基于 RNA 干扰管理甘蓝茎跳甲的有效目标基因。

IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Insect Molecular Biology Pub Date : 2024-07-06 DOI:10.1111/imb.12942
Doga Cedden, Gözde Güney, Xavier Debaisieux, Stefan Scholten, Michael Rostás, Gregor Bucher
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引用次数: 0

摘要

卷心菜茎跳甲(CSFB,Psylliodes chrysocephala)是油菜的主要害虫。出于对环境的担忧,欧盟禁止使用新烟碱类杀虫剂,而且出现了对除虫菊酯产生抗性的种群,这使得对 CSFB 的控制极具挑战性。为了寻找解决方案,我们最近发现 RNA 干扰(RNAi)在 CSFB 的防治中具有潜力。然而,以前测试过的 RNAi 介导的害虫控制目标基因(后称目标基因)表现出中度和缓慢的致死效应。在本研究中,我们利用对蓖麻蒺藜进行全基因组 RNAi 筛选的结果,口服了 27 种双链 RNA(dsRNA),以确定 CSFB 成虫的高效靶基因。我们使用 500 ng dsRNA 筛选出了 10 个中度有效(死亡率大于 50%)和 4 个高效靶基因(8-13 天内死亡率达 100%)。后者主要包括蛋白酶体亚基。基因表达测量证实了靶基因的沉默,剂量反应研究显示,单次暴露于 dsRNA 后 14 天内的半数致死剂量(LD50)低至约 20 毫微克。四种高效的 dsRNA 还能抑制叶片损伤(高达约 75%),其中一种还能影响运动。研究人员在非目标生物(如蜜蜂等有益生物)中对有希望的目标基因序列进行了硅学目标预测,从而设计出环境友好型 dsRNA。总之,这项研究为开发基于 dsRNA 的 CSFB 杀虫剂提供了宝贵的见解。
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Effective target genes for RNA interference-based management of the cabbage stem flea beetle.

The cabbage stem flea beetle (CSFB, Psylliodes chrysocephala) is a key pest of oilseed rape. The ban on neonicotinoids in the European Union due to environmental concerns and the emergence of pyrethroid-resistant populations have made the control of CSFB extremely challenging. In search of a solution, we have recently shown that RNA interference (RNAi) has potential in the management of CSFB. However, the previously tested target genes for RNAi-mediated pest control (subsequently called target genes) exhibited moderate and slow-acting lethal effects. In this study, 27 double-stranded RNAs (dsRNAs) were orally delivered to identify highly effective target genes in CSFB adults by leveraging the findings of a genome-wide RNAi screen in Tribolium castaneum. Our screen using 500 ng of dsRNA identified 10 moderately effective (> 50% mortality) and 4 highly effective target genes (100% mortality in 8-13 days). The latter mainly included proteasome subunits. Gene expression measurements confirmed target gene silencing and dose-response studies revealed LD50 values as low as ~20 ng in 14 days following a single exposure to dsRNA. Four highly effective dsRNAs also inhibited leaf damage (up to ~75%) and one affected locomotion. The sequences of promising target genes were subjected to in silico target prediction in non-target organisms, for example, beneficials such as honeybees, to design environmentally friendly dsRNAs. Overall, the study provides valuable insights for the development of dsRNA-based insecticides against CSFB.

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来源期刊
Insect Molecular Biology
Insect Molecular Biology 生物-昆虫学
CiteScore
4.80
自引率
3.80%
发文量
68
审稿时长
6-12 weeks
期刊介绍: Insect Molecular Biology has been dedicated to providing researchers with the opportunity to publish high quality original research on topics broadly related to insect molecular biology since 1992. IMB is particularly interested in publishing research in insect genomics/genes and proteomics/proteins. This includes research related to: • insect gene structure • control of gene expression • localisation and function/activity of proteins • interactions of proteins and ligands/substrates • effect of mutations on gene/protein function • evolution of insect genes/genomes, especially where principles relevant to insects in general are established • molecular population genetics where data are used to identify genes (or regions of genomes) involved in specific adaptations • gene mapping using molecular tools • molecular interactions of insects with microorganisms including Wolbachia, symbionts and viruses or other pathogens transmitted by insects Papers can include large data sets e.g.from micro-array or proteomic experiments or analyses of genome sequences done in silico (subject to the data being placed in the context of hypothesis testing).
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