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

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.

SUBMITTER: Houghton KM

PROVIDER: S-EPMC7470311 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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

Houghton Karen M KM Stewart Lucy C LC

Access microbiology 20191202 3

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 effe ...[more]

PMID: 32974564

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