# Membrane perturbation by the last-resort antibiotic polymyxin B drives biphasic regulation of horizontal gene transfer

## TL;DR

The antibiotic polymyxin B can both inhibit and promote the spread of antibiotic resistance genes depending on its concentration.

## Contribution

This study reveals a concentration-dependent biphasic effect of polymyxin B on horizontal gene transfer.

## Key findings

- Subinhibitory concentrations of polymyxin B inhibit plasmid conjugation by up to 65.4%.
- Bactericidal concentrations of polymyxin B promote plasmid conjugation up to 15.9-fold.
- High concentrations of polymyxin B increase cellular energy and oxidative stress, facilitating gene transfer.

## Abstract

Although it is increasingly recognized that anthropogenic chemicals modulate horizontal gene transfer (HGT), the nature of these interactions is often more complex than a simple promotion or inhibition. The potential for a single chemical to exert opposing, concentration-dependent effects represents a critical and less-explored frontier in microbial ecology. Here, we investigate the last-resort antibiotic polymyxin B, a membrane-targeting peptide, and reveal a concentration-dependent, biphasic regulation of plasmid conjugation. Subinhibitory concentrations (0.125–0.5 mg/l) consistently inhibited the transfer of antibiotic resistance genes (ARGs) by up to 65.4%, whereas bactericidal concentrations (≥ 1 mg/l) strongly promoted it by up to 15.9-fold. This regulatory switch is driven by distinct physiological states: low-level exposure triggers defensive responses including reduced membrane permeability, whereas high-level exposure causes catastrophic membrane damage, inducing a synergistic stress response involving oxidative damage (> two-fold ROS increase) and a surge in cellular energy (up to 83.0% ATP increase) that facilitates HGT. High-concentration polymyxin B also promotes plasmid transfer in complex microbial communities derived from activated-sludge biofilms. Our findings reveal a new paradigm for the interaction between chemical stressors and microbial evolution, demonstrating that the ecological impact of contaminants on HGT cannot be predicted by monotonic models and highlighting the role of environmental hotspots in shaping the dissemination of antibiotic resistome.