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【目的】揭示铜绿丽金龟Anomala corpulenta Motschulsky气味结合蛋白(odorant binding protein,OBP)基因的表达特征及其与特异信息化合物的结合特性。【方法】通过RT-q PCR测定了铜绿丽金龟AcorOBP1基因的时空表达谱;用原核表达系统对AcorOBP1进行表达纯化,然后利用荧光竞争结合实验测定了AcorOBP1蛋白与56种候选化合物的结合特征。【结果】AcorOBP1基因在铜绿丽金龟卵、1龄幼虫和成虫等特定发育时期表达,在成虫触角中的表达量显著高于其他部位(P<0.05)。在被测的56种化合物中,AcorOBP1与53种化合物具有结合活性,其中结合能力较强的为小叶女贞Ligustrum quihoui挥发物水杨酸甲酯和癸醇,解离常数分别为2.10±0.08和5.04±0.59μmol/L。【结论】铜绿丽金龟AcorOBP1结合谱广,该蛋白可能在铜绿丽金龟对寄主植物小叶女贞的定位过程中发挥重要作用。
【Objective】 The purpose of this study was to reveal the expression characteristics of odorant binding protein (OBP) gene and its binding properties to specific information compounds in Anomala corpulenta Motschulsky. 【Method】 The spatial-temporal expression profiles of AcorOBP1 gene were determined by RT-q PCR. AcorOBP1 was expressed and purified by prokaryotic expression system. Then, the binding characteristics of AcorOBP1 protein and 56 candidate compounds were determined by fluorescence competitive binding assay. 【Result】 The results showed that AcorOBP1 gene was expressed in specific developmental stages of eggs, first-instar larvae and adults of Acacia Platycodon. The expression level of AcorOBP1 in adult antennae was significantly higher than that in other sites (P <0.05). Of the 56 tested compounds, AcorOBP1 has binding activity with 53 compounds, among which Ligustrum quihoui volatiles methyl salicylate and decyl alcohol with higher binding capacity have dissociation constants of 2.10 ± 0.08 and 5.04 ± 0.59μmol / L. 【Conclusion】 AcorOBP1 has a broad spectrum of binding activity, and this protein may play an important role in the localization of Acorus scolytus to the host plant Ligustrum lucidum.