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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). 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). doi: 10.14135/j.cnki.1006-3080.20210221002
WANG Xiao-hui, WANG Ze-jian, XIAO Ci-ying, LIU Ze-bo, GUO Mei-jin, ZHUANG Ying-ping. Optimization of Callus Suspension Culture Medium for enhancing Resveratrol Biosynthesis in Grape (Vitis vinifera)[J]. Journal of East China University of Science and Technology. doi: 10.14135/j.cnki.1006-3080.20210221002
Citation: WANG Xiao-hui, WANG Ze-jian, XIAO Ci-ying, LIU Ze-bo, GUO Mei-jin, ZHUANG Ying-ping. Optimization of Callus Suspension Culture Medium for enhancing Resveratrol Biosynthesis in Grape (Vitis vinifera)[J]. Journal of East China University of Science and Technology. 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: 国家重大专项项目(No.2020YFA0906800)资助项目
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: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 Grape (Vitis vinifera)

  • 摘要: 建立和优化了一种醉金香葡萄悬浮细胞制备白藜芦醇的最优化营养配置工艺。以细胞生长和白藜芦醇生物合成量为响应值,通过单因素试验、Plackett-Burman、响应面试验详细考察了不同碳源、氮源、前体、诱导剂和激素等因素对醉金香葡萄细胞白藜芦醇合成的影响。研究结果获得了醉金香葡萄悬浮细胞的最佳培养条件,在此条件下,葡萄细胞悬浮培养得到单位细胞白藜芦醇最高表达量为3026.64±56 μg/g,与对照相比提高了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

    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.  The effects of different precursors and inducers on cell growth and production of Vitis vinifera

    图  4  MeJA添加时间对醉金香葡萄细胞生长及产物合成的影响

    Figure  4.  the 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.  Different hormone types and combinations on cell growth and production of Vitis vinifera L

    图  6  P-B各实验组中残糖浓度和醉金香葡萄细胞生物量情况

    Figure  6.  Residual sugar concentration and cell biomass of Vitis vinifera L. in P-B experimental groups

    图  7  酵母提取物、茉莉酸甲酯和2,4-D含量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 contents on resveratrol synthesis

    表  1  Plackett-Burman培养组分和水平表头设计

    Table  1.   The culture components and levels in Plackett-Burman design

    FactorCodelevel
    low(−1)high(1)
    B5 (g/L)X115
    Sucrose (g/L)X22437.5
    PVP (g/L)X313
    Yeast extract (mg/L)X41001000
    Phe (mg/L)X510100
    MeJA (μmol/L)X610100
    6-BA (mg/L)X70.21
    2,4-D (mg/L)X815
    Blank termX9−11
    Blank termX10−11
    Blank termX11−11
    下载: 导出CSV

    表  2  Central-Composite设计的因素水平

    Table  2.   The factors and levels in central-composite design

    FactorCode values
    −1.68−1011.68
    Yeast extract (mg/L)631.82700800900968.18
    MeJA (μmol/L)63.1870809096.82
    2,4-D (mg/L)3.263.64.14.64.94
    下载: 导出CSV

    表  3  Plachett-Burman设计实验结果

    Table  3.   The experiment results in Plackett–Burman design

    RunVariable levelRes(μg/g)
    X1X2X3X4X5X6X7X8X9X10X11
    11−1111−1−1−11−111550
    21−1−1−11−111−1111001.85
    3−1111−1−1−11−111460.65
    4−1−11−111−1111−11780
    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
    10−111−1111−1−1−111850
    1111−1111−1−1−11−11875
    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.   The regression results for the Plackett-Burman design

    FactorSum of
    Squares
    dfMean squareF valueProb > FSignificantceImportance ranking
    Model4.37×10694.86×1052056.560.0005Significant
    E-MeJA3.52×10613.52×10614903.65<0.00011
    F-2,4-D2.78×10512.78×1051175.570.00082
    C-Yeast extract2.39×10512.39×1051011.370.0013
    H-PVP1.77×105134550.4750.960.00134
    G-6-BA80508.52180508.52340.730.0295
    下载: 导出CSV

    表  5  最陡爬坡试验设计及结果

    Table  5.   design and results in steepest ascent path

    Groupρ(Yeast extract)/
    (mg·L−1)
    C(MeJA)/
    (μmol·L−1)
    ρ(2,4-D)/
    (mg·L−1)
    Res/
    (μg·g−1)
    15505531690
    2600603.221740
    3650653.441860
    4700703.662310
    5750753.882760
    6800804.13100
    7850854.322940
    8900904.542900
    9950954.762790
    10110052730
    下载: 导出CSV

    表  6  CCD响应面设计及结果

    Table  6.   The design and results of the central composite experiment

    Runρ(Yeast extract)/
    (mg·L−1)
    C(MeJA)/
    (μmol·L−1)
    ρ(2,4-D)/
    (mg·L−1)
    Res/
    (μg·g−1)
    ABCY
    1700703.62460.2
    2900703.61985.6
    3700903.62020.5
    4900903.62360.4
    5700704.62530.7
    6900704.61898
    7700904.62010.4
    8900904.62254.6
    9631.82804.12550.02
    10968.18804.12650
    1180063.184.11899.5
    1280096.824.12750.1
    13800803.262080.4
    14800804.942120.5
    15800804.13271.02
    16800804.13250.7
    17800804.13240.2
    18800804.13308.2
    19800804.13299.42
    20800804.13280.4
    下载: 导出CSV

    表  7  CCD试验方差分析

    Table  7.   The variance analysis of central-composite design

    SourceSum of squaresMean squareF ValueProb > F
    Model5.15×1065.74×10517.45<0.0001
    A-Yeast extract9.23×1039.23×1030.280.6074
    B-MeJA1.06×1051.06×1053.220.1028
    C-2,4-D314.72314.729.59×10−30.9239
    AB3.58×1053.58×10510.90.008
    AC8051.88051.80.250.631
    BC1220.181220.180.0370.8509
    A29.65×1059.65×10529.40.0003
    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 Error3545.11709.02
    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

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