蜕皮是昆虫生长受限的关键解决方案。甲壳素的周期性合成和降解,昆虫表皮的主要组成部分之一,是昆虫生长所必需的。MicroRNA(miRNA)与蜕皮调控有关,然而,它们在几丁质合成途径和20-羟基蜕皮激素信号之间的相互作用中的参与仍然知之甚少.在这项研究中,可溶性海藻糖酶(Tre1)和磷酸乙酰葡糖胺变位酶(PAGM)分别被鉴定为保守miR-8-3p和miR-2a-3的靶标。miR-8-3p-SfTre1和miR-2a-3-SfPAGM的表达谱在不同发育阶段表现出相反的模式,表明它们之间存在负向调节关系。这种关系通过体外双荧光素酶报告系统得到证实。通过注射模拟物过表达miR-8-3p和miR-2a-3抑制其各自靶基因的表达并增加死亡率,在蜕皮前导致死亡,蜕皮和死亡现象。它们还导致几丁质含量和几丁质合成途径中关键基因的表达水平降低(SfTre1,SfTre2,SfHK,SfG6PI,SFGFAT,SFGNA,SfPAGM,SfUAP,SfCHS1,SfCHS1a,和SfCHS1b)。相反,注射miRNA抑制剂导致这些基因的表达水平上调.20E治疗后,miR-8-3p和miR-2a-3的表达水平显著降低,而其相应的靶基因显著增加。这些结果表明,miR-8-3p和miR-2a-3通过靶向SfTre1和SfPAGM在糠疹蜕皮中起调节作用,分别。这些发现为后续新控制策略的开发提供了新的潜在目标。
Molting is a key solution to growth restriction in insects. The periodic synthesis and degradation of chitin, one of the major components of the insect epidermis, is necessary for insect growth. MicroRNA (miRNA) have been implicated in molting regulation, yet their involvement in the interplay interaction between the chitin synthesis pathway and 20-hydroxyecdysone signaling remains poorly understood. In this study, soluble trehalase (Tre1) and phosphoacetylglucosamine mutase (PAGM) were identified as targets of conserved miR-8-3p and miR-2a-3, respectively. The expression profiles of miR-8-3p-SfTre1 and miR-2a-3-SfPAGM exhibited an opposite pattern during the different developmental stages, indicating a negative regulatory relationship between them. This relationship was confirmed by an in vitro dual-luciferase reporter system. Overexpression of miR-8-3p and miR-2a-3 by injection of mimics inhibited the expression of their respective target genes and increased mortality, leading to death in the pre-molting, and molting death phenomena. They also caused a decrease in chitin content and expression levels of key genes in the chitin synthesis pathway (SfTre1, SfTre2, SfHK, SfG6PI, SfGFAT, SfGNA, SfPAGM, SfUAP, SfCHS1, SfCHS1a, and SfCHS1b). Conversely, the injection of miRNA inhibitors resulted in the upregulation of the expression levels of these genes. Following 20E treatment, the expression levels of miR-8-3p and miR-2a-3 decreased significantly, while their corresponding target genes increased significantly. These results indicate that miR-8-3p and miR-2a-3 play a regulatory role in the molting of Sogatella furcifera by targeting SfTre1 and SfPAGM, respectively. These findings provide new potential targets for the development of subsequent new control strategies.