气味受体基因Orco在中华蜜蜂雄蜂触角中的表达及定位分析

doi:10.3864/j.issn.0578-1752.2015.04.17

Abstract

Abstract: 【Objective】 Insect odorant receptors (Ors) adopt a ligand-gated ion channel signal transduction mechanism primarily in olfaction recognition process. This ion channel is formed by a unique Orco protein with a common Ors which are combined to a heterodimer. Odorant receptors coreceptor (Orco) is an unique family, which shares highly conserved gene sequence among different insects and plays a key role in regulating insect behavior. The objective of this study is to research the expression character of Orco in drones of Apis cerana cerana based on the former relevant study in worker bees. 【Method】 Five different tissues of antennae, heads (without antennae), thoraces, abdomens and legs from 1-day-old drones, and the larvae, pupae and antennae of adults at different developmental stages were collected. Total RNA of each sample was extracted, and the expression profiles of the Orco in different tissues and at different developmental stages were examined by real-time quantitative PCR (qPCR). Antennae from 1-day-old drones were dissected and the frozen slides were prepared and the digoxigenin-labelled oligonucleotide probes were synthesized. The cellular localization in antennae was detected by in situ hybridization. 【Result】 qPCR results showed that the expression of Orco mRNA in different tissues were antenna>head>leg>abdomen>thorax, and it was about 100 times more abundant in antenna than in head. Orco transcripts were detected at all development stages of drones. The expression levels were very low in larval and pupal periods, increased after eclosion, then maintained a higher level during and after sexual maturity stage of drones. Combining with the former data got from the qPCR of worker bees, it showed that the expression level of Orco was significantly higher in drones than in workers. Localization test showed that Orco were largely expressed in neuronal cells of sensilla trihodea and sensilla placodea in antennae. 【Conclusion】 This study displayed the odorant receptor gene Orco was predominantly expressed in antennae of drones and the expression reached its peak when drones grow to sexual maturity. In addition, it was speculated combining the results of previous research that Orco expression biased in male sexually of A. cerana cerana.

Key words: Apis cerana cerana, odorant receptor, expression profiling, localization analysis

ZHAO Hui-ting, GAO Peng-fei, ZHANG Gui-xian, TIAN Song-hao, YANG Shan-shan, MENG Jiao, JIANG Yu-suo. Expression and Localization Analysis of the Odorant Receptor Gene Orco in Drones Antennae of Apis cerana cerana[J].Scientia Agricultura Sinica, 2015, 48(4): 796-803.

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References

[1] Vosshall L B, Amrein H, Morozov P S, Rzhetsky A, Axel R. A spatial map of olfactory receptor expression in the Drosophila antenna. Cell, 1999, 96: 725-736.

[2] Vogt R G, Miller N E, Litvack R, Fandino R A, Sparks J, Friedman R, Dickens J C. The insect SNMP gene family. Insect Biochemistry and Molecular Biology,2009, 39: 448-456.

[3] 杨冠煌. 引入西方蜜蜂对中蜂的危害及生态影响. 昆虫学报, 2005, 48(3): 401-406.

Yang G H. Harm of introducing the western honeybee Apis mellifera L. to the Chinese honeybee Apis cerana F. and its ecological impact. Acta Entomologica Sinica, 2005, 48(3): 401-406. (in Chinese)

[4] Benton R, Sachse S, Michnick S W, Vosshall L B. A typical membrane topology and heteromeric function of Drosophila odorant receptors in vivo. PloS Biology, 2006, 4(2): 240-257.

[5] Sato K, Pellegrino M, Nakagawa T, Nakagawa T, Vosshall L B, Touhara K. Insect olfactory receptors are heteromeric ligand-gated ion channels. Nature, 2008,452(7190): 1002-1006.

[6] Wicher D, Schfer R, Bauernfeind R, Stensmyr M C, Heller R, Heinemann S H, Hansson B S. Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide-activated cation channels. Nature, 2008, 452(7190): 1007-1011.

[7] Leal W S. Odorant reception in insects: roles of receptors, binding proteins, and degrading enzymes. Annual Review of Entomology, 2013, 58: 373-391.

[8] Touhara K, Vosshall L B. Sensing odorants and pheromones with chemosensory receptors. Annual Review of Physiology, 2009, 71: 307-332.

[9] Neuhaus E M, Gisselmann G, Zhang W, Dooley R, Stortkuhl K, Hatt H. Odorant receptor heterodimerization in the olfactory system of Drosophila melanogaster. Nature Neuroscience, 2005, 8(1): 15-17.

