A Rapid and Stable Agrobacterium-Mediated Transformation Method of a Medicinal Plant Chelone glabra L.
文献类型： 外文期刊
作者： Gao, Zhenrui 1 ; Li, Ying 1 ; Chen, Jinhua 2 ; Chen, Zhixing 2 ; Cui, Min-Long 1 ;
作者机构： 1.Chinese Acad Sci, Inst Appl Ecol, Key Lab Pollut Ecol & Environm Engn, Shenyang 110016, Peoples R China
2.Yunnan Acad Appl Technol, Kunming 650051, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Zhejiang Acad Agr Sci, Inst Virol & Biotechnol, Hangzhou 310021, Zhejiang, Peoples R China
关键词： Chelone glabra L;Leaf explants;Agrobacterium-mediated transformation;Stable GUS expression
期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）
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收录情况： SCI
摘要： Transformation approach is a useful tool for the study of gene function, the mechanism of molecular regulation, and increase usefulness of components by reverse genetic approach in plants. In this study, we developed a stable and rapid method for Agrobacterium-mediated transformation of a medicinal plant Chelone glabra L. using leaf explants. Stable transformants were obtained using Agrobacterium tumefaciens strains GV2260 and GV3101 that harbored the binary vector pBI121 and contained the neomycin phosphotransferase gene (NPT II) as a selectable marker and a reporter gene beta-glucuronidase (GUS). Putative transformants were identified by kanamycin selection and a histochemical assay. PCR and Southern blot analysis confirmed the integration of the GUS gene into transformed genomes as well as detected stable expression of the beta-glucuronidase gene (GUS) by RT-PCR. Resulting transformed plants had morphologically normal phenotypes. This method requires two changes of medium and few leaf explants as well as the transformation efficiency of 2-8 % after 2-3 months of inoculation. This method can provide a quick and economical transformation method for reverse genetic approach to change the secondary metabolic pathway to increase useful components in C. glabra.
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