【作者】 张莹；

【导师】 黄永红； 陈珊；

【作者基本信息】 东北师范大学 ， 微生物学， 2015， 硕士

【摘要】 微生物提高原油采收率技术(MEOR),是世界上应用于油田采油的一种第三次采油方法,目前已成为一种重要并迅速发展的采油技术,它利用微生物自身的有益活动或其代谢物,提高剩余油采收率。随着这项技术的发展,人们已经从各大油藏中分离得到多种具有一定驱油效果的驱油菌株。但是,由于自然界中可培养的微生物种类在总菌群数量中只占有及其微小的部分,这就使得传统的分离培养技术只能反映很少量的所检测的油藏微生物的菌群特征。近年来随着微生物分子生态学迅速发展,尤其是基于16S r DNA克隆文库、实时荧光原位杂交及DGGE(变性梯度凝胶电泳)等技术手段的不断完善及发展,微生物分子生态技术已广泛应用到油藏微生物群落结构和种群动态研究中。微生物群落结构和功能的解析,以及群落的变化得到了充分的研究。使人们更加全面、客观、准确且及时的认识油藏微生物生物群落结构和种群动态变化规律,可以从分子水平上揭示油藏微生物的多样性,促进对油藏微生物多样性的认识,并深刻影响微生物采油的应用方向。本研究利用基因克隆文库的方法分析了大庆油田聚驱后油藏(采油三厂和采油四厂)、低渗透油藏(采油十厂)典型区块采出液微生物菌群结构特征。分别构建了这些区块细菌和古菌的16Sr DNA基因克隆文库,并利用NCBI数据库进行序列比对分析和MEGA5.1进行系统进化分析。结果表明,采油三厂优势细菌菌属为Uncultured bacterium clone,占总菌群的82%。优势古菌菌属为来源于高热温泉环境,对地下高温环境具有很强的适应性的Crenarchaeotal sp.,占总菌群的36%。采油四厂细菌优势细菌菌属为Uncultured bacterium,占总菌群的60.5%之间。优势古菌菌属为Uncultured bacterium,占总菌群的34%。采油十厂优势细菌菌属为具有降解乙腈,降解4-氯酚等特性Acinetobacter sp.,占43.5%。优势古菌菌属为源于碱性土壤环境,具有产氢及纤维素酶等作用的Pantoea sp.,占总菌群的57.75%。在本文中,利用16Sr DNA基因克隆文库分析了油藏中微生物的群落组成以及优势种群,分析主要的采油功能菌,为微生物提高原油采收率(MEOR)菌群的选择提供指导。同时这些微生物分子生态学技术的应用为确定油藏微生物提高原油采收率技术的可行性,以及进一步指导微生物驱油过程中菌种的配伍、开发和应用。进而优化微生物微生物提高原油采收率技术提供了可靠的依据。更多还原

【Abstract】 Microbial enhanced oil recovery(MEOR) is a third oil recovery method in the world used in oil production, now it has become an important and rapid development of production technology,it uses the beneficial microbial activities by itself or its metabolites to enhance oil recovery. At present, people can use the way of pure culture,some microorganisms in oilfield were isolated and cultured,and have been isolated from the major reservoir with oil flooding some flooding effect strain. However,due to the nature of microbial species can develop only rarely in the possession and total number of bacteria in part,which makes the traditional isolation technology can only reflect the characteristics of a small amount of bacteria detected by microbial reservoirs.In recent years,with the rapid development of microbial molecular ecology,especially the 16 S r DNA clone library, real-time fluorescence in situ hybridization,based on DGGE(denaturing gradient gel electrophoresis) technology improvement and development,microbial molecular ecology technology has been widely used to study the community structure and population dynamics of microorganisms in reservoir.Analysis of microbial community structure and function,and the changes in community has been studied.To make people more comprehensive,objective,accurate and timely understanding of the reservoir microbial community structure and population dynamics,can reveal the diversity of oil reservoir microorganism at the molecular level,to promote understanding of the diversity of the reservoir microorganisms,and deeply affected the microbial recovery and application direction.In this study,using the method of gene clone library analysis of Daqing oilfield after polymer flooding reservoir(three production plants and four production plants),low permeability reservoir(ten plant oil) produced in typical block structure characteristics of liquid bacteria.These blocks were constructed by bacterial and archaeal 16 Sr DNA gene clone library,and MEGA5.1 sequence analysis and phylogenetic analysis using NCBI database.The results show that,the three production plant belongs to the genus of Uncultured bacterium bacteria clone,total bacteria 82%.The advantage of Archaea belongs to the genus from high spring environment,has strong adaptability to the underground high temperature environment Crenarchaeotal sp.,the total bacteria 36%. Four production plant bacteria bacteria from the Uncultured bacterium,accounting for 60.5% of total bacteria.The advantage of Archaea belongs to the genus of Uncultured bacterium,accounted for more than 34% of the bacteria,followed by alkaline soil from the environment Rhizobium sp.reached 29.75% of the total number of bacteria.The ten production plant belongs to the genus of bacteria with degradation of acetonitrile,degradation characteristics of chlorophenol Acinetobacter sp. accounted for 43.5%.The advantage of archaea,bacteria were derived from alkaline soil environment,with hydrogen production and cellulase effect of Pantoea sp. to total bacteria 57.75%.This paper uses the 16 Sr DNA gene clone library analysis reservoir microbial community composition and dominant species,the main production function analysis for bacteria,microbial enhanced oil recovery(MEOR) provides guidance for flora selection,At the same time,the application of Microbial Molecular Ecology Techniques for determining reservoir microbial enhanced oil recovery technology feasibility,compatibility,development and application of strain and further guidance in the process of microbial enhanced oil recovery.Then the optimization provides a reliable basis for the technology of EOR of microorganism.更多还原