绿盲蝽气味结合蛋白AlucOBP31配体筛选及EAG测定

doi:10.16409/j.cnki.2095-039x.2024.01.010

摘要/Abstract

摘要： 绿盲蝽Apolygus lucorum是重要的农业害虫，了解其化学感受机制对该虫的绿色防控具有重要指导意义。气味结合蛋白（ Odorant-binding proteins，OBPs）在昆虫化学感受过程中发挥关键作用。为深入探究绿盲蝽OBP家族成员AlucOBP31的生物学功能，本研究采用荧光竞争结合试验和触角电位技术（ Electroantennogram，EAG），系统分析了AlucOBP31对候选配体化合物的结合特性及其电生理活性。荧光竞争结合试验结果表明，重组AlucOBP31蛋白能够与16种配体化合物发生不同程度的结合，其中与植物挥发物β-紫罗兰酮的结合能力最强，而与十二醛和顺-丁酸-3-己烯酯的结合能力相对较弱。值得注意的，AlucOBP31能与绿盲蝽性信息素组分4-氧代-反-2-己烯醛结合，提示其可能参与绿盲蝽的性信息素识别过程。此外，AlucOBP31还对槲皮素、芦丁水合物、棉酚和单宁酸等4种非挥发性苦味物质具有强结合能力，暗示其可能在绿盲蝽的味觉识别中发挥一定作用。EAG试验进一步验证了AlucOBP31配体的电生理活性。绿盲蝽雌、雄成虫的触角对12种AlucOBP31挥发性配体均产生了明显的EAG反应，其中对顺-丁酸-3-己烯酯和4-氧代-反-2-己烯醛的EAG响应值显著高于其它配体，并且雄性触角对这些配体的EAG反应普遍强于雌性，提示AlucOBP31介导的化学感受过程可能存在性别差异。综上所述，本研究系统分析了绿盲蝽AlucOBP31的结合特性及相应配体的电生理活性，发现该蛋白能够结合寄主植物挥发物、绿盲蝽性信息素组分以及多种非挥发性苦味物质，表明AlucOBP31可能在绿盲蝽的嗅觉和味觉识别过程中发挥多重生物学功能。这一发现为进一步探究AlucOBP31与其配体在绿盲蝽寄主选择、交配等重要生命活动中的生物学意义提供了研究基础，加深了我们对绿盲蝽化学感受机制的认知，也为绿盲蝽的绿色防控提供了新的分子靶标。

关键词: 绿盲蝽, 气味结合蛋白AlucOBP31, 配体结合, EAG, 化学感受

Abstract: The mirid bug,Apolygus lucorum,is a major agricultural pest,and understanding its chemical sensing mechanism is crucial for pest control.Odorant-binding proteins (OBPs) play a key role in the insect chemoreception process.To investigate the function of AlucOBP31,a member of the OBP family in A.lucorum,we conducted fluorescence competitive binding assays and electroantennogram (EAG) recordings to systematically analyze the binding characteristics of AlucOBP31 and electrophysiological activities of candidate ligands.The results of the fluorescence competitive binding assays showed that the recombinant AlucOBP31 protein could bind to 16 compounds to varying degrees.AlucOBP31 exhibited the strongest binding affinity to the plant volatile β-ionone,while its binding affinities to dodecyl aldehyde and cis-3-hexenyl butyrate were relatively weak.Interestingly,AlucOBP31 could also bind to (2E)-4-oxo-2-hexenal,a sex pheromone component of A.lucorum,suggesting its potential involvement in sex pheromone recognition.Furthermore,AlucOBP31 demonstrated strong binding to four non-volatile bitter substances,including quercetin,rutin hydrate,gossypol hydrate,and tannic acid,implying its possible role in taste recognition of A.lucorum.EAG experiments further verified the electrophysiological activities of AlucOBP31 binding ligands.The antennae of both male and female A.lucorum adults exhibited significant EAG responses to 12 volatile ligands bound by AlucOBP31,with the EAG response values to cis-3-hexenyl butyrate and (2E)-4-oxo-2-hexenal being significantly higher than those to other ligands.Intriguingly,the EAG responses of male antennae to these ligands were generally significantly stronger than those of females,suggesting potential sex-specific differences in the AlucOBP31-mediated chemoreception process.In conclusion,this study systematically analyzed the binding characteristics and electrophysiological activities of AlucOBP31 in A.lucorum,revealing that this protein can widely bind to host plant volatiles,sex pheromone components of mirid bug,and various non-volatile bitter substances.These findings indicate that AlucOBP31 may play multiple biological functions in the olfactory and gustatory recognition of A.lucorum.Our result provides a research foundation for further exploring the biological significance of AlucOBP31 and its ligands in important life activities of A.lucorum,such as host selection and mating.It deepens our understanding of the chemical sensing mechanism in A.lucorum and offers new molecular targets for the green control of this pest.

Key words: Apolygus lucorum, odorant binding protein AlucOBP31, ligand binding, EAG, chemoreception

中图分类号:

S476

武娟, 张瑶瑶, 李仔博, 刘靖涛, 安兴奎, 刘顺, 李瑞军, 张永军. 绿盲蝽气味结合蛋白AlucOBP31配体筛选及EAG测定[J]. 中国生物防治学报, 2024, 40(2): 319-326.

WU Juan, ZHANG Yaoyao, LI Zibo, LIU Jingtao, AN Xingkui, LIU Shun, LI Ruijun, ZHANG Yongjun. Ligands Screening of Odorant Binding Protein AlucOBP31 and Electroantennogram Determination of Apolygus lucorum[J]. Chinese Journal of Biological Control, 2024, 40(2): 319-326.

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