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首页> 外文期刊>Applied Microbiology >Cultivation-Independent Detection of Autotrophic Hydrogen-Oxidizing Bacteria by DNA Stable-Isotope Probing
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Cultivation-Independent Detection of Autotrophic Hydrogen-Oxidizing Bacteria by DNA Stable-Isotope Probing

机译：DNA稳定同位素探测独立于培养的自养氢氧化细菌的检测

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Knallgas bacteria are a physiologically defined group that is primarily studied using cultivation-dependent techniques. Given that current cultivation techniques fail to grow most bacteria, cultivation-independent techniques that selectively detect and identify knallgas bacteria will improve our ability to study their diversity and distribution. We used stable-isotope probing (SIP) to identify knallgas bacteria in rhizosphere soil of legumes and in a microbial mat from Obsidian Pool in Yellowstone National Park. When samples were incubated in the dark, incorporation of ~(13)CO_(2) was H_(2) dependent. SIP enabled the detection of knallgas bacteria that were not detected by cultivation, and the majority of bacteria identified in the rhizosphere soils were betaproteobacteria predominantly related to genera previously known to oxidize hydrogen. Bacteria in soil grew on hydrogen at concentrations as low as 100 ppm. A hydB homolog encoding a putative high-affinity NiFe hydrogenase was amplified from ~(13)C-labeled DNA from both vetch and clover rhizosphere soil. The results indicate that knallgas bacteria can be detected by SIP and populations that respond to different H_(2) concentrations can be distinguished. The methods described here should be applicable to a variety of ecosystems and will enable the discovery of additional knallgas bacteria that are resistant to cultivation.
机译：纳氏细菌是一个生理学上明确的群体，主要使用依赖于培养的技术进行研究。鉴于当前的培养技术无法使大多数细菌繁殖，因此有选择地检测和识别克纳尔氏菌的独立于培养的技术将提高我们研究其多样性和分布的能力。我们使用稳定同位素探测（SIP）来识别豆类根际土壤和黄石国家公园黑曜石池微生物垫中的纳氏气细菌。在黑暗中孵育样品时，〜（13）CO_（2）的掺入依赖于H_（2）。 SIP能够检测出通过耕种无法检测到的克纳尔氏菌，而且在根际土壤中鉴定出的大多数细菌都是β变形杆菌，主要与以前已知能氧化氢的属有关。土壤中的细菌在低至100 ppm的氢浓度下生长。紫v和三叶草根际土壤中〜（13）C标记的DNA扩增了编码推定的高亲和力NiFe氢化酶的hydB同源物。结果表明，可以通过SIP检测到knallgas细菌，并且可以区分对不同H_（2）浓度有反应的种群。此处描述的方法应适用于各种生态系统，并将能够发现对栽培有抵抗力的其他纳氏气菌。

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