Characterization of the archaeal community in a minerotrophic fen and terminal restriction fragment length polymorphism-directed isolation of a novel hydrogenotrophic methanogen
Characterization of the archaeal community in a minerotrophic fen and terminal restriction fragment length polymorphism-directed isolation of a novel hydrogenotrophic methanogen
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DOI：
10.1128/aem.02222-07
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发表时间：
2008-04-01
影响因子：
4.4
通讯作者：
Zinder, Stephen H.
中科院分区：
文献类型：
--
作者：
Cadillo-Quiroz, Hinsby;Yashiro, Erica;Zinder, Stephen H.
营养化沼泽泥炭地广泛分布于北纬地区，由于其有机质周转迅速，单位面积大气甲烷的潜在来源比沼泽泥炭地要大。然而，对沼泽中古细菌群落组成的研究很少，特别是在微营养点。几个基于16S rrna的引物集被用来获得纽约州中部一个矿化沼泽中古细菌群落的广泛特征。该遗址中发现了广泛的古细菌多样性：11个euryarchaeal群和2个crenarchaeal群，其中大部分是未培养的。所有文库和末端限制性片段长度多态性（T-RFLP）分析结果显示，Methanomicrobiales目新聚类El和Methanosaetaceae为共优势类群。考虑到El群的丰度和潜在的氢营养化甲烷贡献，我们将其作为低离子强度培养基（PM1）培养的目标。最初的尝试产生了甲烷螺旋菌为主的培养物。然而，通过结合T-RFLP分析作为处理和重复的快速选择工具，我们能够选择以El为主的富集。进一步稀释至10(-9)，并使用T-RFLP进行追踪，得到了命名为E1-9c的菌株。E1-9c是一种新型球虫，嗜氢、中温、微嗜酸产甲烷菌，对Na2S浓度高度敏感
Minerotrophic fen peatlands are widely distributed in northern latitudes and, because of their rapid turnover of organic matter, are potentially larger sources of atmospheric methane than bog peatlands per unit area. However, studies of the archaeal community composition in fens are scarce particularly in minerotrophic sites. Several 16S rRNA-based primer sets were used to obtain a broad characterization of the archaeal community in a minerotrophic fen in central New York State. A wide archaeal diversity was observed in the site: 11 euryarchaeal and 2 crenarchaeal groups, most of which were uncultured. The El group, a novel cluster in the order Methanomicrobiales, and Methanosaetaceae were the codominant groups in all libraries and results of terminal restriction fragment length polymorphism (T-RFLP) analysis. Given its abundance and potential hydrogenotrophic methane contribution, the El group was targeted for culture attempts with a low-ionic-strength medium (PM1). Initial attempts yielded Methanospirillum-dominated cultures. However, by incorporating a T-RFLP analysis as a quick selection tool for treatments and replicates, we were able to select an enrichment dominated by El. Further dilutions to 10(-9) and tracking with T-RFLP yielded a strain named E1-9c. E1-9c is a novel coccoid hydrogenotrophic, mesophilic, slightly acidophilic methanogen and is highly sensitive to Na2S concentrations (requires