Zengler Karsten, Toledo Gerardo, Rappe Michael, Elkins James, Mathur Eric J, Short Jay M, Keller Martin
Diversa Corporation, San Diego, CA 92121, USA.
Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15681-6. doi: 10.1073/pnas.252630999. Epub 2002 Nov 18.
The recent application of molecular phylogeny to environmental samples has resulted in the discovery of an abundance of unique and previously unrecognized microorganisms. The vast majority of this microbial diversity has proved refractory to cultivation. Here, we describe a universal method that provides access to this immense reservoir of untapped microbial diversity. This technique combines encapsulation of cells in gel microdroplets for massively parallel microbial cultivation under low nutrient flux conditions, followed by flow cytometry to detect microdroplets containing microcolonies. The ability to grow and study previously uncultured organisms in pure culture will enhance our understanding of microbial physiology and metabolic adaptation and will provide new sources of microbial metabolites. We show that this technology can be applied to samples from several different environments, including seawater and soil.
分子系统发育学在环境样本中的最新应用，已促使大量独特且此前未被识别的微生物被发现。事实证明，绝大多数这种微生物多样性难以培养。在此，我们描述了一种通用方法，可用于获取这一巨大的未开发微生物多样性库。该技术将细胞包裹于凝胶微滴中，以便在低营养通量条件下进行大规模平行微生物培养，随后通过流式细胞术检测含有微菌落的微滴。在纯培养中培养和研究此前未培养的生物体的能力，将增进我们对微生物生理学和代谢适应性的理解，并将提供微生物代谢产物的新来源。我们表明，这项技术可应用于来自多种不同环境的样本，包括海水和土壤。
