African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12481

Full Length Research Paper

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

Xinglin Li1*, Dandan Wang1, Jie Gao1, Tianjian Xiao1, Xiaojuan Wang2, Liming Zhang1, Dong Liu1, Limei Li1, Fuping Lu 1 and Wen-yuan Gao3          
1Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin, 300457, People’s Republic of China. 2College of Pasture Sciences and Technology, Lanzhou University, Lanzhou, 730000, People’s Republic of China. 3College of Pharmaceuticals and Biotechnology, Tianjin University, Tianjin, 300072, People’s Republic of China.
Email: [email protected]

  •  Accepted: 06 April 2011
  •  Published: 29 June 2011

Abstract

To find out a simpler method that can directly transfer the aim gene into plant genomes, the purple medic seeds irradiated by 60Co with 0.375 Gy were transformed by linear DNA containing a β-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 T0 and Tgenerations, while GUS staining rate was higher than 50%; whereas five T1 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.

 

Key words: DNA transformation, irradiated seeds, purple medic, salt screening.