昆虫学报 ›› 2019, Vol. 62 ›› Issue (12): 1369-1378.doi: 10.16380/j.kcxb.2019.12.003

• 研究论文 • 上一篇    下一篇

小地老虎化学感受蛋白AipsCSP8的表达及配体结合特性分析

苏旭1,2, 张永军2, 耿亭3, 李静1,*, 谷少华4,*   

  1. (1. 河北农业大学植物保护学院, 河北保定 071000; 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; 3. 农业部廊坊作物有害生物科学观测实验站, 河北廊坊 065000; 4. 中国农业大学昆虫学系, 北京 100193)
  • 出版日期:2019-12-20 发布日期:2019-12-12

Expression and ligand binding characteristics of chemosensory protein AipsCSP8 of Agrotis ipsilon (Lepidoptera: Noctuidae)

SU Xu1,2, ZHANG Yong-Jun2, GENG Ting3, LI Jing1,*, GU Shao-Hua4,*   

  1. (1. College of Plant Protection, Agricultural University of Hebei, Baoding, Hebei 071000, China; 2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 3. Langfang Experimental Station of Crop Pests Science, Ministry of Agriculture, Langfang, Hebei 065000, China; 4. Department of Entomology, China Agricultural University, Beijing 100193, China)
  • Online:2019-12-20 Published:2019-12-12

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摘要：  【目的】本研究旨在解析小地老虎Agrotis ipsilon化学感受蛋白AipsCSP8的表达谱及配体结合特征。【方法】利用qRT-PCR方法测定了AipsCSP8在小地老虎成虫头、胸、腹、足、翅、性腺(附腺)、触角、喙和下唇须以及雌雄虫羽化前后触角中的表达水平。体外重组表达AipsCSP8蛋白，通过荧光竞争结合实验测定重组蛋白AipsCSP8对24种性信息素、22种植物挥发物和10种化学农药的结合特性。【结果】qRT-PCR检测结果显示，AipsCSP8在小地老虎雄成虫的足和下唇须中表达量最高，在附腺和触角中的表达量次之；在雌成虫的触角中表达量最高，在足、性腺、下唇须中的表达量次之。另外，雄、雌虫触角中AipsCSP8表达量分别在成虫羽化前1 d和羽化当天达到顶峰。荧光竞争结合实验结果表明，重组蛋白AipsCSP8对植物挥发物邻苯二甲酸-丁酯(解离常数Ki=14.6 μmol/L)、苯乙醛(Ki=17.2 μmol/L)以及化学农药阿维菌素(Ki=12.9 μmol/L)均有较强的结合能力。【结论】化学感受蛋白AipsCSP8可能参与小地老虎嗅觉行为及味觉识别，并可能在小地老虎感受化学农药的过程中发挥作用。

关键词: 小地老虎, 化学感受蛋白, 时空表达谱, 原核表达, 气味结合特征, 化学感受

Abstract: 【Aim】 The aim of this study is to explore the gene expression profiles and ligand binding characteristics of the chemosensory protein AipsCSP8 of Agrotis ipsilon. 【Methods】 The expression levels of AipsCSP8 in the head, thorax, abdomen, leg, wing, pheromone gland (accessory gland), antenna, proboscis and labial palp of adults of A. ipsilon and the antennae of adults before and after emergence were determined by qRT-PCR. The recombinant protein AipsCSP8 was harvested by in vitro prokaryotic expression system, and its binding characteristics to 24 sex pheromones, 22 plant volatiles and 10 chemical pesticides were determined by fluorescence binding assay. 【Results】 The qRT-PCR results indicated that the expression level of AipsCSP8 was the highest in the leg and labial palp of male adults, followed by in the accessory gland and the antenna of male adults. While in female adults, the expression level of AipsCSP8 was the highest in the antenna, followed by in the leg, pheromone gland and labial palp. In addition, the expression levels of AipsCSP8 in the antennae of males and females reached the peak on the 1st day before emergence and on the day of emergence, respectively. The results of fluorescence competitive binding assay showed that AipsCSP8 had strong binding capabilities with dibutyl phthalate, phenylacetaldehyde and avermectin, with the dissociation constant Ki values of 14.6, 17.2 and 12.9 μmol/L, respectively. 【Conclusion】 AipsCSP8 may be involved in the olfactory behavior and taste recognition of A. ipsilon and play a role in the perception of chemical pesticides.

Key words:  Agrotis ipsilon; chemosensory protein, spatio-temporal expression profile, prokaryotic expression, odor binding characteristics, chemoreception

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