Hidden Microbial Diversity Of The Sea: Testing A Novel Approach To Cultivating Presently Uncultivable Microbes

该项目的目标是通过开发培养目前无法培养的微生物的方法，获得大部分微生物生物多样性。不可培养的微生物占大多数环境中所有物种的99%，不可培养的微生物之谜最近被美国微生物学会确认为微生物学基础和应用研究的主要挑战。我们提出了一种与传统的从环境样品中培养微生物的方法截然不同的方法。我们建议在微生物的自然栖息地进行培养。我们设计了一种生长室，通过防止细胞迁移的薄膜将其与环境隔开，从而保持内部空间的纯净性，同时使内部空间和外部环境之间能够完全化学接触。我们提出的在自然环境中培养的微生物将暴露在完整的外部环境条件和成分中。我们对自然环境的模拟可能会导致更广泛的有机体的培养，包括那些以前被认为是“不可培养的”。一旦我们在野外培养的生长室中实现了微生物的生长，我们将使用一系列的分子技术来鉴定和验证已培养的菌落中以前不可培养的成分。即使我们的方法只允许我们培养出几个新的分离物，这个项目，通过证明我们的概念，可能会成为解决微生物生态学主要问题之一的一个重要里程碑。

The objective of this project is to gain access to the bulk of microbial biodiversity by developing approaches to cultivate presently uncultivable microorganisms. Uncultivable microorganisms make up 99% of all species from most environments sampled, and the riddle of uncultivable microorganisms has been recently recognized as the main challenge for basic and applied research in microbiology by the American Society for Microbiology.We propose an approach that is radically different from those traditionally employed for culturing microorganisms from environmental samples. We propose to culture the microorganisms in their natural habitat. We have designed a growth chamber that is separated from the environment by membranes that prevent cell migration and thus maintain purity of the inner space and yet enable full chemical contact between the inner space and the outside environment. Microorganisms incubated in the natural environment as we propose will be exposed to the complete suit of the outside environmental conditions and components. Our simulation of the natural environment may result in cultivation of a wider range of organisms, including those that have previously been considered as "uncultivable".Once we achieve microbial growth in the field-incubated growth chamber, we will use an array of molecular techniques to identify and verify the previously uncultivable component among the grown colonies. Even if our approach allows us to grow just a few novel isolates, this project, by the virtue of proving our concept, may become an important milestone toward solving one of the major problems of microbial ecology.