Bacterial c-di-GMP signaling gene affects mussel larval metamorphosis through outer membrane vesicles and lipopolysaccharides.

Biofilms serve as crucial cues for settlement and metamorphosis in marine invertebrates. Within bacterial systems, c-di-GMP functions as a pivotal signaling molecule regulating both biofilm formation and dispersion. However, the molecular mechanism of how c-di-GMP modulates biofilm-induced larval metamorphosis remains elusive. Our study reveals that the deletion of a c-di-GMP related gene in Pseudoalteromonas marina led to an increase in the level of bacterial c-di-GMP by knockout technique, and the mutant strain had an enhanced ability to produce more outer membrane vesicles (OMVs) and lipopolysaccharides (LPS). The mutant biofilms had higher induction activity for larval metamorphosis in mussels Mytilus coruscus, and OMVs play a major role in the induction activity. We further explored the function of LPS in OMVs. Extracted LPS induced high larval metamorphosis rate, and LPS content were subject to c-di-GMP and LPS-biosynthesis gene. Thus, we postulate that the impact of c-di-GMP on biofilm-induced metamorphosis is mediated through OMVs and LPS.

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

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.

Molecular cloning, tissue expression, and transcriptional regulation of fabp1 and fabp2 in javelin goby (Synechogobius hasta) in response to starvation stress.

Fatty acid binding proteins (FABPs) are intracellular lipid chaperones with low molecular weight, which are widely distributed in a variety of tissues, participating in fatty acid transport, cell proliferation, and angiogenesis. In this study, full-length sequences of two fabp genes (fabp1 and fabp2) from javelin goby (Synechogobius hasta) were cloned via RACE PCR, followed by bioinformatic analyses and gene expression evaluation. The fabp1 and fabp2 cDNA sequences were 493 and 626 bp in length, encoding 126 and 132 amino acids, respectively. Phylogenetic analysis revealed that both genes from S. hasta were clustered with those of other fish species in accordance with their known taxonomic relationships. fabp1 and fabp2 mRNA showed distinct expression patterns in different tissues, with fabp1 being most expressed in the liver and fabp2 in the intestine. Furthermore, the expression of fabp1 in the liver was significantly up-regulated during starvation, whereas fabp2 mRNA level in the intestine initially increased and then decreased, indicating that the transcriptional responses of the two genes could be influenced by malnourishment/starvation. Changes in the transcriptional levels of fabp1 and fabp2 also suggested that glycogen was catabolized in the liver of S. hasta at the beginning of starvation prior to lipid depletion, whereas lipids served as fuel reserves in the intestine during short-term starvation. In conclusion, this study provides fundamental insights into the role of Fabps in S. hasta lipid metabolism.