棉蚜唾液蛋白C002基因克隆及在棉蚜不同寄主专化型中的表达分析
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摘要
C002是蚜虫取食过程中的一种十分重要的水溶性唾液蛋白。本研究从棉蚜的转录组数据中,克隆得到棉蚜C002基因的c DNA序列(Gen Bank登录号:AHX71991.1)。C002基因的开放阅读框全长为641 bp,编码213个氨基酸,推测其蛋白质的分子质量为24.1 k Da,等电点为9.06,具有由24个氨基酸组成的信号肽。运用荧光定量PCR技术,分析棉花型棉蚜和黄瓜型棉蚜的时间表达谱,结果表明,C002基因在两种寄主专化型棉蚜生长发育的早期阶段表达量均较高。将棉花型棉蚜和黄瓜型棉蚜分别转接到西葫芦后,C002基因的表达量在转接前后无显著变化,说明C002基因在棉蚜适应寄主过程中并没有起到关键性作用。
C002 is a watery saliva protein which is thought to be crucial for aphis feeding. In this study,the C002 gene sequence of cotton aphid was cloned from the transcript data of cotton aphid(Gen Bank accession: AHX71991. 1). The open reading frame of the C002 was 641 bp,and encoded 213 amino acids. Its predicted molecular mass was 24. 1 k Da. The isoelectric point was 9. 06. It had a signal peptide sequence with 24 amino acids. The temporal expression pattern of the C002 of the cotton specialized aphid and cucumber specialized aphid were studied by q PCR analysis. The results showed that the expression of C002 gene were higher in the early stage of growth and development of the two host biotypes of cotton aphid. The cotton specialized aphid and cucumber specialized aphid are transferred to zucchini,the expression level of C002 gene did not change significantly before and after the transfer. Theresults showed that the C002 gene didn't play a key role in the process of adapting to the host.
C002 is a watery saliva protein which is thought to be crucial for aphis feeding. In this study,the C002 gene sequence of cotton aphid was cloned from the transcript data of cotton aphid(Gen Bank accession: AHX71991. 1). The open reading frame of the C002 was 641 bp,and encoded 213 amino acids. Its predicted molecular mass was 24. 1 k Da. The isoelectric point was 9. 06. It had a signal peptide sequence with 24 amino acids. The temporal expression pattern of the C002 of the cotton specialized aphid and cucumber specialized aphid were studied by q PCR analysis. The results showed that the expression of C002 gene were higher in the early stage of growth and development of the two host biotypes of cotton aphid. The cotton specialized aphid and cucumber specialized aphid are transferred to zucchini,the expression level of C002 gene did not change significantly before and after the transfer. Theresults showed that the C002 gene didn't play a key role in the process of adapting to the host.
引文
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Miles PW.Aphid saliva[J].Biological Reviews of the Cambridge Philosophical Society,1999,74(1):41-85.
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Mutti NS,Louis J,Pappan LK,et al.A protein from the salivary glands of the pea aphid,Acyrthosiphon pisum is essential in feeding on a host plant[J].Proceedings of the National Academy of Sciences,2008,105(29):9965-9969.
Shaposhnikov GK.Morphological divergence and convergence in an experiment with aphids(Homoptera;Aphidoidea)[J].Entomological Review,1965,44(1):1-12.
Yin HD,Huang CH,Xue K,et al.Roles of insect salivary components in insect-plant interactions[J].Acta Entomological Sinica,2006,49(5):843-849.[殷海娣,黄翠虹,薛堃,等.昆虫唾液成分在昆虫与植物关系中的作用[J].昆虫学报,2006,49(5):843-849]
Zheng CL,Liu XD,Zhai BP.The fitness of host biotypes of the cotton aphid(Aphis gossypii Glover)to host plants and their reciprocal transfer pathways[J].Acta Ecologica Sinica,2007,27(5):1879-1886.[郑彩玲,刘向东,翟保平.棉花型和黄瓜型棉蚜(Aphis gossypii Glover)的寄主适应性及转移通道[J].生态学报,2007,27(5):1879-1886]
Drès M.Host races in plant-feeding insects and their importance insympatric speciation[J].Philosophical Transactions of the Royal Society of London,2002,357(1420):471-492.
