Rhizobial motility preference in root colonization of Medicago truncatula

O_LITunnel-like infection thread (IT) structures support root colonization by symbiotic nitrogen-fixing rhizobia bacteria in most legume species. These tip-grown structures are key to direct rhizobia from root hairs to developing nodules, where they are hosted to fix nitrogen. Rhizobia likely progress inside ITs by combining growth and motility, by modes not yet defined. Here, we tackled this question by combining mathematical modeling, live cell imaging, and bacterial mutant phenotyping in Medicago truncatula. C_LIO_LIModeling the motion of fluorescently-labeled Sinorhizobium meliloti inside early root hair IT compartments estimated slow movement (2 to 6 {micro}m/h), compatible with passive rather than active motility. Consistent with this model, flagella-less fliF and fliF-fliRdel S. meliloti mutants were impaired in active swimming motility in vitro, yet could colonize host roots and nodules in planta. In contrast, mutation in the rhizobactin 1021 siderophore rhbE biosynthesis gene affected both surface motility in vitro, and host root and nodule colonisation. This mutation also promoted the formation of branched ITs in root hairs, which ultimately resulted in impaired nodule development and infection. C_LIO_LIOur findings support the model estimation and suggest that S. meliloti prioritises flagella-independent surface translocation, partially by secreting rhizobactin 1021 surfactants to reach developing nodules in M. truncatula. C_LI