College of Plant Protection, Shanxi Agricultural University/Shanxi Key Laboratory of Integrated Pest Management in Agriculture, Taiyuan, China.
College of Plant Protection, Hunan Agricultural University, Changsha, China.
Pest Manag Sci. 2023 Jul;79(7):2482-2492. doi: 10.1002/ps.7437. Epub 2023 Mar 18.
Recently, RNA interference (RNAi)-based biopesticide, a species-specific pest control alternative, has been deregulated and commercialized in the US and Canada. The hawthorn spider mite, Amphitetranychus viennensis Zacher, is a major pest for rosaceous plants, which has been controlled primarily by synthetic pesticides. To address the emerging resistance issues in A. viennensis, we initiated a project to develop RNAi-based biopesticides.
In this study, we (i) developed a dietary RNAi system for A. viennensis using leaf disc, (ii) assessed the suitability of multiple control genes to distinguish sequence-specific silencing from non-specific effects within this RNAi system, and (iii) screened for the target gene candidates. As a result, β-Glucuronidase (GUS), an enzyme derived from E. coli and a broadly used reporter for plants is the appropriate control for A. viennensis RNAi, while green fluorescent protein (GFP), is not suitable due to its significantly higher mortality than the other controls. For target gene screening, suppression was confirmed for all the candidates, including two housekeeping genes (Vacuolar-type H + -ATPase subunit A (V-ATPase A) and Glyceraldehyde 3-phosphate dehydrogenase, (GAPDH)), and three genes associated with development (ATP-dependent RNA Helicase DDX3Y (Belle), CREB-binding protein (CBP), and Farnesoic acid O-methyltransferase (FaMet)). Knocking down of V-ATPase A resulted in the highest mortality (~ 90%) and reduced fecundity (over 90%) than other candidates. As for the genes associated with development, suppression of Belle and CBP, led to approximately 65% mortality, as well as 86% and 40% reduction in fecundity, respectively. Silencing of FaMet, however, had negligible biological impacts on A. viennensis.
The combined efforts not only establish an effective dsRNA delivery method, but also provide potential target genes for RNAi-based biopesticides against A. viennensis, a devastating invasive pest for fruit trees and woody ornamental plants throughout Asia and Europe. © 2023 Society of Chemical Industry.
最近，基于 RNA 干扰 (RNAi) 的生物农药作为一种针对特定物种的害虫防治替代方法已在美国和加拿大获得监管和商业化。山楂叶螨 (Amphitetranychus viennensis Zacher) 是蔷薇科植物的主要害虫，主要通过合成农药进行防治。为了解决山楂叶螨日益严重的抗药性问题，我们启动了一个基于 RNAi 的生物农药项目。
在这项研究中，我们 (i) 利用叶盘开发了山楂叶螨的饮食 RNAi 系统，(ii) 评估了多个对照基因在该 RNAi 系统中区分序列特异性沉默和非特异性影响的适用性，以及 (iii) 筛选了靶基因候选物。结果表明，β-葡萄糖醛酸酶 (GUS)，一种来自大肠杆菌的酶，是植物中广泛使用的报告基因，是山楂叶螨 RNAi 的合适对照，而绿色荧光蛋白 (GFP) 则因死亡率明显高于其他对照而不适用。对于靶基因筛选，所有候选物的抑制均得到确认，包括两个管家基因（液泡型 H + -ATP 酶亚基 A (V-ATPase A) 和甘油醛 3-磷酸脱氢酶 (GAPDH)）和三个与发育相关的基因（ATP 依赖性 RNA 解旋酶 DDX3Y (Belle)、CREB 结合蛋白 (CBP) 和法呢酸 O-甲基转移酶 (FaMet)）。敲低 V-ATPase A 导致的死亡率 (~90%) 和生殖力降低 (超过 90%) 均高于其他候选物。对于与发育相关的基因，Belle 和 CBP 的抑制导致约 65%的死亡率，以及分别 86%和 40%的生殖力降低。然而，FaMet 的沉默对山楂叶螨几乎没有生物学影响。
这些综合努力不仅建立了一种有效的 dsRNA 传递方法，还为针对山楂叶螨的基于 RNAi 的生物农药提供了潜在的靶基因，山楂叶螨是一种在亚洲和欧洲对果树和木本观赏植物具有破坏性的入侵害虫。© 2023 化学工业协会。