Feng GL,Zhao ZW,Li M,et al.Relationship between esterase activities of the cotton aphid(Aphis gossypii)and overwintering host plants[J].Acta Entomological Sinica,2001,44(3):304-310.[冯国蕾,赵章武,李梅,等.不同寄主植物与棉蚜酯酶活性的关系[J].昆虫学报,2001,44(3):304-310]
Guildmond JA,Tigges WT,Vroker PW.Host race of Aphis gossypii(Homoptera:Aphidiae)on cucumber and chrysanthemum[J].Environmental Entomology,1994,23(5):1235-1240.
Inaizumi M.Life cycle of Aphis gossypii Glover(Homoptera,Aphididae)with special reference to biotype differentiation on various host plants[J].Japanese Journal of Entomology,1981,49:219-240.
Li XF,Fan J,Sun YW,et al.Cloning and RNA interference analysis of the Sitobion avenae salivary protein C002 gene[J].Chinese Journal of Applied Entomology,2014,51(6):1479-1487.[李雪峰,范佳,孙永伟,等.麦长管蚜唾液蛋白C002的基因克隆与RNA干扰研究[J].应用昆虫学报,2014,51(6):1479-1487]
Liu XD,Zhang LJ,Zhang XX,et al.Studies on cotton aphid Aphis gossypii selectivity to host and its host-type[J].Acta Ecologica Sinica,2002,22(8):1281-1285.[刘向东,张立建,张孝羲,等.棉蚜对寄主的选择及寄主专化型研究[J].生态学报,2002,22(8):1281-1285]
Liu XD,Zhang XX,Zhai BP.Host biotypes and their formation causes in aphids[J].Acta Entomological Sinica,2004,47(4):499-506.[刘向东,张孝羲,翟保平.蚜虫寄主专化型及其成因[J].昆虫学报,2004,47(4):499-506]
Livak KJ,Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT,Method[J].Methods,2001,25(4):402-408.
Meng L,Li BP.Researches on biotypes of cotton aphid in Xinjiang[J].Cotton Science,2001,13(1):30-35.[孟玲,李保平.新疆棉蚜生物型的研究[J].棉花学报,2001,13(1):30-35]
Miles PW.Aphid saliva[J].Biological Reviews of the Cambridge Philosophical Society,1999,74(1):41-85.
Mutti NS,Park Y,Reese JC,et al.RNAi knockdown of a salivary transcript leading to lethality in the pea aphid,Acyrthosiphon pisum[J].Journal of Insect Science,2006,6(38):1-7.
Mutti NS,Louis J,Pappan LK,et al.A protein from the salivary glands of the pea aphid,Acyrthosiphon pisum is essential in feeding on a host plant[J].Proceedings of the National Academy of Sciences,2008,105(29):9965-9969.
Shaposhnikov GK.Morphological divergence and convergence in an experiment with aphids(Homoptera;Aphidoidea)[J].Entomological Review,1965,44(1):1-12.
Yin HD,Huang CH,Xue K,et al.Roles of insect salivary components in insect-plant interactions[J].Acta Entomological Sinica,2006,49(5):843-849.[殷海娣,黄翠虹,薛堃,等.昆虫唾液成分在昆虫与植物关系中的作用[J].昆虫学报,2006,49(5):843-849]
Zheng CL,Liu XD,Zhai BP.The fitness of host biotypes of the cotton aphid(Aphis gossypii Glover)to host plants and their reciprocal transfer pathways[J].Acta Ecologica Sinica,2007,27(5):1879-1886.[郑彩玲,刘向东,翟保平.棉花型和黄瓜型棉蚜(Aphis gossypii Glover)的寄主适应性及转移通道[J].生态学报,2007,27(5):1879-1886]
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