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Journal of Bacteriology, January 2008, p. 662-671, Vol. 190, No. 2
0021-9193/08/$08.00+0     doi:10.1128/JB.01473-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Influence of Quorum Sensing and Iron on Twitching Motility and Biofilm Formation in Pseudomonas aeruginosa{triangledown}

Glenn M. Patriquin,1 Ehud Banin,2 Christie Gilmour,1 Rivka Tuchman,2 E. Peter Greenberg,3 and Keith Poole1*

Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada K7L 3N6,1 The Mina and Everard Goodman Faculty of Life Sciences, Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat-Gan 52900, Israel,2 Department of Microbiology, University of Washington School of Medicine, Seattle, Washington 98195-72423

Received 12 September 2007/ Accepted 2 November 2007

Reducing iron (Fe) levels in a defined minimal medium reduced the growth yields of planktonic and biofilm Pseudomonas aeruginosa, though biofilm biomass was affected to the greatest extent and at FeCl3 concentrations where planktonic cell growth was not compromised. Highlighting this apparently greater need for Fe, biofilm growth yields were markedly reduced in a mutant unable to produce pyoverdine (and, so, deficient in pyoverdine-mediated Fe acquisition) at concentrations of FeCl3 that did not adversely affect biofilm yields of a pyoverdine-producing wild-type strain. Concomitant with the reduced biofilm yields at low Fe concentrations, P. aeruginosa showed enhanced twitching motility in Fe-deficient versus Fe-replete minimal media. A mutant deficient in low-Fe-stimulated twitching motility but normal as regards twitching motility on Fe-rich medium was isolated and shown to be disrupted in rhlI, whose product is responsible for synthesis of the N-butanoyl homoserine lactone (C4-HSL) quorum-sensing signal. In contrast to wild-type cells, which formed thin, flat, undeveloped biofilms in Fe-limited medium, the rhlI mutant formed substantially developed though not fully mature biofilms under Fe limitation. C4-HSL production increased markedly in Fe-limited versus Fe-rich P. aeruginosa cultures, and cell-free low-Fe culture supernatants restored the twitching motility of the rhlI mutant on Fe-limited minimal medium and stimulated the twitching motility of rhlI and wild-type P. aeruginosa on Fe-rich minimal medium. Still, addition of exogenous C4-HSL did not stimulate the twitching motility of either strain on Fe-replete medium, indicating that some Fe-regulated and RhlI/C4-HSL-dependent extracellular product(s) was responsible for the enhanced twitching motility (and reduced biofilm formation) seen in response to Fe limitation.

* Corresponding author. Mailing address: Queen's University, Department of Microbiology and Immunology, Rm. 737, Botterell Hall, Kingston, Ontario, Canada K7L 3N6. Phone: (613) 533-6677. Fax: (613) 533-6796. E-mail: poolek{at}queensu.ca

{triangledown} Published ahead of print on 9 November 2007.

Journal of Bacteriology, January 2008, p. 662-671, Vol. 190, No. 2
0021-9193/08/$08.00+0     doi:10.1128/JB.01473-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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