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  • ISSN 1006-3080
  • CN 31-1691/TQ

醉金香葡萄愈伤细胞悬浮培养基优化促进白藜芦醇的合成

王晓惠 王泽建 肖慈英 刘泽波 郭美锦 庄英萍

王晓惠, 王泽建, 肖慈英, 刘泽波, 郭美锦, 庄英萍. 醉金香葡萄愈伤细胞悬浮培养基优化促进白藜芦醇的合成[J]. 华东理工大学学报(自然科学版), 2022, 48(2): 203-212. doi: 10.14135/j.cnki.1006-3080.20210221002
引用本文: 王晓惠, 王泽建, 肖慈英, 刘泽波, 郭美锦, 庄英萍. 醉金香葡萄愈伤细胞悬浮培养基优化促进白藜芦醇的合成[J]. 华东理工大学学报(自然科学版), 2022, 48(2): 203-212. doi: 10.14135/j.cnki.1006-3080.20210221002
WANG Xiaohui, WANG Zejian, XIAO Ciying, LIU Zebo, GUO Meijin, ZHUANG Yingping. Optimization of Callus Suspension Culture Medium for Enhancing Resveratrol Biosynthesis in Vitis vinifera Grape[J]. Journal of East China University of Science and Technology, 2022, 48(2): 203-212. doi: 10.14135/j.cnki.1006-3080.20210221002
Citation: WANG Xiaohui, WANG Zejian, XIAO Ciying, LIU Zebo, GUO Meijin, ZHUANG Yingping. Optimization of Callus Suspension Culture Medium for Enhancing Resveratrol Biosynthesis in Vitis vinifera Grape[J]. Journal of East China University of Science and Technology, 2022, 48(2): 203-212. doi: 10.14135/j.cnki.1006-3080.20210221002

醉金香葡萄愈伤细胞悬浮培养基优化促进白藜芦醇的合成

doi: 10.14135/j.cnki.1006-3080.20210221002
基金项目: 国家重点研发专项(2020YFA0906800)
详细信息
    作者简介:

    王晓惠(1996—),女,山东人,硕士生,主要研究方向为植物细胞培养。E-mail:wxhui1010@163.com

    通讯作者:

    庄英萍,E-mail:ypzhuang@ecust.edu.cn

  • 中图分类号: TS201.3

Optimization of Callus Suspension Culture Medium for Enhancing Resveratrol Biosynthesis in Vitis vinifera Grape

  • 摘要: 建立和优化了一种醉金香葡萄悬浮细胞合成白藜芦醇的工艺。以细胞生长和白藜芦醇生物合成量为响应值,通过单因素实验、Plackett-Burman、响应面实验考察了不同碳源、氮源、前体、诱导剂和激素等因素对醉金香葡萄细胞悬浮培养基合成白藜芦醇的影响。结果表明,在醉金香葡萄悬浮细胞的最佳培养条件下,单位葡萄细胞中白藜芦醇的最高表达量为(3026.64±56)μg/g,与对照组相比提高了51.13倍,为后续实现利用醉金香葡萄细胞悬浮培养生产白藜芦醇提供了理论基础。

     

  • 图  1  不同碳源底物对醉金香葡萄细胞生长及产物合成的影响

    Figure  1.  Effects of substrates with different carbon sources on cell growth and production of Vitis vinifera

    图  2  不同氮源底物对醉金香葡萄细胞生长及产物合成的影响

    Figure  2.  Effects of substrates with different nitrogen sources on cell growth and production of Vitis vinifera

    图  3  不同前体和诱导剂对醉金香葡萄细胞生长及产物合成的影响

    Figure  3.  Effects of different precursors and inducers on cell growth and production of Vitis vinifera

    图  4  茉莉酸甲酯添加时间对醉金香葡萄细胞生长及产物合成的影响

    Figure  4.  Effect of adding time of MeJA on the cell growth and product synthesis of Vitis vinifera

    图  5  不同激素类型和激素组合对醉金香葡萄细胞生长及产物合成的影响

    Figure  5.   Effect of different hormone types and combinations on cell growth and production of Vitis vinifera

    图  6  Plackett-Burman各实验组中白藜芦醇含量和醉金香葡萄细胞生长情况

    Figure  6.  Resveratrol biomass and cell of Vitis vinifera in Plackett-Burman experimental groups

    图  7  酵母提取物、茉莉酸甲酯和2,4-D交互作用对白藜芦醇合成影响的响应面和等高线

    Figure  7.  Response surface and contour of the interaction of yeast extract, methyl jasmonate and 2,4-D on resveratrol synthesis

    表  1  Plackett-Burman培养组分和水平

    Table  1.   Culture components and levels in Plackett-Burman

    FactorCodeLevel
    Low(−1)High(1)
    B5 X1 1 5
    Sucrose X2 24 37.5
    PVP X3 1 3
    Yeast extract X4 100 1000
    Phe X5 10 100
    MeJA X6 10 100
    6-BA X7 0.2 1
    2,4-D X8 1 5
    Blank term X9 −1 1
    Blank term X10 −1 1
    Blank term X11 −1 1
    下载: 导出CSV

