【作者】 罗雁婕； 杨振国； 谢道燕； 丁伟； 达爱斯； 倪婧； 柴建萍； 黄平； 江秀均；

【Author】 Yan-Jie Luo;Zhen-Guo Yang;Dao-Yan Xie;Wei Ding;Ai-Si Da;Jing Ni;Jian-Ping Chai;Ping Huang;Xiu-Jun Jiang;Institute of Sericulture and Apiculture, Yunnan Academy of Agricultural Sciences;College of Plant Protection,Southwest University;

【机构】 云南省农业科学院蚕桑蜜蜂研究所； 西南大学植物保护学院；

【摘要】 朱砂叶螨是世界性害螨且对包括炔螨特在内的许多常用杀螨剂产生了抗药性。本研究在室内采用炔螨特对朱砂叶螨敏感品系进行抗性选育,得到抗性倍数达37.78倍的抗性品系(PRR34)。通过与敏感品系比较,明确了抗炔螨特品系对7中常用杀虫杀螨剂的交互抗性,通过特异性酶抑制剂和酶活测定初步明确谷胱甘肽S-转移酶参与了朱砂叶螨对炔螨特的抗性。克隆得到朱砂叶螨谷胱甘肽S-转移酶基因(TcGSTd1、TcGSTd2,、TcGSTm1、TcGSTm2、TcGSTm3,TcGSTm4和TcGSTm5),聚类分析其主要为GSTs中的delt和mu亚族。q RT-PCR结果显示,朱砂叶螨抗炔螨特品系(PRR34)相对敏感品系,其GSTs在幼螨、若螨和成螨中的表达上调,但卵期则下调。上述研究结果表明,在抗性品系(PRR34)中,GSTs介导了朱砂叶螨对炔螨特的抗性,TcGSTd1、TcGSTd2和TcGSTm2在成螨阶段发挥了重要的作用。该研究首次探讨了GSTs介导的朱砂叶螨对炔螨特的抗药性。更多还原

【Abstract】 The carmine spider mite(CSM) Tetranychus cinnabarinus has become a serious pest in China and has developed resistance to acaricide propargite as it is used to control mites worldwide including T. cinnabarinus. In this study, a resistant colony of T. cinnabarinus, PRR34(37.78-fold resistant ratio), was established after 34 generations of propargite selection, and cross-resistance patterns of 7 other acaricides were determined in comparison with a susceptible strain(SS). The contribution of detoxification enzymes to propargite tolerance were investigated using biological, biochemical and molecular approaches. Enzyme inhibitor synergist tests suggested glutathione S-transferases(GST) involvement in propargite-resistance of PRR34, and GST activity against 1-chloro-2,4-dinitrobenzene(CDNB) was correlated with the development of resistance. Eight novel GST genes(TcGSTd1,TcGSTd2, TcGSTm1, TcGSTm2, TcGSTm3, TcGSTm4 and TcGSTm5) were cloned, and phylogenetic analysis showed that the eight GST genes were most closely related to GST family delta and mu from T. urticae. Quantitative RT-PCR revealed that the expression level of GSTs in PPR34 strain increased in larvae, nymphs and adults, while decreased in eggs compared with that of SS.Collectively, these results support a role of GSTs in mediating resistance to propargite in the PRR34 strain. TcGSTd1,TcGSTd2 and TcGSTm2 genes might play significant roles in propargite resistance of CSM, especially at adult stage. This is the first attempt to define specific genes involved in GST mediated propargite resistance of T. cinnabarinus at the transcriptional level.更多还原