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大豆咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因克隆及结构分析

郭子雯 司修洋 焦莉苹 刘大伟

郭子雯,司修洋,焦莉苹,等. 大豆咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因克隆及结构分析 [J]. 福建农业学报,2023,38(5):616−623 doi: 10.19303/j.issn.1008-0384.2023.05.013
引用本文: 郭子雯,司修洋,焦莉苹,等. 大豆咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因克隆及结构分析 [J]. 福建农业学报,2023,38(5):616−623 doi: 10.19303/j.issn.1008-0384.2023.05.013
GUO Z W, SI X Y, JIAO L P, et al. Cloning and Bioinformatics of CCoAOMT Relating to Resistance of Soybean to Cyst Nematodes [J]. Fujian Journal of Agricultural Sciences,2023,38(5):616−623 doi: 10.19303/j.issn.1008-0384.2023.05.013
Citation: GUO Z W, SI X Y, JIAO L P, et al. Cloning and Bioinformatics of CCoAOMT Relating to Resistance of Soybean to Cyst Nematodes [J]. Fujian Journal of Agricultural Sciences,2023,38(5):616−623 doi: 10.19303/j.issn.1008-0384.2023.05.013

大豆咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因克隆及结构分析

doi: 10.19303/j.issn.1008-0384.2023.05.013
基金项目: 黑龙江省自然科学基金(LH2019C034)
详细信息
    作者简介:

    郭子雯(1999−),女,硕士研究生,研究方向:植物病原线虫学(E-mail:1057138000@qq.com

    通讯作者:

    刘大伟(1983−),男,博士,副教授,研究方向:植物病原线虫学(E-mail:liudawei353@163.com

  • 中图分类号: S435

Cloning and Bioinformatics of CCoAOMT Relating to Resistance of Soybean to Cyst Nematodes

  • 摘要:   目的  对大豆中咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)基因进行克隆和生物信息学分析,为研究木质素代谢途径关键基因CCoAOMT在大豆抗胞囊线虫的作用机制奠定基础。  方法  以高抗大豆胞囊线虫(Heterodera glycines Ichinohe)3号生理小种品种灰皮支黑豆根系为材料,利用RT-PCR技术进行克隆。  结果  克隆得到1条大豆 CCoAOMT 基因的cDNA全长序列,将其命名为 GmCCoAOMT ,GenBank登录号为MW480860。该基因全长为848 bp,含有1个741 bp的开放阅读框,编码246个氨基酸组成的蛋白,分子量为27.6 kDa,等电点为5.67;亚细胞定位预测显示,CCoAOMT蛋白可能位于细胞质,该蛋白含有1个保守的AdoMet_MTases结构域,属于AdoMet_MTases超级家族;在与其他豆科植物CCoAOMT编码的蛋白序列比对及系统进化树分析中,GmCCoAOMT编码的蛋白序列与其他豆科植物具有较高的相似性。  结论  获得大豆CCoAOMT基因的cDNA全长序列,通过构建进化树确定大豆与豇豆、木豆、菜豆和赤豆的CCoAOMT蛋白进化关系较近,而与白羽扇豆和鹰嘴豆进化关系较远。
  • 图  1  根系总RNA的电泳检测

    Figure  1.  Electrophoresis of total RNA in roots

    图  2  CCoAOMT基因的RT-PCR结果

    M: DNA marker;1、2: RT-PCR 产物

    Figure  2.  RT-PCR on CCoAOMT

    M: DNA marker;1 and 2: RT-PCR products.

    图  3  大豆CCoAOMT基因全长序列及其编码的氨基酸序列

    Figure  3.  Full-length cDNA and amino acid sequence of CCoAOMT in soybean

    图  4  大豆CCoAOMT蛋白保守结构域的预测

    Figure  4.  Predicted conservative domain of GmCCoAOMT protein

    图  5  大豆CCoAOMT蛋白亲疏水性分析

    Figure  5.  Hydrophobicity of GmCCoAOMT protein

    图  6  大豆CCoAOMT蛋白拓扑结构分析

    Figure  6.  Topological structure of GmCCoAOMT protein

    图  7  大豆与其他豆科植物CCoAOMT蛋白氨基酸多重序列比对

    Figure  7.  Multiple alignment of amino acid sequences between CCoAOMTs in soybean and other legumes

    图  8  大豆与其他植物CCoAOMT蛋白的进化树

    Figure  8.  Phylogenetic tree of CCoAOMT proteins in soybean and other plants

    图  9  大豆CCoAOMT蛋白的二级结构预测

    Figure  9.  Predicted secondary structure of soybean CCoAOMT protein

    图  10  大豆CCoAOMT蛋白的三维建模

    Figure  10.  Predicted tertiary structure of soybean CCoAOMT protein

    表  1  大豆和其他豆科植物CCoAOMT蛋白的氨基酸组成和理化性质对比

    Table  1.   Amino acid composition and physicochemical properties of CCoAOMTs in soybean and other plants

    序号
    Serial number
    物种
    Species
    登录号
    Accession No.
    氨基酸数量
    Number of
    amino acids
    相对分子量
    Relative molecular
    mass/kDa
    理论等电点
    Theoretical isoelectric
    point
    比例
    Percentage/%
    BAACARAL
    1大豆 Glycine maxMW480860 246 27.6 5.6714.213.87.384.6
    2菜豆 Phaseolus vulgarisAGV54319.124627.85.3213.814.68.583.4
    3虎爪豆 Mucuna pruriens RDX78299.124828.05.5114.114.27.684.8
    4木豆 Cajanus cajanXP_020208653.124627.85.7215.114.26.984.1
    5密花豆 Spatholobus suberectusTKY55699.124627.95.4713.814.28.184.6
    6相思子 Abrus precatoriusXP_027347195.124627.95.5014.714.67.383.7
    7白羽扇豆 Lupinus albus KAE9597637.124728.05.4014.114.68.184.2
    8豇豆 Vigna unguiculataQCD81613.124627.85.4815.115.07.383.7
    9赤豆 Vigna angularisXP_017427550.124627.85.9014.213.48.583.0
    10红车轴草 Trifolium pratense PNY11893.124827.95.4312.412.97.286.0
    11鹰嘴豆 Cicer arietinumNP_001351681.124728.05.6915.014.68.184.6
    BA:碱性氨基酸;AC:酸性氨基酸;AR:芳香族氨基酸;AL:脂肪族氨基酸。
    BA: Basic amino acids; AC: acidic amino acids; AR: aromatic amino acids; AL: aliphatic amino acids.
    下载: 导出CSV
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出版历程
  • 收稿日期:  2022-11-10
  • 修回日期:  2023-03-13
  • 网络出版日期:  2023-05-24
  • 刊出日期:  2023-05-28

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