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中国水仙FT基因的表达及功能分析

李婷婷 潘秋宇 吴菁华

李婷婷,潘秋宇,吴菁华. 中国水仙FT基因的表达及功能分析 [J]. 福建农业学报,2023,38(12):1420−1427 doi: 10.19303/j.issn.1008-0384.2023.12.005
引用本文: 李婷婷,潘秋宇,吴菁华. 中国水仙FT基因的表达及功能分析 [J]. 福建农业学报,2023,38(12):1420−1427 doi: 10.19303/j.issn.1008-0384.2023.12.005
LI T T, PAN Q Y, WU J H. Expressions and Functions of Flowering Locus Ts in Narcissus tazetta var. chinensis Roem [J]. Fujian Journal of Agricultural Sciences,2023,38(12):1420−1427 doi: 10.19303/j.issn.1008-0384.2023.12.005
Citation: LI T T, PAN Q Y, WU J H. Expressions and Functions of Flowering Locus Ts in Narcissus tazetta var. chinensis Roem [J]. Fujian Journal of Agricultural Sciences,2023,38(12):1420−1427 doi: 10.19303/j.issn.1008-0384.2023.12.005

中国水仙FT基因的表达及功能分析

doi: 10.19303/j.issn.1008-0384.2023.12.005
基金项目: 福建省自然科学基金项目(2021J01087); 福建省科技计划星火项目(2021S0068)
详细信息
    作者简介:

    李婷婷(1994 —),女,硕士,主要从事园艺植物遗传育种相关研究,E-mail:15236174388@qq.com

    通讯作者:

    吴菁华(1978 —),女,博士,副教授,主要从事园艺植物遗传育种相关研究,E-mail:wjhham@163.com

  • 中图分类号: S682.21

Expressions and Functions of Flowering Locus Ts in Narcissus tazetta var. chinensis Roem

  • 摘要:   目的  开花基因座T(Flowering Locus TFT)基因广泛参与植物的生长发育,在调控开花、地下茎发育、种子萌发和逆境胁迫等过程中发挥重要作用。揭示FT基因在中国水仙(Narcissus tazetta var. chinensis Roem)中的表达模式和功能可为中国水仙花期调控提供理论依据。  方法  基于中国水仙转录组数据,对FT 基因进行筛选,获得4个中国水仙FT基因。利用荧光定量PCR技术分析它们在中国水仙不同组织和花芽分化不同时期的表达模式,并在拟南芥(Arabidopsis thaliana)中超量表达。利用荧光定量PCR技术分析转NtFT1NtFT4基因拟南芥中SUPPRESSOR OF OVEREXPRESSION OF CO 1SOC1)、LEAFY(LFY)和APETALA 1AP1) 基因表达水平。  结果  克隆了4个中国水仙FT同源基因NtFT1NtFT2NtFT3NtFT4,NtFT3外,中国水仙NtFT都具有FT的保守基序;系统进化分析显示,NtFT1属于FT-like I进化枝,NtFT2、NtFT3和NtFT4同属于FT-like II类进化枝。不同的NtFT基因在中国水仙组织器官和花芽分化不同时期的表达模式存在差异:NtFT1NtFT3在花中表达量最高,NtFT2在叶片中表达量最高,NtFT4在鳞片中的表达量最高;NtFT1在花芽分化过程中均呈先上升后下降的趋势,NtFT2整个主芽分化过程中变化幅度不大,NtFT3NtFT4在整个花芽分化期表达量都较低。异位转化拟南芥结果显示,与野生型拟南芥相比,过表达NtFT1NtFT2的拟南芥提早开花,过表达NtFT3拟南芥开花时间与野生型植株无明显差异,转NtFT4基因的拟南芥植株推迟开花。过表达NtFT1拟南芥中SOC1LFYAP1基因表达量上升。  结论  在中国水仙中存在多个FT基因,在调控开花的功能上存在差异,NtFT1促进开花,NtFT4抑制开花。
  • 图  1  NtFT与其他物种FT氨基酸序列的多重比较

    Figure  1.  Multiple comparison on amino acid sequences of NtFT and FTs of other species

    图  2  NtFT与其他植物FT 蛋白的系统进化树分析

    Figure  2.  Phylogenetic trees of NtFT and FTs of other species

    图  3  NtFT基因在不同组织中的qRT-PCR表达分析

    不同小写字母表示同一组织部位不同基因间差异显著(P<0. 05)。

    Figure  3.  qRT-PCR on NtFTs in different tissues

    Datas with different lowercase letters indicate significant differences between different gene in same tissue (P<0.05).

    图  4  NtFT在中国水仙花芽不同分化阶段的表达分析

    不同小写字母表示同一发育时间不同基因差异显著(P<0. 05)。

    Figure  4.  qRT-PCR on NtFTs from N. tazetta during flower buds at differentiation stages

    Datas with different lowercase letters indicate significant differences between genes at same development time (P<0.05).

