摘要Fungal pathogens pose escalating challenges to global food security,as resistance has emerged against nearly all major fungicides used in agriculture.RNA-based antifungals offer a sustainable and environmen-tally friendly alternative for disease control,but their deployment is hindered by RNA instability under envi-ronmental conditions,especially in soil.In this study,we engineered two plant-beneficial soil bacteria-Ba-cillus subtilis(Gram-positive)and Pseudomonas putida(Gram-negative)-to produce double-stranded RNAs(dsRNAs)targeting fungal genes in the foliar and postharvest pathogen Botrytis cinerea and the soil-borne pathogen Verticillium dahliae.We found that both bacterial species secrete RNA through extracel-lular vesicles(EVs)and that these RNAs are transported into fungal cells,demonstrating cross-kingdom RNA trafficking from bacteria to fungi.Application of dsRNA-containing bacterial EVs to plant leaves sup-pressed B.cinerea infection.In addition,direct treatment with dsRNA-producing bacteria protected both Arabidopsis thaliana and tomato plants from infections byB.cinerea and V.dahliae.Our findings establish beneficial bacteria as a scalable platform for continuous production and delivery of antifungal RNAs,enabling a cost-effective strategy for sustainable crop protection.