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园艺学报 ›› 2013, Vol. 40 ›› Issue (5): 839-.

• 果树 • 上一篇    下一篇

葡萄赤霉素受体基因VvGID1A 的分离、亚细胞定位及表达分析

王西成,吴伟民,房经贵,钱亚明,王 晨,宋长年,赵密珍   

  1. 1 江苏省农业科学院园艺研究所,南京 210014;2 南京农业大学园艺学院,南京 210095
  • 出版日期:2013-05-25 发布日期:2013-05-25

Isolation,Subcellular Localization and Expression Analysis of Gibberellin
Receptor Gene VvGID1A from Grapevine

 WANG  Xi-Cheng, WU  Wei-Min, FANG  Jing-Gui, QIAN  Ya-Ming, WANG   Chen, SONG  Chang-Nian, ZHAO  Mi-Zhen   

  1. 1Institute of Horticulture,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;2 College of Horticulture,
    Nanjing Agricultural University,Nanjing 210095,China
  • Online:2013-05-25 Published:2013-05-25

摘要: 以‘藤稔’葡萄(Vitis vinifera × V. labrusca‘Fujiminori’)的花序、茎尖、叶片以及果实为
试材,应用RT-PCR 结合RACE 技术克隆得到GID1A 同源基因全长cDNA 序列,命名为VvGID1A(基因
登录号:JQ669511),全长1 681 bp,含有1 个1 035 bp 的开放阅读框,编码344 个氨基酸,该氨基酸序
列还包含两个激素敏感酯酶家族保守的结构域HGG 和GXSXG。系统进化分析表明,VvGID1A 与蒺藜苜
蓿、棉花和拟南芥之间的一致性分别为70.25%、70.08%和68.21%。荧光定量RT-PCR 结果表明,VvGID1A
在葡萄花序、老叶和卷须中表达水平高于茎尖和幼叶,而在GA 处理果实中的表达水平低于未处理的对照
样品。瞬时表达载体的构建及洋葱表皮细胞转化后,洋葱表皮细胞的瞬时表达结果显示,该基因的表达
产物定位在细胞核上。

关键词: Vitis vinifera, gibberellin, VvGID1A, subcellular localization, expression level

Abstract: A full length of GID1A gene was isolated from grapevine‘Fujiminori’(Vitis vinifera ×
V. labrusca)by RT-PCR and RACE(rapid amplification of cDNA ends)method. Sequence analysis
indicated that VvGID1A was 1 681 bp in full length,and an opening reading frame(ORF)of 1 035 bp
encoding 344 amino acids. Subsequently,amino acid sequence analysis revealed that VvGID1A included
two conserved domains of hormone sensitive lipase(HSL):HGG and GXSXG. Phylogenetic analysis
showed that VvGID1A share 70.25%,70.08% and 68.21% identity with GID1 in Medicago truncatula,
Gossypium hirsutum and Arabidopsis thaliana. Real-time fluorescent quantitative PCR indicated that the
expression of VvGID1A gene in flower,old leaf and tendril is higher than that in stem tip and young leaf.
Furthermore,the expression of VvGID1A gene was lower in gibberellins treated fruit than in the control.
After construction of transient expression vector and onion epidermal cell transformation,subcellular
localization assays showed that the VvGID1A protein was located in the nucleus.

Key words: Vitis vinifera, gibberellin, VvGID1A, subcellular localization, expression level

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