BACKGROUNDReproduction is the basis of insect population growth and evolution, and encompasses ovarian development, reproductive behavior, and fecundity. Bactrocera dorsalis is a globally significant agricultural pest that is subject to quarantine, with mated females that can lay over 3000 eggs. The post‐transcriptional regulation of ovarian development remains unclear. Here, miR‐31b is shown to be involved in regulating Bactrocera dorsalis ovarian development.RESULTSCRISPR/Cas9 was used to generate miR‐31b loss‐of‐function mutations in Bactrocera dorsalis. The removal of miR‐31b resulted in severely impaired ovarian development in adults, with phenotypes that included dramatically reduced egg production and hatching rates. The relationship between miR‐31b and its target gene arylsulfatase B (ARSB) was subsequently identified using the methods of bioinformatics, transcriptomic sequencing, quantitative polymerase chain reaction (qPCR), RNA pull‐down and dual‐luciferase reporter assay. Finally, miR‐31b was confirmed to bind the target gene arylsulfatase B to affect metabolism and thereby further hindered ovarian development of Bactrocera dorsalis.CONCLUSIONOverall, these results provide new insights into molecular mechanisms at the post‐transcriptional level in regulating ovarian development and insect reproduction, consequently providing potential targets to control arthropod pests through the reproductive strategy. © 2024 Society of Chemical Industry.

中文翻译：

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miR-31b 靶向芳基硫酸酯酶 B 以调节 Bactrocera dorsalis 的卵巢发育


背景繁殖是昆虫种群增长和进化的基础，包括卵巢发育、生殖行为和繁殖力。Bactrocera dorsalis 是一种具有全球意义的农业害虫，需要检疫，交配的雌性可以产下 3000 多个卵。卵巢发育的转录后调控仍不清楚。在这里，miR-31b 被证明参与调节 Bactrocera dorsalis 卵巢发育。结果CRISPR/Cas9 用于在 Bactrocera dorsalis 中产生 miR-31b 功能丧失突变。去除 miR-31b 导致成人卵巢发育严重受损，其表型包括产蛋量和孵化率显著降低。随后使用生物信息学、转录组测序、定量聚合酶链反应 （qPCR）、RNA 下拉和双荧光素酶报告基因测定等方法鉴定 miR-31b 与其靶基因芳基硫酸酯酶 B （ARSB） 之间的关系。最后，证实 miR-31b 与靶基因芳基硫酸酯酶 B 结合以影响代谢，从而进一步阻碍背侧 Bactrocera 的卵巢发育。结论总体而言，这些结果为转录后水平调节卵巢发育和昆虫繁殖的分子机制提供了新的见解，从而为通过繁殖策略控制节肢动物害虫提供了潜在的靶点。© 2024 化工学会.