College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China.
Department of Entomology, University of Kentucky, Lexington, KY 40546, USA.
Int J Mol Sci. 2023 Nov 14;24(22):16311. doi: 10.3390/ijms242216311.
Oogenesis in insects is a carefully orchestrated process, facilitating the formation of female gametes, which is regulated by multiple extrinsic and intrinsic factors, including As a member of the serine protease family, is a homolog of , a maternally required protease defining embryonic dorsoventral polarity in . In this study, we used CRISPR/Cas9-mediated mutagenesis to functionally characterize in the Asian corn borer, a devastating maize pest throughout Asia and Australia. Building on previous knowledge, we hypothesized that knockout of would disrupt embryonic development in females. To examine this overarching hypothesis, we (1) cloned and characterized from , (2) designed target sites on exons 1 and 4 to construct a CRISPR/Cas9 mutagenesis system, and (3) documented phenotypic impacts among mutants. As a result, we (1) examined the temporal-spatial expression profiles of , which has an open reading frame of 5648 bp in length and encodes a protein of 1873 amino acids; (2) established mutants; and (3) documented recessive, female-specific sterility among mutants, including absent or deformed oviducts and reduced fertility in female but not male mutants. Overall, the combined results support our initial hypothesis that is required for embryonic development, specifically ovarian maturation, in females. Given its substantial impacts on female sterility, provides a potential target for the Sterile Insect Technique (SIT) to manage in general and the species complex in particular.
昆虫的卵子发生是一个精心协调的过程，有助于形成雌性配子，这个过程受到多种外在和内在因素的调节，包括 。作为丝氨酸蛋白酶家族的一员，是 的同源物， 是一种母体必需的蛋白酶，定义了 在 中的背腹极性。在这项研究中，我们使用 CRISPR/Cas9 介导的诱变来对亚洲玉米螟中的 进行功能表征，亚洲玉米螟是一种在亚洲和澳大利亚造成严重玉米虫害的害虫。基于先前的知识，我们假设 敲除会破坏 雌性胚胎的发育。为了检验这个总体假设，我们 (1) 从 中克隆和表征了 ，(2) 在exon 1 和 4 上设计了靶位点来构建一个 CRISPR/Cas9 诱变系统，和 (3) 记录了 突变体之间的表型影响。结果，我们 (1) 检测了 的时空表达谱， 有一个 5648bp 的开放阅读框，编码一个 1873 个氨基酸的蛋白质；(2) 建立了 突变体；和 (3) 记录了 突变体中的隐性、雌性特异性不育，包括输卵管缺失或畸形，以及雌性而非雄性突变体的生育能力降低。总的来说，综合结果支持我们最初的假设，即 对于 雌性的胚胎发育，特别是卵巢成熟，是必需的。鉴于它对雌性不育的巨大影响， 为利用不育昆虫技术 (SIT) 来管理 总体上和特别是物种复合体 提供了一个潜在的目标。
