基于自主核酸提取方法分析单油井采出液微生物  

Analyses of Oil-water Microorganisms in an Exploiting Single Well Based on Home-made DNA Extraction Method

作　　者：熊驷骏 万云洋[1] 穆红梅 李文宏 费佳佳[1] 徐飞艳 陈践发[1] Xiong Sijun;Wan Yunyang;Mu Hongmei;Li Wenhong;Fei Jiajia;Xu Feiyan;Chen Jianfa(State Key Laboratory of Petroleum Resources and Prospecting,Research Centre for Geomicrobial Resources and Application,Beijing Key Labaratory of Oil and Gas Pollution Control,Unconventional Petroleum Research Institute,China University of Petroleum-Beijing;Exploration&Development Research Institute of PetroChina Changqing Oilfild Company)

机构地区：[1]中国石油大学(北京)油气资源与探测国家重点实验室,地层微生物资源与应用研究中心,油气污染防治北京市重点实验室,非常规油气科学技术研究院 [2]中国石油集团长庆油田分公司勘探开发研究院

出　　处：《油气田环境保护》2022年第2期16-22,共7页Environmental Protection of Oil & Gas Fields

基　　金：中国石油战略合作科技专项(ZLZX2020010805,ZLZX2020020405);中国石油大学(北京)前瞻导向项目(ZX20190209);国家科技重大专项(2016ZX05050011,2016ZX05040002);地层微生物资源与应用研究中心多学科联合创新创业实验室(第四批)资助。

摘　　要：近年来基于培养和非培养方法研究油藏微生物的报道已经不少,但鲜有基因组总DNA提取技术与微生物检出性的报道。文章通过对长庆油田Q2YG138140油井采出液进行现场分类分级处理,将原样本即时分为油相及水相(分成4种孔径(10.00,1.00,0.22,0.10μm)膜滤)作为研究对象,结合自主原油基因组总DNA提取技术对该油井采出液样本进行菌群分析。结果表明:油相样本共检出11门、25纲、43目、66科和78属,水相样本共检出17门、35纲、64目、105科和150属。该自主提取方法对油藏常见多种菌门及菌属基因组DNA的提取均具有较好的适用性。油相中,检出11属为“其它”,相对丰度10.02%;水相中,检出26属为“其它”,相对丰度17.34%,6属为未分类,相对丰度0.40%,该方法针对未知微生物和生物量相对较低的极端环境微生物仍均具有一定的检出能力。4种滤膜(10.00,1.00,0.22,0.10μm)分别检出10门、8门、10门和13门,59属、65属、79属和82属。0.10μm滤膜检出的菌群多样性明显多,进一步表明该自主提取方法适用于油水微生物的多样性分析。Recently, based on culture and non-culture methods there have been some reports on the study of oil-reservoirs microorganisms, but scarcely reports discuss on the relationship of extraction methods of genomic total DNA and microbial detectability. In this paper, the produced fluids from oil well Q2 YG138140 in Changqing Oilfield were immediately divided into oil phase and water phase on site, and then the later was further successively filtered through four pore-diameter(10.00,1.00,0.22 and 0.10 μm) filter membranes. The total genomic DNA of two-phase samples was extracted by filtration membrane in turn combined with home-made DNA extraction method. The sequence showed: 11 phyla, 25 classes, 43 orders, 66 families and 78 genera were detected in oil phase samples, and 17 phyla, 35 classes, 64 orders, 105 families and 150 genera were detected in water phase samples, indicating that this home-made extraction method has good applicability to the extraction of genomic DNA of common microorganisms in oil reservoir. In the oil phase, 11 genera were detected as “other” with the 10.02% relative abundance;In the aqueous phase, 26 genera were “other” with the 17.34% relative abundance, and 6 genera were “unclassified” with the 0.40% relative abundance, indicating that this method still had a certain detection ability for unknown microorganisms and for extreme environmental microorganisms with relatively low biomass. Four pore-diameter(10.00,1.00,0.22,and 0.10 μm) filtration membranes samples were detected in 10,8,10 and 13 phyla, 59,65,79 and 82 genera, respectively, of which the diversity of microorganisms on 0.10 μm was significantly increased. Hence, this result further indicates that this home-made extraction method will be suitable for the extraction of oil-water microorganisms and subsequent diversity analysis.

关 键 词：采出液 油相 水相 微生物 DNA提取 高通量测序 

分 类 号：TE35[石油与天然气工程—油气田开发工程]