桃蚜鞣化激素基因克隆与表达分析

doi:10.19303/j.issn.1008-0384.2017.011.015

福建农业学报 ›› 2017, Vol. 32 ›› Issue (11): 1244-1250.doi: 10.19303/j.issn.1008-0384.2017.011.015

桃蚜鞣化激素基因克隆与表达分析

张超¹, 杨洪^1,2, 杨茂发^1,2, 喻会平³, 张伟⁴

1. 贵州山地农业病虫害重点实验室, 贵州大学昆虫研究所, 贵州贵阳 550025;
2. 贵州大学烟草学院, 贵州贵阳 550025;
3. 贵州省烟草公司毕节市公司, 贵州毕节 551700;
4. 四川省农业厅植物保护站, 四川成都 610041

收稿日期:2017-07-06 修回日期:2017-10-21 出版日期:2017-11-28 发布日期:2017-11-28
通讯作者: 杨洪(1975-),男,博士,教授,从事昆虫生态与害虫综合治理(E-mail:agr.hyang@gzu.edu.cn) E-mail:agr.hyang@gzu.edu.cn
作者简介:张超(1992-),男,硕士,研究方向:昆虫生态与害虫综合治理(E-mail:zc0178sfp@163.com)
基金资助:
贵州省烟草公司毕节分公司科技项目（[2016]82）；贵州省烟草公司科技项目（[2015]11）；贵州大学研究生创新基金项目（研农2017018）；昆虫进化生物学和害虫控制国际科技合作基地项目（黔科合平台人才[2016]5802）

Cloning and Expression of Bursicon Gene of Myzus persicae (Sulzer)

ZHANG Chao¹, YANG Hong^1,2, YANG Mao-fa^1,2, YU Hui-ping Wei³, ZHANG Wei⁴

1. Guizhou Key Laboratory for Agricultural Pest Management of Mountainous Region, Institute of Entomology of Guizhou University, Guiyang, Guizhou 550025, China;
2. College of Tobacco Science of Guizhou University, Guiyang, Guizhou 550025, China;
3. Bijie Branch of Guizhou Tobacco Corporation, Bijie, Guizhou 551700, China;
4. Plant Protection Station of Sichuan Province, Chengdu, Sichuan 610041, China

Received:2017-07-06 Revised:2017-10-21 Online:2017-11-28 Published:2017-11-28

摘要/Abstract

摘要： Bursicon是调节昆虫表皮硬化及展翅的激素类蛋白，它在昆虫表皮硬化和翅伸展过程中起着关键作用。本研究克隆获得桃蚜Bursicon基因（burs-α和burs-β）全长，分别命名为Mpburs-α（GenBank登录号：MF083566）和Mpburs-β（GenBank登录号：MF083567）。Mpburs-α开放阅读框长483 bp，编码160个氨基酸。其相对分子量为17.59 kDa，分子式为C₇₅₉H₁₂₀₄N₂₁₀O₂₃₂S₁₉；理论等电点为7.96，Mpburs-β开放阅读框长417 bp，编码138个氨基酸。其相对分子量为15.45 kDa，分子式为C₆₇₂H₁₀₇₂N₁₈₀O₂₁₂S₁₂；理论等电点为4.84。分子系统进化树分析表明，桃蚜与豌豆蚜Acyrthosiphon pisum和麦双尾蚜Diuraphis noxia具有较近的亲缘关系。RT-qPCR结果表明：Mpburs-α和Mpburs-β在桃蚜各龄期均有表达，以1龄若蚜期表达量最高；成蚜有翅个体中表达量显著高于无翅个体。推测Bursicon在桃蚜翅的形成中发挥着重要作用。研究结果可为续研究Bursicon在桃蚜蜕皮后表皮的骨化、翅的形成方面相关的功能机制奠定基础。

关键词: 桃蚜, 鞣化激素基因, 克隆, 表达分析

Abstract: Bursicon regulates the epidermal sclerosis and wing-spreading in insects. It plays a key role in epidermis hardening of insects after molting. The full-length sequences of the bursicon genes (burs-α and burs-β) of M. persicae (Sulzer) were obtained through cloning for this study, and named Mpburs-α (GenBank accession number:MF083566) and Mpburs-β (GenBank accession number:MF083567), respectively. Mpburs-α contained a 483 bp open reading frame (ORF) and was encoded 160 amino acid residues. Its molecular mass was predicted to be 17.59 kDa with a formula of C₇₅₉H₁₂₀₄N₂₁₀O₂₃₂S₁₉ and an isoelectric point of 7.96. Mpburs-β contained a 417 bp ORF and was encoded 138 amino acids. Its molecular mass to be 15.45 kDa with a formula of C₆₇₂H₁₀₇₂N₁₈₀O₂₁₂S₁₂ and an isoelectric point of 4.8. The phylogenetic tree analysis showed a relatively close genetic relationship of M. persicae with Acyrthosiphon pisum (Harris) and Diuraphis noxia (Mordvilko). The qRT-PCR results indicated that Mpburs-α and Mpburs-β expressed during the entire developmental stage of M. persicae and arrived at a peak expression level in the 1^st instar nymph. The expression levels of the two genes in the winged adult aphids were significantly higher than those in the wingless adult aphids suggesting an important role played by bursicon in the wing development for the insect. The finding might lead to further studies on the functions of bursicon in the molting epidermal ossification and wing formation of M. persicae.

Key words: Myzus persicae(Sulzer), bursicon, gene cloning, expression analysis

中图分类号:

Q966

张超, 杨洪, 杨茂发, 喻会平, 张伟. 桃蚜鞣化激素基因克隆与表达分析[J]. 福建农业学报, 2017, 32(11): 1244-1250.

ZHANG Chao, YANG Hong, YANG Mao-fa, YU Hui-ping Wei, ZHANG Wei. Cloning and Expression of Bursicon Gene of Myzus persicae (Sulzer)[J]. Fujian Journal of Agricultural Sciences, 2017, 32(11): 1244-1250.

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桃蚜鞣化激素基因克隆与表达分析

Cloning and Expression of Bursicon Gene of Myzus persicae (Sulzer)

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