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

Characterization of bacterial diversity in the Shengli-S12 oil reservoir by cul-ture-dependent and culture-independent methods

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。

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).