Cultivating previously uncultured soil bacteria using a soil substrate membrane system

Belinda C. Ferrari*, Tristrom Winsley, Michael Gillings, Svend Binnerup

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    64 Citations (Scopus)

    Abstract

    Most bacteria are recalcitrant to traditional cultivation in the laboratory. The soil substrate membrane system provides a simulated environment for the cultivation of previously undescribed soil bacteria as microcolonies. The system uses a polycarbonate membrane as a solid support for growth and soil extract as the substrate. Diverse microcolonies can be visualized using total bacterial staining combined with fluorescence in situ hybridization (FISH) after 7-10-d incubation. Molecular typing shows that the majority of microcolony-forming bacteria recovered using this protocol were resistant to growth using standard methods. The protocol takes <4 h of bench time over the 10-d period.

    Original languageEnglish
    Pages (from-to)1261-1269
    Number of pages9
    JournalNature Protocols
    Volume3
    Issue number8
    DOIs
    Publication statusPublished - 2008

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