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

  • X Li
  • D Wang
  • J Gao
  • T Xiao
  • X Wang
  • L Zhang
  • D Liu
  • L Li
  • F Lu
  • WY Gao
Keywords: DNA transformation, irradiated seeds, purple medic, salt screening.

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 T1 generations, 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.

Published
2013-09-19
Section
Articles

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eISSN: 1684-5315