小菜蛾谷氨酸门控氯离子通道基因A309V点突变的功能解析

项目名称： 小菜蛾谷氨酸门控氯离子通道基因A309V点突变的功能解析

项目编号： No.31301693

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： 王兴亮

作者单位： 南京农业大学

项目金额： 23万元

中文摘要： 阿维菌素的大面积推广应用曾使难以治理的小菜蛾得到有效控制，然而小菜蛾对阿维菌素选择压力的适应性进化使该杀虫剂的可持续防控效能面临重大挑战，我国部分地区小菜蛾田间种群已对阿维菌素产生高水平抗性。明确小菜蛾对阿维菌素产生高水平抗性的分子机制，是开展小菜蛾抗性监测与治理的重要基础性工作。本项目以对阿维菌素极高抗的小菜蛾近等基因系为材料，以发现的与抗性连锁的小菜蛾谷氨酸氯离子通道（PxGluCl）基因A309V点突变为切入点，将目的基因导入爪蟾卵母细胞进行功能表达，并进行神经电生理和配基结合分析，以明确PxGluCl基因A309V点突变对阿维菌素结合能力及神经敏感性的影响，从而从功能上明确PxGluCl是阿维菌素的作用靶点以及PxGluCl基因A309V突变致使小菜蛾对阿维菌素产生高水平抗性的分子机理。

中文关键词： 小菜蛾；谷氨酸门控氯离子通道；基因突变；抗药性；电生理学

英文摘要： Abamectin is one of the most effectively and widely used insecticide for diamondback moth (Plutella xylostella) management. However, the sustainable control efficacy of abamectin has been threatened due to strong potential of resistance evolution in this pest.The diamondback moth has evolved high levels of resistance to abamectin in some locations of China. It is therefore essential to investigate the molecular mechanisms of abamectin resistance to facilitate implementing effective resistance monitoring and management strategies in diamondback moth. In this study, we will make use of an isogenic abamectin-resistant strain (Roth-Abm) to investigate molecular mechanisms of abamectin resistance. Our previous work showed that the A309V point mutation in the glutamate-gated chloride channel (PxGluCl)is genetically linked with abamectin resistance in the Roth-Abm strain. To characterize the effect of the mutation on biophysical properties of PxGluCl channel, the wild-type and mutant alleles of PxGluCl will be expressed in Xenopus oocytes, and electrophysiological and radioligand binding studies will be carried out. Our work is expected to demonstrate that PxGluCl is the major target-site of abamectin, and provide functional evidence that the A309V mutation is responsible for high levels of resistance to abamectin in t

英文关键词： Diamondback moth；Glutamate-gated chloride channel；Gene mutation；Resistance；Electrophysiology