial diversity of ShengLi oil reservoir. These results may expand our understanding of the microbial diversity of oil reservoir and provide useful information for MEOR(microbial enhancement of oil recovery).
Keywords: 16S rRNA gene analyses; petroleum reservoirs; bacterial diversity
生物谷推荐全文下载：
胜利油田单12 高温油藏非培养和富集培养样品的细菌组成特性
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