Cultivating the uncultured

  1. Karsten Zengler*,
  2. Gerardo Toledo*,
  3. Michael Rappé,
  4. James Elkins*,
  5. Eric J. Mathur*,
  6. Jay M. Short*, and
  7. Martin Keller*,
  1. *Diversa Corporation, San Diego, CA 92121; and Department of Microbiology, Oregon State University, Corvallis, OR 97331
  1. Communicated by John Abelson, California Institute of Technology, Pasadena, CA (received for review September 10, 2002)

Abstract

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.

Footnotes

  • To whom correspondence should be addressed. E-mail: mkeller{at}diversa.com.

  • The sequences reported in this paper have been deposited in the GenBank database (accession nos. ).

  • Abbreviations:
    1. GMD, gel microdroplet

    2. SSW, Sargasso Sea water

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