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Volume 151, Issue 9

The SBW25 wrinkly spreader biofilm requires attachment factor, cellulose fibre and LPS interactions to maintain strength and integrity Free

Abstract

The wrinkly spreader (WS) isolate of SBW25 forms a substantial biofilm at the air–liquid interface. The biofilm is composed of an extracellular partially acetylated cellulose-fibre matrix, and previous mutagenesis of WS with mini-Tn had identified both the regulatory and cellulose-biosynthetic operons. One uncharacterized WS mutant, WS-5, still expressed cellulose but produced very weak biofilms. In this work, the mini-Tn insertion site in WS-5 has been identified as being immediately upstream of the operon. Like Tol-Pal mutants of other Gram-negative bacteria, WS-5 showed a ‘leaky-membrane’ phenotype, including the serendipitous ability to utilize sucrose, increased uptake of the hydrophilic dye propidium iodide, and the loss of lipopolysaccharide (LPS) expression. WS-5 cells were altered in relative hydrophobicity, and showed poorer recruitment and maintenance in the biofilm than WS. The WS-5 biofilm was also less sensitive to chemical interference during development. However, growth rate, cellulose expression and attachment were not significantly different between WS and WS-5. Finally, WS-5 biofilms could be partially complemented with WS-4, a biofilm- and attachment-deficient mutant that expressed LPS, resulting in a mixed biofilm with significantly increased strength. These findings show that a major component of the WS air–liquid biofilm strength results from the interactions between LPS and the cellulose matrix of the biofilm – and that in the WS biofilm, cellulose fibres, attachment factor and LPS are required for biofilm development, strength and integrity.

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Keyword(s): A–L, air–liquid , CR, Congo red , DOC, deoxycholic acid , EPS, exopolysaccharide , Hr, relative hydrophobicity , MDM, maximum deformation mass and PI, propidium iodide
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2005-09-01
2024-04-16
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The Pseudomonas fluorescens SBW25 wrinkly spreader biofilm requires attachment factor, cellulose fibre and LPS interactions to maintain strength and integrity
Microbiology 151, 2829 (2005); https://doi.org/10.1099/mic.0.27984-0
/content/journal/micro/10.1099/mic.0.27984-0
/content/journal/micro/10.1099/mic.0.27984-0
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