Cultivating previously uncultured soil bacteria using a soil substrate membrane system.
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.
Ferrari
Belinda C
BC
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia. b.ferrari@unsw.edu.au
Winsley
Tristrom
T
Gillings
Michael
M
Binnerup
Svend
S
D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques / Q000295:instrumentation; D003470:Culture Media; D004269:DNA, Bacterial / Q000737:chemistry; D017404:In Situ Hybridization, Fluorescence; D012988:Soil Microbiology
D003470:Culture Media; D004269:DNA, Bacterial