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Despite the importance of outer membrane vesicles (OMVs) in benthic animal settlement, the underlying molecular mechanism remains elusive. Here, the impact of OMVs and OMVs synthesis-related  tolB  gene in  Mytilus coruscus  plantigrade settlement was tested. The OMVs were extracted from  Pseudoalteromonas marina  through density gradient centrifugation, and a  tolB  knockout strain, achieved by homologous recombination, was utilized for the investigation. Our results demonstrated that OMVs could significantly enhance  M. coruscus  plantigrades settlement. Deleting the  tolB  resulted in downregulation of c-di-GMP, accompanied by a reduction of OMV production, a decline in bacterial motility and increasing biofilm-forming ability. Enzyme treatment resulted in a 61.11% reduction in OMV-inducing activity and a 94.87% reduction in LPS content. Thus, OMVs regulate mussel settlement  via  LPS, and c-di-GMP is responsible for the OMV-inducing capacity. These findings provide new insights into the interactions between bacteria and mussels.

Outer membrane vesicles induce the mussel plantigrade settlement via regulation of c-di-GMP