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采用 DNA-SIP 技术揭示了两个具有鲜明对比的北美泥炭地生态系统中的活性甲烷营养菌。

Active methanotrophs in two contrasting North American peatland ecosystems revealed using DNA-SIP.

机构信息

Department of Geography, University of Toronto Mississauga, Mississauga, ON L5L 1C6, Canada.

出版信息

Microb Ecol. 2012 Feb;63(2):438-45. doi: 10.1007/s00248-011-9902-z. Epub 2011 Jul 5.

Abstract

The active methanotroph community was investigated in two contrasting North American peatlands, a nutrient-rich sedge fen and nutrient-poor Sphagnum bog using in vitro incubations and (13)C-DNA stable-isotope probing (SIP) to measure methane (CH(4)) oxidation rates and label active microbes followed by fingerprinting and sequencing of bacterial and archaeal 16S rDNA and methane monooxygenase (pmoA and mmoX) genes. Rates of CH(4) oxidation were slightly, but significantly, faster in the bog and methanotrophs belonged to the class Alphaproteobacteria and were similar to other methanotrophs of the genera Methylocystis, Methylosinus, and Methylocapsa or Methylocella detected in, or isolated from, European bogs. The fen had a greater phylogenetic diversity of organisms that had assimilated (13)C, including methanotrophs from both the Alpha- and Gammaproteobacteria classes and other potentially non-methanotrophic organisms that were similar to bacteria detected in a UK and Finnish fen. Based on similarities between bacteria in our sites and those in Europe, including Russia, we conclude that site physicochemical characteristics rather than biogeography controlled the phylogenetic diversity of active methanotrophs and that differences in phylogenetic diversity between the bog and fen did not relate to measured CH(4) oxidation rates. A single crenarchaeon in the bog site appeared to be assimilating (13)C in 16S rDNA; however, its phylogenetic similarity to other CO(2)-utilizing archaea probably indicates that this organism is not directly involved in CH(4) oxidation in peat.

摘要

我们采用体外培养和(13)C-DNA 稳定同位素探针(SIP)技术,研究了北美两种截然不同的泥炭地(富营养化的莎草湿地和贫营养化的泥炭藓沼泽)中的活性甲烷营养菌群落。该技术可以测量甲烷(CH(4))氧化速率,并标记活性微生物,随后对细菌和古菌 16S rDNA 和甲烷单加氧酶(pmoA 和 mmoX)基因进行指纹图谱分析和测序。沼泽中的 CH(4)氧化速率略高,但具有显著意义,甲烷营养菌属于 α-变形菌纲,与在欧洲沼泽中检测到的或从欧洲沼泽中分离到的甲基单胞菌属、甲基球菌属、甲基杆菌属或甲基囊泡菌属的其他甲烷营养菌相似。沼泽中具有同化(13)C 的生物的系统发育多样性更大,包括来自 α-和 γ-变形菌纲的甲烷营养菌,以及其他与在英国和芬兰的沼泽中检测到的细菌相似的潜在非甲烷营养菌。根据我们研究点与包括俄罗斯在内的欧洲地区细菌之间的相似性,我们得出结论,场地理化特性而非生物地理学控制了活性甲烷营养菌的系统发育多样性,沼泽和湿地之间的系统发育多样性差异与测量的 CH(4)氧化速率无关。沼泽点中的单个泉古菌似乎在 16S rDNA 中同化(13)C;然而,它与其他 CO2 利用古菌的系统发育相似性可能表明该生物不直接参与泥炭中的 CH(4)氧化。

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