Involvement of GOBP2 in the perception of a sex pheromone component in both larval and adult Spodoptera litura revealed using CRISPR/Cas9 mutagenesis
Involvement of GOBP2 in the perception of a sex pheromone component in both larval and adult Spodoptera litura revealed using CRISPR/Cas9 mutagenesis
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使用 CRISPR/Cas9 诱变揭示了 GOBP2 参与斜纹夜蛾幼虫和成虫性信息素成分的感知。
DOI：
10.1016/j.ibmb.2022.103719
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发表时间：
2022-01-11
影响因子：
3.8
通讯作者：
Dong, Shuang-Lin
中科院分区：
文献类型：
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作者：
Han, Wei-Kang;Yang, Yi-Lin;Dong, Shuang-Lin
一般气味结合蛋白（General odorant-binding protein, GOBPs）长期以来被认为与植物气味感知有关。在本研究中，我们利用体内CRISPR/Cas9技术，对重要的夜行害虫斜纹夜蛾（Spodoptera litura）进行了功能表征，发现GOBP2也参与性信息素成分的感知。首先，将GOBP2 sgRNA和Cas9蛋白注射到新产卵的昆虫卵中，在G0个月的时间内实现了35.6%的目标诱变。然后经过三代筛选，获得GOBP2纯合子敲除菌株（GOBP2(-/-)）。敲除雄性和雌性飞蛾对性信息素成分的EAG反应显著降低，敲除雌性飞蛾对植物气味成分的EAG反应也显著降低。在食物选择行为分析中，GOBP2（-/-）幼虫对添加主要性信息素成分Z9， e11 -十四烯乙酸酯（Z9, E11-14:Ac）的人工饲料失去了偏好，而WT幼虫对信息素的偏好较高。y管嗅觉测定和直接信息素刺激试验表明，与WT相比，gop2（-/-）雄性成虫对Z9、E11-14:Ac的选择率、发笔展示率和交配尝试率显著降低。在产卵试验中，与WT雌性相比，GOBP2（-/-）雌性对大豆的偏好显著降低。我们的研究表明，GOBP2在成虫和幼虫阶段对性信息素的感知中发挥了重要作用，为性信息素感知提供了新的认识，并为性信息素行为调控提供了潜在的靶点。
General odorant-binding proteins (GOBPs) are long considered responsible for the perception of plant odorants. In this study with the important noctuid pest Spodoptera litura, we functionally characterized that GOBP2 is also involved in the perception of sex pheromone components using in vivo CRISPR/Cas9 technique. First, the GOBP2 sgRNA and Cas9 protein were injected into the newly laid insect eggs, resulting in a 35.6% target mutagenesis in G0 moths. Then, the homozygous GOBP2 knockout strain (GOBP2(-/-)) was obtained after the screening of three generations. The knockout male and female moths displayed a significant reduction in EAG responses to the sex pheromone components, and the knockout females also displayed a significant reduction to plant odorants. In the behavioral assay of food choice, GOBP2(-/-) larvae lost the preference to artificial diet added with the major sex pheromone component Z9, E11-tetradecadienyl acetate (Z9, E11-14:Ac), whereas the WT larvae highly preferred the pheromone diet. Y-tube olfactometer assay and direct pheromone stimulation assay showed that GOBP2(-/-) male adults reduced significantly than WT males in percentages of choice, hair pencil displaying and mating attempt to Z9, E11-14:Ac. In the oviposition test, GOBP2(-/-) females showed significantly reduced preference for the soybean plants compared to the WT females. Our study demonstrated that GOBP2 plays an important role in perceiving sex pheromones in adult and larval stages, providing new insight into sex pheromone perception and a potential target for sex pheromone-based behavioral regulation in the pest.