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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
  • [1] NOVAES E, KIRST M, CHIANG V, et al. Lignin and biomass: A negative correlation for wood formation and lignin content in trees [J]. Plant Physiology, 2010, 154(2): 555−561. doi: 10.1104/pp.110.161281
    [2] SIMMONS B A, LOQUÉ D, RALPH J. Advances in modifying lignin for enhanced biofuel production [J]. Current Opinion in Plant Biology, 2010, 13(3): 312−319. doi: 10.1016/j.pbi.2010.03.001
    [3] 李雪平, 高志民, 彭镇华, 等. 绿竹咖啡酰辅酶A-O-甲基转移酶基因的克隆与分析 [J]. 分子植物育种, 2008, 6(3):587−592. doi: 10.3969/j.issn.1672-416X.2008.03.030

    LI X P, GAO Z M, PENG Z H, et al. Cloning and characterization of CCoAOMT gene from Bambusa oldhamii [J]. Molecular Plant Breeding, 2008, 6(3): 587−592.(in Chinese) doi: 10.3969/j.issn.1672-416X.2008.03.030
    [4] VAN ACKER R, VANHOLME R, STORME V, et al. Lignin biosynthesis perturbations affect secondary cell wall composition and saccharification yield in Arabidopsis thaliana [J]. Biotechnology for Biofuels, 2013, 6(1): 46. doi: 10.1186/1754-6834-6-46
    [5] 方东鹏, 靳立梅, 董利东, 等. 野生大豆接种大豆疫霉菌后木质素含量的变化 [J]. 大豆科学, 2015, 34(1):99−102.

    FANG D P, JIN L M, DONG L D, et al. Change of lignin content in wild soybeans inoculated with Phytophthora sojae [J]. Soybean Science, 2015, 34(1): 99−102.(in Chinese)
    [6] 颜清上, 王连铮, 陈品三. 中国小黑豆抗源对大豆孢囊线虫4号生理小种抗病的生化反应 [J]. 作物学报, 1997, 23(5):529−537. doi: 10.3321/j.issn:0496-3490.1997.05.003

    YAN Q S, WANG L Z, CHEN P S. Biochemical responses of resistance to race 4 of Heterodera glycines in Chinese black soybean [J]. Acta Agronomica Sinica, 1997, 23(5): 529−537.(in Chinese) doi: 10.3321/j.issn:0496-3490.1997.05.003
    [7] 吴晓宇, 胡尚连, 曹颖, 等. 慈竹CCoAOMT基因的克隆及生物信息学分析 [J]. 南京林业大学学报(自然科学版), 2012, 36(3):17−22.

    WU X Y, HU S L, CAO Y, et al. Cloning of CCoAOMT gene in Neosinocalamus affinis and its bioinformatics analysis [J]. Journal of Nanjing Forestry University (Natural Sciences Edition), 2012, 36(3): 17−22.(in Chinese)
    [8] YE Z H. Association of caffeoyl coenzyme A 3-O-methyltransferase expression with lignifying tissues in several dicot plants [J]. Plant Physiology, 1997, 115(4): 1341−1350. doi: 10.1104/pp.115.4.1341
    [9] BOERJAN W, RALPH J, BAUCHER M. Lignin biosynthesis [J]. Annual Review of Plant Biology, 2003, 54: 519−546. doi: 10.1146/annurev.arplant.54.031902.134938
    [10] JOSHI C P, CHIANG V L. Conserved sequence motifs in plant S-adenosyl-L-methionine-dependent methyltransferases [J]. Plant Molecular Biology, 1998, 37(4): 663−674. doi: 10.1023/A:1006035210889
    [11] 王宇光, 吕笑言, 季美超, 等. 甜菜M14品系咖啡酰辅酶A-O-甲基转移酶提高植物抗逆性功能分析 [J]. 中国农学通报, 2018, 34(34):30−35. doi: 10.11924/j.issn.1000-6850.casb18060037

