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水稻OsPLATZ14基因启动子的克隆及表达分析

陈睿 陈建民 吴明基 杨绍华 胡昌泉

陈睿,陈建民,吴明基,等. 水稻 OsPLATZ14基因启动子的克隆及表达分析 [J]. 福建农业学报,2019,34(10):1137−1143. doi: 10.19303/j.issn.1008-0384.2019.10.004
引用本文: 陈睿,陈建民,吴明基,等. 水稻 OsPLATZ14 基因启动子的克隆及表达分析 [J]. 福建农业学报,2019,34(10):1137−1143. doi: 10.19303/j.issn.1008-0384.2019.10.004
CHEN R, CHEN J M, WU M J, et al. Cloning and Expression Analysis of OsPLATZ14 Promoter in Rice ( Oryza sativa L.) [J]. Fujian Journal of Agricultural Sciences,2019,34(10):1137−1143. doi: 10.19303/j.issn.1008-0384.2019.10.004
Citation: CHEN R, CHEN J M, WU M J, et al. Cloning and Expression Analysis of OsPLATZ14 Promoter in Rice ( Oryza sativa L.) [J]. Fujian Journal of Agricultural Sciences,2019,34(10):1137−1143. doi: 10.19303/j.issn.1008-0384.2019.10.004

水稻OsPLATZ14基因启动子的克隆及表达分析

doi: 10.19303/j.issn.1008-0384.2019.10.004
基金项目: 福建省自然科学基金项目(2017J01055);福建省科技计划公益类专项(2016R1018-8)
详细信息
    作者简介:

    陈睿(1980−),女,硕士,副研究员,研究方向:水稻遗传育种(E-mail:candy_chenrui@163.com

  • 中图分类号: S 511

Cloning and Expression Analysis of OsPLATZ14 Promoter in Rice (Oryza sativa L.)

  • 摘要:   目的  研究pOsPLATZ14的结构及时空表达模式,对深入研究OsPLATZ14基因功能、了解PLATZ转录因子在水稻生长发育进程的作用具有重要意义。  方法  利用BDGP、FPROM及Cister预测分析pOsPLATZ14大小,以水稻日本晴基因DNA为模板扩增pOsPLATZ14;应用PlantCARE分析序列中的顺序作用元件,构建pOsPLATZ14::GUS载体,转化水稻获得转基因植株;通过GUS组织化学染色法分析pOsPLATZ14在水稻中的时空表达特征。  结果  经PCR扩增获得pOsPLATZ14长度为1 899 bp,该区域含有光信号、逆境响应及激素应答等多种顺式作用元件。pOsPLATZ14驱动的GUS报告基因在水稻种子萌发期,苗期根、茎、叶,抽穗期的根、茎、叶、花穗、小花、叶夹角、茎结合部位、根茎过渡区及成熟期种子均有明显表达。  结论  pOsPLATZ14为组成型启动子,其下游调控基因OsPLATZ14可能在水稻生长发育过程中起重要作用。
  • 图  1  表达载体pOsPLATZ14::GUS示意图

    Figure  1.  Schematic diagram of pOsPLATZ14 :: GUS

    图  2  OsPLATZ14启动子扩增片段电泳图

    注:Marker:DL5000。

    Figure  2.  Electrophoretogram of pOsPLATZ14 amplified fragment

    Note:Marker:DL5000.

    图  3  重组质粒pOsPLATZ14::GUS酶切验证

    注:Marker为λ-EcoT14 I digest DNA Marker,1为BamHI、NcoI双酶切pOsPLATZ14::GUS产物。

    Figure  3.  The recombinant plasmid digested by restriction enzymes

    Note: Marker: λ-EcoT14 I digest DNA Marker; 1: pOsPLATZ14 :: GUS plasmid digested by BamHI and NcoI.

    图  4  转基因水稻T0代PCR检测

    注:Marker为DL5000,1-6为转基因阳性株,7为质粒pOsPLATZ14::GUS,8为转基因阴性株,9为日本晴。

    Figure  4.  PCR analysis on DNA in T0 transgenic rice plant

    Note: Marker: DL5000; 1-6: transgene positive line; 7: pOsPLATZ14 :: GUS ; 8: transgene negative line; 9: Nipponbare.

    图  5  pOsPLATZ14::GUS转基因水稻的组织化学染色

    注:a-b为根、c-d为茎、e为茎结合部、f为叶、g为叶夹角、h为雄蕊、i为雌蕊、j为种子、k为小花、l为根茎结合部、m为萌发期种子、n为花穗。

    Figure  5.  Histochemical staining of pOsPLATZ14::GUS transgenic rice

    Note:a-b: root; c-d: stem; e: stem-stem junction; f: leaf; g: leaf angle; h: stamen; i: pistil; j: seed; k: floret; l: root-stem junction; m: germinating seed; n: flower spike.

    表  1  OsPLATZ14基因启动子序列顺式作用元件

    Table  1.   The cis-acting elements in promoter sequence of OsPLATZ14

    位点名称 Site name 信号序列 Signal Sequence 数量 Amount 位点功能 Function of Site
    TATA-box ATATAA/ TATA 2 主要顺式调控元件 Common cis-acting elements
    CAAT-box CAAT/CCAAT/ CAAAT 10 主要顺式调控元件 Common cis-acting elements
    3-AF1 binding site TAAGAGAGGAA 1 光响应元件 Light response element
    GATA-motif GATAGGA 1 光响应元件 Light response element
    Sp1 GGGCGG 1 光响应元件 Light response element
    MRE AACCTAA 1 光响应元件 Light response element
    ARE AAACCA 2 缺氧胁迫响应元件 Anaerobic stress response element
    MBS CAACTG 1 干旱胁迫响应元件 Drought stress response element
    CAT-box GCCACT 1 分生组织特异性元件 Meristem expression element
    ABRE ACGTG 1 脱落酸响应元件 abscisic acid response element
    P-box CCTTTTG 1 赤霉素响应元件 Gibberellin response element
    TCA-element TCAGAAGAGG 1 水杨酸响应元件 Salicylic acid response element
    TGA-element AACGAC 1 生长素响应元件 Auxin response element
    TGACG-motif TGACG 1 茉莉酸响应元件 MeJA response element
    CCAAT-box CAACGG 1 MYB结合位点 MYB binding site
    AAGAA-motif GAAAGAA 3 功能未知元件 Function unknown component
    ERE ATTTCATA 1 功能未知元件 Function unknown component
    MYB CAACCA 1 功能未知元件 Function unknown component
    MYC CATTTG 3 功能未知元件 Function unknown component
    Myb CAACTG 1 功能未知元件 Function unknown component
    STRE AGGGG 2 功能未知元件 Function unknown component
    TATA TATAAAAT 1 功能未知元件 Function unknown component
    TCA TCATCTTCAT 1 功能未知元件 Function unknown component
    Unnamed_1 CGTGG 1 功能未知元件 Function unknown component
    Unnamed_4 CTCC 11 功能未知元件 Function unknown component
    WUN-motif AAATTACT 1 功能未知元件 Function unknown component
    as-1 TGACG 1 功能未知元件 Function unknown component
    下载: 导出CSV
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  • 收稿日期:  2019-09-18
  • 修回日期:  2019-10-18
  • 刊出日期:  2019-10-01

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