Cloning,expression and binding specificity analysis of odorant binding protein 3 of the lucerne plant bug,Adelphocoris lineolatus(Goeze)  

作　　者：GU ShaoHua SUN Yang REN LiYan ZHANG XueYing ZHANG YongJun WU KongMing GUO YuYuan 

机构地区：[1]State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences,Beijing 100193, China [2]College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China

出　　处：《Chinese Science Bulletin》2010年第34期3911-3921,共11页

基　　金：supported by the National Key Basic Research Program of China(2007CB109202);the National High-Technology Research and Development Program of China(2008BAD5B05);National Natural Science Foundation of China(31071694)

摘　　要：Credible evidence shows that odorant binding proteins(OBPs)are required for insect olfaction perception and play a key role in transporting hydrophobic odorants across the sensillum lymph to the olfactory receptors(ORs).In the present study,a novel OBP(AlinOBP3)gene from the lucerne plant bug,Adelphocoris lineolatus,was cloned and expressed.The expression pattern of AlinOBP3 was evaluated by qPCR,which indicated that AlinOBP3 was dominantly expressed in antennae.The binding properties of AlinOBP3 with 9 cotton volatiles and 5 sex pheromone analogs were measured by fluorescence competitive binding assays with the fluorescence probe 1-NPN.The results revealed that of 9 cotton volatiles,Myrcene,β-Ocimene and α-Phellandrene can bind with AlinOBP3.α-Phellandrene especially bound to AlinOBP3 with a high binding affinity,with a dissociation constant of 56.68 μmol/L.Of the 5 sex pheromone analogs,Hexyl butyrate had the strongest binding affinity with AlinOBP3,with a dissociation constant as 59.53 μmol/L.Butyl butyrate,trans-2-Hexenyl butyrate and Ethyl butyrate had medium binding affinities with AlinOBP3,with dissociation constants of 227.39,108.77 and 143.47 μmol/L,respectively.The results suggest that AlinOBP3 might be a pheromone binding protein(PBP)with a dual-function for the perception of sex pheromones and plant volatiles.Credible evidence shows that odorant binding proteins （OBPs） are required for insect olfaction perception and play a key role in transporting hydrophobic odorants across the sensillum lymph to the olfactory receptors （ORs）. In the present study, a novel OBP （AlinOBP3） gene from the lucerne plant bug, Adelphocoris lineolatus, was cloned and expressed. The expression pattern of AlinOBP3 was evaluated by qPCR, which indicated that AlinOBP3 was dominantly expressed in antennae. The binding properties of AlinOBP3 with 9 cotton volatiles and 5 sex pheromone analogs were measured by fluorescence competitive binding assays with the fluorescence probe 1-NPN. The results revealed that of 9 cotton volatiles, Myrcene, 13-Ocimene and ct-Phellandrene can bind with AlinOBP3. α-Phellandrene especially bound to AlinOBP3 with a high binding affinity, with a dissociation constant of 56.68 μmol/L. Of the 5 sex pheromone analogs, Hexyl butyrate had the strongest binding affinity with AlinOBP3, with a dissociation constant as 59.53 μmol/L. Butyl butyrate, trans-2-Hexenyl butyrate and Ethyl butyrate had medium binding affinities with AlinOBP3, with dissociation constants of 227.39, 108.77 and 143.47 μmol/L, respectively. The results suggest that AlinOBP3 might be a pheromone binding protein （PBP） with a dual-function for the perception of sex pheromones and plant volatiles.

关 键 词：气味结合蛋白 克隆和表达 结合特异性 线纹 结合亲和力 植物挥发物 性信息素 解离常数 

分 类 号：S513[农业科学—作物学] Q966[生物学—昆虫学]