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黄斑星天牛雌虫触角气味结合蛋白基因的鉴定及组织分布

李广伟*,陈秀琳,尚天翠   

  1. (伊犁师范学院生物与地理科学学院, 新疆伊宁 835000)
  • 出版日期:2017-06-10 发布日期:2017-06-10

Identification and tissue distribution of odorant binding proteins of female antennae Anoplophora nobilis Ganglbauer (Coleoptera: Cerambycidae).

LI Guang-wei*, CHEN Xiu-lin, SHANG Tian-cui   

  1. (College of Biology and Geography, Yili Normal University, Yining 835000, Xinjiang, China).
  • Online:2017-06-10 Published:2017-06-10

摘要: 为发掘黄斑星天牛(Anoplophora nobilis Ganglbauer)触角OBPs基因,采用Illumina HiSeq2500测序平台对雌虫触角进行转录组测序,通过基因功能注释及传统测序验证鉴定OBPs基因,半定量PCR检测OBPs在不同组织中的分布情况。结果表明,在黄斑星天牛雌虫触角中共鉴定到16种OBPs基因,包括11种Minus-C OBPs、3种Classical OBPs和2种Plus-C OBPs。每个蛋白序列其N端均有由16~25个氨基酸组成的信号肽序列,不同亚家族OBPs均具有各自典型保守的Cys残基。黄斑星天牛OBPs之间高度分化,序列相似度在8.00%~48.44%。黄斑星天牛与其他昆虫OBPs的系统进化树分析表明,虽然同种或不同物种的OBPs之间高度分化,但大多数OBPs亚家族基因在进化树中仍聚类到独立的进化枝,表明它们具有同源性。16种OBPs在雌虫触角中的表达量差异明显,其RPKM值在12.0~9065.5,其中AnobOBP1、AnobOBP2、AnobOBP3和AnobOBP7在触角中特异性表达,其他OBPs除在触角中表达外,还在头、胸、腹、足、翅的个别组织中表达,表明OBPs除执行嗅觉生理功能外,还参与其他生命代谢活动。本试验为进一步研究黄斑星天牛识别寄主植物挥发物的分子机制提供了基因保障,也为利用信息化学物质治理钻蛀性害虫提供了一种新思路。

Abstract: To explore the OBP genes of antennae Anoplophora nobilis Ganglbauer, the transcriptome of female antennae was generated using Illumina HiSeq2500, the OBPs were identified by GO annotation and traditional DNA sequencing, and the tissue distribution of OBPs was performed by semiquantitative PCR. The results showed that 16 OBPs were identified from female antennae A. nobilis, including 11 Minus-C OBPs, three Classical OBPs and two Plus-C OBPs. Each OBP with a signal peptide at Nterminal consisted of 16-25 amino acids. OBPs in each subfamily have their own typical conservative Cys residues. The AnobOBPs are highly divergent and the sequence similarities range from 8.00% to 48.44%. Phylogenetic analysis showed that although OBPs of the same or different species are highly divergent, the great majority of OBPs in each subfamily is still clustered into independent clade, suggesting that they have a common ancestor. The expression levels of 16 female antennae AnobOBPs were significantly different, with the RPKM values ranging from 12.0 to 9065.5. AnobOBP1, AnobOBP2, AnobOBP3 and AnobOBP7 exclusively expressed in antennae, and the other AnobOBPs expressed in antennae and other tissues, including head, thorax, abdomen, leg and wings, indicating that the OBPs in insects may also participate in other functions in addition to chemosensation. This study established a foundation for determining the chemoreception molecular mechanism of A. nobilis, which would provide a new perspective for controlling the borer pests by using semiochemicals.