Multiple microRNAs control ecdysone signaling in the midgut of Spodoptera litura.
Multiple microRNAs control ecdysone signaling in the midgut of Spodoptera litura.
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多种 microRNA 控制斜纹夜蛾中肠中的蜕皮激素信号传导。
DOI：
10.1111/1744-7917.12745
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发表时间：
2020
期刊：
影响因子：
4
通讯作者：
Huang Li-Hua
中科院分区：
文献类型：
--
作者：
Luo Wei;Huang Li-Xia;Qin Shuang-Kang;Zhang Xian;Feng Qi-Li;Gu Jun;Huang Li-Hua
变态是昆虫最重要的生理过程之一。它受一系列蜕皮激素级联基因的调控。近年来，人们对昆虫体内的microRNAs （miRNAs）进行了大量的研究；然而，它们在变态中的作用在很大程度上是未知的。在本研究中，通过对鳞翅目害虫spodoptera liturd的中肠RNA测序，研究了小RNA群体在幼虫-蛹蜕变过程中的动态。共鉴定出101个mirna，其中75个mirna在变质过程中差异表达。通过四个经典软件程序分析了这些差异表达的mirna与12个蜕膜激素级联基因之间的关系，发现这些mirna与它们的靶标之间存在多对多的调控网络。其中，miR - 14 - 3p及其两个靶点（EcRandE75）被选中进行进一步验证。MiR‐14‐3p在6龄幼虫中的表达量高于预蛹和蛹，这与crrande75这两个蜕皮激素级联基因的表达量相反。荧光素酶报告基因实验证实，这两个基因都受到miR‐14‐3p的调控。有趣的是，在ecr、USP、E75、E74、E78、E93、Hr3、Hr4、Hr39、krh11和ftzf1等不同的ecr级联基因转录变体中，3 '非翻译区几乎是相同的。因此，一个蜕皮激素级联基因的不同转录变体可以被相同的miRNA调控。以上数据表明蜕皮激素信号通路受到miRNA的严格控制。这些发现扩大了我们对昆虫变态机制的认识，也可能为今后害虫的防治提供新的可能性。
Metamorphosis is one of the most important physiological processes in insects. It is regulated by a serial of ecdysone cascade genes. Recently, lots of microRNAs (miRNAs) were investigated in insects; however, their function in metamorphosis is largely unknown. In the present study, the dynamics of a small RNA population was investigated by RNA sequencing from the midgut of a lepidopteran pestSpodoptera lituraduring larval–pupal metamorphosis. A total of 101 miRNAs were identified, and 75 miRNAs were differentially expressed during the metamorphic process. The relationship between these differentially expressed miRNAs and 12 ecdysone cascade genes was analyzed by four classical software programs, and a multiple‐to‐multiple regulatory network was found to exist between these miRNAs and their targets. Among them, miR‐14‐3p and its two targets (EcRandE75) were chosen for further validation. MiR‐14‐3p had higher expression level in the 6th instar larvae as compared with either the prepupae or pupae, which was opposite to that of bothEcRandE75, two ecdysone cascade genes. Luciferase reporter assay confirmed that bothEcRandE75were regulated by miR‐14‐3p. Interestingly, the 3′ untranslated regions are nearly identical to each other among different transcript variants of the ecdysone cascade genes, includingEcR,USP,E75,E74,E78,E93,Hr3,Hr4,Hr39,Krh1andFtzf1. Thus, different transcript variants of one ecdysone cascade gene could be regulated by the same miRNA. The above data suggest that the ecdysone signaling pathway is under the tight control of miRNA. These findings expand our understanding of the mechanism of insect metamorphosis and may also provide a novel possibility for the control of pest insects in the future.