Masculinizer and Doublesex as Key Factors Regulate Sexual Dimorphism in Ostrinia furnacalis

SIMPLE SUMMARY: In animals, sexually dimorphic traits are ubiquitous and play vital roles in reproduction, courtship, and environmental adaptation, especially in insects. In this study, we used the CRISPR/Cas9 genome editing system to generate somatic mutations of the Masculinizer (Masc) and doublesex (dsx) genes in the sex determination pathway of Ostrinia furnacalis. The OfMasc and Ofdsx genes are structural orthologs of the key sex regulation factors in Bombyx mori. Mutation of the OfMasc and Ofdsx genes induced abnormal external genitalia, adult sterility, and sex reversal of sexually dimorphic traits including wing pigmentation, gene expression patterns, and dsx sex-specific splicing. These results demonstrate that the Masc and dsx genes are conserved factors in sexually dimorphic traits, and therefore represent potential target genes in the effort to control O. furnacalis and other lepidopteran pests. ABSTRACT: Sex determination is an important and traditional biological process. In Lepidoptera, Masculinizer (Masc) and doublesex (dsx) are the essential genes for sex determination and play critical roles in sexual differentiation and development. The functions of Masc and dsx have been characterized in several model insect species. However, the molecular mechanism and sex determination functions of Masc and dsx in Ostrinia furnacalis, an agricultural pest, are still unknown. Here, we successfully used the CRISPR/Cas9 genome editing system to knock out OfMasc and Ofdsx. Mutation of OfMasc induced male external genital defects and sterility. Disruptions of the Ofdsx common region caused sex-specific defects in the external genitals and adult sterility. In addition, we found that OfMasc and Ofdsx can regulate the pigmentation genes that control wing pigmentation patterns. These results demonstrate that OfMasc and Ofdsx play key roles in the sex determination of O. furnacalis, and suggest novel genetic control approaches for the management of pests, including O. furnacalis.