A fresh method of DNA transformation to the seeds irradiated by Co-60 without the use of antibiotic selection

作者:Li, Xinglin*; Wang, Dandan; Gao, Jie; Xiao, Tianjian; Wang, Xiaojuan; Zhang, Liming; Liu, Dong; Li, Limei; Lu, Fuping; Gao, Wen-yuan
来源:African Journal of Biotechnology, 2011, 10(31): 5845-5849.

摘要

To find out a simpler method that can directly transfer the aim gene into plant genomes, the purple medic seeds irradiated by Co-60 with 0.375 Gy were transformed by linear DNA containing a beta-glucuronidase (GUS) gene (as an aim gene), a betaine aldehyde dehydrogenase (BADH) gene (as a selectable marker) and two pairs of both CaMV35S promoter and Nos terminator. Subsequently, the seeds were planted and grown in perlite media watered with NaCl solution as a kind of selective compound. The results showed that, positive frequency of PCR identification by the GUS gene or the BADH gene was higher than 53.2 and 89.5% in T-0 and T-1 generations, while GUS staining rate was higher than 50%; whereas five T-1 plants assayed by southern hybridization all showed positive reaction. In conclusion, by this method, transgenic plants may be easily obtained with the antibiotic markers for free; moreover, the plant regeneration-system must not be erected by directly transforming the seeds.