A method of Agrobacterium tumefaciens mediated transformation for perennial ryegrass was developed using the calli of ryegrass derived from mature enrbryos. The calli were inoculated with a disarmed A. tumefaciens str...A method of Agrobacterium tumefaciens mediated transformation for perennial ryegrass was developed using the calli of ryegrass derived from mature enrbryos. The calli were inoculated with a disarmed A. tumefaciens strain EHA105 harboring binary vector p2328. Vector p2328 contained transcription factor DREB1B and neomycin phosphotransferase （npt H） genes which were driven by promoters of rd29B and CaMV35S, respectively. The inoculated calli were selected on paromomycin- or kanamycin-containing media till the established plants being transferred to soil. Six tmnsgenic plants with DREB1B had been obtained from perennial ryegrass strain Tove. PCR and Southern-blotting showed that npt Ⅱ and DREBIB genes were integrated in perennial ryegrass genome. Stress treatment confirmed that transgenic plants with higher drought tolerance were obtained.展开更多
基金Supported by the National Natural Science Foundation of China（30170589）and the National Special Project for Research and Industrialization of Transgenic Plants （J-2002-B-006）. Acknowledgements： We are grateful to Dr. Zhang Xiao-dong of Beijing Academy of Agricultural and Forestry Sciences for donating gene DRB1B and Agrobacterium tumefaciens EHA105. We thank Ms. Yang Hong of Chengdu Institute of Biology, Chinese Academy of Sciences, for conducting much work of tissue culture. We also thank Center for Application of Molecular Biology to International A griculture （CAMBIA）, Australia for permission of use pCAMBIA2301.
文摘A method of Agrobacterium tumefaciens mediated transformation for perennial ryegrass was developed using the calli of ryegrass derived from mature enrbryos. The calli were inoculated with a disarmed A. tumefaciens strain EHA105 harboring binary vector p2328. Vector p2328 contained transcription factor DREB1B and neomycin phosphotransferase （npt H） genes which were driven by promoters of rd29B and CaMV35S, respectively. The inoculated calli were selected on paromomycin- or kanamycin-containing media till the established plants being transferred to soil. Six tmnsgenic plants with DREB1B had been obtained from perennial ryegrass strain Tove. PCR and Southern-blotting showed that npt Ⅱ and DREBIB genes were integrated in perennial ryegrass genome. Stress treatment confirmed that transgenic plants with higher drought tolerance were obtained.
