Use of Ichip for High-Throughput In Situ Cultivation of "Uncultivable" Microbial Species
Use of Ichip for High-Throughput In Situ Cultivation of "Uncultivable" Microbial Species
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DOI：
10.1128/aem.01754-09
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发表时间：
2010-04-01
影响因子：
4.4
通讯作者：
Epstein, S. S.
中科院分区：
文献类型：
--
作者：
Nichols, D.;Cahoon, N.;Epstein, S. S.
最古老的悬而未决的微生物学现象之一是为什么多样性的微生物种群中只有一小部分在人工培养基上生长。“无法培养”的大多数微生物可以说代表着地球上最大的未开发的生物和化学新奇物质库。在此之前，我们曾证明，这个池中的物种可以在原位培养的扩散室中生长，这可能是因为扩散为微生物提供了自然产生的生长因子。在这里，我们利用这种方法并开发了一个新的高通量平台，用于从各种环境中并行培养和分离以前未培养的微生物物种。我们设计并测试了一种由数百个微型扩散室组成的隔离芯片(IChip)，每个扩散室都接种了一个环境细胞。我们表明，iChip中的微生物回收率超过了标准培养的许多倍，而且所培育的物种具有显著的系统发育新颖性。这种新方法允许获得大量和多样化的以前无法接触到的微生物，非常适合基础和应用研究。
One of the oldest unresolved microbiological phenomena is why only a small fraction of the diverse microbiological population grows on artificial media. The "uncultivable" microbial majority arguably represents our planet's largest unexplored pool of biological and chemical novelty. Previously we showed that species from this pool could be grown inside diffusion chambers incubated in situ, likely because diffusion provides microorganisms with their naturally occurring growth factors. Here we utilize this approach and develop a novel high-throughput platform for parallel cultivation and isolation of previously uncultivated microbial species from a variety of environments. We have designed and tested an isolation chip (ichip) composed of several hundred miniature diffusion chambers, each inoculated with a single environmental cell. We show that microbial recovery in the ichip exceeds manyfold that afforded by standard cultivation, and the grown species are of significant phylogenetic novelty. The new method allows access to a large and diverse array of previously inaccessible microorganisms and is well suited for both fundamental and applied research.