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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Novel Cultivation-Based Approach To Understanding the Miscellaneous Crenarchaeotic Group (MCG) Archaea from Sedimentary Ecosystems
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Novel Cultivation-Based Approach To Understanding the Miscellaneous Crenarchaeotic Group (MCG) Archaea from Sedimentary Ecosystems

机译：基于耕种的新方法用于从沉积生态系统中了解杂类古生物群（MCG）古细菌

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The uncultured miscellaneous crenarchaeotic group (MCG) archaea comprise one of the most abundant microbial groups in the Earth's subsurface environment. However, very little information is available regarding the lifestyle, physiology, and factors controlling the distribution of members of this group. We established a novel method using both cultivation and molecular techniques, including a pre-PCR propidium monoazide treatment, to investigate viable members of the MCG in vitro. Enrichment cultures prepared from estuarine sediment were provided with one of a variety of carbon substrates or cultivation conditions and incubated for 3 weeks. Compared with the samples from time zero, there was an order-of-magnitude increase in the number of MCG 16S rRNA genes in almost all cultures, indicating that MCG archaea are amenable to in vitro cultivation. None of the tested substrates or conditions significantly stimulated growth of MCG archaea more than the basal medium alone; however, glycerol (0.02%) had a significantly inhibitory effect (P < 0.05). Diversity analysis of populations resulting from four culture treatments (basal medium, addition of amino acids, H2-CO2 as the gas phase, or initial aerobic conditions) revealed that the majority of viable MCG archaea were affiliated with the MCG-8 and MCG-4 clusters. There were no significant differences in MCG diversity between these treatments, also indicating that some members of MCG-4 and MCG-8 are tolerant of initially oxic conditions. The methods outlined here will be useful for further investigation of MCG archaea and comparison of substrates and cultivation conditions that influence their growth in vitro.
机译：未培养的杂类颅骨古菌群（MCG）古细菌是地球地下环境中最丰富的微生物群之一。但是，关于生活方式，生理学和控制该组成员分布的因素的信息很少。我们建立了一种使用培养和分子技术（包括PCR前单叠氮化丙二胺处理）的新方法，以研究体外MCG的可行成员。向从河口沉积物制备​​的富集培养物提供多种碳底物或培养条件之一，并孵育3周。与从零时开始的样本相比，几乎所有培养物中MCG 16S rRNA基因的数量都有数量级的增加，这表明MCG古细菌适合体外培养。测试的底物或条件中没有一个比单独的基础培养基更能刺激MCG古细菌的生长。然而，甘油（0.02％）具有明显的抑制作用（P <0.05）。四种培养方法（基础培养基，添加氨基酸，H2-CO2作为气相或初始有氧条件）导致的种群多样性分析表明，大多数可行的MCG古细菌与MCG-8和MCG-4相关集群。这些处理之间的MCG多样性没有显着差异，也表明MCG-4和MCG-8的某些成员对最初的高氧条件耐受。此处概述的方法将有助于进一步研究MCG古细菌以及比较影响其体外生长的底物和培养条件。

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