Knockout BR-C induces premature expression of E93 thus triggering adult differentiation under larval morphology

Holometabolan pupal-specifier broad-complex (BR-C) and adult specifier ecdysone-induced protein 93F (E93) are essential for metamorphosis; however, their interaction and effects on programmed cell death and cell differentiation during pupation remain unclear. Here, multiple single-guide RNA (sgRNA)-mediated mosaic knockout of BR-C induced a deformed larva/pupa intermediate phenotype in Spodoptera frugiperda. Quantitative real-time polymerase chain reaction (qPCR) analysis showed that the adult specifier E93 was prematurely expressed in the BR-C mutants during the penultimate and last instar larval stages. Additionally, histological observation and TUNEL assay showed that apoptosis in the fat body and midgut was activated in the larval tissues; astonishingly, the adult midgut appeared in the pupae of BR-C mutants. Overall, the results demonstrated that the premature expression of E93 induced by lack of BR-C triggers adult differentiation during the larval stages, which revealed the inhibitory effect of BR-C on E93 during metamorphosis in S. frugiperda. © 2024 Society of Chemical Industry.