LitR and its quorum-sensing regulators modulate biofilm formation by <i>Vibrio fischeri</i>

Quorum sensing controls numerous processes ranging from the production of virulence factors to biofilm formation. Biofilms, communities of bacteria that are attached to one another and/or a surface, are common in nature, and when they form, they can produce a quorum of bacteria. One model system to study biofilms is the bacterium <i>Vibrio fischeri</i>, which forms a biofilm that promotes the colonization of its symbiotic host. Many factors promote <i>V. fischeri</i> biofilm formation <i>in vitro</i>, including the symbiosis polysaccharide (SYP) and cellulose, but the role of quorum sensing is currently understudied. Recently, a quorum-sensing-dependent transcription factor, LitR, was shown to negatively influence <i>V. fischeri</i> biofilm formation in the context of a biofilm-overproducing strain. To better understand the importance of LitR, we identified conditions in which the impact of LitR on biofilm formation could be observed in an otherwise wild-type strain and then investigated its role and the roles of upstream quorum regulators in biofilm phenotypes. In static conditions, LitR and its upstream quorum regulators, including autoinducer synthases LuxS and AinS, contributed to control over biofilms that were both SYP and cellulose dependent. In shaking liquid conditions, LitR and AinS contributed to control over biofilms that were primarily cellulose dependent. LitR modestly inhibited cellulose transcription in a manner that depended on the transcription factor VpsR. These findings expand our understanding of LitR and the quorum-sensing pathway in the physiology of <i>V. fischeri</i> and illuminate negative control mechanisms that prevent robust biofilm formation by wild-type <i>V. fischeri</i> under laboratory conditions.IMPORTANCEQuorum sensing is a key regulatory mechanism that controls diverse phenotypes in numerous bacteria, including <i>Vibrio fischeri</i>. In many microbes, quorum sensing has been shown to control biofilm formation, yet in <i>V. fischeri</i>, the link between quorum sensing and biofilm formation has been understudied. This study fills that knowledge gap by identifying roles for the quorum sensing-controlled transcription factor, LitR, and its upstream quorum-sensing regulators, including the autoinducer synthases AinS and LuxS, in inhibiting biofilm formation under specific conditions. It also determined that LitR inhibits the transcription of genes required for cellulose biosynthesis. This work thus expands our understanding of the complex control over biofilm regulation.