Bacteria colonize surfaces as heterogeneous structures called biofilms. Intercellular communication using acyl homoserine lactones has been implicated in biofilm formation in some systems. Here, we investigate cell structure in biofilms and associated physiochemical properties of wild type and quorum sensing mutants of Pseudomonas chlororaphis O6 (PcO6), a root-colonizing bacterium. The wild type strain generates multilayered biofilms under conditions where the quorum sensing mutant, deficient in the GacS sensor kinase, does not mature beyond a monolayer structure. However, this gacS mutant rapidly evolves to form a small colony variant (gacS-SCV) that again produces a multilayered biofilm structure although AHSL production is not restored to wild type level. Biofilms formed by the gacS-SCV (114±12°) mutant were the most hydrophobic displaying a higher average ethylene glycol contact angle than those formed by the wild type (28±7°) and gacS (18±6°). Tapping mode atomic force microscopy revealed elongated cell structure in both of the mutant biofilm cells. Digital pulsed force mode adhesion mapping showed that the average adhesion followed the order gacS>gacS-SCV, wild-type. Certain of these gacS mutant cells displayed strong interactions of the AFM tip with cell boundaries, the role of which in biofilm formation is currently under investigation.

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