CRISPR/Cas9 mutagenesis abolishes odorant-binding protein BdorOBP56f-2 and impairs the perception of methyl eugenol in Bactrocera dorsalis (Hendel)
CRISPR/Cas9 mutagenesis abolishes odorant-binding protein BdorOBP56f-2 and impairs the perception of methyl eugenol in Bactrocera dorsalis (Hendel)
复制标题
CRISPR/Cas9 诱变消除了气味结合蛋白 BdorOBP56f-2 并损害了橘小实蝇 (Hendel) 对甲基丁香酚的感知
DOI：
10.1016/j.ibmb.2021.103656
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发表时间：
2021-09-29
影响因子：
3.8
通讯作者：
Jiang, Hongbo
中科院分区：
文献类型：
--
作者：
Chen, Xiaofeng;Lei, Yibo;Jiang, Hongbo
嗅觉是许多昆虫行为的基础，如觅食、寄主定位、交配和躲避捕食者。在昆虫嗅觉的第一步，气味结合蛋白（OBP）结合疏水性气味物质并将其运输到气味受体。甲基丁香酚（ME）是一种对东方实蝇（Bactrocera dorsalis（Hendel））成熟雄性有很强引诱作用的物质。潜在的分子机制尚不清楚，但有体外证据表明BdorOBP 56 f-2参与ME感知。我们使用微量热电泳来证实BdorOBP 56 f-2在体外以强亲和力直接结合ME。然后，我们使用CRISPR/Cas9敲除BdorOBP 56 f-2基因，使我们能够建立纯合突变体B。背线在突变体中，对ME的触角电位反应和行为吸引力显着降低，提供了体内证据表明BdorOBP 56 f-2是有效的ME感知所必需的。我们的研究结果提供了深入了解ME感知的分子机制在B。为其他骨关节的功能分析提供了理论依据。
Olfaction underpins many insect behaviors, such as foraging, host location, mating, and predator avoidance. In the first step of insect olfaction, odorant-binding proteins (OBPs) bind hydrophobic odorants and transport them to odorant receptors. Methyl eugenol (ME) is a powerful attractant for mature males of the oriental fruit fly Bactrocera dorsalis (Hendel), one of the most destructive fruit pests. The underlying molecular mechanism is unclear, but there is in vitro evidence that BdorOBP56f-2 is involved in ME perception. We used microscale thermophoresis to confirm that BdorOBP56f-2 directly binds ME with strong affinity in vitro. We then used CRISPR/Cas9 to knock out the BdorOBP56f-2 gene, allowing us to establish a homozygous mutant B. dorsalis line. The electroantennogram response and behavioral attraction to ME were significantly reduced in the mutant, providing in vivo evidence that BdorOBP56f-2 is necessary for efficient ME perception. Our results offer insight into the molecular mechanism of ME perception in B. dorsalis and provide a theoretical basis for the functional analysis of other OBPs.