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Volume 2, Issue 3

Temperature-gradient incubation isolates multiple competitive species from a single environmental sample Open Access

Abstract

High-throughput sequencing has allowed culture-independent investigation into a wide variety of microbiomes, but sequencing studies still require axenic culture experiments to determine ecological roles, confirm functional predictions and identify useful compounds and pathways. We have developed a new method for culturing and isolating multiple microbial species with overlapping ecological niches from a single environmental sample, using temperature-gradient incubation. This method was more effective than standard serial dilution-to-extinction at isolating methanotrophic bacteria. It also highlighted discrepancies between culture-dependent and -independent techniques; 16S rRNA gene amplicon sequencing of the same sample did not accurately reflect cultivatable strains using this method. We propose that temperature-gradient incubation could be used to separate out and study previously ‘unculturable’ strains, which co-exist in both natural and artificial environments.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): 16S rRNA , Culture , Environmental sample , Isolation , Methanotroph and Temperature gradient
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2019-12-02
2024-05-11
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Temperature-gradient incubation isolates multiple competitive species from a single environmental sample
Access Microbiology 2, e000081 (2020); https://doi.org/10.1099/acmi.0.000081
/content/journal/acmi/10.1099/acmi.0.000081
/content/journal/acmi/10.1099/acmi.0.000081
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