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桃蚜鞣化激素基因克隆与表达分析

张超 杨洪 杨茂发 喻会平 张伟

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. doi: 10.19303/j.issn.1008-0384.2017.011.015
引用本文: 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. doi: 10.19303/j.issn.1008-0384.2017.011.015
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. doi: 10.19303/j.issn.1008-0384.2017.011.015
Citation: 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. 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

doi: 10.19303/j.issn.1008-0384.2017.011.015
基金项目: 

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

详细信息
    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)

    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)

  • 中图分类号: Q966

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

  • 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-αburs-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。
  • 图  1  桃蚜burs-α全长序列及氨基酸序列

    注:3个糖基化位点的氨基酸用方框标识,3个蛋白激酶磷酸化位点的氨基酸残基用下划波浪线标识,信号肽用下划直线标识。

    Figure  1.  Full-length amino acid sequence of burs-α in M. persicae

    图  2  桃蚜burs-β全长序列及氨基酸序列

    注:2个蛋白激酶磷酸化位点的氨基酸残基用下划波浪线标识,信号肽用下划直线标识。

    Figure  2.  Full-length amino acid sequence of burs-β in M. persicae

    图  3  桃蚜与豌豆蚜Bursicon基因三级结构预测

    注:A为桃蚜burs-α; B为豌豆蚜burs-α; C为桃蚜burs-β; D为豌豆蚜burs-β。

    Figure  3.  Tertiary structures of bursicon in M. persicae and A. pisum

    图  4  桃蚜burs-α蛋白和其他昆虫的系统发育树

    Figure  4.  Phylogenetic trees on burs-α amino acid sequence of M. persicae and other insects

    图  5  桃蚜burs-β蛋白和其他昆虫的系统发育树

    Figure  5.  Phylogenetic trees on burs-β amino acid sequence of M. persicae and other insects

    图  6  桃蚜不同龄期Mpburs-α基因的表达水平

    注:不同字母表示差异显著(P<0.05),图 7同。

    Figure  6.  Expression levels of Mpburs-α in different developmental stages of M. persicae

    图  7  桃蚜不同龄期Mpburs-β基因的表达水平

    Figure  7.  Expression levels of Mpburs-β in different developmental stages of M. persicae

    表  1  桃蚜burs-α和burs-β基因扩增及实时荧光定量PCR引物信息

    Table  1.   Primers used for amplification and qRT-PCR of burs-α and burs-β in M. persicae

    扩增区域引物名称序列(5′-3′)
    中间片段Mpburs-αFGTCTACCATCAACCAGGAAT
    Mpburs-αRGCGGCATAGTTGGACATT
    Mpburs-βFACCCAGAAGAATGTGAGAC
    Mpburs-βRGGTCAATCTTACGCCATC
    5′末端扩增5′Mpburs-αR1:GCGATACCGAAGCTTCTCGTTC
    5′Mpburs-αR2:CTGCTGCAGCGACCTGTACATG
    5′Mpburs-αR3:GGCGTTACTTGGCAGTCGTCGC
    5′Mpburs-βR1:GCAATGTGTAAGAGCGATCACTC
    5′Mpburs-βR2:CCACTAGGAGTAATGACACTAG
    5′Mpburs-βR3:CTTGTTCACTGCTATGTCACC
    3′末端扩增3′Mpburs-αF1TGGTCGTGACCGCTAGGTCTAGC
    3′Mpburs-αF2GAACGAGAAGCTTCGGTATCGC
    3′Mpburs-αF3CACTGGAATGCATGTGCCGC
    3’ Mpburs-βF1GAACTTGGACGGCTTCAGAGGAC
    3′Mpburs-βF2GGAGCGTGCAATTCACAAGTAC
    3′Mpburs-βF3GAGAGAGAGAGTGATCGCTC
    荧光定量yg-Mpburs-αFGTCATCCATGTTCTACAATAC
    yg-Mpburs-αRCTTCTCGTTCACCACTCTC
    yg-Mpburs-βFTGGTGACATAGCAGTGAAC
    yg-Mpburs-βRGGATCGTAGCAATGTGTAAG
    内参基因actin-FTGGTATCGTCTTGGATTCTG
    actin-RTTAGGTAGTCGGTGAGATCA
    下载: 导出CSV
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  • 收稿日期:  2017-07-06
  • 修回日期:  2017-10-21
  • 刊出日期:  2017-11-28

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