背景:Grapholitamolesta是世界范围内一种重要且有害的水果害虫,有广泛的喂养宿主。胰蛋白酶,昆虫肠道中不可或缺的水解消化蛋白酶,对消化至关重要,成长和发展。我们分析了胰蛋白酶编码基因的特征,筛选由纳米载体介导的RNAi的最佳剂量,并调查了G.molesta幼虫生长发育的各种指标。
结果:肠道含量(GC)和RNA酶A降解的双链RNA(dsRNA),在较高浓度下降解速率较快。星形聚阳离子(SPc)纳米材料通过阴离子-阳离子结合保护dsGFP免受降解,并且不会通过琼脂糖凝胶迁移。胰蛋白酶编码基因的关键保守基序相似,与其他鳞翅目昆虫具有很高的同源性。用0.05μgdsRNA的SPc纳米材料介导的RNA干扰实现了约70%的干扰效率。连续干扰效率稳定。胰蛋白酶活性,8日龄幼虫的体重,p的重量和出苗率显着降低,幼虫期明显延长。
结论:所研究的胰蛋白酶基因是G的生长发育的关键靶基因。我们研究了在昆虫功能研究中饲喂SPc纳米材料的效率和便利性。我们的结果为开发有效的胰蛋白酶靶向农药提供了有价值的数据。©2024化学工业学会。
BACKGROUND: Grapholita molesta is an important and harmful fruit pest worldwide, with widespread feeding hosts.
Trypsin, an indispensable hydrolytic digestive protease in the insect gut, is crucial in digestion, growth and development. We analyzed the characteristics of the
trypsin-encoding genes, screened for the optimal dose of RNAi mediated by nanocarriers, and investigated various indices of larval growth and development of G. molesta.
RESULTS: Gut content (GC) and RNase A degraded double-stranded RNA (dsRNA), with a faster degradation rate at higher concentrations. Star polycation (SPc) nanomaterials protected dsGFP from degradation by anion-cation binding and did not migrate through agarose gel. The key conserved motifs of the
trypsin-encoding genes were similar, exhibiting high homology with those in other lepidopteran insects. An interference efficiency of ≈70% was achieved with SPc nanomaterial-mediated RNA interference with 0.05 μg dsRNA. The efficiency of continuous interference was stable.
Trypsin activity, body weight of 8-day-old larvae, pupal weight and emergence rate were significantly reduced, and the larval stage was significantly prolonged.
CONCLUSIONS: The investigated trypsin gene is a key target gene in the growth and development of G. molesta. We investigated the efficiency and convenience of feeding SPc nanomaterials in a functional study of insects. Our results provide valuable data for the development of efficient
trypsin-targeting pesticides. © 2024 Society of Chemical Industry.