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口服细菌表达的双链RNA可使一种极具入侵性的树木致死害虫——翡翠灰螟的基因沉默并导致其死亡。

Oral Ingestion of Bacterially Expressed dsRNA Can Silence Genes and Cause Mortality in a Highly Invasive, Tree-Killing Pest, the Emerald Ash Borer.

作者信息

Leelesh Ramya Shanivarsanthe, Rieske Lynne K

机构信息

Department of Entomology, University of Kentucky, Lexington, KY 40546-0091, USA.

School of Life Sciences, University of Bedfordshire, Luton LU13JU, UK.

出版信息

Insects. 2020 Jul 14;11(7):440. doi: 10.3390/insects11070440.

Abstract

RNA interference (RNAi) is a naturally occurring process inhibiting gene expression, and recent advances in our understanding of the mechanism have allowed its development as a tool against insect pests. A major challenge for deployment in the field is the development of convenient and efficient methods for production of double stranded RNA (dsRNA). We assessed the potential for deploying bacterially produced dsRNA as a bio-pesticide against an invasive forest pest, the emerald ash borer (EAB). EAB feeds on the cambial tissue of ash trees ( spp.), causing rapid death. EAB has killed millions of trees in North America since its discovery in 2002, prompting the need for innovative management strategies. In our study, bacterial expression and synthesis of dsRNA were performed with strain HT115 using the L4440 expression vector. EAB-specific dsRNAs ( and ) over-expressed in were toxic to neonate EAB after oral administration, successfully triggering gene silencing and subsequent mortality; however, a non-specific dsRNA control was not included. Our results suggest that ingestion of transformed expressing dsRNAs can induce an RNAi response in EAB. To our knowledge, this is the first example of an effective RNAi response induced by feeding dsRNA-expressing bacteria in a forest pest.

摘要

RNA干扰(RNAi)是一种天然存在的抑制基因表达的过程,而我们对其机制理解的最新进展使其得以发展成为一种防治害虫的工具。在田间应用的一个主要挑战是开发方便高效的双链RNA(dsRNA)生产方法。我们评估了将细菌产生的dsRNA作为生物杀虫剂来防治一种入侵性森林害虫——翡翠灰螟(EAB)的潜力。翡翠灰螟以白蜡树(白蜡属)的形成层组织为食,导致树木迅速死亡。自2002年被发现以来,翡翠灰螟已在北美杀死了数百万棵树木,这促使人们需要创新的管理策略。在我们的研究中,使用L4440表达载体,利用HT115菌株进行dsRNA的细菌表达和合成。在大肠杆菌中过表达的EAB特异性dsRNAs(……此处原文缺失具体序列信息)经口服后对初孵出的EAB有毒性,成功引发基因沉默并导致后续死亡;然而,未设置非特异性dsRNA对照。我们的结果表明,摄入表达dsRNAs的转化大肠杆菌可在翡翠灰螟中诱导RNAi反应。据我们所知,这是在森林害虫中通过喂食表达dsRNA的细菌诱导有效RNAi反应的首个实例。

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