Antifungal efficacy of microencapsulated miR166 and miR159 oligoDNAs through whey protein concentrate (WPC) as coated protein against Verticillium dahliae

Verticillium dahliae Kleb. is one of the most destructive fungal pathogens, infecting susceptible plants through their roots and disrupting vascular tissues. As soil-borne diseases pose a persistent threat to agriculture, the development of sustainable biocontrol strategies has gained significant attention. Among these, biomolecule-based approaches—particularly the use of oligonucleotides—have emerged as an innovative and eco-friendly alternative for plant disease management. In this study, we evaluated the antifungal potential of microencapsulated double-stranded oligoDNAs (corresponding to microRNA166 and microRNA159) produced via electrospraying with a whey protein concentrate (WPC) polymer. The encapsulated oligoDNAs were tested against V. dahliae by mixing 10 µL of fungal spores (1×10⁶ spores/mL) with 5 µL of the microcapsule solution, followed by culturing on potato dextrose agar (PDA). Our findings demonstrate that both oligoDNA 159 and oligoDNA 166 significantly inhibit fungal growth. Notably, encapsulation with WPC enhanced their antifungal efficacy, suggesting that this technology improves oligoDNA stability and enables controlled release. These results highlight the potential of oligonucleotides as effective biocontrol agents against fungal pathogens. Furthermore, encapsulation presents a promising strategy to optimize their delivery and application in sustainable agriculture. This study provides compelling evidence for the use of microencapsulated oligoDNAs in fungal disease management, offering a viable, environmentally safe alternative to conventional chemical treatments. Future research should explore field applications and long-term effects to validate their practical use in crop protection.