BACKGROUNDThe doublesex (dsx) gene is a crucial component of the insect sex determination pathway, playing a pivotal role in sexual differentiation and development. While the functions of dsx have been extensively characterized in several insect species, its molecular mechanisms and specific roles in sex determination remain largely unexplored in the lepidopteran rice stem borer, Chilo suppressalis (Walker), a significant agricultural pest.RESULTSIn this study, we identified two male‐specific and five female‐specific Csdsx transcripts in C. suppressalis. Phylogenetic analyses and multiple sequence alignments revealed that Csdsx is highly conserved among lepidopterans. To investigate its functional roles, we employed CRISPR/Cas9‐mediated mutagenesis to generate C. suppressalis mutants. Specifically, target sites were designed on exons 1, 3, 4, and 5 to disrupt the common region (exon 1), female‐specific exons (exon 3 and exon 4), and male‐specific regions of Csdsx (exon 5). As anticipated, mutants exhibited abnormal development of both external and internal genitalia during the pupal and adult stages. Furthermore, mutations induced sex‐specific sterility. Notably, knockout of sex‐specific Csdsx isoforms also resulted in abnormal wing patterns and antennal morphology.CONCLUSIONThis study demonstrates the conserved role of Csdsx in sexual differentiation in C. suppressalis and highlights its potential as a molecular target for environmentally friendly pest management strategies against this agricultural pest. © 2025 Society of Chemical Industry.

中文翻译：

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Doublesex 调节水稻螟 Chilo suppressalis 的性别特异性器官的发育

背景 doublesex （dsx） 基因是昆虫性别决定途径的重要组成部分，在性分化和发育中起关键作用。虽然 dsx 的功能已在几种昆虫物种中得到广泛表征，但其分子机制和在性别决定中的特定作用在鳞翅目稻螟 Chilo suppressalis （Walker） 中仍未得到探索，这是一种重要的农业害虫。结果在这项研究中，我们在 C. suppressalis 中鉴定了 2 个雄性特异性和 5 个雌性特异性 Csdsx 转录本。系统发育分析和多序列比对表明 Csdsx 在鳞翅目中高度保守。为了研究其功能作用，我们采用 CRISPR/Cas9 介导的诱变来产生抑制梭菌突变体。具体来说，在外显子 1 、 3 、 4 和 5 上设计靶位点以破坏 Csdsx 的共同区域 （外显子 1） 、女性特异性外显子 （外显子 3 和外显子 4） 和男性特异性区域 （外显子 5）。正如预期的那样，突变体在蛹和成虫阶段表现出外生殖器和内生殖器的异常发育。此外，突变诱导性别特异性不育。值得注意的是，性别特异性 Csdsx 亚型的敲除也导致异常的翅膀图案和触角形态。结论该研究表明 Csdsx 在 C. suppressalis 的性分化中的保守作用，并强调了其作为针对该农业害虫的环境友好型害虫管理策略的分子靶点的潜力。© 2025 化工学会.