Istituto per la Protezione Sostenibile Delle Piante, Consiglio Nazionale Delle Ricerche, IPSP-CNR, Strada delle Cacce 73, 10135 Torino, Italy.
Environmental Research and Innovation Department (ERIN), Luxembourg Institute of Science and Technology (LIST), 41 Rue du Brill, 4422 Luxembourg, Luxembourg.
Int J Mol Sci. 2022 Jan 11;23(2):765. doi: 10.3390/ijms23020765.
(Hemiptera: ) is the natural vector of Flavescence dorée phytoplasma, a quarantine pest of grapevine with severe impact on European viticulture. RNA interference (RNAi) machinery components are present in transcriptome and injection of ATP synthase β dsRNAs into adults caused gene silencing, starting three days post injection (dpi) up to 20 dpi, leading to decrease cognate protein. Silencing of this gene in the closely related leafhopper previously showed female sterility and lack of mature eggs in ovaries. Here, alteration of developing egg morphology in ovaries as well as overexpression of hexamerin transcript (amino acid storage protein) and cathepsin L protein (lysosome proteinase) were observed in dsATP-injected females. To evaluate RNAi-specificity, was used as dsRNA-receiving model-species. Different doses of two sets of dsRNA-constructs targeting distinct portions of ATP synthase β gene of both species induced silencing, lack of egg development, and female sterility in , indicating that off-target effects must be evaluated case by case. The effectiveness of RNAi in provides a powerful tool for functional genomics of this non-model species and paves the way toward RNAi-based strategies to limit vector population, despite several technical and regulatory constraints that still need to be overcome to allow open field application.
(半翅目: )是 Flavescence dorée 植原体的天然载体,该植原体是葡萄藤的检疫害虫,对欧洲葡萄种植业有严重影响。RNA 干扰(RNAi)机制组件存在于 转录组中,将 ATP 合酶βdsRNA 注入成虫会导致基因沉默,从注射后 3 天(dpi)开始,持续到 20 dpi,导致同源蛋白减少。先前在密切相关的叶蝉中沉默该基因导致雌性不育和卵巢中无成熟卵。在这里,在 dsATP 注射的雌虫中观察到卵巢中正在发育的卵形态发生改变,以及六聚体转录本(氨基酸储存蛋白)和组织蛋白酶 L 蛋白(溶酶体蛋白酶)的过度表达。为了评估 RNAi 的特异性, 被用作 dsRNA 接收模型物种。两种针对两种物种的 ATP 合酶β基因不同部分的 dsRNA 构建体的不同剂量均诱导了沉默、卵发育缺乏和 雌性不育,表明必须逐个评估脱靶效应。在 中 RNAi 的有效性为该非模型物种的功能基因组学提供了有力工具,并为基于 RNAi 的策略来限制载体种群铺平了道路,尽管仍需要克服一些技术和监管限制,以允许在野外应用。