Q型烟粉虱化学感受蛋白基因BtabCSP1的克隆与分析

doi:10.3864/j.issn.0578-1752.2012.18.022

摘要/Abstract

摘要： 【目的】克隆Q型烟粉虱（Bemisia tabaci）化学感受蛋白基因，分析其序列特征和表达特性，为深入研究该类基因功能奠定基础。【方法】根据桃蚜化学感受蛋白OS-D2a（ABM5558）的氨基酸序列，借助生物信息学搜索烟粉虱EST序列拼接了1个烟粉虱化学感受蛋白基因（BtabCSP1），以烟粉虱mRNA为模板，采用RT-PCR扩增克隆烟粉虱化学感受蛋白cDNA。采用半定量RT-PCR对不同发育阶段的烟粉虱样品中BtabCSP1的表达模式进行研究。【结果】BtabCSP1完整阅读框全长为2 626 bp，包含2个外显子和1个内含子，编码131个氨基酸残基，其中含有1个CX6CX18CX2结构域，为化学感受蛋白的典型特征。BtabCSP1在供试样品中均有表达，并且在2龄、3龄和伪蛹阶段表达量更高。聚类分析结果表明，烟粉虱与果蝇、桃蚜等物种遗传关系较远，化学感受蛋白基因在物种进化过程中较为活跃。【结论】从Q型烟粉虱若虫中克隆了1个化学感受蛋白基因 BtabCSP1，该基因在烟粉虱不同发育阶段中均能表达，推测其对烟粉虱的生长发育具有重要的调控作用。

关键词: Q型烟粉虱, 化学感受蛋白, 基因克隆, 特征分析

Abstract: 【Objective】 The objective of this study is to clone the full length cDNA of the gene encoding chemosensory proteins from Bemisia tabaci Q biotype, and to analyze its expression pattern at different developmental stages.【Method】Based on the amino acid sequence of aphids (Myzus persicae) chemosensory proteins OS-D2a (ABM5558) reported previously, a pair of primers were designed based on simulation of gene splicing and RT-PCR was used to clone the chemosensory protein gene BtabCSP1. 【Result】Sequencing and structural analysis showed that BtabCSP1 contains a 2 626 bp long complete reading frame (with 2 exons and 1 intron) and encodes 131 amino acid residues with a CX6CX18CX2 domain, the typical characteristics of a chemosensory protein gene. As displayed by semi-quantitative RT-PCR amplification from total RNA of B. tabaci of various stages, BtabCSP1 was expressed in all samples and the expressions were more apparent in the 2nd instar, 3rd instar and 4th instar (pupal stage) nymphs. The amino acid (encoded by BtabCSP1) sequence cluster analysis showed that B. tabaci was distantly related to fruit flies and aphids and chemosensory protein gene was active in the process of species evolution.【Conclusion】BtabCSP1 was isolated from B. tabaci Q biotype, and the expression pattern suggested the close relationship between the expression of this gene and the development of B. tabaci.

Key words: Bemisia tabaci Q biotype, CSP, gene cloning, characteristics analysis

白润娥, 李静静, 唐雅菲, 熊大斌, 李帅良, 闫凤鸣. Q型烟粉虱化学感受蛋白基因BtabCSP1的克隆与分析[J]. 中国农业科学, 2012, 45(18): 3892-3898.

BAI Run-E, LI Jing-Jing, TANG Ya-Fei, XIONG Da-Bin, LI Shuai-Liang, YAN Feng-Ming. Cloning and Characteristics Analysis of Chemosensory Protein Gene BtabCSP1 in Bemisia tabaci Q Biotype[J]. Scientia Agricultura Sinica, 2012, 45(18): 3892-3898.

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参考文献

[1]Pilpel Y, Lancet D. Olfaction: Good reception in fruitfly antennae. Nature, 1999, 398: 285-287.

[2]Getchell T V, Margolis F L, Getchell M L. Perireceptor and receptor event s in vertebrate olfaction. Progress in Neurobiology, 1984, 23(4): 317-345.

[3]Yang G, Winberg G, Ren H, Zhang S. Expression, purification and functional analysis of an odorant binding protein AaegOBP22 from Aedes aegypti. Protein Expression and Purification, 2011, 75(2): 165-171.

[4]Angeli S, Ceron F, Scaloni A, Monti M, Monteforti G, Minnocci A, Petacchi R, Pelosi P. Purification, structural characterization, cloning and immunocytochemical localization of chemoreception proteins from Schistocerca gregaria. European Journal of Biochemistry, 1999, 262(3): 745-754.

[5]Pelosi P, Zhou J J, Ban L P, Calvello M. Soluble proteins in insect chemical communication. Cellular and Molecular Life Sciences, 2006, 63(14): 1658-1676.

[6]Robertson H M, Martos R, Sears C R, Todres E Z, Walden K K O, Nardi J B. Diversity of odourant binding proteins revealed by an expressed sequence tag project on male Manduca sexta moth antennae. Insect Molecular Biology, 1999, 8(4): 501-518.

[7]Sabatier L, Jouanguy E, Dostert C, Zachary D, Dimarcq J L, Bulet P, Imler J L. Pherokine-2 and -3. Two Drosophila molecules related to pheromone/odor-binding proteins induced by viral and bacterial infections. European Journal of Biochemistry, 2003, 270: 3398-3407.

