微生物學(xué)報(bào)Acta Microbiologica Sinica
48(8)：1082~1087; 4 August 2008
基金項(xiàng)目: 國(guó)家十五攻關(guān)課題(2003BA613A-08-1); 中科院知識(shí)創(chuàng)新項(xiàng)目(KSCX2-YW-G-011)
*通訊作者,。Tel: +86-10-64807618; Fax: +86-10-64807616; E-mail:[email protected]
作者簡(jiǎn)介: 袁三青(1981. ), 男, 湖北荊門人, 碩士研究生, 主要從事油藏微生物生態(tài)的研究。E-mail: [email protected]
袁三青1,，薛燕芬1*,，汪衛(wèi)東2，李希明2,，馬延和1
(1 中國(guó)科學(xué)院微生物研究所,，北京 100101)
(2 勝利油田采油工藝研究院，東營(yíng) 257000)
摘要：【目的】通過比較分析油藏樣品的微生物群落結(jié)構(gòu)特點(diǎn),，認(rèn)識(shí)油藏微生物的生態(tài)功能?！痉椒ā坷? 種油藏微生物研究中常用的富集培養(yǎng)方法,，對(duì)勝利油田單12 區(qū)塊S12-4 油井產(chǎn)出水樣品進(jìn)行了選擇性富集培養(yǎng),，運(yùn)用構(gòu)建16S rRNA 基因文庫(kù)的方法分析了富集樣品和非培養(yǎng)樣品的細(xì)菌多樣性?！窘Y(jié)果】通過16S rRNA 基因序列比對(duì)發(fā)現(xiàn),，非培養(yǎng)樣品、異養(yǎng)菌富集樣品,、烴降解菌富集樣品和硫酸鹽還原菌富集樣品中的優(yōu)勢(shì)菌分別為Pseudomonas 屬,，Thermotoga 屬，Thermaerobacter 屬和Thermotoga 屬的成員,。多樣性分析結(jié)果表明,，非培養(yǎng)樣品的微生物多樣性最豐富，同時(shí)非培養(yǎng)樣品和富集樣品的微生物群落結(jié)構(gòu)存在很大的差異,，富集樣品中的微生物包括優(yōu)勢(shì)菌在油藏原位環(huán)境中含量很低,。【結(jié)論】細(xì)菌組成差異的比較結(jié)果,，對(duì)油藏微生物的生態(tài)功能研究和微生物驅(qū)油潛力評(píng)估具有重要意義,。
關(guān)鍵詞：油藏；非培養(yǎng),；富集培養(yǎng),；16S rRNA 基因文庫(kù)
中圖分類號(hào)：X172 文獻(xiàn)標(biāo)識(shí)碼：A 文章編號(hào)：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
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勝利油田單12 高溫油藏非培養(yǎng)和富集培養(yǎng)樣品的細(xì)菌組成特性
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