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Applied and Environmental Microbiology, November 2004, p. 6686-6694, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6686-6694.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Morphotypic Conversion in Listeria monocytogenes Biofilm Formation: Biological Significance of Rough Colony Isolates

Ian R. Monk,1,{dagger} Gregory M. Cook,1 Brian C. Monk,2 and Philip J. Bremer3*

Department of Microbiology,1 Department of Oral Sciences,2 Department of Food Science, University of Otago, Dunedin, New Zealand3

Received 18 April 2004/ Accepted 4 July 2004

Adherence to a stainless steel surface selected isolates of Listeria monocytogenes with enhanced surface colonization abilities and a change in phenotype from the common smooth colony morphology to a succession of rough colony morphotypes. Growth in broth culture of the best-adapted, surface-colonizing rough colony morphotype gave a smooth colony revertant. Comparative analysis revealed that the smooth and rough variants had similar phenotypic and biochemical characteristics (e.g., identical growth rates and tolerances to antibiotics and environmental stressors). Rough colony isolates, however, failed to coordinate motility or induce autolysis. The defect in autolysis of rough colony isolates, which involved impaired cellular localization of several peptidoglycan-degrading enzymes, including cell wall hydrolase A (CwhA), suggested a link to a secretory pathway defect. The genetic basis for the impairment was studied at the level of the accessory secretory pathway component SecA2. DNA sequencing of the secA2 gene in smooth and rough colony isolates found no mutations in the coding or promoter regions. Analysis of SecA2 expression with an integrated secA2-FLAG tag construct found the protein to be upregulated in the rough and revertant backgrounds compared to the parental smooth colony isolate. A compensatory mechanism involving the SecA2 secretion pathway components is postulated to control smooth to rough interconversion of L. monocytogenes. Such phenotypic variation may enhance the ability of this opportunistic pathogen to colonize environments as diverse as processing surfaces, food products, and animal hosts.

* Corresponding author. Mailing address: Department of Food Science, University of Otago, P.O. Box 56, Dunedin, New Zealand. Phone: 64-3-4795469. Fax: 64-3-4797567. E-mail: phil.bremer{at}stonebow.otago.ac.nz.

{dagger} Present address: Alimentary Pharmabiotic Centre, Department of Microbiology, University College Cork, Cork, Ireland.

Applied and Environmental Microbiology, November 2004, p. 6686-6694, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6686-6694.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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