地黄基因RghBNG转化地黄过表达研究

用RT-PCR技术从地黄[Rehmannia glutinosa(Gaertn.)Libosch.]幼叶中克隆地黄基因RghBNG,使用Pst I和HindⅢ分别酶切地黄基因RghBNG和植物表达载体p CAMBIA302-GFP,经过重组,构建其过表达载体,转化农杆菌菌株GV3101,获得其工程菌;采用农杆菌介导法,将该基因转入地黄幼叶细胞,再生转化地黄植株。经过潮霉素抗性筛选、PCR和qRT-PCR检测,获得过表达基因RghBNG地黄植株。这些结果为进一步阐明地黄基因RghBNG功能和地黄遗传改良奠定了基础。

The Transformation of Rehmannia glutinosa RghBNG Gene into Rehmannia glutinosa (Gaertn.) Libosch. For Overexpression

The RghBNG gene was cloned from Rehmannia glutinosa(Gaertn.) Libosch.leaves by RT-PCR.Pst Ⅰ and Hind Ⅲ as upstream and downstream enzyme,respectively,were used to digest RghBNG and the plant expression vector pCAMBIA-1302 in order to construct plant overexpression vector pCAMBIA1302-RghBNG-GFP.The overexpression vector was introduced into Agrobacterium tumefaciens strain GV3101 so that its engineered bacteria were obtained.RghBNG was transformed into Rehmannia glutinosa(Gaertn.) Libosch.leaf cells using Agrobacterium-mediated transformation,and hygromycin-resistant transgenic Rehmannia glutinosa (Gaertn.) Libosch.plantlets were obtained via tissue culture,which were confirmed by PCR and qRT-PCR analyses.These results will build up the foundation for further verification of RghBNG functions and its application in Rehmannia glutinosa (Gaertn.) Libosch.genetic improvement.