Profiling of RNAi sensitivity after foliar dsRNA exposure in different European populations of Colorado potato beetle reveals a robust response with minor variability,Pesticide Biochemistry and Physiology

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Profiling of RNAi sensitivity after foliar dsRNA exposure in different European populations of Colorado potato beetle reveals a robust response with minor variability
Pesticide Biochemistry and Physiology ( IF 4.2 ) Pub Date : 2020-06-01 , DOI: 10.1016/j.pestbp.2020.104569
Sonja G Mehlhorn ₁ , Sven Geibel ₂ , Gregor Bucher ₃ , Ralf Nauen ₂

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In recent years, substantial effort was spent on the exploration and implementation of RNAi technology using double-stranded RNA (dsRNA) for pest management purposes. However, only few studies investigated the geographical variation in RNAi sensitivity present in field-collected populations of the targeted insect pest. In this baseline study, 2nd instar larvae of 14 different European populations of Colorado potato beetle (CPB), Leptinotarsa decemlineata, collected from nine different countries were exposed to a foliarly applied diagnostic dose of dsactin (dsact) to test for possible variations in RNAi response. Only minor variability in RNAi sensitivity was observed between populations. However, the time necessary to trigger a dsRNA-mediated phenotypic response varied significantly among populations, indicated by significant differences in mortality figures obtained five days after treatment. An inbred German laboratory reference strain D01 and a Spanish field strain E02 showed almost 100% mortality after foliar exposure to 30 ng dsactin (equal to 0.96 g/ha), whereas another Spanish strain E01 was least responsive and showed only 30% mortality. Calculated LD50-values for foliarly applied dsact against strains D01 (most sensitive) and E01 (least sensitive) were 9.22 and 68.7 ng/leaf disc, respectively. The variability was not based on target gene sequence divergence or knock-down efficiency. Variability in expression of the core RNAi machinery genes dicer (dcr2a) and argonaute (ago2a) was observed but did not correlate with sensitivity. Interestingly, RT-qPCR data collected for all strains revealed a strong correlation between the expression level of dcr2a and ago2a (r 0.93) as well as ago2a and stauC (r 0.94), a recently described dsRNA binding protein in Coleopterans. Overall, this study demonstrates that sensitivity of CPB to sprayable RNAi slightly varies between strains but also shows that foliar RNAi as a control method works against all tested CPB populations collected across a broad geographic range in Europe. Thus, underpinning the potential of RNAi-based CPB control as a promising component in integrated pest management (IPM) and resistance management programs.

中文翻译：

对不同欧洲科罗拉多马铃薯甲虫种群的叶面 dsRNA 暴露后 RNAi 敏感性的分析揭示了一种具有微小变异性的稳健反应

近年来，人们在利用双链 RNA (dsRNA) 进行害虫管理的 RNAi 技术的探索和实施上投入了大量精力。然而，只有少数研究调查了现场采集的目标害虫种群中存在的 RNAi 敏感性的地理变异。在这项基线研究中，从 9 个不同国家收集的 14 个不同欧洲科罗拉多马铃薯甲虫 (CPB) Leptinotarsa decemlineata 种群的 2 龄幼虫暴露于叶面施用诊断剂量的 dsactin (dsact) 以测试 RNAi 反应的可能变化. 在种群之间仅观察到 RNAi 敏感性的微小变异。然而，触发 dsRNA 介导的表型反应所需的时间在人群中差异很大，由治疗后五天获得的死亡率数字的显着差异表明。近交德国实验室参考菌株 D01 和西班牙田间菌株 E02 在叶面暴露于 30 ng dsactin（等于 0.96 g/ha）后表现出几乎 100% 的死亡率，而另一个西班牙菌株 E01 反应最差，死亡率仅为 30%。针对菌株 D01（最敏感）和 E01（最不敏感）的叶面施用 dsact 的计算 LD50 值分别为 9.22 和 68.7 ng/叶盘。变异性不是基于目标基因序列差异或击倒效率。观察到核心 RNAi 机器基因 dicer (dcr2a) 和 argonaute (ago2a) 表达的变异性，但与敏感性无关。有趣的是，为所有菌株收集的 RT-qPCR 数据显示 dcr2a 和 ago2a (r 0.93) 以及 ago2a 和 stauC (r 0.94) 的表达水平之间存在很强的相关性，这是最近描述的鞘翅目中的 dsRNA 结合蛋白。总体而言，这项研究表明 CPB 对可喷洒的 RNAi 的敏感性在菌株之间略有不同，但也表明叶面 RNAi 作为一种控制方法适用于在欧洲广泛的地理范围内收集的所有测试 CPB 种群。因此，支持基于 RNAi 的 CPB 控制作为综合虫害管理 (IPM) 和抗性管理计划中一个有前途的组成部分的潜力。这项研究表明，CPB 对可喷洒的 RNAi 的敏感性在菌株之间略有不同，但也表明叶面 RNAi 作为一种控制方法适用于在欧洲广泛的地理范围内收集的所有测试 CPB 种群。因此，支持基于 RNAi 的 CPB 控制作为综合虫害管理 (IPM) 和抗性管理计划中一个有前途的组成部分的潜力。这项研究表明，CPB 对可喷洒的 RNAi 的敏感性在菌株之间略有不同，但也表明叶面 RNAi 作为一种控制方法适用于在欧洲广泛的地理范围内收集的所有测试 CPB 种群。因此，支持基于 RNAi 的 CPB 控制作为综合虫害管理 (IPM) 和抗性管理计划中一个有前途的组成部分的潜力。

更新日期：2020-06-01

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