Extreme parallel evolution of flagellar motility in Pseudomonas fluorescens is facilitated by silent mutations

Research output: Contribution to conferenceAbstractpeer-review


There is a growing need for accurate evolutionary forecasting of microbes, but we must first understand how possible evolutionary paths can be constrained by silent genetic features. Here we show that synonymous sequence variation determines extreme parallel evolution during the evolutionary rescue of flagellar motility. An immotile variant of the soil microbe, Pseudomonas fluorescens, swiftly recovers flagellum-dependent motility through parallel de novo mutation. This typically manifests within 96 h under strong selection through repeatable mutation within the nitrogen pathway’s histidine kinase gene, ntrB. We found that evolution was parallel to nucleotide resolution in over 95% of cases in minimal medium (M9), with lineages repeatedly fixing an identical mutation (ntrB A289C). This repeatable de novo mutation was robust to nutrient condition despite evidence for antagonistic pleiotropy across nutrient regimes. Competition assays against alternative motile alleles revealed some evidence for selection enforcing repeated fixation of ntrB mutants, but there was no evidence for clonal interference driving parallel evolution to nucleotide resolution. Instead, the introduction of 6 synonymous substitutions surrounding the mutational hotspot reduced parallel evolution from >95% to 0% at the site. In a reciprocal experiment, we introduced 6 synonymous substitutions into a homologous strain that did not ancestrally evolve in parallel and observed that parallel evolution at the site rose from 0% to 80%. We argue that these silent mutations facilitate extremely localised mutation rate heterogeneity. Our results reveal that unique quirks in DNA structure at specific loci can strongly bias evolutionary outcomes.
Original languageEnglish
Publication statusPublished - 27 Apr 2021
EventMicrobiology Society Annual Conference 2021 - Online, UK United Kingdom
Duration: 26 Apr 202130 Apr 2021


ConferenceMicrobiology Society Annual Conference 2021
Country/TerritoryUK United Kingdom


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