[Characterization of the Structure of the Prokaryotic Complex of Antarctic Permafrost by Molecular Genetic Techniques].

Mikrobiologiia Pub Date : 2016-01-01
N A Manucharova, E V Trosheva, E M Kol'tsova, E V Demkina, E V Karaevskaya, E M Rivkina, A V Mardanov, G I El'-Registan
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Abstract

A prokaryotic mesophilic organotrophic community responsible for 10% of the total microbial number determined by epifluorescence microscopy was reactivated in the samples ofAntarctic permafrost retrieved from the environment favoring long-term preservation of microbial communities (7500 years). No culturable forms were obtained without resuscitation procedures (CFU = 0). Proteobacteria, Actinobacteria, and Firmicutes were the dominant microbial groups in the complex. Initiation of the reactivated microbial complex by addition of chitin (0.1% wt/vol) resulted in an increased share of metabolically active biomass (up to 50%) due to the functional domination of chitinolytics caused by the target resource. Thus, sequential application of resuscitation procedures and initiation of a specific physiological group (in this case, chitinolytics) to a permafrost-preserved microbial community made it possible to reveal a prokaryotic complex capable of reversion of metabolic activity (FISH data), to determine its phylogenetic structure by metagenomic anal-ysis, and to isolate a pure culture of the dominant microorganism with high chitinolytic activity.

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[用分子遗传学技术表征南极冻土原核复合体的结构]。
从有利于微生物群落长期保存(7500年)的环境中提取的南极永久冻土层样品中,一个原核嗜中温有机营养群落被重新激活,占总微生物数量的10%。未经复苏程序未获得可培养形式(CFU = 0)。变形菌门,放线菌门和厚壁菌门是复合体中的优势微生物群。通过添加几丁质(0.1% wt/vol)启动再活化的微生物复合物,由于目标资源引起的几丁质降解的功能优势,导致代谢活性生物量的份额增加(高达50%)。因此,复苏程序的顺序应用和特定生理群(在这种情况下,几丁质分解)对永久冻土保存的微生物群落的启动,使得有可能揭示能够逆转代谢活性的原核复合物(FISH数据),通过元基因组分析确定其系统发育结构,并分离出具有高几丁质分解活性的优势微生物的纯培养物。
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