A possible mechanism of farnesol tolerance in biofilms implemented by activating the PKC signalling pathway and stabilizing ROS levels No Access

Abstract

Biofilms are the natural growth state for most microorganisms. biofilms are composed of multiple cell types (round budding yeast-form cells, oval pseudohyphal cells, and elongated hyphal cells) encased in an extracellular matrix. biofilms are notorious for resistance to antimicrobial treatments, a property that might be determined by complex mechanisms. Exogenous farnesol exerts a certain antifungal activity against with medical implications. Different from other microbes, biofilms can tolerate exogenous farnesol at high concentration with some cells still surviving and even maintaining proliferation, but the mechanism is unclear.

The study hypothesizes that resists farnesol by activating the PKC signalling pathway.

The study aims to discuss the molecular mechanism of resistance to farnesol.

The ROS levels, the genes and proteins of the PKC pathway were compared between the farnesol-tolerant and non-tolerant groups using ROS levels assay, -RT PCR and Western blot, respectively. Further, the mutant strains (Δ/Δ and Δ/Δ) were constructed, then the survival rates and ROS levels of biofilms exposed to farnesol were compared between mutant and wild strains. The morphological changes were observed using TEM.

The survival rates of biofilms decreased rapidly under the lower concentration of farnesol (<0.05), and kept stable (>0.05) as the concentration rose up to 200 µM. The gene expression of the PKC pathway increased, while ROS levels remained stable and even decreased in the farnesol-tolerant biofilms, compared with those in the farnesol-nontolerant biofilms after farnesol treatment (<0.05); and were significantly upregulated by during the development of biofilm tolerance to farnesol. The cell wall and cytoplasm of Δ/Δ and Δ/Δ were damaged, and the ROS level increased (<0.05); meanwhile, the survival rate of biofilms decreased compared with that of wild-type strain under the same farnesol concentrations (<0.05). ROS inhibitors reversed these changes in Δ/Δ and Δ/Δ when the mutant strains exposed to farnesol.

biofilms can tolerate high concentrations of farnesol by activating and of the PKC pathway and stabilizing ROS levels. The and are two key genes regulated by in the process of biofilm tolerance to farnesol.

Funding
This study was supported by the:
  • The Foundation of the Priority Academic Program Development of Jiangsu Higher Education Institutions (Award PAPD, 2018-87)
    • Principle Award Recipient: XinWei
  • National Natural Sciences Foundation of China (Award 81970945)
    • Principle Award Recipient: XinWei
  • National Natural Sciences Foundation of China (Award 81371156)
    • Principle Award Recipient: XinWei
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/content/journal/jmm/10.1099/jmm.0.001476
2022-01-12
2024-03-29
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