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Applied and Environmental Microbiology, May 2009, p. 3352-3354, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02407-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cultivation of Fastidious Bacteria by Viability Staining and Micromanipulation in a Soil Substrate Membrane System{triangledown}

B. C. Ferrari1* and M. R. Gillings2

School of Biotechnology and Biomolecular Sciences, University of New South Wales, Randwick, New South Wales 2052, Australia,1 Department of Biological Sciences, Macquarie University, North Ryde, New South Wales 2019, Australia2

Received 21 October 2008/ Accepted 10 March 2009

Soil substrate membrane systems allow for microcultivation of fastidious soil bacteria as mixed microbial communities. We isolated established microcolonies from these membranes by using fluorescence viability staining and micromanipulation. This approach facilitated the recovery of diverse, novel isolates, including the recalcitrant bacterium Leifsonia xyli, a plant pathogen that has never been isolated outside the host.

* Corresponding author. Mailing address: School of Biotechnology and Biomolecular Sciences, University of New South Wales, Randwick, NSW 2052, Australia. Phone: 612 9385 2032. Fax: 612 9385 1438. E-mail: b.ferrari{at}unsw.edu.au

{triangledown} Published ahead of print on 20 March 2009.

Applied and Environmental Microbiology, May 2009, p. 3352-3354, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02407-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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