    WANG Y G, LV X Y, JI M C, et al. Stress tolerance improvement by BvM14-CCoAOMT gene in sugar beet M14 strain [J]. Chinese Agricultural Science Bulletin, 2018, 34(34): 30−35.(in Chinese) doi: 10.11924/j.issn.1000-6850.casb18060037
    [12] SALEKDEH G H, SIOPONGCO J, WADE L J, et al. A proteomic approach to analyzing drought- and salt-responsiveness in rice [J]. Field Crops Research, 2002, 76(2/3): 199−219.
    [13] RICCARDI F, GAZEAU P, DE V D, et al. Protein changes in response to progressive water deficit in maize. Quantitative variation and polypeptide identification [J]. Plant Physiology, 1998, 117(4): 1253−1263. doi: 10.1104/pp.117.4.1253
    [14] KÜHNL T, KOCH U, HELLER W, et al. Elicitor induced: Caffeoyl-CoA 3-O-methyltransferase from carrot cell suspension cultures [J]. Plant Science, 1989, 60(1): 21−25. doi: 10.1016/0168-9452(89)90039-3
    [15] PAKUSCH A E, KNEUSEL R E, MATERN U. S-adenosyl-l-methionine: Trans-caffeoyl-coenzyme A 3-O-methyltransferase from elicitor-treated parsley cell suspension cultures [J]. Archives of Biochemistry and Biophysics, 1989, 271(2): 488−494. doi: 10.1016/0003-9861(89)90299-3
    [16] 魏建华, 宋艳茹. 木质素生物合成途径及调控的研究进展 [J]. 植物学报, 2001, 43(8):771−779.

    WEI J H, SONG Y R. Recent advances in study of lignin biosynthesis and manipulation [J]. Journal of Integrative Plant Biology, 2001, 43(8): 771−779.(in Chinese)
    [17] DO C T, POLLET B, THÉVENIN J, et al. Both caffeoyl Coenzyme A 3-O-methyltransferase 1 and caffeic acid O-methyltransferase 1 are involved in redundant functions for lignin, flavonoids and sinapoyl malate biosynthesis in Arabidopsis [J]. Planta, 2007, 226(5): 1117−1129. doi: 10.1007/s00425-007-0558-3
    [18] 吕萌, 倪志勇, 王娟, 等. 棉花甲基化酶基因COMT和CCoAOMT的组织特异性表达分析 [J]. 核农学报, 2010, 24(4):713−719. doi: 10.11869/hnxb.2010.04.0713

    LÜ M, NI Z Y, WANG J, et al. Tissue specificity express analysis of methyltransferase gene comt and ccoaomt in cotton [J]. Journal of Nuclear Agricultural Sciences, 2010, 24(4): 713−719.(in Chinese) doi: 10.11869/hnxb.2010.04.0713
    [19] 吕笑言, 王宇光, 金英. 甜菜BvM14-CCoAOMT基因的克隆、表达及生物信息学分析 [J]. 黑龙江大学自然科学学报, 2018, 35(3):317−323. doi: 10.13482/j.issn1001-7011.2018.05.113

    LÜ X Y, WANG Y G, JIN Y. Cloning, expression and bioinformatics analyses of BvM14-CCoAOMT gene [J]. Journal of Natural Science of Heilongjiang University, 2018, 35(3): 317−323.(in Chinese) doi: 10.13482/j.issn1001-7011.2018.05.113
    [20] 林海燕, 罗勇, 陈丝, 等. 茶树CCoAOMT基因克隆及启动子的结构分析 [J]. 分子植物育种, 2019, 17(6):1804−1813. doi: 10.13271/j.mpb.017.001804

    LIN H Y, LUO Y, CHEN S, et al. Cloning of CCoAOMT gene and structure analysis of its promoter in Camellia sinensis [J]. Molecular Plant Breeding, 2019, 17(6): 1804−1813.(in Chinese) doi: 10.13271/j.mpb.017.001804
    [21] 刘大伟, 陈立杰, 段玉玺. 大豆胞囊线虫胁迫下不同抗性大豆杂交后代根系蛋白质组分析 [J]. 华北农学报, 2013, 28(5):29−33. doi: 10.7668/hbnxb.2013.05.005

    LIU D W, CHEN L J, DUAN Y X. Proteomic analysis of soybean with different resistance differentially expressed proteins induced by Heterodera glycines [J]. Acta Agriculturae Boreali-Sinica, 2013, 28(5): 29−33.(in Chinese) doi: 10.7668/hbnxb.2013.05.005
    [22] CHEN C J, CHEN H, ZHANG Y, et al. TBtools: An integrative toolkit developed for interactive analyses of big biological data [J]. Molecular Plant, 2020, 13(8): 1194−1202. doi: 10.1016/j.molp.2020.06.009
    [23] ARTIMO P, JONNALAGEDDA M, ARNOLD K, et al. ExPASy: SIB bioinformatics resource portal [J]. Nucleic Acids Research, 2012, 40(W1): W597−W603. doi: 10.1093/nar/gks400
    [24] KYTE J, DOOLITTLE R F. A simple method for displaying the hydropathic character of a protein [J]. Journal of Molecular Biology, 1982, 157(1): 105−132. doi: 10.1016/0022-2836(82)90515-0
    [25] YU C S, LIN C J, HWANG J K. Predicting subcellular localization of proteins for Gram-negative bacteria by support vector machines based on n-peptide compositions [J]. Protein Science:a Publication of the Protein Society, 2004, 13(5): 1402−1406. doi: 10.1110/ps.03479604
    [26] KUMAR S, STECHER G, TAMURA K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets [J]. Molecular Biology and Evolution, 2016, 33(7): 1870−1874. doi: 10.1093/molbev/msw054
    [27] 于明革, 杨洪强, 翟衡. 植物木质素及其生理学功能 [J]. 山东农业大学学报(自然科学版), 2003, 34(1):124−128.

