Native plasmid-encoded mercury resistance genes are functional and demonstrate natural transformation in environmental bacterial isolates

Plasmid-mediated horizontal gene transfer(HGT) is a major driver of genetic diversity in bacteria. We experimentally validated the function of a putative mercury resistance operon present on an abundant 8 kilobase pair native plasmidfound in groundwater samples without detectable levels of mercury. Phylogenetic analyses of the plasmid-encoded mercury reductases from the studied groundwater site show them to be distinct from those reported in proximal metal-contaminated sites. We synthesized the entire native plasmidand demonstrated that the plasmidwas sufficient to confer functional mercury resistance in Escherichia coli. Given the possibility that natural transformationis a prevalent HGT mechanism in the low cell density environments of groundwaters, we also assayed bacterial strains from this environment for competence. We used the native plasmid-encoded metal resistance to design a screen and identified 17 strains positive for natural transformation. We selected 2 of the positive strains along with a model bacterium to fully confirm HGT via natural transformation. From an ecological perspective, the role of the native plasmidpopulation in providing advantageous traits combined with the microbiome9s capacity to take up environmental DNAenables rapid adaptation to environmental stresses.