基于高通量测序技术研究页岩气区上方微生物多样性

顾磊; 许科伟; 汤玉平; 杨帆; 王国建; 杜伟; 高波; 腾格尔

doi:10.11781/sysydz202003443

基于高通量测序技术研究页岩气区上方微生物多样性

doi: 10.11781/sysydz202003443

顾磊^1,,
许科伟^1, ,,
汤玉平¹,
杨帆¹,
王国建¹,
杜伟²,
高波²,
腾格尔^{1, 3}

1.
中国石化石油勘探开发研究院无锡石油地质研究所, 无锡 214126
2.
中国石化石油勘探开发研究院非常规资源研究所, 北京 100083
3.
中国地质调查局油气资源调查中心, 北京 100083

基金项目:

国家科技重大专项 Z017ZX05036-002

国家自然科学基金 U1663202

国家自然科学基金 41690133

国家自然科学基金 41872126

详细信息

作者简介:
顾磊(1987—)，男，博士，副研究员，从事油气微生物勘探开发研究。E-mail：gulei.syky@sinopec.com

通讯作者:
许科伟(1982—)，男，博士，高级工程师，从事油气微生物勘探开发研究。E-mail：xukw.syky@sinopec.com

中图分类号: TE132.2
计量
- 文章访问数: 651
- HTML全文浏览量: 126
- PDF下载量: 103
- 被引次数: 0
出版历程
- 收稿日期: 2020-01-27
- 修回日期: 2020-04-29
- 刊出日期: 2020-05-28

Microbial diversity above a shale gas field using high-throughput sequencing

GU Lei^1
,,
XU Kewei^{1
, ,},
TANG Yuping¹,
YANG Fan¹,
WANG Guojian¹,
DU Wei²,
GAO Bo²,
BORJIGIN Tenger^{1, 3}

1.
Wuxi Research Institute of Petroleum Geology, SINOPEC, Wuxi, Jiangsu 214126, China
2.
Research Institute of Unconventional Resources, SINOPEC Petroleum Exploration & Production Research Institute, Beijing 100083, China
3.
Oil and Gas Survey, China Geological Survey, Beijing 100083, China

摘要

摘要: 微生物油气勘探技术近年来已成为油气地表勘探技术中新兴的勘探检测技术。为进一步了解油气田上方的微生物信息，对页岩气区上方微生物多样性进行了研究。采用高通量测序技术研究了川东南丁山地区油气微生物物种丰度、群落多样性等特征。鉴定出了3种主要油气微生物：甲基嗜氧反硝化菌（Methylophilales）、甲基孢囊菌（Methylocystis）、甲基球菌（Methylococcus），值得进一步做生化研究，可以作为潜在油气微生物指示菌。3种微生物发育丰度与丁山地区逸散区域分布具有较好的匹配关系。高通量解析技术能够在丁山地区油气微生物研究中有很好的应用，得出了基岩样品中微生物分布的特征，并与焦石坝地区油气微生物进行了初步的差异性分析，鉴定出的潜在油气指示菌值得进一步研究。
- 油气微生物勘探 /
- 高通量测序 /
- 群落结构 /
- 微生物多样性
Abstract: In recent years, microbial analysis has become a new detection technology in oil and gas surface exploration. The microbial diversity above a shale gas field was studied in order to further understand microbial information. High-throughput sequencing technology was used to study the abundance and diversity of hydrocarbon species in a shale gas field in the Dingshan area of the southeastern Sichuan Basin. Three main microorganisms were identified, including Methylophilales, Methylocystis, and Methylococcus, which are worthy of further biochemical research and can be used as potential oil and gas indicating bacteria. The developmental abundance of three kinds of microorganisms has a good matching relationship with the distribution of the fugitive area in Dingshan. The high-throughput analytical technology can be applied well in the study of oil and gas microorganisms in Dingshan area, and the distribution characteristics of microorganisms in bedrock samples were obtained. Furthermore, a preliminary analysis of the differences between oil and gas microbes in Dingshan and Jiaoshiba areas was made, indicating that the identified potential oil and gas indicating bacteria deserve further study.
- microbial exploration for oil and gas /
- high-throughput sequencing /
- community structure /
- microbial diversity

HTML全文

图 1 川东南丁山和焦石坝剖面样品采集部署

Figure 1. Sample distribution in Dingshan and Jiaoshiba areas, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 2 川东南丁山与焦石坝地区页岩富集区微生物属水平物种的相对丰度

Figure 2. Relative abundance at genus level of microbes in shale gas fields in Dingshan and Jiaoshiba areas, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 3 川东南丁山与焦石坝地区页岩富集区物种Rank-Abundance曲线

Figure 3. Species rank vs. relative abundance in shale gas fields in Dingshan and Jiaoshiba areas, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 4 川东南丁山与焦石坝地区物种属水平丰度聚热图

Figure 4. Heat distribution at genus level of microbes in shale gas fields in Dingshan and Jiaoshiba areas, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 5 川东南丁山地区上方土壤中甲基类油气微生物分布

剖面SS’位置见图 1a

Figure 5. Distribution of methyl hydrocarbons in soil above Dingshan area, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 6 川东南焦石坝地区上方土壤中甲基类油气微生物分布

