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Experimental evolution of the plant pathogen  Ralstonia solanacearum , where bacteria were maintained on plant lineages for more than 300 generations, revealed that several independent single mutations in the  efpR  gene from populations propagated on beans were associated with fitness gain on bean. In the present work, novel allelic  efpR  variants were isolated from populations propagated on other plant species, thus suggesting that mutations in  efpR  were not solely associated to a fitness gain on bean, but also on additional hosts. A transcriptomic profiling and phenotypic characterization of the  efpR  deleted mutant showed that EfpR acts as a global catabolic repressor, directly or indirectly down-regulating the expression of multiple metabolic pathways. EfpR also controls virulence traits such as exopolysaccharide production, swimming and twitching motilities and deletion of  efpR  leads to reduced virulence on tomato plants after soil drenching inoculation. We studied the impact of the single mutations that occurred in  efpR  during experimental evolution and found that these allelic mutants displayed phenotypic characteristics similar to the deletion mutant, although not behaving as complete loss-of-function mutants. These adaptive mutations therefore strongly affected the function of  efpR , leading to an expanded metabolic versatility that should benefit to the evolved clones. Altogether, these results indicated that EfpR is a novel central player of the  R .  solanacearum  virulence regulatory network. Independent mutations therefore appeared during experimental evolution in the evolved clones, on a crucial node of this network, to favor adaptation to host vascular tissues through regulatory and metabolic rewiring.

plos pathogens

Enhanced in planta Fitness through Adaptive Mutations in EfpR, a Dual Regulator of Virulence and Metabolic Functions in the Plant Pathogen Ralstonia solanacearum