Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, MA, USA.
Bioinformatics Section, National Institute of Neurological Disorders and Stroke, National Institute of Health, Bethesda, MD, USA.
Methods Mol Biol. 2022;2391:171-184. doi: 10.1007/978-1-0716-1795-3_14.
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.
尖孢镰刀菌是一种跨物种的真菌病原体，不仅会导致严重的植物血管疾病，还可能伺机感染人类。在这里，我们描述了两种高通量筛选方法，即 Resazurin 细胞活力测定法和光密度测定法，用于筛选具有抗真菌特性的天然产物，以抑制临床分离的尖孢镰刀菌。在用茉莉酸甲酯处理或与尖孢镰刀菌共培养后，作为非生物诱导剂和生物诱导剂，我们分别鉴定出了三个产生抑制真菌生长物质的细胞系。我们的程序验证了结合高通量方法发现新型抗病原物先导化合物的强大潜力。
