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靶向兰尼碱受体的 dsRNA 的离体递送用于控制烟粉虱。

Ex vivo delivery of dsRNA targeting ryanodine receptors for control of Tuta absoluta.

机构信息

School of Natural and Environmental Sciences, Newcastle University, Newcastle upon Tyne, UK.

出版信息

Pest Manag Sci. 2024 Dec;80(12):6400-6408. doi: 10.1002/ps.8368. Epub 2024 Aug 16.

DOI:10.1002/ps.8368
PMID:39148493
Abstract

BACKGROUND

RNA interference (RNAi) is an endogenous eukaryote viral defence mechanism representing a unique form of post-transcriptional gene silencing. Owing to its high specificity, this technology is being developed for use in dsRNA-based biopesticides for control of pest insects. Whilst many lepidopteran species are recalcitrant to RNAi, Tuta absoluta, a polyphagous insect responsible for extensive crop damage, is sensitive. Ryanodine receptors (RyRs) are intracellular calcium channels regulating calcium ion (Ca) release. The chemical pesticide class of diamides functions agonistically against lepidopteran RyR, resulting in uncontrolled Ca release, feeding cessation and death. Resistance to diamides has emerged in T. absoluta, derived from RyR point mutations.

RESULTS

RNAi was used to target RyR transcripts of T. absoluta. Data presented here demonstrate the systemic use of exogenous T. absoluta RyR-specific (TaRy) dsRNA in tomato plants (Solanum lycopersicum) to significantly downregulate expression of the target gene, resulting in significant insect mortality and reduced leaf damage. Using a leaflet delivery system, daily dosing of 3 μg TaRy dsRNA for 72 h resulted in 50% downregulation of the target gene and 50% reduction in tomato leaf damage. Corrected larval mortality and adult emergence were reduced by 38% and 33%, respectively. TaRy dsRNA demonstrated stability in tomato leaves ≤72 h after dosing.

CONCLUSIONS

This work identifies TaRy as a promising target for RNAi control of this widespread crop pest. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

摘要

背景

RNA 干扰 (RNAi) 是一种内源性真核生物病毒防御机制,代表了一种独特的转录后基因沉默形式。由于其高度特异性,该技术正被开发用于基于双链 RNA 的生物杀虫剂,以控制害虫。虽然许多鳞翅目昆虫对 RNAi 有抗性,但多寄主昆虫桃蚜对 RNAi 敏感,它会造成严重的作物损害。Ryanodine 受体 (RyR) 是调节钙离子 (Ca) 释放的细胞内钙通道。化学农药类二酰胺类化合物通过与鳞翅目 RyR 激动剂作用,导致 Ca 释放失控、摄食停止和死亡。桃蚜对二酰胺类化合物的抗性是由 RyR 点突变引起的。

结果

使用 RNAi 靶向桃蚜的 RyR 转录本。本文介绍了外源性桃蚜 RyR 特异性 (TaRy) dsRNA 在番茄植株中的系统性使用,该方法显著下调了靶基因的表达,导致显著的昆虫死亡率和叶片损伤减少。使用小叶递送系统,每天 3μg TaRy dsRNA 剂量处理 72 小时,可使靶基因下调 50%,番茄叶片损伤减少 50%。校正的幼虫死亡率和成虫羽化率分别降低了 38%和 33%。TaRy dsRNA 在给药后≤72 小时内保持在番茄叶片中的稳定性。

结论

这项工作鉴定 TaRy 作为控制这种广泛分布的作物害虫的 RNAi 的一个有前途的靶标。

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