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PsOr1,一种基于 RNA 干扰的害虫管理的潜在靶标。

PsOr1, a potential target for RNA interference-based pest management.

机构信息

Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, China.

出版信息

Insect Mol Biol. 2011 Feb;20(1):97-104. doi: 10.1111/j.1365-2583.2010.01049.x. Epub 2010 Sep 21.

Abstract

Insect pests cause billions of dollars in agricultural losses, and attempts to kill them have resulted in growing threats from insecticide resistance, dietary pesticide pollution and environmental destruction. New approaches to control refractory insect pests are therefore needed. The host-plant preferences of insect pests rely on olfaction and are mediated via a seven transmembrane-domain odorant receptor (Or) family. The present study reports the cloning and characterization of PsOr1, the first candidate member of the Or gene family from Phyllotreta striolata, a devastating beetle pest that causes damage worldwide. PsOr1 is remarkably well conserved with respect to other insect orthologues, including DmOr83b from Drosophila melanogaster. These insect orthologues form an essential non-conventional Or sub-family and may play an important and generalized role in insect olfaction. We designed double-stranded (ds) RNA directly against the PsOr1 gene and exploited RNA interference (RNAi) to control P. striolata. The chemotactic behavioural measurements showed that adult beetles were unable to sense the attractant or repellent odour stimulus after microinjection of dsRNA against PsOr1. Reverse Transcription (RT)-PCR analysis showed specific down-regulation of mRNA transcript levels for this gene. Furthermore, host-plant preference experiments confirmed that silencing PsOr1 by RNAi treatment impaired the host-plant preferences of P. striolata for cruciferous vegetables. These results demonstrate that this insect control approach of using RNAi to target PsOr1 and its orthologues might be effective in blocking host-plant-seeking behaviours in diverse insect pests. The results also support the theory that this unique receptor type plays an essential general role in insect olfaction.

摘要

害虫造成数十亿美元的农业损失,而试图杀死它们的行为导致了杀虫剂抗药性、饮食性农药污染和环境破坏等日益严重的威胁。因此,需要采取新的方法来控制难治性害虫。害虫对寄主植物的偏好依赖于嗅觉,并通过七跨膜结构域气味受体(Or)家族来介导。本研究报告了来自 Phyllotreta striolata 的 Or 基因家族的第一个候选成员 PsOr1 的克隆和特性,Phyllotreta striolata 是一种破坏性的甲虫害虫,在全球范围内造成损害。与其他昆虫同源物(包括来自 Drosophila melanogaster 的 DmOr83b)相比,PsOr1 非常保守。这些昆虫同源物形成一个重要的非传统 Or 亚家族,可能在昆虫嗅觉中发挥重要而普遍的作用。我们设计了针对 PsOr1 基因的双链 RNA(dsRNA),并利用 RNA 干扰(RNAi)来控制 P. striolata。趋化行为测量表明,成年甲虫在注射针对 PsOr1 的 dsRNA 后无法感知引诱剂或驱避剂气味刺激。逆转录(RT)-PCR 分析显示该基因的 mRNA 转录水平特异性下调。此外,寄主植物偏好实验证实,通过 RNAi 处理沉默 PsOr1 会损害 P. striolata 对十字花科蔬菜的寄主植物偏好。这些结果表明,这种使用 RNAi 靶向 PsOr1 及其同源物来控制昆虫的方法可能有效地阻止不同害虫寻找寄主植物的行为。这些结果还支持了这样一种理论,即这种独特的受体类型在昆虫嗅觉中发挥着至关重要的普遍作用。

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