Liposome encapsulation and EDTA formulation of dsRNA targeting essential genes increase oral RNAi-caused mortality in the Neotropical stink bug Euschistus heros
Liposome encapsulation and EDTA formulation of dsRNA targeting essential genes increase oral RNAi-caused mortality in the Neotropical stink bug Euschistus heros
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DOI：
10.1002/ps.5167
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发表时间：
2019-02-01
影响因子：
4.1
通讯作者：
Christiaens, Olivier
中科院分区：
文献类型：
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作者：
Castellanos, Nathaly L.;Smagghe, Guy;Christiaens, Olivier
背景新热带臭虫Euschistus heros是大豆田的主要害虫。基于RNA干扰(RNAi)的高度物种特异性农药的开发可以提供一种新的可持续和环境友好的防治策略。结果评价了RNAi作为害虫防治工具的潜力。首先，使用显微注射方法进行靶基因选择。在测试的15个候选基因中，有7个在血淋巴注射27.5 ng dsRNA后显示出95%的死亡率。随后，用不同的制剂口服dsRNA：裸dsRNA、脂质体包裹的dsRNA和EDTA形成的dsRNA。脂质体包裹的针对vATPase A和肌肉肌动蛋白的dsRNA在14天后导致显著的死亡率(分别为45%和42%)，而EDTA形成的dsRNA只对其中一个靶基因产生了影响。对采集唾液中dsRNA稳定性的体外分析表明，英雄E.heros唾液具有很强的dsRNA降解能力，这可以解释dsRNA制剂的必要性。结论结果表明，与EDTA或脂质体包裹的dsRNA一起持续摄入dsRNA可以防止dsRNA被酶降解，并表明这些制剂在dsRNA传递中具有巨大的潜力，可以将RNAi用作功能基因组学工具或用于害虫防治臭虫。(三)2018年化学工业学会
BACKGROUNDThe Neotropical stink bug Euschistus heros is a major pest in soybean fields. Development of highly species-specific pesticides based on RNA interference (RNAi) could provide a new sustainable and environmentally friendly control strategy.RESULTSHere, the potential of RNAi as a pest control tool against E. heros was assessed. First, target gene selection using a microinjection approach was performed. Seven of the 15 candidate genes tested exhibited >95% mortality after hemolymph injection of 27.5ng dsRNA. Subsequently, dsRNA was administered orally using different formulations: naked dsRNA, liposome-encapsulated-dsRNA and dsRNA formulated with EDTA. Liposome-encapsulated dsRNA targeting vATPase A and muscle actin led to significant mortality after 14days (45% and 42%, respectively), whereas EDTA-formulated dsRNA did so for only one of the target genes. Ex vivo analysis of the dsRNA stability in collected saliva indicated a strong dsRNA-degrading capacity by E. heros saliva, which could explain the need for dsRNA formulations.CONCLUSIONThe results demonstrate that continuous ingestion of dsRNA with EDTA or liposome-encapsulated dsRNA can prevent dsRNA from being degraded enzymatically and suggest great potential for using these formulations in dsRNA delivery to use RNAi as a functional genomics tool or for pest management of stink bugs. (c) 2018 Society of Chemical Industry