Laboratory of Agrozoology, Department Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
Pest Manag Sci. 2022 Sep;78(9):3815-3822. doi: 10.1002/ps.6732. Epub 2021 Dec 8.
N-glycosylation is one of the most common and important post-translational modifications in the eukaryotic cell. The study of protein N-glycosylation in several model insects confirmed the importance of this process in insect development, immunity, survival and fertility. The Colorado potato beetle (Leptinotarsa decemlineata) (CPB) is a common pest of Solanaceae crops. With the infamous title of champion of insecticide resistance, novel pest control strategies for this insect are needed. Luckily this pest insect is reported as very sensitive for the post-genomic technology of RNA interference (RNAi).
In this project, we investigated the importance of N-glycosylation in the survival and development of CPB using RNAi-mediated gene silencing of N-glycosylation-related genes (NGRGs) during the different transition steps from the larva, through the pupa to the adult stage. High mortality was observed in the larval stage with the silencing of early NGRGs, as STT3a, DAD1 and GCS1. With dsRNA against middle NGRGs, abnormal phenotypes at the ecdysis process and adult formation were observed, while the silencing of late NGRGs did not cause mortality.
The lethal phenotypes observed on silencing of the genes involved in the early processing steps of the N-glycosylation pathway suggest these genes are good candidates for RNAi-mediated control of CPB. Next to the gene-specific mechanism of RNAi for biosafety and possible implementation in integrated pest management, we believe these early NGRGs provide a possible alternative to the well-known target genes Snf7 and vacuolar ATPases that are now used in the first commercial RNAi-based products and thus they may be useful in the context of proactive resistance management. © 2021 Society of Chemical Industry.
N-糖基化是真核细胞中最常见和最重要的翻译后修饰之一。在几种模式昆虫中对蛋白质 N-糖基化的研究证实了这一过程在昆虫发育、免疫、生存和繁殖中的重要性。科罗拉多马铃薯甲虫（Leptinotarsa decemlineata）（CPB）是茄科作物的常见害虫。这种昆虫以臭名昭著的杀虫剂抗性冠军而闻名，因此需要新的害虫防治策略。幸运的是，这种害虫对 RNA 干扰（RNAi）的后基因组技术非常敏感。
在本项目中，我们通过 RNAi 介导的基因沉默，研究了 N-糖基化相关基因（NGRGs）在幼虫、蛹到成虫不同发育阶段的生存和发育中的重要性。早期 NGRGs（如 STT3a、DAD1 和 GCS1）的沉默导致幼虫死亡率很高。用针对中间 NGRGs 的 dsRNA 处理，会观察到蜕皮过程和成虫形成的异常表型，而晚期 NGRGs 的沉默则不会导致死亡。
沉默 N-糖基化途径早期加工步骤相关基因观察到的致死表型表明这些基因是 CPB RNAi 介导控制的良好候选基因。除了 RNAi 的基因特异性机制对生物安全性的影响以及在综合虫害管理中的潜在应用外，我们还认为这些早期 NGRGs 为 Snf7 和液泡型 ATP 酶等知名目标基因提供了一种替代方案，这些基因目前用于第一批商业化的 RNAi 产品，因此在积极的抗性管理方面可能会很有用。 © 2021 化学工业学会。
