Tomato yellow leaf curl virus (TYLCV), a major plant virus, infects multiple plant species, severely threatening global food security. Arbutin, a natural product used in cosmetics to reduce pigmentation, also exhibits antibacterial and anti-inflammatory properties. However, its potential in plant protection remains undocumented. This study reveals arbutin's ability to inhibit TYLCV infection. In Nicotiana benthamiana, 100 μg/mL arbutin inhibited viral gene accumulation by up to 76.8%, surpassing ningnanmycin (65.8%) and ribavirin (39.5%). Besides, microscale thermophoresis indicated that arbutin bound strongly to the TYLCV coat protein (CP). Molecular docking indicated that arbutin interacted with ARG58, VAL65, and CYS69. RT-qPCR and Western blot experiments confirmed the crucial roles of these amino acids, especially VAL65, in viral infection. Transcriptome analysis revealed that mutating VAL65 affected plant-pathogen interaction pathways and MAPK signaling in host defense mechanisms. This study unveils arbutin's novel antiviral function, providing crucial insights for developing new biopesticides against plant viruses.
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