Levi Ofri, Zuchman Rina, Sleman Nour, Koren Roni, Khamaisi Hazem, Horwitz Benjamin A
Faculty of Sciences and Technology, Tel-Hai Academic College, Upper Galilee, Israel.
MIGAL - Galilee Research Institute, Kiryat Shmona, Israel.
mBio. 2025 Aug 25:e0190925. doi: 10.1128/mbio.01909-25.
Fungal pathogens of plants must overcome host-imposed stressors, including antimicrobial small molecules. Ferulic acid (FA), a plant-derived phenolic compound, induces fungal stress and cell death. To uncover genetic determinants of FA sensitivity, we performed a genome-wide CRISPR interference (CRISPRi) screen in . We confirmed that FA impairs yeast growth and triggers stress granule marker sequestration, establishing a relevant selection condition. The CRISPRi screen identified 194 genes involved in the FA-induced stress response and 12 whose repression enhanced resistance. Among them, ERG9, encoding squalene synthase, was most strongly enriched, and its repression conferred FA resistance alongside upregulation of HMG1, implicating the ergosterol biosynthesis pathway. Proteomic profiling of FA-resistant strains further revealed conserved upregulation of ergosterol biosynthetic enzymes. FA also synergized with fluconazole, a known ergosterol-targeting antifungal, and enhanced susceptibility in azole-resistant strains, suggesting interference with ergosterol metabolism. , FA exhibited dose-dependent antifungal activity, significantly reducing lesion formation in maize. These findings establish FA as a promising antifungal agent that targets conserved lipid biosynthesis pathways and overcomes resistance mechanisms, supporting its potential as a sustainable therapeutic and agricultural fungicide.IMPORTANCEFungal infections are a growing threat to human health and agriculture, with rising antifungal resistance limiting treatment options. In this study, we used a genome-wide screening approach to identify ferulic acid (FA), a naturally occurring compound found in plants, as a promising antifungal agent. FA targets the same cellular pathway as many current antifungal drugs and works especially well when combined with fluconazole, a commonly used treatment. Remarkably, FA is also effective against drug-resistant strains, offering hope for new ways to treat difficult infections. In addition to its medical potential, FA protects maize from fungal pathogens, highlighting its usefulness as a sustainable and environmentally friendly crop protectant. These results suggest that FA could be developed into a versatile antifungal agent with applications in both clinical and agricultural settings, helping address the urgent need for new strategies to overcome antifungal resistance.
植物真菌病原体必须克服宿主施加的应激源，包括抗菌小分子。阿魏酸（FA）是一种植物来源的酚类化合物，可诱导真菌应激和细胞死亡。为了揭示对FA敏感性的遗传决定因素，我们在……中进行了全基因组CRISPR干扰（CRISPRi）筛选。我们证实FA会损害酵母生长并触发应激颗粒标记物隔离，从而建立了相关的选择条件。CRISPRi筛选鉴定出194个参与FA诱导应激反应的基因以及12个其抑制作用增强抗性的基因。其中，编码角鲨烯合酶的ERG9富集程度最高，其抑制作用与HMG1的上调一起赋予了对FA的抗性，这表明麦角固醇生物合成途径与之相关。对FA抗性菌株的蛋白质组分析进一步揭示了麦角固醇生物合成酶的保守上调。FA还与一种已知的靶向麦角固醇的抗真菌药物氟康唑协同作用，并增强了对唑类抗性菌株的易感性，表明其对麦角固醇代谢有干扰作用。此外，FA表现出剂量依赖性的抗真菌活性，显著减少了玉米中的病斑形成。这些发现确立了FA作为一种有前景的抗真菌剂，它靶向保守的脂质生物合成途径并克服抗性机制，支持了其作为可持续治疗剂和农业杀菌剂的潜力。
重要性
真菌感染对人类健康和农业构成的威胁日益增加，抗真菌耐药性的上升限制了治疗选择。在本研究中，我们使用全基因组筛选方法鉴定出阿魏酸（FA），一种在植物中天然存在的化合物，作为一种有前景的抗真菌剂。FA与许多当前的抗真菌药物靶向相同的细胞途径，并且与常用治疗药物氟康唑联合使用时效果特别好。值得注意的是，FA对耐药菌株也有效，为治疗难治性感染提供了新方法的希望。除了其医学潜力外，FA还能保护玉米免受真菌病原体侵害，突出了其作为可持续且环境友好型作物保护剂的有用性。这些结果表明，FA可以开发成一种多功能抗真菌剂，应用于临床和农业领域，有助于满足克服抗真菌耐药性新策略的迫切需求。