剖面EE’位置见图 1b

Figure 6. Distribution of methyl hydrocarbons in soil above Jiaoshiba area, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 7 川东南丁山地区页岩气逸散模式^[21]

Figure 7. Shale gas dissipation model in Dingshan area, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

图 8 川东南丁山地区联井剖面热释烃甲烷浓度分布

剖面SS’位置见图 1a

Figure 8. Concentration of pyrolysis hydrocarbon methane in Dingshan area, southeastern Sichuan Basin

下载: 全尺寸图片幻灯片

参考文献(23)

[1]	WAGNER M, WAGNER M, PISKE J, et al. Case histories of microbial prospection for oil and gas, onshore and offshore in northwest Europe[J]. AAPG Special Volumes, 2002(11): 453-479.
[2]	张春林, 庞雄奇, 梅海, 等. 微生物油气勘探技术在岩性气藏勘探中的应用: 以柴达木盆地三湖坳陷为例[J]. 石油勘探与开发, 2010, 37(3): 310-315. https://www.cnki.com.cn/Article/CJFDTOTAL-SKYK201003009.htm ZHANG Chunlin, PANG Xiongqi, MEI Hai, et al. Application of microbial oil surveying to exploration of lithologic gas reservoirs: a case from the Sanhu Depression, Qaidam Basin, NW China[J]. Petroleum Exploration and Development, 2010, 37(3): 310-315. https://www.cnki.com.cn/Article/CJFDTOTAL-SKYK201003009.htm
[3]	TUCKER J, HITZMAN D. Detailed microbial surveys help improve reservoir characterization[J]. Oil and Gas Journal, 1994, 92(23): 65-69.
[4]	索孝东, 石东阳. 油气地球化学勘探技术发展现状与方向[J]. 天然气地球科学, 2008, 19(2): 286-292. https://www.cnki.com.cn/Article/CJFDTOTAL-TDKX200802030.htm SUO Xiaodong, SHI Dongyang. Present situation and development trend of geochemical exploration for oil and gas[J]. Natural Gas Geoscience, 2008, 19(2): 286-292. https://www.cnki.com.cn/Article/CJFDTOTAL-TDKX200802030.htm
[5]	徐凤银, 彭德华, 侯恩科. 柴达木盆地油气聚集规律及勘探前景[J]. 石油学报, 2003, 24(4): 1-6. https://www.cnki.com.cn/Article/CJFDTOTAL-SYXB200304000.htm XU Fengyin, PENG Dehua, HOU Enke. Hydrocarbon accumulation and exploration potential in Qaidam Basin[J]. Acta Petrolei Sinica, 2003, 24(4): 1-6. https://www.cnki.com.cn/Article/CJFDTOTAL-SYXB200304000.htm
[6]	张春林, 庞雄奇, 梅海, 等. 烃类微渗漏与宏渗漏的识别及镇巴长岭-龙王沟地区勘探实践[J]. 天然气地球科学, 2009, 20(5): 794-800. https://www.cnki.com.cn/Article/CJFDTOTAL-TDKX200905028.htm ZHANG Chunlin, PANG Xiongqi, MEI Hai, et al. Identification of microseepage from macroseepage and exploration practice in Changling-Longwanggou area of Zhenba block[J]. Natural Gas Geoscience, 2009, 20(5): 794-800. https://www.cnki.com.cn/Article/CJFDTOTAL-TDKX200905028.htm
[7]	梅博文, 袁志华, 王修垣. 油气微生物勘探法[J]. 中国石油勘探, 2002, 7(3): 42-53. https://www.cnki.com.cn/Article/CJFDTOTAL-KTSY200203007.htm MEI Bowen, YUAN Zhihua, WANG Xiuyuan. Microbial prospection for oil and gas[J]. China Petroleum Exploration, 2002, 7(3): 42-53. https://www.cnki.com.cn/Article/CJFDTOTAL-KTSY200203007.htm
[8]	袁志华, 张玉清, 赵青, 等. 中国油气微生物勘探技术新进展: 以大庆卫星油田为例[J]. 中国科学(D辑地球科学), 2008, 38(S2): 139-145. https://www.cnki.com.cn/Article/CJFDTOTAL-JDXK2008S2020.htm YUAN Zhihua, ZHANG Yuqing, ZHAO Qing, et al. Progress of microbial oil surveying to exploration of oil and gas in China: a case from Daqing Oilfield[J]. Science in China (Series D Earth Sciences), 2008, 38(S2): 139-145. https://www.cnki.com.cn/Article/CJFDTOTAL-JDXK2008S2020.htm
[9]	梅海, 林壬子, 梅博文, 等. 油气微生物检测技术: 理论、实践和应用前景[J]. 天然气地球科学, 2008, 19(6): 888-893. https://www.cnki.com.cn/Article/CJFDTOTAL-TDKX200806026.htm MEI Hai, LIN Renzi, MEI Bowen, et al. Microbial oil gas detection technologies: theory, practice and application prospect[J]. Natural Gas Geoscience, 2008, 19(6): 888-893. https://www.cnki.com.cn/Article/CJFDTOTAL-TDKX200806026.htm
