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Related Experiment Video

Updated: Feb 5, 2026

Design and Use of a Low Cost, Automated Morbidostat for Adaptive Evolution of Bacteria Under Antibiotic Drug Selection
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Temporal dynamics of bacteria-plasmid coevolution under antibiotic selection.

Michael J Bottery1, A Jamie Wood2,3, Michael A Brockhurst4

  • 1Department of Biology, University of York, Wentworth Way, York, YO10 5DD, UK. michael.bottery@york.ac.uk.

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Bacteria evolved a repeatable genetic pathway to reduce the cost of antibiotic resistance. This involved chromosomal mutations before altering the plasmid-borne resistance gene, demonstrating constrained bacterial evolution.

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Area of Science:

  • Microbiology
  • Evolutionary Biology
  • Genetics

Background:

  • Horizontally acquired genes, like antibiotic resistance genes on plasmids, can impose a metabolic cost on bacteria.
  • Previous research indicated that Escherichia coli with the tetracycline-resistance plasmid RK2 evolved reduced tetracycline resistance cost through mutations on both the chromosome and plasmid when exposed to tetracycline.

Purpose of the Study:

  • To investigate the temporal dynamics and repeatability of intragenomic coevolution between bacteria and plasmids under selection pressure.
  • To understand the specific order of adaptive mutations that reduce the cost of horizontally acquired antibiotic resistance.

Main Methods:

  • Utilizing whole-genome sequencing to analyze adaptive mutations in three independently evolving populations of Escherichia coli.
  • Applying tetracycline selection pressure to drive the evolutionary process.

Main Results:

  • The order of adaptive mutations was highly repeatable across all three populations.
  • The initial adaptive mutation consistently occurred in the chromosomal gene ompF, leading to a shortened lag phase and increased tetracycline resistance.
  • Subsequent mutations affected the plasmid-encoded tetracycline efflux pump, followed by additional chromosomal resistance mutations.

Conclusions:

  • Reducing the metabolic cost of horizontally acquired tetracycline resistance is contingent upon the prior evolution of chromosomally encoded resistance.
  • The trajectory of bacteria-plasmid coevolution in this system is constrained, following a single, repeatable adaptive path.