昆虫学报 ›› 2017, Vol. 60 ›› Issue (4): 363-371.doi: 10.16380/j.kcxb.2017.04.001

• 研究论文 •    下一篇

铜绿丽金龟气味结合蛋白AcorOBP1的表达和结合特性分析

王超群1,2,#, 赵莹2,#, 曹雅忠2, 魏红爽2, 李克斌2, 张帅2, 彭宇1,*, 尹姣2,*   

  1. (1. 湖北大学生命科学学院, 湖北省生物资源绿色转化协同创新中心, 武汉430062; 2. 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京100193)
  • 出版日期:2017-04-20 发布日期:2017-04-20

Expression and binding characterization of odorant binding protein 1 (AcorOBP1) in Anomala corpulenta (Coleoptera: Scarabaeoidea)

WANG Chao-Qun1,2,#, ZHAO Ying2,#, CAO Ya-Zhong2, WEI Hong-Shuang2, LI Ke-Bin2, ZHANG Shuai2, PENG Yu1,*, YIN Jiao2,*   

  1. (1. Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, College of Life Sciences, Hubei University, Wuhan 430062, China; 2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences,Bejing 100193, China)
  • Online:2017-04-20 Published:2017-04-20

摘要: 【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-qPCR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±0.59 μmol/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。

关键词: 铜绿丽金龟, 气味结合蛋白, 表达谱, 荧光竞争结合, 解离常数

Abstract: 【Aim】 This study aims to reveal the expression profiles of the odorant binding protein 1 (AcorOBP1) gene in Anomala corpulenta Motschulsky and its binding specificity to specific semiochemicals.【Methods】 The spatio-temporal expression profiles of AcorOBP1 in A.corpulenta was assayed by RT-qPCR. Prokaryotic expression system was used to express and purify AcorOBP1. The binding characteristics of AcorOBP1 to 56 candidate compounds were assayed by fluorescence competitive binding experiment. 【Results】 AcroOBP1 was expressed in eggs, 1st instar larvae and adults of A. corpulenta, and its  expression level in adult antenna was extremely significantly higher than those in other adult tissues (P<0.05). In the ligand-binding experiment, 53 of the 56 compounds tested were able to bind to AcorOBP1. Particularly, methyl salicylate and decyl alcohol, which are volatile components of the wax leaf privet Ligustrum quihoui, showed outstanding binding affinities to AcorOBP1 with the dissociation constants of 2.10±0.08 and 5.04±0.59 μmol/L,respectively. 【Conclusion】 AcorOBP1 shows binding affinity to a wide range of test compounds and may play a vital role in the localization of host plant L. quihoui by A. corpulenta.

Key words: Anomala corpulenta, odorant binding protein, expression profile, fluorescence competitive binding, dissociation constant