Microminutinin, a Fused Bis-Furan Coumarin from Murraya euchrestifolia, Exhibits Strong Broad-Spectrum Antifungal Activity by Disrupting Cell Membranes and Walls.

Plant fungal diseases pose a serious threat to crop production and safety, and natural products are one of the important directions for the development of new green fungicides. This study found that the extract of Murraya euchristifolia had significant antifungal activity, and a main antifungal coumarin (1) was isolated by bioassay-guided fractionation. The structure of 1 was identified by NMR and MS spectroscopic data as a fused bis-furan coumarin (microminutinin) which was first isolated from the Murraya genus and exhibited strong broad-spectrum antifungal activity against eight plant pathogenic fungi from different families and genera. The EC50 value of 1 (11.33 μg/mL) against Pestalotiopsis theae (the most sensitive to 1) was slightly higher than that (7.03 μg/mL) of the positive drug (80% carbendazim WP), indicating that 1 has the potential to serve as a lead compound for botanical fungicides. The bioassay results against P. theae in vivo indicated that 1 also has the potential for field application. Scanning electron microscopy and optical microscopy revealed that 1 disrupted the morphological structure of mycelium, causing hyphae to twist, shrink, and even crack and severely reducing hyphal branching. Furthermore, propidium iodide staining proved that microminutinin destroyed the integrity of the cell membrane, causing leakage of cellular components. In addition, calcofluor white staining and chitin content changes illustrated that microminutinin disrupted the cell wall structure. This research provides compound sources and a theoretical basis for the development of botanical fungicides.