[10] Tsitoura P, Andronopoulou E, Tsikou D, Agalou A, Papakonstantinou M P, Kotzia G A, Labropoulou V, Swevers L, Georgoussi Z, Koatas L. Expression and membrane topology of Anopheles gambiae odorant receptors in Lepidopteran insect cells. PLoS One, 2010, 5(11): e15428.

[11] Sakurai T, Nakagawa T, Mitsuno H, Mori H, Endo Y, Tanoue S, Yasukochi Y, Touhara K, Nishioka T. Identification and functional characterization of a sex pheromone receptor in the silkmoth Bombyx mori. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(47): 16653-16658.

[12] Robertson H M, Gadau J, Wanner K W. The insect chemoreceptor superfamily of the parasitoid jewel wasp Nasonia vitripennis. Insect Molecular Biology, 2010, 19(Suppl.1): 121-136.

[13] Robertson H M, Wanner K W. The chemoreceptor superfamily in the honeybee Apis melifera: expansion of the odorant, but not gustatory, receptor family. Genome Research, 2006, 16: 1395-1403.

[14] Krieger J, Klink O, Mohl C, Raming K, Breer H. A candidate olfactory receptor subtype highly conserved across different insect orders. Journal of Comparative Physiology A, 2003, 189(7): 519-526.

[15] 张林雅, 谢冰花, 倪翠侠, 赵磊, 李红亮, 商晗武. 中华蜜蜂Orco嗅觉受体基因的克隆、表达及亚细胞定位. 昆虫学报, 2012, 55(11): 1246-1254.

Zhang L Y, Xie B H, Ni C X, Zhao L, Li H L, Shang H W. Cloning, expression and subcellualr localization of the olfactory co-receptor Orco gene in the Chinese honeybee, Apis cerana cerana (Hymentoptera: Apidae). Acta Entomologica Sinica, 2012, 55(11): 1246-1254. (in Chinese)

[16] Zhao H T, Gao P F, Zhang G X, Ma W H, Jiang Y S. Molecular identification and expressive characterization of an olfactory co- receptor gene in Apis cerana cerana. Journal of Insect Science, 2013, 13: Article 80.

[17] Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-^△△^Ct method. Methods, 2001, 25(4): 402-408.

[18] 孔畅仪, 王桂荣, 刘杨, 严善春. 小菜蛾三个普通气味受体基因的克隆及表达谱. 中国农业科学, 2014, 47(9): 1735-1742.

Kong C Y, Wang G R, Liu Y, Yan S C. Gene cloning and expression analysis of three odorant receptors in the diamondback moth (Plutella xylostella). Scientia Agricultura Sinica, 2014, 47(9): 1735-1742. (in Chinese)

[19] Miller R, Tu Z. Odorant receptor c-terminal motifs in divergent insect species. Journal of Insect Science, 2008, 8: Article 53.

[20] Olafson P U. Molecular characterization and immunolocalization of the olfactory co-receptorOrcofrom two blood-feeding muscid flies, the stable fly (Stomoxys calcitrans, L.) and the horn fly (Haematobia irritans irritans, L.). Insect Molecular Biology, 2013, 22(2): 131-142.

[21] 陈茜, 吴仲南, 杜永均, 诸葛启钏. 斜纹夜蛾嗅觉受体基因Ⅱ的表达谱分析. 昆虫学报, 2011, 54(8): 881-886.

Chen X, Wu Z N, Du Y J, ZhuGe Q C. Expression profiling of olfactory receptor gene Ⅱin the tobacco cutworm, Spodoptera litura (Lepidoptera: Notuidae). Acta Entomologica Sinica, 2011, 54(8): 881-886. (in Chinese)

[22] 申建梅, 胡黎明, 宾淑英, 林进添. 瓜实蝇嗅觉受体基因的克隆及表达谱分析. 昆虫学报, 2011, 54(3): 265-271.

Shen J M, Hu L M, Bin S Y, Lin J T. Cloning and expression profiling of an olfactory receptor gene in Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae). Acta Entomologica Sinica, 2011, 54(3): 265-271. (in Chinese)

[23] 赵海明, 易欣, 胡美英, 安国栋, 王会东, 黄林. 橘小实蝇非典型气味受体Orco基因的克隆与表达谱分析. 华中农业大学学报, 2013, 32(1): 40-47.

Zhao H M, Yi X, Hu M Y, An G D, Wang H D, Huang L. Gene cloning and expression profiling of atypical odorant receptors Orco gene in Bactrocera dorsalis (Hendel). Journal of Huazhong Agricultural University, 2013, 32(1): 40-47. (in Chinese)

[24] 张帅, 张永军, 苏宏华, 高希武, 郭予元. 中红侧沟茧蜂非典型气味受体的克隆及组织特异性表达. 中国农业科学, 2009, 42(5): 1639-1645.

