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Applied and Environmental Microbiology, December 2003, p. 7210-7215, Vol. 69, No. 12
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.12.7210-7215.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Laboratory Cultivation of Widespread and Previously Uncultured Soil Bacteria

Shayne J. Joseph,1 Philip Hugenholtz,2 Parveen Sangwan,1 Catherine A. Osborne,1 and Peter H. Janssen1*

Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia,1 Department of Environmental Science, Policy and Management, Division of Ecosystem Sciences, University of California Berkeley, Berkeley, California 94720-31102

Received 17 June 2003/ Accepted 16 September 2003

Most soil bacteria belong to family-level phylogenetic groups with few or no known cultivated representatives. We cultured a collection of 350 isolates from soil by using simple solid media in petri dishes. These isolates were assigned to 60 family-level groupings in nine bacterial phyla on the basis of a comparative analysis of their 16S rRNA genes. Ninety-three (27%) of the isolates belonged to 20 as-yet-unnamed family-level groupings, many from poorly studied bacterial classes and phyla. They included members of subdivisions 1, 2, 3, and 4 of the phylum Acidobacteria, subdivision 3 of the phylum Verrucomicrobia, subdivision 1 of the phylum Gemmatimonadetes, and subclasses Acidimicrobidae and Rubrobacteridae of the phylum Actinobacteria. In addition, members of 10 new family-level groupings of subclass Actinobacteridae of the phylum Actinobacteria and classes Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria of the phylum Proteobacteria were obtained. The high degree of phylogenetic novelty and the number of isolates affiliated with so-called unculturable groups show that simple cultivation methods can still be developed further to obtain laboratory cultures of many phylogenetically novel soil bacteria.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia. Phone: 61 (3) 8344-5706. Fax: 61 (3) 9347-1540. E-mail: pjanssen{at}unimelb.edu.au.

Applied and Environmental Microbiology, December 2003, p. 7210-7215, Vol. 69, No. 12
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.12.7210-7215.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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