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双股 RNA 注射和喂食对摩门蟋蟀(直翅目:螽斯科)的不同反应。

Differential responses to double-stranded RNA injection and feeding in Mormon cricket (Orthoptera: Tettigoniidae).

机构信息

Department of Entomology, Texas A&M University, College Station, TX, USA.

Department of Pharmaceutical Sciences, The University of Oklahoma Health Sciences, Center, Oklahoma City, OK, USA.

出版信息

J Insect Sci. 2023 Jul 1;23(4). doi: 10.1093/jisesa/iead063.

Abstract

The Mormon cricket, Anabrus simplex, is a flightless katydid, one of the major devastating rangeland pests in several states of the western United States. During the past few years, their sudden and periodic outbreaks into massive migratory bands caused significant economic losses to the rangeland forage and agricultural crops, particularly grain crops. Current population management methods rely heavily on broad-spectrum chemical insecticides, which could be toxic to nontargets, and even the targeted species might develop resistance in the long run. Therefore, we assessed the potential of RNA interference (RNAi)-based alternative management strategies that could supplement the current methods. In insects, RNAi efficiency varies with the method of double-stranded RNA (dsRNA) delivery. We tested 2 different methods of dsRNA delivery: injection and oral feeding of dsRNA. The results showed that Mormon crickets are sensitive to injection of dsRNA in a dose-dependent manner, but refractory to the oral feeding of dsRNA. Further, we confirmed the high nuclease activity in the insect midgut. In order to protect the dsRNA from the dsRNase activity and facilitate its uptake in the midgut, we encapsulated dsRNA inside poly lactic-co-glycolic acid (PLGA) nanoparticles and studied its release kinetics and RNAi efficiency by oral feeding. The release kinetics clearly suggested that the PLGA nanoparticle permeates from the insect digestive system to the hemolymph; however, it failed to induce an efficient RNAi response of the targeted genes. In conclusion, our findings suggest the different responses to dsRNA delivery methods in Mormon crickets, and further investigations involving dsRNA stability and its uptake mechanism are required to use RNAi as an alternative Mormon cricket population management strategy.

摘要

刺胫蝗,又名摩门蟋蟀,是一种不能飞行的螽斯,也是美国西部几个州的主要破坏性牧场害虫之一。在过去的几年里,它们突然周期性地大量迁徙,给牧场草料和农业作物,特别是粮食作物造成了巨大的经济损失。目前的种群管理方法主要依赖于广谱化学杀虫剂,这些杀虫剂可能对非目标物有毒,甚至目标物种也可能长期产生抗药性。因此,我们评估了基于 RNA 干扰 (RNAi) 的替代管理策略的潜力,这些策略可以补充当前的方法。在昆虫中,RNAi 的效率因双链 RNA (dsRNA) 传递方法而异。我们测试了两种不同的 dsRNA 传递方法:dsRNA 的注射和口服。结果表明,摩门蟋蟀对 dsRNA 的注射剂量呈依赖性敏感,但对 dsRNA 的口服摄入具有抗性。此外,我们证实昆虫中肠具有高核酸酶活性。为了保护 dsRNA 免受 dsRNase 活性的影响,并促进其在中肠中的摄取,我们将 dsRNA 包裹在聚乳酸-共-羟基乙酸 (PLGA) 纳米颗粒中,并通过口服喂养研究其释放动力学和 RNAi 效率。释放动力学清楚地表明,PLGA 纳米颗粒从昆虫消化系统渗透到血淋巴中;然而,它未能诱导靶向基因的有效 RNAi 反应。总之,我们的研究结果表明,刺胫蝗对 dsRNA 传递方法的反应不同,需要进一步研究 dsRNA 的稳定性及其摄取机制,以便将 RNAi 用作替代摩门蟋蟀种群管理策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5555/10393271/87719aadd29e/iead063_fig1.jpg

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