Investigation and characterisation of diamide insecticide resistance conferred by target-site mutations in the ryanodine receptor of lepidopteran pest

二胺类杀虫剂，如氟苯二胺和氯虫腈，作为昆虫虫胺受体（RyR）的构象敏感激活剂，从昆虫肌肉的细胞内储存中释放钙，并引起不可逆的瘫痪。这些化合物是二胺制剂的主要活性成分，用于控制各种食草昆虫，特别是鳞翅目昆虫，如小菜蛾，十字花科作物上臭名昭著的全球害虫。这种常见的害虫最近通过RyR跨膜结构域的特定突变对二胺产生了广泛的抗性。目前正在开发其他二胺类杀虫剂，如环虫酰胺、溴虫酰胺和四虫酰胺，这些杀虫剂将进一步增加对一些目标害虫的选择压力。因此，通过详细的生化和分子研究来研究这些耐药机制是一个关键的优先事项。该项目将利用最先进的方法，包括重组蛋白的细胞系表达、放射性配体结合研究、电生理记录、钙成像和CRISPR/Cas9引导果蝇转基因，描述它们对二胺类杀虫剂作用和RyR功能的影响，并将成为了解二胺类杀虫剂与RyR等位基因变异结合的基本关键。这一知识将进一步影响未来对二胺类杀虫剂的耐药性管理策略和新型杀虫RyR调节剂的发现管道。您将接受广泛适用的标准生化、分子和遗传技术方面的培训，并将从学习更专业的技术（如昆虫转基因）中受益。您还将被要求在位于德国蒙海姆的行业合作伙伴拜耳作物科学公司实习3个月。

Diamide insecticides, such as flubendiamide and chlorantraniliprole, act as conformation sensitive activators of insect ryanodine receptors (RyR), releasing calcium from intracellular stores in insect muscle, and causing an irreversible paralysis. These compounds are the main active ingredients in diamide formulations used to control a diverse range of herbivorous insects, particularly lepidopterans such as the diamondback moth, Plutella xylostella, a notorious global pest on cruciferous crops. This common pest has recently developed widespread resistance to diamides through specific mutations in the trans-membrane domain of the RyR. Additional diamide insecticides such as cyclaniliprole, broflanilide and tetraniliprole are currently under development and will further increase the selection pressure on a number of target pests. It is therefore a key priority to investigate these resistance mechanisms by means of detailed biochemical and molecular studies. This project will characterise their impact on diamide insecticide action and RyR function, using state of the art approaches including cell-line expression of recombinant proteins, radioligand binding studies, electrophysiological recordings, calcium imaging and CRISPR/Cas9 guided drosophila transgenesis, and will be key to providing a fundamental understanding of diamide insecticide binding to allelic RyR variants. This knowledge will further impact future resistance management strategies for diamides and the discovery pipeline for novel insecticidal RyR modulators. You will be trained in standard biochemical, molecular and genetic techniques that are widely applicable and will also benefit from learning more specialised techniques such as insect transgenesis. You will also be required to spend 3 months on placement with the industry partner Bayer CropScience in Monheim, Germany.