[10]	张春林, 庞雄奇, 梅海, 等. 微生物油气勘探技术的实践与发展[J]. 新疆石油地质, 2010, 31(3): 320-322. https://www.cnki.com.cn/Article/CJFDTOTAL-XJSD201003037.htm ZHANG Chunlin, PANG Xiongqi, MEI Hai, et al. Application and progress of microbial oil survey technique (MOST)[J]. Xinjiang Petroleum Geology, 2010, 31(3): 320-322. https://www.cnki.com.cn/Article/CJFDTOTAL-XJSD201003037.htm
[11]	WHITTAKER R H. Evolution and measurement of species diversity[J]. Taxon, 1972, 21(2/3): 213-251.
[12]	LUNDBERG D S, YOURSTONE S, MIECZKOWSKI P, et al. Practical innovations for high-throughput amplicon sequencing[J]. Nature Methods, 2013, 10(10): 999-1002.
[13]	LOZUPONE C, KNIGHT R. UniFrac: a new phylogenetic method for comparing microbial communities[J]. Applied and Environmental Microbiology, 2005, 71(12): 8228-8235.
[14]	LOZUPONE C, LLADSER M E, KNIGHTS D, et al. UniFrac: an effective distance metric for microbial community comparison[J]. The ISME Journal, 2011, 5(2): 169-172.
[15]	LOZUPONE C A, HAMADY M, KELLEY S T, et al. Quantitative and qualitative β diversity measures lead to different insights into factors that structure microbial communities[J]. Applied and Environmental Microbiology, 2007, 73(5): 1576-1585.
[16]	AVERSHINA E, FRISLI T, RUDI K. De novo semi-alignment of 16S rRNA gene sequences for deep phylogenetic characterization of next generation sequencing data[J]. Microbes and Environments, 2013, 28(2): 211-216.
[17]	CAPORASO J G, LAUBER C L, WALTERS W A, et al. Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample[J]. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(S1): 4516-4522.
[18]	YOUSSEF N, SHEIK C S, KRUMHOLZ L R, et al. Comparison of species richness estimates obtained using nearly complete fragments and simulated pyrosequencing-generated fragments in 16S rRNA gene-based environmental surveys[J]. Applied and Environmental Microbiology, 2009, 75(16): 5227-5236.
[19]	HESS M, SCZYRBA A, EGAN R, et al. Metagenomic discovery of biomass-degrading genes and genomes from cow rumen[J]. Science, 2011, 331(6016): 463-467.
[20]	顾磊, 许科伟, 汤玉平, 等. 基于高通量测序技术研究玉北油田上方微生物多样性[J]. 应用与环境生物学报, 2017, 23(2): 276-282. https://www.cnki.com.cn/Article/CJFDTOTAL-YYHS201702015.htm GU Lei, XU Kewei, TANG Yuping, et al. Microbial diversity in Yubei Oil field determined by high-throughput sequencing[J]. Chinese Journal of Applied & Environmental Biology, 2017, 23(2): 276-282. https://www.cnki.com.cn/Article/CJFDTOTAL-YYHS201702015.htm
[21]	黄仁春, 魏祥峰, 王强. 四川盆地东南缘丁山地区页岩气成藏富集的关键控制因素[J]. 海相油气地质, 2017, 22(2): 22-30. https://www.cnki.com.cn/Article/CJFDTOTAL-HXYQ201702004.htm HUANG Renchun, WEI Xiangfeng, WANG Qiang. Key factors of shale gas accumulation in Dingshan area of southeastern Sichuan Basin[J]. Marine Origin Petroleum Geology, 2017, 22(2): 22-30. https://www.cnki.com.cn/Article/CJFDTOTAL-HXYQ201702004.htm
[22]	姚威, 许锦, 夏文谦, 等. 四川盆地涪陵地区茅一段酸解气、吸附气特征及气源对比[J]. 天然气工业, 2019, 39(6): 45-50. https://www.cnki.com.cn/Article/CJFDTOTAL-TRQG201906008.htm YAO Wei, XU Jin, XIA Wenqian, et al. Characteristic analysis between acidolysis gas and absorbed gas and its application to gas-source correlation in Mao 1 Member, Fuling area, Sichuan Basin[J]. Natural Gas Industry, 2019, 39(6): 45-50. https://www.cnki.com.cn/Article/CJFDTOTAL-TRQG201906008.htm
[23]	黄文明, 刘树根, 马文辛, 等. 川东南-鄂西渝东地区下古生界页岩气勘探前景[J]. 地质通报, 2011, 30(2): 364-371. https://www.cnki.com.cn/Article/CJFDTOTAL-ZQYD2011Z1024.htm HUANG Wenming, LIU Shugen, MA Wenxin, et al. Shale gas exploration prospect of Lower Paleozoic in southeastern Sichuan and western Hubei-eastern Chongqing areas, China[J]. Geolo-gical Bulletin of China, 2011, 30(2): 364-371. https://www.cnki.com.cn/Article/CJFDTOTAL-ZQYD2011Z1024.htm