Abstract
Shared enzymatic targets between azoles used in agriculture as fungicides and clinical antifungals are hypothesized to have led to the drift of resistant fungal isolates from field to clinic, reducing efficacy of azoles in infectious disease treatment. Recent discovery of a novel herbicide, tetflupyrolimet, and a potential clinical antifungal, olorofim, both targeting dihydroorotate dehydrogenase (DHODH), raises concern that widespread use of tetflupyrolimet may similarly cause clinical resistance to olorofim in Aspergillus isolates migrating from agricultural settings. Using minimum inhibitory concentration analysis on Aspergillus subspecies, enzymatic assays of recombinant DHODH, and a series of DHODH knockout yeast strains complemented with various DHODH orthologs, we demonstrate that tetflupyrolimet is highly selective for plant DHODH over Aspergillus DHODH. Based on the inability of tetflupyrolimet to inhibit Aspergillus DHODH or hinder growth of various Aspergillus species, the data demonstrate that risk of olorofim resistance developing in Aspergillus due to agrochemical use of the herbicide tetflupyrolimet is minimal.
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All data generated or analyzed during this study are included in this published article and its Supplementary Information files.
References
Brown, G. D. et al. The pathology of human fungal infections. Nat. Rev. Microbiol. 22, 687–704 (2024).
Jain, A., Sarsaiya, S., Wu, Q., Lu, Y. & Shi, J. A review of plant leaf fungal diseases and its environment speciation. Bioengineered 10, 409–421 (2019).
Boyer, J., Feys, S., Zsifkovits, I., Hoenigl, M. & Egger, M. Treatment of invasive Aspergillosis: How it’s going, where it’s heading. Mycopathologia 188, 667–681 (2023).
Verweij, P. E. et al. Dual use of antifungals in medicine and agriculture: How do we help prevent resistance developing in human pathogens? Drug Resist. Updat. 65, 100885 (2022).
Fisher, M. C. et al. Tackling the emerging threat of antifungal resistance to human health. Nat. Rev. Microbiol. 20, 557–571 (2022).
Earle, K. et al. Pathogenicity and virulence of Aspergillus fumigatus. Virulence 14, 2172264 (2023).
Hui, S. T., Gifford, H. & Rhodes, J. Emerging antifungal resistance in fungal pathogens. Curr. Clin. Microbiol. Rep. 11, 43–50 (2024).
Snelders, E. et al. Triazole fungicides can induce cross-resistance to medical triazoles in Aspergillus fumigatus. PLoS One. 7, e31801 (2012).
Kang, S. E. et al. Evidence for the agricultural origin of resistance to multiple antimicrobials in Aspergillus fumigatus, a fungal pathogen of human. G3 12, jkab427 (2022).
Oliver, J. D. et al. F901318 represents a novel class of antifungal drug that inhibits dihydroorotate dehydrogenase. Proc. Natl. Acad. Sci. U S A. 113, 12809–12814 (2016).
Selby, T. P. et al. Bioisosteric tactics in the discovery of tetflupyrolimet: a new mode-of-action herbicide. J. Agric. Food Chem. 71, 18197–18204 (2023).
Kang, I. H., Emptage, R. P., Kim, S. I. & Gutteridge, S. A novel mechanism of herbicide action through disruption of pyrimidine biosynthesis. Proc. Natl. Acad. Sci. U S A. 120, e2313197120 (2023).
van Rhijn, N. et al. Aspergillus fumigatus strains that evolve resistance to the agrochemical fungicide ipflufenoquin in vitro are also resistant to olorofim. Nat. Microbiol. 9, 29–34 (2024).
Pinder, C., Lebedinec, R., Levine, T. P., Birch, M. & Oliver, J. D. Characterisation of putative class 1A DHODH-like proteins from Mucorales and dematiaceous mould species. PLoS One. 18, e0289441 (2023).
Reis, R. A. G., Calil, F. A., Feliciano, P. R., Pinheiro, M. P. & Nonato, M. C. The dihydroorotate dehydrogenases: Past and present. Arch. Biochem. Biophys. 632, 175–191 (2017).
Higashimura, N., Hamada, A. & Banba, S. Novel fungicide quinofumelin shows selectivity for fungal dihydroorotate dehydrogenase over the corresponding human enzyme. J. Pestic Sci. 48, 17–21 (2023).
Banba, S. Exploring quinone-binding sites as targets for pesticides. Pest Manag Sci. 81, 6062–6075 (2025).
CLSI. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi. 3rd ed. CLSI standard M38. Wayne, PA: Clinical and Laboratory Standards Institute (2017).
Geitz, R. D. & Schiestl, R. H. High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat. Protoc. 2, 31–34 (2007).
Acknowledgements
Yeast strains complemented with plant DHODH were assembled with assistance of Keelin Reilly.
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SIK, VT, BA, SS, and RPE designed, performed, and analyzed experiments. QL and JAS contributed critical reagents. SIK, VT, BA, and RPE prepared the figures. RPE wrote the manuscript with methods assistance from SIK. All authors reviewed the manuscript.
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JAS, VT, and BA were formerly employed by FMC. All other authors are currently employed by FMC.
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Kim, SI., Turlapati, V., Agashe, B. et al. Tetflupyrolimet shows selectivity for plant dihydroorotate dehydrogenase over Aspergillus orthologs. Sci Rep (2026). https://doi.org/10.1038/s41598-026-43966-y
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DOI: https://doi.org/10.1038/s41598-026-43966-y
