蜕皮激素调控绿盲蝽水溶性海藻糖酶合成的分子机制

项目名称： 蜕皮激素调控绿盲蝽水溶性海藻糖酶合成的分子机制

项目编号： No.31301668

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 谭永安

作者单位： 江苏省农业科学院

项目金额： 23万元

中文摘要： 绿盲蝽为多种经济作物的致灾性害虫，水溶性海藻糖酶（Tre-1）是其体内主要水解酶系，并在几丁质合成、飞行等多项生理活动中发挥重要作用。前期研究表明，蜕皮激素(20-E)有诱导Tre-1表达的功能，但其转录信号的传导机制尚不清晰。因此，本项目在已获得绿盲蝽Tre-1基因及重组蛋白的基础上，拟开展以下研究：首先克隆20-E受体基因EcR和USP，明确其在绿盲蝽体内表达谱；再分别通过RNA干扰EcR和USP基因抑制其表达量及微注射20-E后使其表达量上调后，利用双跟踪标定定量 PCR法，逐一分析Tre-1酶活、mRNA及蛋白表达量的差异性；最后构建Tre-1基因启动子控制的荧光素酶基因报告质粒，通过分析荧光素酶活力差异来阐明20-E对Tre-1基因启动子转录活性的调节功能。项目的研究结果，有望揭示蜕皮激素调控绿盲蝽Tre-1合成的分子机制，为发展海藻糖酶抑制剂类杀虫剂提供前期研究基础。

中文关键词： 绿盲蝽；水溶性海藻糖酶；蜕皮激素；调控；合成

英文摘要： Apolygus lucorum is the most important pest for the cotton and other industrial crops in china,trehalose is the principal sugar circulating in the blood or haemolymph, and the souble trehalase(Tre-1) is the mainly enzyme capable of hydrolyzing trehalose to its monomeric unit, glucose, and also play an important role in the synthesis of chitin, flight and other physiological activity.Previous studies had showned that,20-hydroxyecdysone(20-E) could induce the high expression of Tre-1,but its transcriptional signal conduction mechanism is still not clear.Therefore, this project would carry out the following research in the basis of obtained Apolygus lucorum Tre-1 gene and recombinant protein, to : firstly, cloned 20-E receptor gene EcR and USP, clear the vivo expression; and then through the RNA interference of EcR gene and USP gene inhibits the expression and the expression of 20-E micro injection the up-regulated after calibration,analyze Tre-1 enzyme activity, mRNA and protein expression differences by using the dual-spike-inqPCR method; finally, construction of Tre-1 gene promoter luciferase gene reporter plasmid control, to elucidate the regulatory function of the transcription activity of 20-E promoter of Tre-1 gene through the analysis of luciferase activity difference.The results of the project, expected to

英文关键词： Apolygus lucorum；soluble trehalase；20-E；regulation；synthetise