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[北方湿地中甲烷营养γ-变形菌的丰度和多样性]

[Abundance and diversity of methanotrophic Gammaproteobacteria in northern wetlands].

作者信息

Danilova O V, Dedysh S N

出版信息

Mikrobiologiia. 2014 Mar-Apr;83(2):204-14.

Abstract

Numeric abundance, identity and pH preferences of methanotrophic Gammaproteobacteria (type I methanotrophs) inhabiting the northern acidic wetlands were studied. The rates of methane oxidation by peat samples from six-wetlands of European Northern Russia (pH 3.9-4.7) varied from 0.04 to 0.60 μg CH4 g(-1) peat h(-1). The number of cells revealed by hybridization with fluorochrome-labeled probes M84 + M705 specific for type I methanotrophs was 0.05-2.16 x 10(5) cells g(-1) dry peat, i.e. 0.4-12.5% of the total number of methanotrophs and 0.004-0.39% of the total number of bacteria. Analysis of the fragments of the pmoA gene encoding particulate methane monooxygenase revealed predominance of the genus Methylocystis (92% of the clones) in the studied sample of acidic peat, while the proportion of the pmoA sequences of type I methanotrophs was insignificant (8%). PCR amplification of the 16S rRNA gene fragments of type I methanotrophs with TypeIF-Type IR primers had low specificity, since only three sequences out of 53 analyzed belonged to methanotrophs and exhibited 93-99% similarity to those of Methylovulum, Methylomonas, and Methylobacter species. Isolates of type I methanotrophs obtained from peat (strains SH10 and 83A5) were identified as members of the species Methylomonaspaludis and Methylovulum miyakonense, respectively. Only Methylomonaspaludum SH10 was capable of growth in acidic media (pH range for growth 3.8-7.2 with the optimum at pH 5.8-6.2), while Methylovulum miyakonense 83A5 exhibited the typical growth characteristics of neutrophilic methanotrophs (pH range for growth 5.5-8.0 with the optimum at pH 6.5-7.5).

摘要

对栖息于俄罗斯北部酸性湿地的甲烷氧化γ-变形菌(I型甲烷营养菌)的数量丰度、种类及pH偏好进行了研究。来自俄罗斯欧洲部分北部六个湿地(pH 3.9 - 4.7)的泥炭样品的甲烷氧化速率在0.04至0.60 μg CH4 g(-1)泥炭 h(-1)之间。用针对I型甲烷营养菌的荧光标记探针M84 + M705杂交显示的细胞数量为0.05 - 2.16×10(5)个细胞 g(-1)干泥炭,即占甲烷营养菌总数的0.4 - 12.5%,占细菌总数的0.004 - 0.39%。对编码颗粒性甲烷单加氧酶的pmoA基因片段的分析表明,在所研究的酸性泥炭样品中,甲基孢囊菌属占主导(92%的克隆),而I型甲烷营养菌的pmoA序列比例微不足道(8%)。用TypeIF-Type IR引物对I型甲烷营养菌的16S rRNA基因片段进行PCR扩增特异性较低,因为在分析的53个序列中只有3个属于甲烷营养菌,与甲基卵菌属、甲基单胞菌属和甲基杆菌属的序列相似度为93 - 99%。从泥炭中获得的I型甲烷营养菌分离株(菌株SH10和83A5)分别被鉴定为沼泽甲基单胞菌和宫古甲基卵菌的成员。只有沼泽甲基单胞菌SH10能够在酸性培养基中生长(生长的pH范围为3.8 - 7.2,最适pH为5.8 - 6.2),而宫古甲基卵菌83A5表现出嗜中性甲烷营养菌的典型生长特征(生长的pH范围为5.5 - 8.0,最适pH为6.5 - 7.5)。

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