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Microcolony cultivation on a soil substrate membrane system selects for previously uncultured soil bacteria

机译:在土壤基质膜系统上的小菌落栽培选择了以前未培养的土壤细菌

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摘要

Traditional microbiological methods of cultivation recover less than 1% of the total bacterial species, and the culturable portion of bacteria is not representative of the total phylogenetic diversity. Classical cultivation strategies are now known to supply excessive nutrients to a system and therefore select for fast-growing bacteria that are capable of colony or biofilm formation. New approaches to the cultivation of bacteria which rely on growth in dilute nutrient media or simulated environments are beginning to address this problem of selection. Here we describe a novel microcultivation method for soil bacteria that mimics natural conditions. Our soil slurry membrane system combines a polycarbonate membrane as a growth support and soil extract as the substrate. The result is abundant growth of uncharacterized bacteria as microcolonies. By combining microcultivation with fluorescent in situ hybridization, previously "unculturable" organisms belonging to cultivated and noncultivated divisions, including candidate division TM7, can be identified by fluorescence microscopy. Successful growth of soil bacteria as microcolonies confirmed that the missing culturable majority may have a growth strategy that is not observed when traditional cultivation indicators are used.
机译:传统的微生物培养方法只能回收不到总细菌种类的1%,并且细菌的可培养部分不能代表总的系统发育多样性。现在已知经典的栽培策略会向系统提供过多的营养,因此会选择能够集落或形​​成生物膜的快速生长的细菌。依靠稀营养培养基或模拟环境中的细菌生长的新型细菌培养方法正开始解决这一选择问题。在这里,我们描述了一种模拟自然条件的土壤细菌的新型微培养方法。我们的土壤浆膜系统结合了聚碳酸酯膜作为生长支持物和土壤提取物作为基质。结果是未定殖的细菌作为微菌落大量繁殖。通过将微培养与荧光原位杂交相结合,可以通过荧光显微镜鉴定属于已培养和未培养区(包括候选区TM7)的先前“不可培养的”生物。土壤细菌作为小菌落的成功生长证实,缺少的可培养多数菌株可能具有使用传统栽培指标时未观察到的生长策略。

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