Ecological filtering and phylogeographic structuring of Psychrilyobacter within two closely related limpet species from the Southern Ocean

The ecological interdependence between macroorganisms and their microbial communities promotes stable associations over time, potentially leading to their evolutionary co-diversification. The detection of intricate eco-evolutionary interactions between animals and their microbiota is challenging, primarily due to complex bacterial communities related to poorly resolved host population structure. Strikingly, co-diversification in invertebrates, characterized by generally less complex microbiota, remains largely unexplored. Here, we compared the bacterial communities associated with two distinct lineages of Nacella limpets, a dominant shallow water patellogastropod of the Southern Ocean shores with a well-described population structure. Our goals were to elucidate the uniqueness of Nacella microbiota, resulting from an ecological filter that selectively favors certain bacterial taxa. Additionally, we aimed to depict the genetic structure of bacterial symbiont seeking evidence of co-diversification with Nacella. We sequence the V4-V5 regions of the bacterial 16S rRNA gene in three distinct microenvironments associated with Nacella: rock substrate, radula, and whole intestine. These samples were collected from two populations of Nacella deaurata and Nacella concinna, located in the West Antarctic Peninsula and Falkland/Malvinas Islands, respectively. We assessed ecological filtering patterns in the limpet microbiota, uncovering unique bacterial communities in both radulas and intestines, with specifically enriched bacterial taxa compared to the surrounding environment. By examining microdiversity patterns of core bacterial taxa, we revealed a deep phylogeographic structure of Psychrilyobacter in Nacella intestines. We highlight the Southern Ocean limpets of the Nacella genus as a novel and promising model for studying co-diversification between marine mollusks and their resident microbiota.