Recovery of transgenic orchid plants with hygromycin selection by particle bombardment to protocorms |
| |
Authors: | Yu Zhihua Chen Minyong Nie Lin Lu Huafei Ming Xiaotian Zheng Honghong Qu Li-Jia Chen Zhangliang |
| |
Institution: | (1) The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing, 100871, China;(2) De Montfort University, Leicester, LE1 9BH, UK |
| |
Abstract: | Protocorms of orchid (Dendrobium hybrid) were transformed by microprojectile bombardment with a helium-pressured PDS 1000 particle gun. Gold particles coated with plasmid DNA containing ß-glucuronidase (GUS) and hygromycin phosphotransferase (Hpt) marker genes were used. Potentially transformed tissues were identified by active growth on MS medium supplemented with 50mg l-1 hygromycin. After 4–6 months of continuous selection, 15 hygromycin-resistant lines were recovered. Integration of transgenes into the genome of the transformed protocorms and plantlets were confirmed by GUS histochemical assay and Southern blot hybridization. The transgenic protocorms have gone through propagation for more than 8 months and maintained their transgenic characters. These results indicate that we have established a system for orchid transformation in a relatively high frequency and the transgenes are stably expressed in the transgenic plants. |
| |
Keywords: | ?-glucuronidase dendrobium hygromycin phosphotransferase orchid transformation |
本文献已被 SpringerLink 等数据库收录! |