    表  2  Central-Composite设计的因素水平

    Table  2.   Factors and levels of central-composite design

    Code value$\rho $/(mg·L−1
    Yeast extractMeJA2,4-D
    −1.68 631.82 63.18 3.26
    −1 700 70 3.6
    0 800 80 4.1
    1 900 90 4.6
    1.68 968.18 96.82 4.94
    下载: 导出CSV

    表  3  Plachett-Burman设计试验结果

    Table  3.   Experiment results in Plackett–Burman design

    GroupVariable levelResveratrol
    biomass/(μg·g−1)
    X1X2X3X4X5X6X7X8X9X10X11
    11−1111−1−1−11−111550.00
    21−1−1−11−111−1111001.85
    3−1111−1−1−11−111460.65
    4−1−11−111−1111−11780.00
    51−111−1111−1−1−1467.95
    6−11−111−1111−111035.35
    7−1−1−1−1−1−1−1−111−1301.20
    811−1−1−11−11−1−11366.50
    9111−1−1−11−111−1580.00
    10−111−1111−1−1−111850.00
    1111−1111−1−1−11−11875.00
    12−1−1−11−111−1111415.30
    1300000000000915.35
    X1—B5;X2—Sucrose;X3—Yeast extract;X4—Phe;X5—MeJA; X6—2,4-D;X7—6-BA;X8—PVP;X9~X11 —Error
    下载: 导出CSV

    表  4  Plachett-Burman设计各因数效应分析

    Table  4.   Regression results for the Plackett-Burman design

    FactorSum of squaresDegree of freedomMean squareF valuePImportance ranking
    Model 4.37×106 9 4.86×105 2056.56 0.0005
    MeJA 3.52×106 1 3.52×106 14903.65 <0.0001 1
    2,4-D 2.78×105 1 2.78×105 1175.57 0.0008 2
    Yeast extract 2.39×105 1 2.39×105 1011.37 0.001 3
    PVP 1.77×105 1 3.46×104 750.96 0.0013 4
    6-BA 8.05×104 1 8.05×104 340.73 0.029 5
    下载: 导出CSV

    表  5  最陡爬坡实验设计及结果

    Table  5.   Design and result in steepest ascent path

    Groupρ(Yeast extract)/
    (mg·L−1)
    c(MeJA)/
    (μmol·L−1)
    ρ(2,4-D)/
    (mg·L−1)
    Resveratrol
    biomass/(μg·g−1)
    1 550 55 3.00 1690
    2 600 60 3.22 1740
    3 650 65 3.44 1860
    4 700 70 3.66 2310
    5 750 75 3.88 2760
    6 800 80 4.10 3100
    7 850 85 4.32 2940
    8 900 90 4.54 2900
    9 950 95 4.76 2790
    10 1000 100 5.00 2730
    下载: 导出CSV

    表  6  CCD响应面设计及结果

    Table  6.   Design and result of the central composite experiment

    Groupρ(Yeast extract)/
    (mg·L−1)
    c(MeJA)/
    (μmol·L−1)
    ρ(2,4-D)/
    (mg·L−1)
    Resveratrol biomass/(μg·g−1)
    1700703.62460.2
    2900703.61985.6
    3700903.62020.5
    4900903.62360.4
    5700704.62530.7
    6900704.61898.0
    7700904.62010.4
    8900904.62254.6
    9631.82804.12550.0
    10968.18804.12650.0
    1180063.184.11899.5
    1280096.824.12750.1
    13800803.262080.4
    14800804.942120.5
    15800804.13271.0
    16800804.13250.7
    17800804.13240.2
    18800804.13308.2
    19800804.13299.4
    20800804.13280.4
    下载: 导出CSV

    表  7  CCD试验方差分析

    Table  7.   Variance analysis of central-composite design

    SourceSum of squaresMean squareFP
    Model5.15×1065.74×10517.45<0.0001
    Yeast extract9.23×1039.23×1030.280.6074
    MeJA1.06×1051.06×1053.220.1028
    2,4-D314.72314.729.59×10−30.9239
    AB3.58×1053.58×10510.900.0080
    AC8051.808051.800.250.6310
    BC1220.181220.180.0370.8509
    A29.65×1059.65×10529.400.0003
    B21.83×1061.83×10655.67<0.0001
    C22.73×1062.73×10683.24<0.0001
    Residual3.28×10532810.75
    Lack of fit3.25×1056.49×10491.55<0.0001
    Pure error355.11709.02
    Cor total5.48×106
    R2= 0.9401; Adj R2=0.8862
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-02-21
  • 网络出版日期:  2021-06-29
  • 刊出日期:  2022-04-22

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