Mutagenesis of the odorant receptor co-receptor (Orco) reveals severe olfactory defects in the crop pest moth Helicoverpa armigera.
Mutagenesis of the odorant receptor co-receptor (Orco) reveals severe olfactory defects in the crop pest moth Helicoverpa armigera.
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DOI：
10.1186/s12915-022-01411-2
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发表时间：
2022-09-30
期刊：
影响因子：
5.4
通讯作者：
Wang, Chen-Zhu
中科院分区：
文献类型：
--
作者：
Fan, Xiao-Bin;Mo, Bao-Tong;Li, Guo-Cheng;Huang, Ling-Qiao;Guo, Hao;Gong, Xin-Lin;Wang, Chen-Zhu
气味受体作为气味门控离子通道，在昆虫嗅觉中起着至关重要的作用。它们是由气味受体共受体(ORCO)和配体选择性OR形成的异多聚体复合体。其他类型的嗅觉受体蛋白，如离子型受体(IRS)和一些味觉受体(GRs)，也参与昆虫的嗅觉系统。ORCO作为ORs的一个必需亚单位是高度保守的，为系统地评估OR依赖的嗅觉反应提供了机会。在这里，我们利用CRISPR/CAS9基因编辑技术成功地建立了一种臭名昭著的农作物害虫棉铃虫的纯合子突变体(Orco−/−)。然后我们比较了野生型(WT)和Orco−/−成虫和幼虫的嗅觉反应特征。Orco−/−雄性不育，而Orco−/−雌性可育。Orco−/−雌虫的寿命比WT雌虫长。Orco−/−成虫触角中大多数ORS、IR等嗅觉相关基因的表达没有受到明显影响，但也有部分表达上调或下调。此外，在触角叶(包括雄蛾的大肾小球复合区)水平上，Orco−/−蛾的神经解剖表型没有变化。利用EAG和SSR技术，我们发现Orco−/−蛾对性信息素和多种寄主植物气味缺乏电生理反应。在风洞实验中，Orco−/−雄性对性信息素的迎风飞行行为严重减少。Orco−/−雌虫对寄主植物(青椒)的产卵选择性完全消失，对青椒的趋化性也丧失；Orco−/−幼虫对青椒的趋化性丧失。我们的研究表明，OR介导的嗅觉对于棉铃虫的信息素通讯、产卵选择和幼虫趋化性是必不可少的，这表明可以通过抑制或沉默Orco的表达来扰乱害虫的交配搜索和寄主寻找行为。网上版载有补充材料，可在10.1186/s12915-022-01411-2查阅。
Odorant receptors (ORs) as odorant-gated ion channels play a crucial role in insect olfaction. They are formed by a heteromultimeric complex of the odorant receptor co-receptor (Orco) and a ligand-selective Or. Other types of olfactory receptor proteins, such as ionotropic receptors (IRs) and some gustatory receptors (GRs), are also involved in the olfactory system of insects. Orco as an obligatory subunit of ORs is highly conserved, providing an opportunity to systematically evaluate OR-dependent olfactory responses. Herein, we successfully established a homozygous mutant (Orco−/−) of Helicoverpa armigera, a notorious crop pest, using the CRISPR/Cas9 gene editing technique. We then compared the olfactory response characteristics of wild type (WT) and Orco−/− adults and larvae. Orco−/− males were infertile, while Orco−/− females were fertile. The lifespan of Orco−/− females was longer than that of WT females. The expressions of most Ors, Irs, and other olfaction-related genes in adult antennae of Orco−/− moths were not obviously affected, but some of them were up- or down-regulated. In addition, there was no change in the neuroanatomical phenotype of Orco−/− moths at the level of the antennal lobe (including the macroglomerular complex region of the male). Using EAG and SSR techniques, we discovered that electrophysiological responses of Orco−/− moths to sex pheromone components and many host plant odorants were absent. The upwind flight behaviors toward sex pheromones of Orco−/− males were severely reduced in a wind tunnel experiment. The oviposition selectivity of Orco−/− females to the host plant (green pepper) has completely disappeared, and the chemotaxis toward green pepper was also lost in Orco−/− larvae. Our study indicates that OR-mediated olfaction is essential for pheromone communication, oviposition selection, and larval chemotaxis of H. armigera, suggesting a strategy in which mate searching and host-seeking behaviors of moth pests could be disrupted by inhibiting or silencing Orco expression. The online version contains supplementary material available at 10.1186/s12915-022-01411-2.
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