目录
摘要Ⅰ
关键词Ⅰ
AbstractⅡ
引言
1 研究背景1
2 材料与方法4
2.1 供试昆虫4
2.2 供试试剂6
2.3 二化螟鱼尼丁受体基因片段的克隆7
2.3.1 二化螟总RNA的提取和cDNA的合成7
2.3.2 PCR扩增7
2.3.3 PCR产物的回收、连接、转化及测序7
2.4 二化螟田间品系突变位点的检测7
2.4.1 DNA模板的制备7
2.4.2 PCR产物的制备7
2.4.3 突变位点的突变频率计算8
3 结果与分析8
3.1 二化螟鱼尼丁受体基因突变鉴定8
3.2 田间二化螟鱼尼丁受体基因突变类型及其频率检测8
4 讨论10
5 展望11
致谢12
参考文献13
二化螟鱼尼丁受体基因突变频率的田间检测
摘 要
二化螟Chilo suppressalis（Walker）是我国稻区内分布广且为害严重的鳞翅目螟蛾科的害虫之一，双酰胺类杀虫剂自2008年后成为防治二化螟的主要药剂，虽然其防治效果优异，但由于不合理使用，导致浙、湘、赣三省二化螟对其快速产生了高水平抗性。双酰胺类杀虫剂的作用靶标较为单一，靶标基因的点突是此类药剂产生抗性的重要机制。因此，及早检测二化螟田间种群鱼尼丁受体点突变的频率，对预测该地区氯虫苯甲酰胺的抗性发展和二化螟的抗药性治理具有重要指导作用。本研究采用PCR产物直接测序的方法对2019年6个二化螟田间种群鱼尼丁受体的基因突变频率进行检测，结果显示浙江苍南种群和瑞安种群I4758M突变位点的突变频率均为100%；象山种群I4758M突变位点的突变频率为72.4%；余姚种群I4758M突变位点和Y4667C突变位点的突变频率分别为54.6%和34.4%；湖南攸县种群I4758M突变位点和Y4667C突变位点的突变频率分别达到了41.65%和12.5%，Y4667D突变位点的突变频率则为33.4%；江西南城种群Y4667D突变位点的突变频率为30%，新 *51今日免费论文网|www.51jrft.com +Q: @351916072@ 
发现的Y4891F突变位点的突变在南城种群中频率较高，达到了75%。
MUTATION FREQUENCY DETECTION OF RYANODINE RECEPTORS GENE IN FIELD POPULATION OF Chilo suppressalis(Walker)
ABSTRACT
The rice stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is one of the most widespread rice pests in China. The diamide insecticides are main pesticides for control Hepidoptera pests, which showed excellent efficacy on rice stem borer. High level resistance had been monitored in Zhejiang province in 2013 because of the overuse of chlorantraniliprole and flubendimide. The target of diamide insecticides is Ryanodine receptor (RyRs), and the point mutation of the target gene has become an important factor affecting the resistance of pests to these insecticides. Therefore, earlier detection of the point mutations frequency of ryanodine receptors gene will provide important information on predicting the resistance development of chlorantraniliprole and the resistance management of C. suppressalis. In this study, the mutation frequency of RyRs gene was detected by direct sequencing of PCR products in field populations of C. suppressalis from 6 locations in 2019. The results revealed that the mutation frequency of I4758M mutation in Cangnan population and Ruian population was 100%; the mutation frequency of I4758M mutation in Xiangshan population was lower, but still reached 72.4%; the mutation frequency of I4758M mutation and Y4667C mutation made up 54.6% and 34.4% respectively of Yuyao population; the mutation frequency of I4758M mutation and Y4667C mutation in Youxian population reached 41.65% and 12.5% respectively, and the mutation frequency of Y4667D mutation was 33.4%; the mutation frequency of Y4667D mutation accounted for 30% of Nancheng population. The mutation frequency of the newly discovered Y4891F mutation is higher, reaching 75%.

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