页岩气返排水中的反硝化厌氧甲烷氧化微生物——群落结构与关键影响因子

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摘要首次探究了高温高压高盐的页岩气返排水中DAMO (denitrifying anaerobic methane oxidation)微生物存在的证据及其与关键环境因子之间的关联性.结果表明,在所研究的7个页岩气井场返排水中均检测到了DAMO古菌与细菌的存在,且古菌的丰度远高于细菌,其16S rRNA基因拷贝数分别为4.32×10⁴~5.83×10⁵copies/L和1.16×10³~6.00×10³copies/L.DAMO古菌与细菌的功能基因mcrA与pomA同样在所有的样品中检出.页岩气返排水中共发现了4个DAMO细菌代表性OTU与1个DAMO古菌代表性OTU,虽然返排水中盐度很高,但检出的DAMO细菌与古菌与已报道的淡水环境中的DAMO微生物亲缘性更近.RDA相关性分析表明,返排水常规理化因子对DAMO微生物的影响并非体现在微生物的丰度上,而是体现在微生物的功能上,其中pmoA基因丰度与TOC含量呈正显著相关,而mcrA基因丰度则与NO₃^-、NH₄⁺、PO₄^3-和Cl^-浓度呈正相关,Cl^-影响最大.研究结果拓展了关于DAMO微生物环境分布的认识,可为缓解返排水中逸散甲烷所导致的温室效应的控制提供理论基础,也有助于加深对深部页岩微生物过程的认知.

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关键词 ：反硝化厌氧甲烷氧化, 页岩气返排水, 高盐, 环境因子, 群落结构

Abstract：In this study, we investigated the presence of denitrifying anaerobic methane oxidizing microorganisms in high temperature, high pressure and high salt shale gas flowback water for the first time, along with their correlation with key environmental factors. The results indicated that DAMO archaea and bacteria were detected in all the seven shale gas flowback water, and the archaea exhibiting significantly higher abundance compared to bacteria with 16S rRNA gene copy ranging from 4.32×10⁴～ 5.83×10⁵copies/L and 1.16×10³~6.00×10³ copies/L, respectively. Additionally, the functional genes mcrA and pomA of DAMO archaea and bacteria were also detected in all the samples. Four representative OTUs of DAMO bacteria and one representative OTU of DAMO archaea were identified in the shale gas flowback water. Despite the high salinity, the DAMO bacteria and archaea detected in flowback water shared a closer genetic relationship to those reported in freshwater environments. RDA correlation analysis revealed that the influence of physicochemical factors in flowback water on DAMO microorganisms was more evident in microbial function rather than microbial abundance. The abundance of pmoA gene was positively and significantly correlated with TOC content, while the abundance of mcrA gene was positively correlated with the concentrations of NO₃^-, NH₄^-, PO₄^3-, and Cl^-, with Cl^- having the greatest impact. These results of the study expanded the understanding of the environmental distribution of DAMO microorganisms, providing a theoretical foundation for controlling greenhouse effect caused by methane emissions in flowback water, and contributing to a deeper comprehension of microbial processes in deep shale formations.

Key words： denitrifying anaerobic methane oxidation shale gas flowback water high salinity environmental factors community structure

收稿日期: 2024-02-19

PACS:

X172

基金资助:国家自然科学基金资助项目(52070025,52200086);国家重点研发计划项目(2019YFC1805503);煤矿灾害动力学与控制国家重点实验室开放项目(2011DA105287-MS202213)

通讯作者: 丁阿强,副教授,aqding@cqu.edu.cn E-mail: aqding@cqu.edu.cn

作者简介: 李欣悦(1999-),女,重庆人,硕士研究生,主要从事反硝化厌氧甲烷氧化过程及其微生物研究.1937222582@qq.com.

引用本文:

李欣悦, 卢培利, 刘俊, 李仕康, 侯争鸣, 丁阿强. 页岩气返排水中的反硝化厌氧甲烷氧化微生物——群落结构与关键影响因子[J]. 中国环境科学, 2024, 44(9): 5303-5312. LI Xin-yue, LU Pei-li, LIU Jun, LI Shi-kang, HOU Zheng-ming, DING A-qiang. Denitrifying anaerobic methane oxidation microorganisms in shale gas flowback water ― Community structure and key influencing factors. CHINA ENVIRONMENTAL SCIENCECE, 2024, 44(9): 5303-5312.

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http://www.zghjkx.com.cn/CN/ 或 http://www.zghjkx.com.cn/CN/Y2024/V44/I9/5303

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