Discovery of 5-hydroxymethyl-2-furancarboxylic acid as a potential agent for controlling Verticillium dahliae and its action mechanism research

Abstract Background An endophytic fungal strain ( Penicillium simplicissimum CEF-818) with excellent biocontrol potential was isolated from cotton plants in our previous study, but its mechanism of suppressing cotton Verticillium wilt (caused by Verticillium dahliae ) remains unclear. This study aimed to clarify the biocontrol efficacy of CEF-818 and the underlying mechanism. Results CEF-818 showed strong inhibitory activity against V. dahliae. A bioactive compound was isolated and identified as 5-hydroxymethyl-2-furancarboxylic acid (HFA) via spectroscopic and chromatographic analyses. In vitro biological activity assays indicated that HFA inhibited V. dahliae mycelial growth with a median effective concentration (EC₅₀) of 25.29 µg·mL⁻ 1 . Microscopic observations revealed that HFA disrupted the fungal cell membrane integrity, damaged intracellular organelles (e.g., mitochondria and nuclei), and further inhibited spore germination and microsclerotia formation—two critical processes in V. dahliae' s disease cycle. Transcriptome sequencing and bioinformatics analysis showed that HFA targets membrane-intrinsic components (e.g., membrane transporters and lipid biosynthesis-related proteins) and perturbs key metabolic pathways (e.g., ribosomal biogenesis and valine metabolism), thereby impairing fungal cell viability and pathogenicity. Conclusion This study confirms that HFA effectively inhibits the growth and pathogenicity of V. dahliae. Mechanistically, HFA disrupts the structural integrity of V. dahliae 's cell membrane and induces reactive oxygen species (ROS) accumulation, leading to severe cellular damage and fungal death. These findings provide a scientific basis for developing HFA as a novel bioactive agrochemical fungicide for the sustainable control of cotton Verticillium wilt.