Stable expression and functional characterisation of the diamondback moth ryanodine receptor G4946E variant conferring resistance to diamide insecticides.
Stable expression and functional characterisation of the diamondback moth ryanodine receptor G4946E variant conferring resistance to diamide insecticides.
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DOI：
10.1038/srep14680
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发表时间：
2015-10-01
影响因子：
4.6
通讯作者：
Davies TG
中科院分区：
文献类型：
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作者：
Troczka BJ;Williams AJ;Williamson MS;Field LM;Lüemmen P;Davies TG
二酰胺类化合物，如氟虫双酰胺和氯虫苯甲酰胺，属于一类新的化学杀虫剂，可作为昆虫ryanodine受体（RyRs）的构象敏感激活剂。这两种化合物都被注册用于对抗鳞翅目物种，例如小菜蛾、小菜蛾（Plutella xylostella），小菜蛾是十字花科作物的臭名昭著的全球害虫。最近获得的耐二酰胺杀虫剂在这个物种被认为是由于一个目标位点突变赋予氨基酸取代（G4946E），位于跨膜结构域的RyR，虽然这种突变的确切作用尚未完全确定。为了解决这个问题，我们克隆了编码小菜蛾RyR的全长cDNA，并建立了稳定表达野生型RyR或G4946E变体的克隆Sf9细胞系，以测试重组受体对氟虫双酰胺和氯虫苯甲酰胺的敏感性。我们报告说，这两个二酰胺的疗效显着降低克隆Sf9细胞稳定表达G4946E修饰的RyR，提供明确的功能证据表明，G4946E RyR突变损害二酰胺杀虫剂结合。
Diamides, such as flubendiamide and chlorantraniliprole, belong to a new chemical class of insecticides that act as conformation-sensitive activators of insect ryanodine receptors (RyRs). Both compounds are registered for use against lepidopteran species such as the diamondback moth, Plutella xylostella, a notorious global pest of cruciferous crops. Recently acquired resistance to diamide insecticides in this species is thought to be due to a target-site mutation conferring an amino acid substitution (G4946E), located within the trans-membrane domain of the RyR, though the exact role of this mutation has not yet been fully determined. To address this we have cloned a full-length cDNA encoding the P. xylostella RyR and established clonal Sf9 cell lines stably expressing either the wildtype RyR or the G4946E variant, in order to test the sensitivity to flubendiamide and chlorantraniliprole on the recombinant receptor. We report that the efficacy of both diamides was dramatically reduced in clonal Sf9 cells stably expressing the G4946E modified RyR, providing clear functional evidence that the G4946E RyR mutation impairs diamide insecticide binding.