Abstract

BackgroundBeneficial mutations play an essential role in bacterial adaptation, yet little is known about their fitness effects across genetic backgrounds and environments. One prominent example of bacterial adaptation is antibiotic resistance. Until recently, the paradigm has been that antibiotic resistance is selected by the presence of antibiotics because resistant mutations confer fitness costs in antibiotic free environments. In this study we show that it is not always the case, documenting the selection and fixation of resistant mutations in populations of Escherichia coli B that had never been exposed to antibiotics but instead evolved for 2000 generations at high temperature (42.2°C).ResultsWe found parallel mutations within the rpoB gene encoding the beta subunit of RNA polymerase. These amino acid substitutions conferred different levels of rifampicin resistance. The resistant mutations typically appeared, and were fixed, early in the evolution experiment. We confirmed the high advantage of these mutations at 42.2°C in glucose-limited medium. However, the rpoB mutations had different fitness effects across three genetic backgrounds and six environments.ConclusionsWe describe resistance mutations that are not necessarily costly in the absence of antibiotics or compensatory mutations but are highly beneficial at high temperature and low glucose. Their fitness effects depend on the environment and the genetic background, providing glimpses into the prevalence of epistasis and pleiotropy.

Highlights

  • Beneficial mutations play an essential role in bacterial adaptation, yet little is known about their fitness effects across genetic backgrounds and environments

  • At the end of the experiment, we identified rifampicin resistant clones. Surprised to find their emergence and prevalence in the absence of antibiotics, we have explored the evolutionary context of their appearance, along with the fitness effect of single mutants under different environmental conditions and genetic backgrounds

  • The high temperature adapted lines were founded from a common ancestral strain of E. coli B, which was descended from the strain REL606 after 2000 generations at 37°C in Davis minimal medium supplemented with 25 μg/ml glucose (DM25)

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Summary

Introduction

Beneficial mutations play an essential role in bacterial adaptation, yet little is known about their fitness effects across genetic backgrounds and environments. The paradigm has been that antibiotic resistance is selected by the presence of antibiotics because resistant mutations confer fitness costs in antibiotic free environments. Genetic resistance to antibiotics can result either from sequential accumulation of multiple beneficial mutations - e.g. resistance to fluoroquinolones [15] – or from a single amino acid substitution - e.g. resistance to rifamycins [16]. This last mutational type is typically highly advantageous in the presence of antibiotics, leading to rapid fixation, often within hundreds of generations [17]. We did not detect a significant background-by-mutation interaction, the low P-value (0.053) suggests that the three mutations have differential effects across genetic backgrounds

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