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RNAi 方法的开发以控制鬼脸天牛,Murgantia histrionica。

Development of RNAi methods to control the harlequin bug, Murgantia histrionica.

机构信息

Department of Entomology, University of Kentucky, Lexington, Kentucky.

出版信息

Arch Insect Biochem Physiol. 2020 Aug;104(4):e21690. doi: 10.1002/arch.21690. Epub 2020 May 11.

Abstract

The harlequin bug (HB), Murgantia histrionica, is a major pest of cabbage family plants throughout its range in the United States. RNA interference (RNAi) is a posttranscriptional gene silencing mechanism that is showing promise as a biopesticide due to the ability to target species-specific genes necessary for growth and/or survival with synthetic double-stranded RNA (dsRNA). In the present study, dsRNA stability assays revealed that nucleases present in the saliva of harlequin bugs did not rapidly degrade dsRNA. We tracked the movement and localization of radioactively labeled dsRNA in both mustard plant seedlings and harlequin bug nymphs that fed on treated host plants. Movement of P-labeled-dsRNA from soil to plant and plant to insect was detected. The efficacy of RNAi in inducing mortality in harlequin bug adults and nymphs injected or fed with dsRNA targeting inhibitor of apoptosis (IAP), ATPase N2B (ATPase), serine/threonine-protein phosphatase PP1-β catalytic subunit (PP1), signal recognition particle 54 kDa protein (SRP), and G protein-coupled receptor 161-like (GPCR) genes was evaluated. Injection of dsRNA targeting candidate genes into adults caused between 40% and 75% mortality and induced significant knockdown of target gene expression. Feeding dsRNA targeting the IAP gene to nymphs by plant-mediated and droplet feeding methods induced knockdown of the target gene and caused 40-55% mortality. These findings suggest that RNAi may be a viable approach for managing this pest.

摘要

红猎蝽(HB),Murgantia histrionica,是美国范围内十字花科植物的主要害虫。RNA 干扰(RNAi)是一种转录后基因沉默机制,由于能够针对生长和/或生存所需的特定物种基因合成双链 RNA(dsRNA),因此作为一种生物农药具有广阔的前景。在本研究中,dsRNA 稳定性测定表明,红猎蝽唾液中存在的核酸酶不会迅速降解 dsRNA。我们跟踪了放射性标记的 dsRNA 在芥菜幼苗和取食处理过的宿主植物的猎蝽若虫中的运动和定位。从土壤到植物和植物到昆虫都检测到了 P 标记-dsRNA 的移动。通过注射或喂食 dsRNA 靶向凋亡抑制剂(IAP)、ATPase N2B(ATPase)、丝氨酸/苏氨酸蛋白磷酸酶 PP1-β 催化亚基(PP1)、信号识别颗粒 54 kDa 蛋白(SRP)和 G 蛋白偶联受体 161 样(GPCR)基因,评估了 RNAi 在诱导红猎蝽成虫和若虫死亡中的功效。将针对候选基因的 dsRNA 注射到成虫中会导致 40%至 75%的死亡率,并显著下调靶基因的表达。通过植物介导和液滴喂养方法用靶向 IAP 基因的 dsRNA 喂养若虫会导致靶基因的下调并导致 40-55%的死亡率。这些发现表明 RNAi 可能是一种可行的管理这种害虫的方法。

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