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Journal of Bacteriology, January 2007, p. 119-130, Vol. 189, No. 1
0021-9193/07/$08.00+0     doi:10.1128/JB.00930-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Phenotypic Diversification and Adaptation of Serratia marcescens MG1 Biofilm-Derived Morphotypes{triangledown}

Kai Shyang Koh,1,2 Kin Wai Lam,1,2 Morten Alhede,3 Shu Yeong Queck,1,2,{dagger} Maurizio Labbate,1,2,{ddagger} Staffan Kjelleberg,1,2* and Scott A. Rice1,2

The Centre for Marine Biofouling and Bio-Innovation, The University of New South Wales, Sydney, NSW 2052, Australia,1 The School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, NSW 2052, Australia,2 Center for Biomedical Microbiology, BioCentrum-DTU, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark3

Received 28 June 2006/ Accepted 2 October 2006

We report here the characterization of dispersal variants from microcolony-type biofilms of Serratia marcescens MG1. Biofilm formation proceeds through a reproducible process of attachment, aggregation, microcolony development, hollow colony formation, and dispersal. From the time when hollow colonies were observed in flow cell biofilms after 3 to 4 days, at least six different morphological colony variants were consistently isolated from the biofilm effluent. The timing and pattern of variant formation were found to follow a predictable sequence, where some variants, such as a smooth variant with a sticky colony texture (SSV), could be consistently isolated at the time when mature hollow colonies were observed, whereas a variant that produced copious amounts of capsular polysaccharide (SUMV) was always isolated at late stages of biofilm development and coincided with cell death and biofilm dispersal or sloughing. The morphological variants differed extensively from the wild type in attachment, biofilm formation, and cell ultrastructure properties. For example, SSV formed two- to threefold more biofilm biomass than the wild type in batch biofilm assays, despite having a similar growth rate and attachment capacity. Interestingly, the SUMV, and no other variants, was readily isolated from an established SSV biofilm, indicating that the SUMV is a second-generation genetic variant derived from SSV. Planktonic cultures showed significantly lower frequencies of variant formation than the biofilms (5.05 x 10–8 versus 4.83 x 10–6, respectively), suggesting that there is strong, diversifying selection occurring within biofilms and that biofilm dispersal involves phenotypic radiation with divergent phenotypes.

* Corresponding author. Mailing address: The School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, NSW 2052, Australia. Phone: 61-2-9385-2102. Fax: 61-2-9385-1779. E-mail: S.Kjelleberg{at}unsw.edu.au.

{triangledown} Published ahead of print on 27 October 2006.

{dagger} Present address: Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840.

{ddagger} Present address: Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.

Journal of Bacteriology, January 2007, p. 119-130, Vol. 189, No. 1
0021-9193/07/$08.00+0     doi:10.1128/JB.00930-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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