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Journal of Bacteriology, June 2004, p. 3970-3979, Vol. 186, No. 12
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.12.3970-3979.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Genes Involved in Formation of Structured Multicellular Communities by Bacillus subtilis

Steven S. Branda,1 José Eduardo González-Pastor,2,{dagger} Etienne Dervyn,3 S. Dusko Ehrlich,3 Richard Losick,2 and Roberto Kolter1*

Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115,1 Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138,2 Génétique Microbienne, Institut National de la Recherche Agronomique, 78352 Jouy en Josas, France3

Received 17 October 2003/ Accepted 9 February 2004

The spore-forming bacterium Bacillus subtilis is capable of assembling multicellular communities (biofilms) that display a high degree of spatiotemporal organization. Wild strains that have not undergone domestication in the laboratory produce particularly robust biofilms with complex architectural features, such as fruiting-body-like aerial projections whose tips serve as preferential sites for sporulation. To discover genes involved in this multicellular behavior and to do so on a genome-wide basis, we took advantage of a large collection of mutants which have disruptions of most of the uncharacterized genes in the B. subtilis genome. This collection, which was generated with a laboratory strain, was screened for mutants that were impaired in biofilm formation. This subset of mutated genes was then introduced into the wild strain NCIB 3610 to study their effects on biofilm formation in liquid and solid media. In this way we identified six genes that are involved in the development of multicellular communities. These are yhxB (encoding a putative phosphohexomutase that may mediate exopolysaccharide synthesis), sipW (encoding a signal peptidase), ecsB (encoding an ABC transporter subunit), yqeK (encoding a putative phosphatase), ylbF (encoding a regulatory protein), and ymcA (a gene of unknown function). Further analysis revealed that these six genes play different roles in B. subtilis community development.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115. Phone (617) 432-1776. Fax: (617) 738-7664. E-mail: rkolter{at}hms.harvard.edu.

{dagger} Present address: Centro de Astrobiología (CSIC-INTA), Instituto Nacional de Técnica Aeroespacial, 28850 Torrejón de Ardoz, Madrid, Spain.

Journal of Bacteriology, June 2004, p. 3970-3979, Vol. 186, No. 12
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.12.3970-3979.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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