Expression of a viral ecdysteroid UDP‐glucosyltransferase enhanced the insecticidal activity of the insect pathogenic fungus Beauveria bassiana

Abstract BACKGROUND Entomopathogenic fungi, such as Beauveria bassiana , hold promise as biological control agents against insect pests. However, the efficacy of these fungi can be hindered by insect immune responses. One strategy to enhance fungal virulence is to manipulate host immune by targeting key regulatory molecules like 20‐hydroxyecdysone (20E). RESULTS In this study, we engineered B. bassiana strains to constitutively express the enzyme ecdysteroid UDP‐glucosyltransferase (EGT), which inactivates 20E, a crucial insect molting hormone. The engineered strain Bb::EGT‐1 exhibited robust expression of EGT , leading to a significant reduction in insect 20E levels upon infection. Moreover, infection with Bb::EGT‐1 resulted in accelerated larval mortality. Immune responses analysis revealed repression of insect immune response genes and decreased phenoloxidase (PO) activity in larvae infected with Bb::EGT‐1 . Microbiome analysis indicated alterations in bacterial composition within infected insects, with increased abundance observed during infection with Bb::EGT‐1 . Additionally, the presence of bacteria hindered hyphal emergence from insect cadavers, suggesting a role for microbial competition in fungal dissemination. CONCLUSIONS Constitutive expression of EGT in B. bassiana enhances fungal virulence by reducing insect 20E levels, suppressing immune responses, and altering the insect microbiome. These findings highlighted the potential of engineered fungi as effective biocontrol agents against insect pests and provide insights into the complex interactions between entomopathogenic fungi, their hosts, and associated microbes. © 2024 Society of Chemical Industry.