Abstract
Fusarium oxysporum is a cross-kingdom fungal pathogen that not only causes devastating plant vascular diseases but can also opportunistically infect humans. Here we describe two high-throughput screening assays, a resazurin cell viability assay and an optical density assay, to screen natural products from cultured plant cells with antifungal properties against a clinical isolate of F. oxysporum. After elicitation by applying methyl jasmonate or by co-culture with F. oxysporum, as an abiotic elicitor and a biotic elicitor, respectively, we identified three cell lines that produce materials that inhibit fungal growth. Our procedure validates the powerful potential of combining high-throughput methods for the discovery of novel anti-pathogenic leads.
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Acknowledgments
We thank Katie Webster, Michael Daley, and many undergraduate students for their dedication in maintaining the PCCL cell lines; UMass Institute for Applied Life Sciences Core Facilities for maintaining the PCCL; Jennifer Normanly and Elizabeth Vierling for their consistent leadership in securing various funds to support the PCCL and related activities; and Jennifer Normanly for her involvement in the development of this research and her critical review of this manuscript.
Funding: The establishment of the PCCL has been sponsored by National Science Foundation (NSF/CSBR-1561572), US Department of Agriculture Massachusetts Experiment Station Awards (MAS00496 and MAS00520), the Science and Technology fund from the President’s Office of the University of Massachusetts, and the 2016 UMass Amherst Armstrong Fund for Science. YZ and LJM are supported by the National Institute of Health (R01EY030150) and Burroughs Welcome Foundation (1014893).
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Zhang, Y., Kao, PL., Rampal, A., Mafu, S., Savinov, S., Ma, LJ. (2022). High-Throughput Screening Assays to Identify Plant Natural Products with Antifungal Properties Against Fusarium oxysporum. In: Coleman, J. (eds) Fusarium wilt. Methods in Molecular Biology, vol 2391. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1795-3_14
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DOI: https://doi.org/10.1007/978-1-0716-1795-3_14
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