Unveiling Vacuolar H + -ATPase Subunit a as the Primary Target of the Pyridinylmethyl-Benzamide Fungicide, Fluopicolide
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Abstract
An estimated 240 fungicides are presently in use, but the direct targets for the majority remain elusive, constraining fungicide development and efficient resistance monitoring. In this study, we found that Pcα-actinin knockout did not influence the sensitivity of Phytophthora capsici to fluopicolide, which is a notable oomycete inhibitor. Using a combination of Bulk Segregant Analysis Sequencing and Drug Affinity Responsive Target Stability (DARTS) assays, the vacuolar H + -ATPase subunit a (PcVHA-a) was pinpointed as the target protein of fluopicolide. We also confirmed four distinct point mutations in PcVHA-a responsible for fluopicolide resistance in P. capsici through site-directed mutagenesis. Molecular docking, ATPase activity assays, and a DARTS assay suggested a fluopicolide-PcVHA-a interaction. Sequence analysis and further molecular docking validated the specificity of fluopicolide for oomycetes or fish. These findings support the claim that PcVHA-a is the target of fluopicolide, proposing vacuolar H + -ATPase as a promising target for novel fungicide development.
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Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764 · see citing works
ColabFold - Making protein folding accessible to all
Mirdita, M., Schütze, K., Moriwaki, Y., Heo, L., Ovchinnikov, S., & Steinegger, M. (2021) · doi:10.1101/2021.08.15.456425 · Cited by 9,391 · see citing works
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- Authors
- Dai, T., Yang, J., Zhao, C., Chen, J., Zhang, C., Wang, Z., Peng, Q., Liu, P., Miao, J., & Liu, X.
- Affiliations
- China Agricultural University (CN)
- Northwest A&F University (CN)
- Corresponding author
- Miao, Jianqiang
- Liu, Xili
- Venue
- Journal of Agricultural and Food Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2024-01-09
- Volume
- 72
- Issue
- 3
- Pages
- 1527-1538
- ISSN
- 0021-8561 (ISSN-L)
- 1520-5118
- OpenAlex
- W4390748438
- Cited by
- 27
- References
- 2
- Citations by year
- 2026: 12 · 2025: 8 · 2024: 6 · 2023: 1
- FWCI
- 10.07
- Type
- journal article
- Language
- en
- Open access
- green
- dx.doi.org
- Licence
- Locations
- Journal of Agricultural and Food Chemistry (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- National Key Research and Development Program of China (2022YFD1400900)
- National Natural Science Foundation of China (32001942)
- Innovation Capability Support Plan of Shaanxi Province (2020TD-035)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Ecology, Evolution, Behavior and Systematics › Fungal Plant Pathogen Control
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Pathogens and Resistance
- Physical Sciences › Environmental Science › Pollution › Pesticide and Herbicide Environmental Studies
- Keywords
- fluopicolide, Phytophthora capsici, point mutations, drug target specificity, molecular docking
- MeSH terms
- Benzamides, Fungicides, Industrial, Proton-Translocating ATPases, Phytophthora, Plant Diseases, Molecular Docking Simulation
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