Zhang S, Zhang Y J, Su H H, Gao X W, Guo Y Y. Gene cloning and tissue-specific expression of an olfactory receptor in Microplitis mediator (Hymenoptera: Braconidae). Scientia Agricultura Sinica, 2009, 42(5): 1639-1645. (in Chinese)

[25] Larsson M C, Domingos A I, Jones W D, Chiappe M E, Amrein H, Vosshall L B. Or83b encodes a broadly expressed odorant receptor essential for Drosophila olfaction. Neuron, 2004, 43(5): 703-714.

[26] 李红亮, 张林雅, 庄树林, 倪翠侠, 韩宝瑜, 商晗武. 中华蜜蜂普通气味结合蛋白ASP2的气味结合功能模式分析. 中国农业科学, 2013, 46(1): 154-161.

Li H L, Zhang L Y, Zhuang S L, Ni C X, Han B Y, Shang H W. Interpretation of odorant binding function and mode of general odorant binding protein ASP2 in Chinese honeybee (Apis cerana cerana). Scientia Agricultura Sinica, 2013, 46(1): 154-161. (in Chinese)

[27] Kaissling K E, Renner M. Antennae rezeptoren für queen substance und sterzelduft bei der Honigbiene. Zeitschrift für vergleichende Physiologie, 1968, 59(4): 357-361.

[28] Wanner K W, Anderson A R, Trowell S C, Theilmann D A, Robertson H M, Newcomb R D. Female-biased expression of odourant receptor genes in the adult antennae of the silkworm, Bombyx mori. Insect Molecular Biology, 2007,16(1): 107-119.

[29] Brigaud I, Montagné N, Monsempes C, François M C, Jacquin-Joly E. Identification of an atypical insect olfactory receptor subtype highly conserved within noctuids. The FEBS Journal, 2009, 276(22): 6537-6547.

[30] 张帅, 张永军, 苏宏华, 高希武, 郭予元. 棉铃虫气味受体的克隆与组织特异性表达. 昆虫学报, 2009, 52(7): 728-735.

Zhang S, Zhang Y J, Su H H, Gao X W, Guo Y Y. Cloning and tissue-specific expression of olfactory receptors in Helicoverpa armigera (Hübner). Acta Entomologica Sinica, 2009, 52(7): 728-735. (in Chinese)

[31] Zheng W W, Zhu C P, Peng T, Zhang H Y. Odorant receptor co- receptor Orco is upregulated by methyl eugenol in male Bactrocera dorsalis (Diptera: Tephritidae). Journal of Insect Physiology, 2012, 58: 1122-1127.

[32] 马龙, 王山宁, 路子云, 刘泽文, 张永军, 郭予元. 中红侧沟茧蜂嗅觉受体MmedOr2基因的克隆及组织表达谱. 中国生物防治学报, 2014, 30(1): 13-19.

Ma L, Wang S N, Lu Z Y, Liu Z W, Zhang Y J, Guo Y Y. Cloning and tissue expression analysis of an olfactory receptor MmedOr2 in Microplitis mediator (Hymenoptera: Braconidae). Chinese Journal of Biological Control, 2014, 30(1): 13-19. (in Chinese)

[33] Jordan M D, Anderson A, Begum D, Carraher C, Authier A, Marshall S D G, Kiely A, Gatehouse L N, Greenwood D R, Christie D L, Kralicek A V, Trowell S C, Newcomb R D. Odorant receptors from the light brown apple moth (Epiphyas postvittana) recognize important volatile compounds produced by plants. Chemical Senses, 2009, 34(5): 383-394.

[34] Liu S, Huang Y J, Qiao F, Zhou W W, Gong Z J, Cheng J A, Zhu Z R. Cloning, tissue distribution, and transmembrane orientation of the olfactory co-receptor Orco from two important Lepidopteran rice pests, the leaffolder (Cnaphalocrocis medinalis) and the striped stem borer (Chilo suppressalis). Journal of Integrative Agriculture, 2013, 12(10): 1816-1825.

[35] Esslen J, Kaissling K E. Number and distribution of the sensilla on the antennal flagellum of the honeybee (Apis mellifera L.). Zoomorphologie, 1976, 83(3): 227-251.

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Expression and Localization Analysis of the Odorant Receptor Gene Orco in Drones Antennae of Apis cerana cerana

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