Preferential cultivation of type II methanotrophic bacteria from littoral sediments (Lake Constance)
Preferential cultivation of type II methanotrophic bacteria from littoral sediments (Lake Constance)
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DOI：
10.1016/s0168-6496(03)00260-5
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发表时间：
2004-02-15
影响因子：
4.2
通讯作者：
Schink, B
中科院分区：
文献类型：
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作者：
Bussmann, I;Pester, M;Schink, B
用于培养来自各种环境的甲烷氧化菌的最广泛使用的培养基是Whittenbury在1970年提出的。为了适应和优化淡水沉积物中甲烷氧化菌的培养培养基，采用不同浓度的基质、磷酸盐、硝酸盐和其他无机盐对甲烷氧化菌进行了最大可能数计数。高浓度（> 1 mM）的镁和硫酸盐以及高浓度的硝酸盐（> 500 μ M）显著减少了培养的甲烷氧化菌的数量，而15-1500 μ M范围内的磷酸盐没有影响。氧气和二氧化碳对培养效率也有影响，最佳混合比例为17%O-2和3%CO2;甲烷混合比例（6- 32%）对培养效率没有影响。从沉积物中提取的DNA通过PCR扩增的pmoA基因的克隆文库揭示了I型和II型甲烷氧化菌的存在。尽管如此，甲烷氧化菌的培养，也用改进的培养基，显然有利于甲基弯菌属/甲基孢囊菌属组的II型甲烷氧化菌的生长。虽然改良培养基可以培养出更多的甲烷氧化菌，但它们的多样性并不能反映出分子生物学方法检测到的沉积物样品中甲烷氧化菌的多样性。(C)2003年，欧洲微生物学会联合会。Elsevier B. V.出版，保留所有权利。
Most widely used medium for cultivation of methanotrophic bacteria from various environments is that proposed in 1970 by Whittenbury. In order to adapt and optimize medium for culturing of methanotrophs from freshwater sediment, media with varying concentrations of substrates, phosphate, nitrate, and other mineral salts were used to enumerate methanotrophs by the most probable number method. High concentrations ( > 1 mM) of magnesium and sulfate, and high concentrations of nitrate (> 500 muM) significantly reduced the number of cultured methanotrophs, whereas phosphate in the range of 15-1500 muM had no influence. Also oxygen and carbon dioxide influenced the culturing efficiency, with an optimal mixing ratio of 17% O-2 and 3% CO2; the mixing ratio of methane (6- 32%) had no effect. A clone library of pmoA genes amplified by PCR from DNA extracted from sediment revealed the presence of both type I and type II methanotrophs. Nonetheless, the cultivation of methanotrophs, also with the improved medium, clearly favored growth of type II methanotrophs of the Methylosinus/Methylocystis group. Although significantly more methanotrophs could be cultured with the modified medium, their diversity did not mirror the diversity of methanotrophs in the sediment sample detected by molecular biology method. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.