Inactivation of Sebol Induces Male Infertility in Spodoptera exigua via Arrested Meiosis: A Potential Molecular Target for Genetics-Based Sterile Insect Technique

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The beet armyworm, Spodoptera exigua, is a significant agricultural pest managed primarily through chemical insecticides, which present environmental concerns and promote resistance. Genetics-based sterile insect technique (gSIT) provides an alternative by inducing male sterility without irradiation. In this study, we cloned the S. exigua boule gene ( Sebol ), a 741-nucleotide open reading frame encoding a 246-amino-acid protein with an RNA recognition motif. Phylogenetic analysis confirmed its conservation across Lepidoptera and high similarity to orthologs in Spodoptera litura and Helicoverpa armigera . Sebol is predominantly expressed in adult males, particularly in the testis. CRISPR/Cas9-mediated deletion of 190 bp in exon 1 generated a truncated Sebol protein, resulting in complete male sterility. Transcriptome analysis revealed 2069 differentially expressed genes associated with sperm development, with potential interactions involving syf1 and ZNFX1 . Our findings suggest that Sebol is crucial for spermatogenesis, making it a promising target for gSIT-based pest control.

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