Tan Y-A, Xiao L-B, Zhao J, Xiao Y-F, Sun Y, Bai L-X
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
Entomology and Nematology, Mid-Florida Research and Education Center, University of Florida, FL, USA.
Insect Mol Biol. 2015 Dec;24(6):611-23. doi: 10.1111/imb.12185. Epub 2015 Sep 3.
Ecdysone receptor (EcR) is the hormonal receptor of ecdysteroids and strictly regulates growth and development in insects. However, the action mechanism of EcR is not very clear. In this study, the cDNA of EcR isoform-B was cloned from Apolygus lucorum (AlEcR-B) and its expression profile was investigated. We reduced AlEcR-B mRNA expression using systemic RNA interference in vivo, and obtained knockdown specimens. Examination of these specimens indicated that AlEcR-B is required for nymphal survival, and that reduced expression is associated with longer development time and lower nymphal weight. To investigate the underlying molecular mechanism of the observed suppression effects, we selected trehalase for a detailed study. Transcript encoding soluble trehalase (AlTre-1) was up-regulated by 20-hydroxyecdysone and in agreement with the mRNA expression of AlEcR-B. The expression profile of AlTre-1, soluble trehalase activity and translated protein level in the midgut of surviving nymphs were down-regulated, compared with controls, after the knockdown expression of AlEcR-B. By contrast, membrane-bound trehalase activity, the related gene expression and translated protein level remained at their initial levels. However, trehalose content significantly increased and the glucose content significantly decreased under the same conditions. We propose that AlEcR-B controls normal carbohydrate metabolism by mediating the expression of AlTre-1 to regulate the growth and development in A. lucorum, which provide an extended information into the functions of AlEcR-B.
蜕皮激素受体(EcR)是蜕皮甾类激素的受体,严格调控昆虫的生长和发育。然而,EcR的作用机制尚不完全清楚。在本研究中,从绿盲蝽中克隆了EcR亚型B的cDNA(AlEcR-B),并对其表达谱进行了研究。我们通过体内系统性RNA干扰降低了AlEcR-B mRNA的表达,获得了基因敲低样本。对这些样本的检测表明,AlEcR-B是若虫存活所必需的,其表达降低与发育时间延长和若虫体重降低有关。为了研究观察到的抑制效应的潜在分子机制,我们选择海藻糖酶进行详细研究。编码可溶性海藻糖酶(AlTre-1)的转录本受20-羟基蜕皮激素上调,且与AlEcR-B的mRNA表达一致。在敲低AlEcR-B的表达后,与对照相比,存活若虫中肠中AlTre-1的表达谱、可溶性海藻糖酶活性和翻译后的蛋白质水平均下调。相比之下,膜结合海藻糖酶活性、相关基因表达和翻译后的蛋白质水平保持在初始水平。然而,在相同条件下,海藻糖含量显著增加,葡萄糖含量显著降低。我们认为,AlEcR-B通过介导AlTre-1的表达来控制正常的碳水化合物代谢,从而调节绿盲蝽的生长和发育,这为AlEcR-B的功能提供了更多信息。