[8]Wanner K W, Willis L G, Theilmann D A, Isman M B, Feng Q, Plettner E. Analysis of the insect os-d-like gene family. Journal of Chemical Ecology, 2004, 30(5): 889-911.

[9]Oduol F, Xu J, Niare O, Natarajan R, Vernick K D. Genes identified by an expression screen of the vector mosquito Anopheles gambiae display differential molecular immune response to malaria parasites and bacteria. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(21): 11397-11402.

[10]Nagnan-Le Meillour P, Cain A H, Jacquin-Joly E, François M C, Ramachandran S, Maida R, Steinbrecht R A. Chemosensory proteins from the proboscis of Mamestra brassicae. Chemical Senses, 2000, 25: 541-553.

[11]Gong L A, Zhong G H, Hu M Y, Luo Q, Ren Z Z. Molecular cloning, expression profile and 5' regulatory region analysis of two chemosensory protein genes from the diamondback moth, Plutella xylostella. Journal of Insect Science, 2010, 10: article 143.

[12]Ishida Y, Chiang V, Leal W S. Protein that makes sense in the Argentine ant. Naturwissenschaften, 2002, 89: 505-507.

[13]Zhou S H, Zhang J, Zhang S G, Zhang L. Expression of chemosensory proteins in hairs on wings of Locusta migratoria (Orthoptera: Acrididae). Journal of Applied Entomology, 2008, 132(6): 439-450.

[14]Jacobs S P, Liggins A P, Zhou J J, Pickett J A, Jin X, Field L M. OS-D-like genes and their expression in aphids (Hemiptera: Aphididae). Insect Molecular Biology, 2005, 14(4): 423-432.

[15]丁新红, 陆建林. 烟粉虱的危害及防治. 上海农业科技, 2008(3): 123.

Ding X H, Lu J L. Damage and control of Bemisia tabaci. Shanghai Agricultural Science and Technology, 2008(3): 123. (in Chinese)

[16]褚栋, 刘国霞, 陶云荔, 万方浩, 张友军. 烟粉虱Q型与B型种群动态及其影响因子研究进展. 植物保护学报, 2007, 34(3): 326-330.

Chu D, Liu G X, Tao Y L, Wan F H, Zhang Y J. Population dynamics of biotypes Q and B of Bemisia tabaci (Gennadius) and the influential factors. Acta Phytophylacica Sinica, 2007, 34(3): 326-330. (in Chinese)

[17]Calvello M, Brandazza A, Navarrini A, Dani F R, Turillazzi S, Felicioli A, Pelosi P. Expression of odorant-binding proteins and chemosensory proteins in some Hymenoptera. Insect Biochemistry and Molecular Biology, 2005, 35(4): 297-307.

[18]刘金香, 钟国华, 谢建军, 官珊, 胡美英. 昆虫化学感受蛋白研究进展. 昆虫学报, 2005, 48(3): 418-426.

Liu J X, Zhong G H, Xie J J, Guan S, Hu M Y. Recent advances in chemosensory proteins of insects. Acta Entomologica Sinica, 2005, 48(3): 418-426. (in Chinese)

[19]Mosbah A, Campanacci V, Lartigue A, Tegoni M, Cambillau C, Darbon H. Solution structure of a chemosensory protein from the moth Mamestra brassicae. Biochemical Journal, 2003, 369: 39-44.

[20]Tomaselli S, Crescenzi O, Sanfelice D, Ab E, Wechselberger R, Angeli S, Scaloni A, Boelens R, Tancredi T, Pelosi P, Picone D. Solution structure of a chemosensory protein from the desert locust Schistocerca gregaria. Biochemistry, 2006, 45(35): 10606-10613.

[21]Dani F R, Michelucci E, Francese S, Mastrobuoni G, Cappellozza S, La Marca G, Niccolini A, Felicioli A, Moneti G, Pelosi P. Odorant-binding proteins and chemosensory proteins in pheromone detection and release in the silkmoth Bombyx mori. Chemical Senses, 2011, 36(4): 335-344.

[22]McKenna M P, Hekmat-Scafe D S, Gaines P, Carlson J R. Putative Drosophila pheromone-binding proteins expressed in a subregion of the olfactory system. The Journal of Biological Chemistry, 1994, 269(23): 16340-16347.

[23]Pikielny C W, Hasan G, Rouyer F, Rosbach M. Members of a family of Drosophila putative odorant-binding proteins are expressed in different subsets of olfactory hairs. Neuron, 1994, 12: 35-49.

[24]Jansen S, Chmelík J, Zídek L, Padrta P, Novák P, Zdráhal Z, Picimbon J F, Löfstedt C, Sklenár V. Structure of Bombyx mori chemosensory protein 1 in solution. Archives of Insect Biochemistry and Physiology, 2007, 66: 135-145.

[25]龚亮, 陈永, 程功, 钟国华. 昆虫化学感受蛋白. 昆虫知识, 2009, 46(4): 646-652.

Gong L, Chen Y, Cheng G, Zhong G H. Insect chemosensory proteins. Chinese Bulletin of Entomology, 2009, 46(4): 646-652. (in Chinese)

[26]Maleszka J, Foret S, Saint R, Maleszka R. RNAi-induced phenotypes suggest a novel role for a chemosensory protein CSP5 in the development of embryonic integument in the honeybee (Apis mellifera). Development Genes and Evolution, 2007, 217: 189-196.