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园艺学报 ›› 2019, Vol. 46 ›› Issue (7): 1270-1278.doi: 10.16420/j.issn.0513-353x.2018-0768

• 研究论文 • 上一篇    下一篇

越橘花青苷合成相关基因VcTTG1的克隆与功能鉴定

宋 杨,刘红弟,王海波,张红军*,刘凤之*   

  1. 中国农业科学院果树研究所,农业部园艺作物种质资源利用重点实验室,辽宁省落叶果树矿质营养与肥料高效利用重点实验室,辽宁兴城 125100
  • 出版日期:2019-07-25 发布日期:2019-07-25
  • 基金资助:
    山东农业大学作物生物学国家重点实验室开放课题项目(2018KF08);辽宁省农业领域青年科技创新人才培养计划项目(2015059);中国农业科学院科技创新工程项目;国家自然科学基金项目(31301754);中央级公益性科研院所基本科研业务费专项(1610182019009)

Molecular Cloning and Functional Characterization of Anthocyanin Synthesis Related Genes VcTTG1 of Blueberry

SONG Yang,LIU Hongdi,WANG Haibo,ZHANG Hongjun*,and LIU Fengzhi*   

  1. Research Institute of Pomology,Chinese Academy of Agricultural Sciences/Key Laboratory of Fruit Germplasm Resources Utilization,Ministry of Agriculture,Laboratory of Mineral Nutrition and Efficient Fertilization for Deciduous Fruits,Liaoning Province,Xingcheng,Liaoning 125100,China
  • Online:2019-07-25 Published:2019-07-25

摘要: 以‘Duke’越橘(Vaccinium corymbosum‘Duke’)为试材,从转录组数据库中克隆编码WD40蛋白的基因VcTTG1,分析其表达模式并鉴定其在花青苷合成过程中的作用功能,为进一步探讨越橘花青苷合成调控机理奠定理论基础。结果表明,克隆获得越橘VcTTG1(GenBank登录号为MH717246),ORF为1 044 bp,推测其编码348个氨基酸,含有典型的WD40结构域。系统发生分析表明,VcTTG1与葡萄VvWDR1的同源性最高。VcTTG1在越橘根、枝条、幼叶、花和果实中均有表达,但表达量差异显著,在果实中较高,枝条中较低。在果实中随着VcTTG1表达的升高,花青苷含量呈递增的趋势。在拟南芥中超表达VcTTG1,其花青苷积累在VcTTG1转基因植株中显著增加。酵母双杂交试验结果表明,VcTTG1可与拟南芥bHLH蛋白AtTT8相互作用。由此推测,VcTTG1在调控花青苷合成过程中发挥重要作用。

关键词: 越橘, VcTTG1, 花青苷, 表达分析, 功能鉴定

Abstract: The objective of this study is to isolate a blueberry(Vaccinium corymbosum)cultivar ‘Duke’VcTTG1 gene encoding WD40 protein by PCR technology from transcriptome data and study its expression,to identify its function in anthocyanin biosynthesis. This study laid the foundation for further study of the molecular mechanism of VcTTG1 affecting anthocyanin biosynthesis in blueberry. Sequence analysis showed that the open reading frame(ORF)of VcTTG1(GenBank accession No. MH717246) is 1 044 bp,putatively encoded 348 amino acids. Protein structure analysis showed that VcTTG1 contained conserved WD40 domain. A phylogenetic tree indicated that the blueberry VcTTG1 exhibited the highestsequence similarity to Vitis vinifera VvWDR1. Expression analysis showed that VcTTG1 was expressed in roots,stems,young leaves,flowers and fruits. However,the expression levels varied,with the highest expression level in fruits and the relatively low transcript levels in stems. The content of anthocyanin in fruits was increased with the up-regulation of relative expression of VcTTG1. The VcTTG1 protein could interact with Arabidopsis bHLH protein AtTT8 and favorably contributes to anthocyanin accumulation in transgenic Arabidopsis. It is speculated that VcTTG1 plays a regulatory role in anthocyanin biosynthesis.

Key words: blueberry, VcTTG1, anthocyanin, expression analysis, functional identification

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