论文部分内容阅读
为探讨外源GA介导的无核葡萄果实膨大的机理,本试验根据葡萄基因组序列设计特异性引物,以葡萄果实cDNA一链为模板,利用聚合酶链式反应(PCR)克隆了2个假定的葡萄GID基因。测序比对结果表明其中一个属于GID1ac家族,根据序列相似性,命名为VvGID1-3,另一个属于GID2 F-box基因家族,命名为VvGID2。在烟草中分别超表达这2个基因,结果表明:VvGID1-3和VvGID2转基因植株叶圆片在MS培养基上生长7d后面积都约为对照的140%;VvGID1-3和VvGID2转基因烟草种子的萌发率降低,萌发过程对0.5和1μmol/L的GA3处理表现超敏特征;内源激素水平测定结果显示VvGID1-3转基因烟草中的GA和ABA含量低于对照,VvGID2转基因植株中IAA含量高于对照;在葡萄胚性愈伤组织中分别表达VvGID1-3-GFP和VvGID2-GFP,信号均定位于细胞核中;对VvGID1-3和VvGID2的启动子进行克隆,pVvGID1-3::GUS和pVvGID2::GUS转基因烟草的染色结果表明,信号在幼嫩和快速生长的组织中最强,在成叶中主要在叶脉中表达,在不定根中的表达量很低。
In order to explore the mechanism of exogenous GA-mediated enlargement of fruitless grapes, two hypotheses were cloned by polymerase chain reaction (PCR) based on the sequence of grape genomes. Grape GID gene. Sequencing results showed that one of them belonged to the GID1ac family, named VvGID1-3 according to sequence similarity and the other belonged to the GID2 F-box gene family named VvGID2. Overexpression of these two genes in tobacco showed that both VvGID1-3 and VvGID2 transgenic leaf discs grew about 140% of the control on the MS medium after 7 days of growth. The VvGID1-3 and VvGID2 transgenic tobacco seeds The germination rate was reduced and the germination process had hypersensitivity to GA3 of 0.5 and 1 μmol / L. The results of endogenous hormone levels showed that the content of GA and ABA in VvGID1-3 transgenic tobacco was lower than that in the control, while the content of IAA in VvGID2 transgenic plants was higher than VvGID1-3-GFP and VvGID2-GFP were expressed respectively in embryogenic callus of embryo. The signals were located in the nucleus. The promoters of VvGID1-3 and VvGID2 were cloned, pVvGID1-3 :: GUS and pVvGID2: : The staining results of GUS transgenic tobacco showed that the signal was the strongest in the young and rapidly growing tissues, mainly in the veins in the leaves, and low in the adventitious roots.