极端油藏微生物多样性分析
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摘要
文章通过对哈得油田超高温、超高盐油藏油水样品中微生物种类多样性和优势菌群的研究,考察极端油藏内源微生物的分布特征,探讨生物采油技术在超高温、超高盐油藏条件下的应用可行性。采集哈得油田HD1-1H、HD1-9H、HD1-15H、HD1-21H和HD1-23H 5组水平井油水样,利用第二代高通量测序技术测定油水样品内微生物的16S rDNA V3-V4变异区序列,应用QIIME软件整理和统计样本序列数目和操作分类单元(OTUs)数量,分析物种的丰度、分布和多样性,以及不同井微生物物种多样性丰度的差异。哈得油水样中内源微生物资源丰富,5组油水样属水平的微生物类群数分别为196、199、184、212和220,微生物在分类种类和所占比例上具有显著差异。优势菌群中有多种明确的采油功能微生物,如芽孢杆菌属、假单胞菌属、乳球菌属等。该研究明确了哈德油田具有较丰富的微生物采油物质基础,丰富了对超高温、超高盐极端油藏微生物多样性的认识,拓展了生物采油技术在极端油藏中的应用范围。
To study the diversity of endogenous microbial species and dominant microflora in oil-water samples of ultrahigh temperature and ultra-high salinity reservoirs in Hade Oilfield, the distribution characteristics of endogenous microbes in extreme reservoirs were investigated. Additionally, the feasibility of application of biotechnology enhanced oil recovery in extreme reservoirs was discussed. Five groups of water-oil samples were collected from the horizontal wells of HD1-1 H,HD1-9 H, HD1-15 H, HD1-21 H and HD1-23 H in Hade Oilfield. The second generation, high throughput sequencing technique was used to determine the 16 S rDNA V3-V4, meanwhile the QIIME software was used to analyze the abundance, distribution and diversity of species and the abundance of microbial species abundance in different wells by sorting the number of sample sequences and the number of operational taxonomic units(OTUs). The results showed that the endogenous microbial resources were rich in the Hade samples, and the number of microbial groups in the five samples was 196, 199, 184, 212 and220, respectively. There were significant differences in the classification and proportion of microbes. Among the dominant bacteria, there are many functional microorganisms, such as Bacillus, Pseudomonas and Lactococcus. It is clear that the Hade Oilfield has abundant material base of microbial oil production. This study enriched the understanding of the microbial diversity of extreme reservoirs in ultra-high temperature and salinity, and expanded the application range of biotechnology enhanced oil recovery technology in extreme reservoirs.
To study the diversity of endogenous microbial species and dominant microflora in oil-water samples of ultrahigh temperature and ultra-high salinity reservoirs in Hade Oilfield, the distribution characteristics of endogenous microbes in extreme reservoirs were investigated. Additionally, the feasibility of application of biotechnology enhanced oil recovery in extreme reservoirs was discussed. Five groups of water-oil samples were collected from the horizontal wells of HD1-1 H,HD1-9 H, HD1-15 H, HD1-21 H and HD1-23 H in Hade Oilfield. The second generation, high throughput sequencing technique was used to determine the 16 S rDNA V3-V4, meanwhile the QIIME software was used to analyze the abundance, distribution and diversity of species and the abundance of microbial species abundance in different wells by sorting the number of sample sequences and the number of operational taxonomic units(OTUs). The results showed that the endogenous microbial resources were rich in the Hade samples, and the number of microbial groups in the five samples was 196, 199, 184, 212 and220, respectively. There were significant differences in the classification and proportion of microbes. Among the dominant bacteria, there are many functional microorganisms, such as Bacillus, Pseudomonas and Lactococcus. It is clear that the Hade Oilfield has abundant material base of microbial oil production. This study enriched the understanding of the microbial diversity of extreme reservoirs in ultra-high temperature and salinity, and expanded the application range of biotechnology enhanced oil recovery technology in extreme reservoirs.
引文
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[14]万春黎,杨雪,杜茂安,等.石油污染土壤中细菌群落结构特征[J].石油学报:石油加工, 2010,26(6):928-933.Wan Chunli, Yang Xue, Du Mao'an, et al. Characteristics of bacterial community in long-term of polluted soil[J]. Acta Petrolei Sinica:Petroleum Processing Section, 2010, 26(6):928-933.
[15]夏文杰,董汉平,俞理,等.铜绿假单胞菌WJ-1降解原油特性[J].化工学报, 2011,62(7):2013-2019.Xia Wenjie, Dong Hanping, Yu Li, et al. Oil-degradation characteristics of Pseudomonas aeruginosa WJ-1[J]. CIESC Journal, 2011,62(7):2013-2019.
[16] Salwa M S, Asshifa M N N, Amirul A A, et al. Different feeding strategy for the production of biosurfactant from Pseudomonas aeruginosa USM AR2 in modified bioreactor[J]. Biotechnology&Bioprocess Engineering, 2009, 14(6):763-768.
[17]王娟娟,张宇,付娜,等.超低渗油藏微生物吞吐技术的矿场试验[J].微生物学通报, 2016,43(2):241-253.Wang Juanjuan, Zhang Yu, Fu Na, et al. Microbial huff and puff field trial in ultra-low permeability reservoir[J]. Microbiology China, 2016,43(2):241-253.