    YU M G, YANG H Q, ZHAI H. Lignin and physiological function in plant [J]. Journal of Shandong Agricultural University, 2003, 34(1): 124−128.(in Chinese)
    [28] DÍAZ M L, GARBUS I, ECHENIQUE V. Allele-specific expression of a weeping lovegrass gene from the lignin biosynthetic pathway, caffeoyl-coenzyme A 3-O-methyltransferase [J]. Molecular Breeding, 2010, 26(4): 627−637. doi: 10.1007/s11032-010-9399-z
    [29] MARTZ F, MAURY S, PINÇON G, et al. cDNA cloning, substrate specificity and expression study of tobacco caffeoyl-CoA 3-O-methyltransferase, a lignin biosynthetic enzyme [J]. Plant Molecular Biology, 1998, 36(3): 427−437. doi: 10.1023/A:1005969825070
    [30] 黄春琼, 刘国道, 郭安平. 反义CCoAOMT基因调控烟草木质素的生物合成 [J]. 安徽农业科学, 2008, 36(19):8026−8027, 8043. doi: 10.3969/j.issn.0517-6611.2008.19.034

    HUANG C Q, LIU G D, GUO A P. Lignin biosynthesis regulated by antisense CCoAOMT gene in tobacco [J]. Journal of Anhui Agricultural Sciences, 2008, 36(19): 8026−8027, 8043.(in Chinese) doi: 10.3969/j.issn.0517-6611.2008.19.034
    [31] 陈夏鑫, 陈振林, 帅良, 等. 荸荠CCoAOMT基因的克隆与表达分析 [J]. 分子植物育种, 2020, 18(20):6685−6691. doi: 10.13271/j.mpb.018.006685

    CHEN X X, CHEN Z L, SHUAI L, et al. Cloning and expression analysis of CCoAOMT gene in Chinese water-chestnut [J]. Molecular Plant Breeding, 2020, 18(20): 6685−6691.(in Chinese) doi: 10.13271/j.mpb.018.006685
    [32] 陈凌娜, 郭晓娟, 杨汉奇. 巨龙竹木质素合成关键基因CCoAOMT的克隆及表达分析 [J]. 植物遗传资源学报, 2019, 20(2):476−484.

    CHEN L N, GUO X J, YANG H Q. Cloning and expression analysis of CCoAOMT, a key gene in lignin biosynthesis of Dendrocalamus sinicus [J]. Journal of Plant Genetic Resources, 2019, 20(2): 476−484.(in Chinese)
    [33] 刘鑫, 孙燕铭, 王涵, 等. 梨咖啡酰辅酶A-O-甲基转移酶基因PbCCoAOMT1的功能验证 [J]. 安徽农业大学学报, 2020, 47(5):845−850. doi: 10.13610/j.cnki.1672-352x.20200616.001

    LIU X, SUN Y M, WANG H, et al. Functional verification of pear caffeoyl coenzyme A-O-methyltransferase gene Pb CCo AOMT1 [J]. Journal of Anhui Agricultural University, 2020, 47(5): 845−850.(in Chinese) doi: 10.13610/j.cnki.1672-352x.20200616.001
    [34] 李元玉, 杨颖博, 张国宁, 等. 菘蓝中咖啡酰辅酶A氧甲基转移酶基因克隆与表达特征 [J]. 分子植物育种, 2022, 20(2):349−356. doi: 10.13271/j.mpb.020.000349

    LI Y Y, YANG Y B, ZHANG G N, et al. Cloning and expression characteristics of caffeoyl-CoA oxygen methyltransferase gene from Isatis indigotica fort [J]. Molecular Plant Breeding, 2022, 20(2): 349−356.(in Chinese) doi: 10.13271/j.mpb.020.000349
    [35] 马倩, 闫启, 张正社, 等. 紫花苜蓿CCoAOMT基因家族的鉴定、进化及表达分析 [J]. 草业学报, 2021, 30(11):144−156. doi: 10.11686/cyxb2020429

    MA Q, YAN Q, ZHANG Z S, et al. Identification, evolution and expression analysis of the CCoAOMT family genes in Medicago sativa [J]. Acta Prataculturae Sinica, 2021, 30(11): 144−156.(in Chinese) doi: 10.11686/cyxb2020429
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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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