苹果MdFT基因对番茄的遗传转化

 通过RT2PCR扩增, 从苹果叶片cDNA中克隆了^FT基因的同源基因^MdFT, 构建了花椰菜病毒35S启动子驱动的MdFT植物表达载体35S: : MdFT, 并利用根癌农杆菌介导法将其导入番茄栽培品种‘中蔬四号’; 同时转化拟南芥A tFT基因作为阳性对照。从添加卡那霉素的筛选培养基上再生了抗性植株,PCR扩增证明, 外源基因MdFT和AtFT已经整合到转基因番茄的基因组, 半定量RT-PCR则证明它们已经在转基因番茄中得到异位过量表达。形态鉴定发现, 转基因番茄植株比非转基因对照植株开花早, 表明成功地从苹果中克隆了成花素基因MdFT, 该基因具有通过转基因缩短苹果树童期的潜在价值。

Genetic Transformation of Apple MdFT Gene into Tomato

It is of great importance for field yield of fruit trees to generate flower buds. Recently, FT protein has been proved to be a florigen in higher plants. In this study, full-length cDNA of MdFT gene, a homolog of Arabidopsis AtFT, was amplified from apple leaf cDNAs with RT-PCR. Subsequently, a plant expression construct containing MdFT gene driven by cauliflowermosaic virus 35S promoter was obtained and introduced into tomato cultivar ‘Zhongshu 4 ’with Agrobacterium-mediated transformation. In parallel, Arabidopsis AtFT gene was transformed as a positive control. Finally, transgenic tomato lineswere regenerated from selection medium plus kanamycin. PCR amp lification verified the integration of exogenous genes into the host genome, and semiquantitative RT-PCR disp layed their ectopic overexpression in transgenic tomatoes. Furthermore, morphological observation found that transgenic tomato lines flowered earlier than the non-transgenic control, suggestive of our successful cloning of florigen gene MdFT from apple, which has thepotential as a target gene to shorten juvenility in apple tree.