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产甲烷条件下岩溶湿地沉积物中古菌群落的变化规律
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国家自然科学基金(41662019);广西自然科学基金(2016JJB130197)


Characterization of archaeal communities in a karst wetland under methanogenic conditions
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    摘要:

    【背景】桂林会仙湿地位于喀斯特峰丛洼地和峰丛平原的过渡区,主要由岩溶地下河补给,水质呈现富钙偏碱特征,是一处典型的岩溶湿地环境,湿地沉积物以厌氧环境为主,独特的环境特征使之成为研究新型厌氧微生物的理想场所。氢型产甲烷菌和乙酸型产甲烷菌是环境中有机质厌氧降解的重要参与者,但目前会仙湿地产甲烷菌生理生态功能的研究十分有限。【目的】揭示乙酸盐营养型产甲烷条件和氢气营养型产甲烷条件下岩溶湿地环境古菌群落结构的变化规律和产甲烷菌的主要类群,探讨岩溶环境古菌的功能和相互关系。【方法】以会仙岩溶湿地沉积物为研究材料,分别以乙酸盐和氢气为唯一能源物进行富集培养,结合未添加任何能源底物的对照处理,动态监测产甲烷通量,分别构建了3个共包含146条古菌16S rRNA基因全长序列的文库,以及3个共包含138条甲基辅酶M还原酶基因mcrA基因序列的文库,通过构建系统发育树比较分析不同产甲烷底物培养条件下典型岩溶湿地古菌群落的变化规律。【结果】从乙酸型和氢型产甲烷条件培养物顶空气中都检测到了几乎等浓度的甲烷产生,甲烷八叠球菌是mcrA文库中主要的古菌类群,其中包含了Methanosarcina属古菌、Zoige Cluster I (ZC-I)和ANME-2d Cluster (AD)类群古菌,以及两个相对独立且不包含任何参考序列的分支KT-I和KT-II,可能代表产甲烷菌的新类群;经过两种产甲烷条件的富集培养后,ZC-I、AD和KT-II类群古菌的序列数占比有较为显著的增加(45%–155%);深古菌(Bathyarchaeota)是所有古菌16S rRNA基因文库的优势类群,序列占比为88%–100%,其中MCG-11亚群最为丰富,占所有深古菌的84%,并且在乙酸盐产甲烷条件下增加了17%。【结论】会仙湿地沉积物中蕴涵着丰富的新型古菌序列,沉积物中主要的氢型产甲烷菌和乙酸型产甲烷菌都来自甲烷八叠球菌目,深古菌在岩溶环境和乙酸型产甲烷条件下可能都发挥着重要的作用。

    Abstract:

    [Background] The Huixian wetland, located in the transition zone between the karst peak-cluster plain and karst depression area of Guilin city, is a typical karst wetland with a calcium-rich, alkalescent underground water supply. The sediment of the wetland is mostly free of oxygen, making it an ideal place to study novel species of anaerobic microbes. Hydrogenotrophic and acetoclastic methanogens play significant roles in the degradation of organic matter in the anaerobic environment; however, the physiology and ecology of microbial methanogenesis within the Huixian karst wetland is poorly understood. [Objective] To reveal the variation of archaeal communities, the functional groups of methanogens present, and the potential interactions among archaea from the sediment of the Huixian wetland under hydrogenotrophic and acetoclastic methanogenic conditions. [Methods] Sediment samples were incubated in anaerobic basal medium supplemented with hydrogen or acetate as the sole energy source. During incubation, the methane concentration in the headspace of the incubator was monitored. After 21 days of incubation, 3 libraries of nearly complete archaeal 16S rRNA gene sequences including 146 sequences and 3 mcrA (methyl-coenzyme reductase gene) libraries including 136 sequences from different samples were constructed. The dynamics of the archaeal communities in response to different conditions were studied through phylogenetic analysis. [Results] Methane was detected in the headspace of both H2- and acetate-enriched samples. The mcrA libraries were dominated by Methanosarcinales archaea consisting of four major phylogenetic branches: Zoige cluster I (ZC-I), anaerobic methanotrophic archaea of ANME-2d (AD), and the KT-I and KT-II subgroups. The branches of KT-I and KT-II were comparatively independent in the phylogenetic tree, with no previously reported sequences included, and these branches might represent novel subgroups of Methanosarcinales archaea. The composition of ZC-I, AD and KT-II increased by 45%–155% after incubation under methanogenetic conditions. Bathyarchaeota archaea accounted for 88%–100% of the 16S rRNA gene libraries. Most of the Bathyarchaeota came from the MCG-11 subgroup, which increased by 17% after incubation under acetic methanogenic conditions. [Conclusion] The sediment of the Huixian wetland contains novel archaeal sequences. The dominant hydrogenotrophic and acetoclastic methanogens within the wetland sediment are archaea of the Methanosarcinales order. Bathyarchaeota may play important roles in both in situ karst environments and methanogenic conditions.

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吴麒,陈颖,邱凯瑞,罗倩倩. 产甲烷条件下岩溶湿地沉积物中古菌群落的变化规律[J]. 微生物学通报, 2019, 46(12): 3193-3204

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  • 在线发布日期: 2019-11-26
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