    图  5  NtFT基因在拟南芥的异位表达

    A:过表达NtFT基因拟南芥开花情况;B:过表达 NtFT 基因拟南芥开花性状数据。*、**表示与WT相比差异显著(P<0.05)或极显著(P<0.01)。图6同。

    Figure  5.  Ectopic expressions of NtFTs in A. thaliana

    A: Flowering of A. thaliana with overexpressed NtFT; B: Flowering traits data of A. thaliana with overexpressed NtFT; * and ** indicate significant difference(P<0.05) and (P<0.01), respectively. Same for Fig. 6.

    图  6  转基因拟南芥中AtLFYAtSOC1AtAP1的表达分析

    Figure  6.  Expressions of AtLFY, AtSOC1, and AtAP1 in transgenic plants

    表  1  克隆中国水仙NtFT基因引物

    Table  1.   Primers for cloning NtFT of N. tazetta

    引物名称
    Primer name
    上游引物序列(5′-3′)
    Upstream primer sequence (5′-3′)
    下游引物序列(5′-3′)
    Downstream primer sequence (5′-3′)
    NtFT1 TTTCCGCTTATATCTCTTCTGGGAC TCGGGAAGTAGCAAGACGATCAAAC
    NtFT2 ATGTTGAGAGAGAGGGTACC TCAGCAAAGTCCTGAGAACCTTCTT
    NtFT3 GAAGTAGTCATGTTGAGAGAGAGG GTATCACATATTGCATGGCTTAGG
    NtFT4 GGTTAAGAGACAGAATGCCGAT TTTATGTCATTTATCGTCTG CTAG
    下载: 导出CSV

    表  2  FT基因表达qRT-PCR引物

    Table  2.   Primers for qRT-PCR of FT gene

    引物名称
    Primer name
    上游引物序列(5’-3’)
    Upstream primer sequence (5′-3′)
    上游引物序列(5’-3’)
    Downstream primer sequence (5′-3′)
    NtActin GTTGACCCACCACTAAGAACAATG TGCCCAGAAGTGCTATTCCAG
    QNtFT1 CCAGCCAAAGGTTGAAGTCG CCCTGTGGTTCCTGGTATG
    QNtFT2 CTTATGAGAGCCCTCGAACACC CGCACACAGTTTGTTGAACTTCC
    QNtFT3 CGCACACAGTTTGTTGAACTTCC CACTGGTTGGTGACAGACATACC
    QNtFT4 TGGCAGGATGCGATGCAAGA CACGATGCGATGAATCCCCGA
    下载: 导出CSV

    表  3  pSAK277表达载体构建引物

    Table  3.   Primers for constructing pSAK277 expression vector

    引物名称
    Primer name
    上游引物序列(5′-3′)
    Upstream primer sequence (5′-3′)
    下游引物序列(5′-3′)
    Upstream primer sequence (5′-3′)
    277NtFT1 ACTAGTGGATCCAAAGAATTCATGA
    GTAGGGATCCTTTGGTTATTG
    AGAAGTACTCTCGAGAAGCTT
    TTAGGGGTACATCCTCCGGCCACCA
    277NtFT2 ACTAGTGGATCCAAAGAATTCAGTTGA
    GAGAGAGGGTACCAAGGG
    AGAAGTACTCTCGAGAAGCTTT
    CAGCAAAGTCCTGAGAACCTTCTT
    277NtFT3 ACTAGTGGATCCAAAGAATTC
    ATGTTGAGAG AGAGGGTACC
    AGAAGTACTCTCGAGAAGCTT
    TCACATATTG CATGGCTTAG
    277NtFT4 ACTAGTGGATCCAAAGAATTCAT
    GCCGATACTGGGACAAGT
    AGAAGTACTCTCGAGAAGCTTTC
    AGAACCTTCTTCCTCCGCA
    下载: 导出CSV

    表  4  转基因拟南芥的不同基因qRT-PCR引物

    Table  4.   Primers for qRT-PCR of different genes in transgenic A. thaliana

    引物名称
    Primer name
    上游引物序列(5’-3’)
    Upstream primer sequence (5′-3′)
    上游引物序列(5’-3’)
    Upstream primer sequence (5′-3′)
    Actin GCTGAGAGTTGATGGTGTGCT GGATACCCTTTCGCAGATAGAG
    AtLFY GTTAGGTTTTACGGCGAGCA GCAATCGTCTCCGTTCAGC
    AtAP1 ACCAAATCCAGCATCCTTACA TCAAGAGTCAGTTCGAGATCATTC
    AtSOC1 CTCTCAGTGCTTTGTGATGCT CGATTGAGCATGTTCCTATGCC
    下载: 导出CSV
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出版历程
  • 收稿日期:  2023-09-12
  • 修回日期:  2023-11-02
  • 网络出版日期:  2024-01-06
  • 刊出日期:  2023-12-28

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