Cultivating previously uncultured soil bacteria using a soil substrate membrane system

Nature Protocols
B C FerrariSvend J Binnerup

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.

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Jun 10, 2011·Applied Microbiology and Biotechnology·Katsuhiko FujiiYuki Yamashita
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