[18] Amani H, Kariminezhad H. Study on emulsification of crude oil in water using emulsan biosurfactant for pipeline transportation[J]. Petroleum Science&Technology, 2016,34(3):216-222.
[19]黄海东,王薇,马挺,等.鞘氨醇单胞菌TP-3合成新型生物聚合物Ss的发酵条件优化[J].微生物学通报, 2009,36(2):155-159.Huang Haidong, Wang Wei, Ma Ting, et al. Optimization of culture condition for novel biopolymer Ss production by Sphingomonas sp. strain TP-3[J]. Microbiology China, 2009,36(2):155-159.
[2] Bachmann RT, Johnson AC, Edyvean RGJ. Biotechnology in the petroleum industry:an overview[J]. International Biodeterioration&Biodegradation, 2014, 86(Part C):225-237.
[3]李辉,牟伯中.油藏微生物多样性的分子生态学研究进展[J].微生物学通报, 2008,35(5):803-808.Li Hui, Mu Bozhong. Recent advances in molucular microbial ecology of petroleum reservoirs[J]. Microbiology China,2008,35(5):803-808.
[4]汤玉平,顾磊,许科伟,等.油气微生物勘探机理及应用[J].微生物学通报, 2016,43(11):2386-2395.Tang Yuping, Gu Lei, Xu Kewei, et al. Mechanism and application of microbial oil and gas exploration[J]. Microbiology China, 2016,43(11):2386-2395.
[5] Carrara S, Cavallini A, Leblebici Y, et al. Capacitance DNA bio-chips improved by new probe immobilization strategies[J]. Microelectronics Journal, 2010,41(11):711-717.
[6] Yang YR, Xie BB, Yan JW. Application of next-generation sequencing technology in forensic science[J]. Genomics,Proteomics&Bioinformatics, 2014,12(5):190-197.
[7] Luciani F, Bull R A, Lloyd A R. Next generation deep sequencing and vaccine design:today and tomorrow[J].Trends in Biotechnology, 2012,30(9):443-52.
[8]秦楠,栗东芳,杨瑞馥.高通量测序技术及其在微生物学研究中的应用[J].微生物学报, 2011,51(4):445-457.Qin Nan, Li Dongfang, Yang Ruifu. High throughput sequencing technology and its application in microbiology research[J]. Acta Microbiologica Sinica, 2011,51(4):445-457.
[9] Hong C, Si YX, Xing Y, et al. Illumina MiSeq sequencing investigation on the contrasting soil bacterial community structures in different iron mining areas[J]. Environmental Science and Pollution Research, 2015,22(4):10788-99.
[10] Sambrook J, Russell D W. Molecular Cloning:a Laboratory Manual[M]. 3rd Edition. New York:Cold Spring Harbor Laboratory Press, 2001.
[11] Edgat RC. UPARSE:highly accurate OTU sequences from microbial amplicon reads[J]. Nature Methods, 2013,10(10):996-998.
[12] Cerqueira VS, Hollenbach EB, MABONI F, et al. Biodegradation potential of oily sludge by pure and mixed bacterial cultures[J].BioresourceTechnology, 2011,102:11003-11010.
[13] Youssef N, Elshahed MS, MC-inerney MJ. Microbial process in oil field:culprits, problems, and opportunities[J]. Advances in Applied Microbiology, 2009,66:141-251.
[14]万春黎,杨雪,杜茂安,等.石油污染土壤中细菌群落结构特征[J].石油学报:石油加工, 2010,26(6):928-933.Wan Chunli, Yang Xue, Du Mao'an, et al. Characteristics of bacterial community in long-term of polluted soil[J]. Acta Petrolei Sinica:Petroleum Processing Section, 2010, 26(6):928-933.
[15]夏文杰,董汉平,俞理,等.铜绿假单胞菌WJ-1降解原油特性[J].化工学报, 2011,62(7):2013-2019.Xia Wenjie, Dong Hanping, Yu Li, et al. Oil-degradation characteristics of Pseudomonas aeruginosa WJ-1[J]. CIESC Journal, 2011,62(7):2013-2019.
[16] Salwa M S, Asshifa M N N, Amirul A A, et al. Different feeding strategy for the production of biosurfactant from Pseudomonas aeruginosa USM AR2 in modified bioreactor[J]. Biotechnology&Bioprocess Engineering, 2009, 14(6):763-768.
[17]王娟娟,张宇,付娜,等.超低渗油藏微生物吞吐技术的矿场试验[J].微生物学通报, 2016,43(2):241-253.Wang Juanjuan, Zhang Yu, Fu Na, et al. Microbial huff and puff field trial in ultra-low permeability reservoir[J]. Microbiology China, 2016,43(2):241-253.
[18] Amani H, Kariminezhad H. Study on emulsification of crude oil in water using emulsan biosurfactant for pipeline transportation[J]. Petroleum Science&Technology, 2016,34(3):216-222.
[19]黄海东,王薇,马挺,等.鞘氨醇单胞菌TP-3合成新型生物聚合物Ss的发酵条件优化[J].微生物学通报, 2009,36(2):155-159.Huang Haidong, Wang Wei, Ma Ting, et al. Optimization of culture condition for novel biopolymer Ss production by Sphingomonas sp. strain TP-3[J]. Microbiology China, 2009,36(2):155-159.