Decrypting the matrix: how interspecies interactions influence matrix in multispecies biofilm?

The biofilm matrix contributes to the establishment of microbial cells on very diverse surfaces, stabilizing biofilms and providing cells with protection against multiple hostile conditions. Moreover, the biofilm matrix can also retain nutrients, enzymes or quorum sensing molecules, favoring the establishment of social interactions among biofilm cells. Functional bacterial amyloids are part of the biofilm structural components of various species, and they were previously proven to bind QS molecules and strengthen the matrix. Multiple studies have been conducted to characterize matrix determinants and their regulation in single species biofilms, while these remain scarcely understood in multispecies biofilms. We have previously isolated and characterized a soil-derived consortium composed of Xanthomonas retroflexus, Stenotrophomonas rhizophila, Microbacterium oxydans and Paenibacillus amylolyticus showing enhanced biofilm biomass and differential gene/protein expression specific of the four-species biofilm.

This study aimed at exploring the effect of interspecies interactions on biofilm matrix production in the four-species biofilm. We hypothesize that interspecies interactions may result in differential expression of matrix-encoding genes responsible for biofilm emergent properties.

We searched for matrix determinant homologues in X.retroflexus and combined different techniques for characterizing the matrix identity and expression in mono-, dual- and multispecies biofilms.

The fap amyloid operon, described in Pseudomonas as a biofilm-scaffold contributing element, was deleted in X. retroflexus, replaced in the four-species model and compared to the parental community for biofilm structure and adhesion capability. The fap mutant displayed poor substrate colonization in flow cells in both mono- and multispecies biofilms with relative filamentous structure compared to the parental strain/ consortium. However, adhesion did not significantly change under static conditions. To characterize matrix composition, we tested 78 different lectins in multispecies biofilms and identified five that bound to our samples. Interestingly, some matrix glycoconjugates were only produced in the consortium.

Our data suggest that loss of matrix components, such as the Fap amyloid, and the presence of other species, influences synergistic biofilm properties in the four-species consortium. Ongoing approaches involving localized expression of matrix-encoding genes and matrix proteomes will aid in identifying the mechanisms underlying emergent properties in the four-species biofilm.