马铃薯甲虫气味结合蛋白的序列和基因表达谱分析

doi:10.16380/j.kcxb.2019.04.005

昆虫学报 ›› 2019, Vol. 62 ›› Issue (4): 428-441.doi: 10.16380/j.kcxb.2019.04.005

马铃薯甲虫气味结合蛋白的序列和基因表达谱分析

张方梅^1,2, 刘杨², 李祥瑞^2,*, 张云慧², 程登发², 郭文超³, 吐尔逊·阿合买提³

(1. 信阳农林学院, 河南省豫南农作物有害生物绿色防控院士工作站, 河南信阳 464000; 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. 新疆农业科学院植物保护研究所, 乌鲁木齐 830091)

出版日期:2019-04-20 发布日期:2019-04-08

Sequence analysis and gene expression profiling of odorant binding proteins in the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

ZHANG Fang-Mei¹, LIU Yang², LI Xiang-Rui^2,*, ZHANG Yun-Hui², CHENG Deng-Fa², GUO Wen-Chao³, TURSUN Ahmat³

(1. HenanProvincial South Henan Crop Pest Green Prevention and Control Academician Workstation, Xinyang Agriculture andForestryUniversity,Xinyang,Henan46400,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.InstituteofPlantProtection,XinjiangAcademyof Agricultural Sciences, Urumchi 830091,China)

Online:2019-04-20 Published:2019-04-08

摘要/Abstract

摘要： 【目的】昆虫气味结合蛋白(OBPs)在昆虫嗅觉行为中发挥着重要作用。马铃薯甲虫Leptinotarsa decemlineata是马铃薯上一种最主要的毁灭性害虫。为阐明该虫嗅觉识别分子机制，本研究对马铃薯甲虫26个OBP基因序列特征及组织表达谱进行研究。【方法】基于马铃薯甲虫触角转录组测序数据，利用生物信息学方法及qRT-PCR技术，分别对马铃薯甲虫26个LdecOBPs (LdecOBP1-LdecOBP26)的系统进化及基因的组织表达谱进行分析。【结果】除LdecOBP26基因外，其余25个LdecOBPs基因序列均具有完整的开放阅读框，编码120~255个氨基酸残基，预测的蛋白分子量为13.66~29.38 kD，等电点为4.12~8.42，它们属于两个亚家族，其中13个为Classical-C OBPs, 12个为Minus-C OBPs。除LdecOBP3和LdecOBP26外，其他24个LdecOBPs的N端均由16~23个氨基酸组成的信号肽序列。不同的OBPs亚家族均具有各自典型保守的Cys残基。LdecOBPs之间高度分化，氨基酸序列一致性在3.20%~41.91%。系统进化树分析表明，LdecOBPs与沙葱萤叶甲Galeruca daurica的GdauOBPs亲缘关系最近。基因表达谱分析显示，26个LdecOBPs基因在马铃薯甲虫的不同组织中表达，其中有12个LdecOBPs基因(LdecOBP2, LdecOBP4, LdecOBP6, LdecOBP9, LdecOBP10, LdecOBP12, LdecOBP13, LdecOBP16, LdecOBP20-22和LdecOBP24)在触角中高表达，2个LdecOBPs基因(LdecOBP5和LdecOBP17)在足中高表达，其他12个LdecOBPs基因(LdecOBP1, LdecOBP3, LdecOBP7, LdecOBP8, LdecOBP11, LdecOBP14, LdecOBP15, LdecOBP18, LdecOBP19, LdecOBP23, LdecOBP25和LdecOBP26)在触角、头(去除触角)、胸、腹、足和翅这些组织中均表达。【结论】本研究结果为进一步研究马铃薯甲虫嗅觉识别分子机制奠定了基础。

关键词: 马铃薯甲虫, 气味结合蛋白, 序列分析, 组织表达谱, 系统发育

Abstract: 【Aim】 Odorant-binding proteins (OBPs) play a fundamental role in insect olfaction. The Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), is one of the most notorious insect pests of potato. The sequence properties and tissue expression profiles of 26 putative OBP genes of L. decemlineata were identified to clarify the molecular mechanisms of its olfactory perception. 【Methods】 Based on antennal transcriptome sequencing data of L. decemlineata, the phylogeny and tissue-specific gene expression profiles of 26 LdecOBPs (LdecOBP1-LdecOBP26) in L. decemlineata were analyzed by bioinformatic methods and qRT-PCR, respectively. 【Results】 Except LdecOBP26, the genes of the other 25 LdecOBPs have the full-length open reading frames, encoding 120-255 amino acid residues with the predicted molecular weights of 13.66-29.38 kD and isoelectric points of 4.12-8.42. The 25 LdecOBPs belong to two subfamilies, with 13 Classical-C OBPs and 12 Minus-C OBPs. Except LdecOBP3 and LdecOBP26, each of the other 24 LdecOBPs contains a signal peptide at the N-terminus consisting of 16-23 amino acid residues. These OBPs in each subfamily have their own typical conservative Cys residues. LdecOBPs are highly divergent and their amino acid sequence identities range from 3.20% to 41.91%. Phylogenetic analysis showed that LdecOBPs and GdauOBPs of Galeruca daurica are most closely related. Gene expression profiling showed the 26 LdecOBPs genes were expressed in different adult tissues of L. decemlineata, with 12 LdecOBPs genes (LdecOBP2, LdecOBP4, LdecOBP6, LdecOBP9, LdecOBP10, LdecOBP12, LdecOBP13, LdecOBP16, LdecOBP20-22 and LdecOBP24) highly expressed in antennae, two genes (LdecOBP5 and LdecOBP17) highly expressed in legs, and the other 12 (LdecOBP1, LdecOBP3, LdecOBP7, LdecOBP8, LdecOBP11, LdecOBP14, LdecOBP15, LdecOBP18, LdecOBP19, LdecOBP23, LdecOBP25 and LdecOBP26) expressed in tissues antennae, head (with antennae removed), thorax, abdomen, legs and wings. 【Conclusion】 These results establish a foundation for further research on the molecular mechanisms of chemical communications in L. decemlineata.

Key words: Leptinotarsa decemlineata, odorant-binding protein, sequence analysis, tissue expression profile, phylogeny

张方梅, 刘杨, 李祥瑞, 张云慧, 程登发, 郭文超, 吐尔逊·阿合买提. 马铃薯甲虫气味结合蛋白的序列和基因表达谱分析[J]. 昆虫学报, 2019, 62(4): 428-441.

ZHANG Fang-Mei, LIU Yang, LI Xiang-Rui, ZHANG Yun-Hui, CHENG Deng-Fa, GUO Wen-Chao, TURSUN Ahmat.

Sequence analysis and gene expression profiling of odorant binding proteins in the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) [J]. Acta Entomologica Sinica, 2019, 62(4): 428-441.

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马铃薯甲虫气味结合蛋白的序列和基因表达谱分析

Sequence analysis and gene expression profiling of odorant binding proteins in the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

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