钟其恩,岑永杰,马康,彭亚楠,郑思春,2023,novel-m0287-3p调控斜纹夜蛾中肠蜕皮激素受体的表达[J].环境昆虫学报,(4):988-996
novel-m0287-3p调控斜纹夜蛾中肠蜕皮激素受体的表达
novel-m0287-3p regulates the expression of ecdysone receptor in the midgut of Spodoptera litura
  
DOI:
中文关键词:  斜纹夜蛾  novel-m0287-3p  生长发育  蜕皮激素受体
英文关键词:Spodoptera litura  novel-m0287-3p  growth and development  ecdysone receptor
基金项目:国家自然科学基金(31672362,32070493)
作者单位
钟其恩,岑永杰,马康,彭亚楠,郑思春 1.华南师范大学生命科学学院昆虫科学与技术研究所广州市昆虫发育调控与应用研究重点实验室广州 5106312.梅州市华师昆虫发育生物学与应用技术重点实验室广梅园研发中心广东梅州 514779 
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中文摘要:
      microRNA (miRNA)是一类长度约为23个核苷酸的非编码内源性小分子RNA,参与真核生物基因表达的调控,从而在生物发育和应对环境中起着重要的作用。为了探讨novel miRNA在害虫发育中的作用,本研究对斜纹夜蛾Spodoptera litura miRNA数据库中表达量较高的novel-m0287-3p进行了研究。通过生物信息学分析,确定了novel-m0287-3p是一个新的miRNA,并预测其可能靶向蜕皮激素受体EcRb1,表明novel-m0287-3p可能参与斜纹夜蛾发育的调控。双荧光素酶实验表明,novel-m0287-3p分别结合于EcRb1 3′非编码区的2个位点上。实时荧光PCR结果显示novel-m0287-3p和EcRb1均在斜纹夜蛾的中肠中表达。为了进一步揭示novel-m0287-3p对EcRb1表达的调控以及对幼虫发育的影响,本研究进行了体内实验。将novel-m0287-3p的模拟体类似物注射入斜纹夜蛾幼虫,结果显示斜纹夜蛾体内EcRb1的mRNA水平发生了下调,证明novel-m0287-3p能够抑制幼虫EcRb1的表达。向4龄第1天的斜纹夜蛾幼虫注射novel-m0287-3p后,幼虫的生长受到抑制;而注射斜纹夜蛾6龄幼虫则导致其化蛹滞后。研究结果表明,novel-m0287-3p可通过调控蜕皮激素受体EcRb1的表达,从而影响斜纹夜蛾的生长发育。
英文摘要:
      MicroRNAs (miRNAs) are a class of non-coding endogenous small RNAs with about 23 nucleotides in length, and are involved in the regulation of eukaryotic gene expression and play an important role in biological development and response to environment. In order to explore the role of novel miRNAs in pest development, the novel-m0287-3p with high expression in the Spodoptera litura miRNA database was studied in this study. By bioinformatics analysis, novel-m0287-3p was identified as a novel miRNA, and it was predicted that it might target to ecdysone receptor EcRb1, indicating that novel-m0287-3p may be involved in the developmental regulation of S. litura. Dual-luciferase activity assay showed that novel-m0287-3p bound to two sites in the 3’ non-coding region of EcRb1. Real-time PCR results showed that both novel-m0287-3p and EcRb1 were expressed in the midgut of S. litura. To further verify the regulation of novel-m0287-3p on EcRb1 expression and its effect on larval development, in vivo experiments were performed. novel-m0287-3p agomir was injected into S. litura larvae, and the results showed that the mRNA level of EcRb1 in S. litura was down-regulated, which proved that novel-m0287-3p could inhibit the expression of EcRb1 in larvae. The growth of S. litura was inhibited after injection of novel-m0287-3p into one-day-old 4th instar larvae, and the pupation was delayed when the 6th instar larvae were injected with novel-m0287-3p agomir. The results revealed that novel-m0287-3p could affect the growth and development of S. litura by regulating the expression of ecdysteroid receptor EcRb1.
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