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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1681bp,含有1个1035bp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。
The full length cDNA sequence of GID1A homology gene was cloned by RT-PCR and RACE technology from the inflorescences, shoot tips, leaves and fruits of ’Vitis vinifera × V.labrusca’Fujiminori’ VvGID1A (Gene Accession No: JQ669511), with a total length of 1681 bp, contains a 1035 bp open reading frame encoding 344 amino acids. The amino acid sequence also contains two conserved domains of HGG and GXSXG. Phylogenetic analysis showed that the identity of VvGID1A with Medicago truncatula, cotton and Arabidopsis were 70.25%, 70.08% and 68.21%, respectively. Fluorescence quantitative RT-PCR results showed that VvGID1A expression level in grape inflorescence, old leaf and tendril was higher than that in shoot tip and young leaf, while the expression level in GA-treated fruit was lower than that in untreated control. The transient expression of transient expression vector and onion epidermal cells transformed onion epidermal cells showed that the gene expression product located in the nucleus.