Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi, China.
College of Life Science, Shanxi University, Taiyuan, Shanxi, China.
Insect Sci. 2021 Aug;28(4):941-951. doi: 10.1111/1744-7917.12837. Epub 2020 Jul 20.
Cytochrome P450 monooxygenases (CYPs) play essential physiological functions in insects. CYP303A1 is highly conserved in insect species studied to date, and shows an indispensable role for adult eclosion in both Locusta migratoria and Drosophila melanogaster. However, how CYP303A1 is regulated to control insect developmental processes remains uninvestigated. In this study, we discovered functional binding sites for miR-184 in the coding sequence of LmCYP303A1. The luciferase reporter assay showed that miR-184 could target LmCYP303A1 and regulate its expression in vitro. Furthermore, overexpression of miR-184 through microinjection of agomir to locusts reduced the transcripts of LmCYP303A1 and led to abnormal molting, which is similar to the phenotype of silencing LmCYP303A1 by direct injection of dsLmCYP303A1 to locusts. Meanwhile, down-regulation of miR-184 by injection of antagomir increased the LmCYP303A1 transcript and caused molting defects. These findings suggested that miR-184 could target LmCYP303A1 to regulate the molting process in L. migratoria, which might be considered as a novel target for pest control.
细胞色素 P450 单加氧酶（CYPs）在昆虫中发挥着重要的生理功能。迄今为止，在已研究的昆虫物种中，CYP303A1 高度保守，并且在 Locusta migratoria 和 Drosophila melanogaster 中对成虫蜕皮起着不可或缺的作用。然而，CYP303A1 如何被调控以控制昆虫的发育过程仍未被研究。在本研究中，我们在 LmCYP303A1 的编码序列中发现了 miR-184 的功能结合位点。荧光素酶报告基因检测表明，miR-184 可以靶向 LmCYP303A1，并在体外调节其表达。此外，通过向蝗虫注射 agomir 过表达 miR-184 会降低 LmCYP303A1 的转录本，并导致异常蜕皮，这与直接注射 dsLmCYP303A1 到蝗虫中沉默 LmCYP303A1 的表型相似。同时，通过注射 antagomir 下调 miR-184 会增加 LmCYP303A1 的转录本并导致蜕皮缺陷。这些发现表明，miR-184 可以靶向 LmCYP303A1 来调节 L. migratoria 的蜕皮过程，这可能被认为是害虫防治的一个新靶点。
