Evolutionary loss of the rdar morphotype in Salmonella as a result of high mutation rates during laboratory passage
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Rapid evolution of microbes under laboratory conditions can lead to domestication of environmental or clinical strains. In this work, we show that domestication due to laboratory passage in rich medium is extremely rapid. Passaging of wild-type Salmonella in rich medium led to diversification of genotypes contributing to the loss of a spatial phenotype, called the rdar morphotype, within days. Gene expression analysis of the rdar regulatory network demonstrated that mutations were primarily within rpoS, indicating that the selection pressure for scavenging during stationary phase had the secondary effect of impairing this highly conserved phenotype. If stationary phase was omitted from the experiment, radiation of genotypes and loss of the rdar morphotype was also demonstrated, but due to mutations within the cellulose biosynthesis pathway and also in an unknown upstream regulator. Thus regardless of the selection pressure, rapid regulatory changes can be observed on laboratory timescales. The speed of accumulation of rpoS mutations during daily passaging could not be explained by measured fitness and mutation rates. A model of mutation accumulation suggests that to generate the observed accumulation of sigma 38 mutations, this locus must experience a mutation rate of approximately 10(-4) mutations/gene/generation. Sequencing and gene expression of population isolates indicated that there were a wide variety of sigma 38 phenotypes within each population. This suggests that the rpoS locus is highly mutable by an unknown pathway, and that these mutations accumulate rapidly under common laboratory conditions.
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