›› 2011, Vol. 54 ›› Issue (9): 975-981.doi:

• 研究论文 • 上一篇    下一篇

枫长镰管蚜气味结合蛋白OBP3的cDNA克隆和序列分析及其成虫嗅觉行为反应测定

 范佳, Sophie VANDERMOTEN, Frederic FRANCIS, 刘勇, 陈巨莲, 程登发   

  • 收稿日期:2011-05-03 出版日期:2011-09-20 发布日期:2011-09-20
  • 通讯作者: 陈巨莲, 程登发 E-mail:jlchen@ippcaas.cn; dfcheng@ippcaas.cn
  • 作者简介:范佳,女,1980年生,黑龙江嫩江人,博士研究生,研究方向为昆虫生理与分子生物学,E-mail: jiahangyouxiang@163.com
  • 基金资助:
    国家自然科学基金项目(30971920); 国家重大科技专项(2009ZX08012-007B); 国家小麦产业建设体系专项经费(nycytx-03);中比合作项目(2010DFA32810,PIC SHANDONG)

cDNA cloning and sequence analysis of odorant binding protein OBP3 in Drepanosiphum platanoidis (Homoptera: Drepanosiphidae) and testing of its adult olfactory behavior

 FAN  Jia, Sophie  VANDERMOTEN, Frederic  FRANCIS, LIU  Yong, CHEN  Ju-Lian, CHENG  Deng-Fa   

  • Received:2011-05-03 Online:2011-09-20 Published:2011-09-20
  • Contact: CHEN Ju-Lian, CHENG Deng-Fa E-mail:jlchen@ippcaas.cn; dfcheng@ippcaas.cn
  • About author:E-mail: jiahangyouxiang@163.com

摘要: 枫长镰管蚜Drepanosiphum platanoidis是少数几种不分泌E-β-法尼烯 (E-β-farnesene, EBF)的蚜虫之一。ApisOBP3是首个被鉴定的特异结合EBF的昆虫气味结合蛋白,来自于豌豆蚜Acyrthosiphum pisum。本研究通过基因克隆技术,在枫长镰管蚜体内获得2个表达ApisOBP3类似蛋白的cDNA序列,长度均为426 bp,其中, 5′末端的69 bp为信号肽编码序列。编码的成熟蛋白分别命名为DplaOBP3-1和DplaOBP3-2,两个蛋白均由119个氨基酸残基组成,序列一致性为95.8%。与ApisOBP3的序列一致性分别为95.8%和98.3%。DplaOBP3-1, DplaOBP3-2及ApisOBP3 3个蛋白具有数量和位置完全一致的6个保守的半胱氨酸残基。以测得3D结构的马德拉蜚蠊Leucophaea maderae的信息素结合蛋白LmadPBP为模板比较建模,结果显示两个蛋白在疏水中心均具有与ApisOBP3一致的关键疏水残基及芳香族氨基酸残基Tyr84。利用四臂嗅觉仪测定枫长镰管蚜的嗅觉行为反应,结果显示:枫长镰管蚜残体挥发物及外源EBF对该蚜均有显著的驱避作用(P=0.01)。本研究提示,枫长镰管蚜具有种内报警信息素,该蚜残体挥发物即为其种内报警信息素;枫长镰管蚜尽管并不分泌EBF,但具有表达2种OBP3蛋白的基因,并可识别环境中的EBF且对其作出行为反应。

关键词:

Abstract: ApisOBP3 from Acyrthosiphum pisum was known as its affinity to E-β-farnesene (EBF), and the volatile substances released by Drepanosiphum platanoidis contain no EBF which is a conventional component secreted by most of aphids. In this study, two odorant binding proteins named as DplaOBP3-1 and DplaOBP3-2 sharing high identities (95.8% and 98.32%, respectively) with ApisOBP3 were identified in D. platanoidis. The lengths of coding sequences of both DplaOBP3-1 and DplaOBP3-2 are 426 bp with 69 bp coding for signal peptide at the 5′ end. DplaOBP3-1, DplaOBP3-2 and ApisOBP3 share 6 highly conserved cysteine residues at the same position among sequences. Based on the 3D structure of LmadPBP, a pheromone binding protein from cockroach Leucophaea maderae, we modeled the structures of DplaOBP3-1 and DplaOBP3-2, and all the five hydrophobic amino acids and the aromatic amino acid residue, Tyr84, located in the binding cavity probably participating in interacting with ligands have their counterparts in the structure of ApisOBP3. Further, four-arm olfactometer assay showed that D. platanoidis were significantly repelled by both EBF and crushed aphids (P=0.01). The results suggest that volatiles from fresh crushed bodies could be the intraspecies alarm pheromone in D. platanoidis; D. platanoidis are repelled by allogenetic EBF whereas they never release EBF.

Key words: Drepanosiphum platanoidis, odorant binding protein, alarm pheromone, E-β-farnesene (EBF), cDNA cloning, four-arm olfactometer