红脉穗螟两个气味结合蛋白的序列特征及结合特性分析

Sequence characterization and ligand binding analysis of two odorant-binding proteins in Tirathaba rufivena (Walker)

摘要

昆虫嗅觉系统在寄主识别、交配和产卵等行为中发挥关键作用，其中气味结合蛋白（odorant-binding proteins，OBPs）在嗅觉信号传递过程中起重要作用。本研究针对红脉穗螟（Tirathaba rufivena）两个气味结合蛋白 TrufOBP1 和 TrufOBP5 进行了序列特征及与配体结合特性分析，并通过风洞实验验证其在行为调控中的作用。序列分析表明，TrufOBP1 具有 4 个保守半胱氨酸，属于 Minus-C 型 OBP；TrufOBP5 具有 10 个保守半胱氨酸，属于 Plus-C 型 OBP。荧光竞争结合实验结果表明，TrufOBP1 具有较广的结合谱，与 11 种配体结合，其中包括 9 种槟榔花挥发物和 2 种性信息素；TrufOBP5 结合特异性强，仅与反式−4−甲氧基肉桂酸异辛酯和香兰素结合，推测其可能在寄主植物识别过程中发挥重要作用。风洞实验发现：寄主挥发物中，β−月桂烯、反式−4−甲氧基肉桂酸异辛酯和乙酸叶醇酯可引诱红脉穗螟雌成虫。性信息素组分 2,2,6−三甲基−6−乙烯基四氢−2H−呋喃−3−醇引诱效果最强，此外，香兰素也表现出引诱活性。本研究揭示了 TrufOBPs 在红脉穗螟寄主识别和性信息素感受中的作用，为害虫行为调控提供了理论依据，并为基于嗅觉的绿色防控策略开发提供支持。

Abstract

The insect olfactory system plays a pivotal role in host recognition, mating, and oviposition, with odorant-binding proteins (OBPs) serving critical functions in olfactory signal transduction. An attempt was made to analyze the sequence characteristics and ligand-binding properties of two OBPs (TrufOBP1 and TrufOBP5) in Tirathaba rufivena, a major pest infesting inflorescences and fruit of arecanut (Areca catechu), and to validate their behavioral regulatory roles through wind tunnel experiments. Sequence analysis revealed that TrufOBP1 contains four conserved cysteines and is classified as a Minus-C OBP, whereas TrufOBP5 possesses ten conserved cysteines and is categorized as a Plus-C OBP. Fluorescence competitive binding assays demonstrated that TrufOBP1 exhibited broad ligand specificity and was bound to 11 compounds, including nine volatiles from areca nut (Areca catechu) inflorescences and two sex pheromones. In contrast, TrufOBP5 displayed high binding specificity, and interacted exclusively with octyl p-methoxycinnamate and vanillin, suggesting its potential role in host plant recognition. Wind tunnel experiments identified β-myrcene, octyl p-methoxycinnamate, and 3-hexenyl acetate as attractants for female adults from host volatiles. Among sex pheromone components, 2,2,6-trimethyl-6-vinyltetrahydro-2H-pyran-3-OL exhibited the highest attraction activity, while vanillin also demonstrated significant attractant effects. This study elucidates the functional roles of TrufOBP1 and TrufOBP5 in host recognition and sex pheromone perception inT. rufivena, providing a theoretical foundation for pest behavior regulation and supporting the development of olfaction-based green control strategies.

关键词

红脉穗螟 / 气味结合蛋白 / 荧光竞争结合 / 风洞行为 / 植物挥发物 / 性信息素

Key words

Tirathaba rufivena (Walker) / odorant-binding protein / fluorescent competitive binding / wind tunnel behavior / plant volatiles / sex pheromones

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基金资助

海南省自然科学基金面上项目(322MS012)

海南省科技人才创新项目(KJRC2023D01)