Boosting the efficacy of fungal biocontrol: miRNA339-5p-mediated mosquito immunity regulation.

Abstract

Aedes mosquitoes are vectors for numerous viral diseases, including dengue, zika, chikungunya, and yellow fever. Therefore, underscoring the urgent need for eco-friendly alternatives to combat insecticide resistance and the scarcity of effective vaccines. Entomopathogenic fungi present a sustainable alternative to chemical insecticides; however, their widespread application is limited by their relatively low virulence. Here, we investigated the immunological interactions between Metarhizium anisopliae and Aedes albopictus, demonstrating that fungal infection significantly up-regulated immune-related genes in the Toll and melanization pathways, thereby enhancing antifungal and antibacterial defenses at 48 h post-infection (hpi). Small RNA sequencing identified miR339-5p as a crucial modulator, targeting the immune genes Gram-Negative Binding Protein 1 (GNBP1) and CLIP-domain Serine Protease B15 (CLIPB15), which are critical for Toll and phenoloxidase (PO) pathway activation. The administration of a synthetic miR339-5p mimic increased fungal virulence, resulting in a higher mortality rate among adult mosquitoes and a significant increase in the mortality rate of mosquito larvae within 24 hpi. GNBP1 was found to regulate both Toll and PO pathways, while CLIPB15 specifically modulated the PO system by cleaving prophenoloxidase (PPO). This research highlights the potential of leveraging Ae. albopictus-encoded miR339-5p through advanced genetic engineering techniques to bolster the efficacy of existing fungal-based mosquito control strategies, providing a promising approach in the fight against mosquito-borne diseases. © 2024 Society of Chemical Industry.