A rapid Agrobacterium-mediated transformation of Antirrhinum majus L. by using direct shoot regeneration from hypocotyl explants

Abstract

A rapid genetic transformation of Antirrhinum majus L. was developed using direct shoot regeneration from hypocotyl explants inoculated with Agrobacterirum tumefaciens. We used the binary vectors pBI121 containing the neomycin phosphotransferase (NPT II) gene as a selectable marker and the β-glucuronidase (GUS) gene as a reporter and pIG121-Hm harboring NPT II and the hygromycin phosphotransferase (hpt) gene as selectable markers and GUS as a reporter. Transformed shoots directly emerged at the end of hypocotyl explants inoculated with Agrobacterium strain GV2260/pIG121-Hm or GV3101/ pBI121 after 4–6 weeks of culturing on selection medium. The transformed shoots were rooted after 3 weeks of culturing on rooting medium. The transformation efficiency reached to 1% of inoculated explants. Putative transformants were identified using kanamycin selection and GUS activity. The PCR and Southern blot analysis confirmed the integration of GUS into the genomes of transformants. Using the developed protocol, transgenic A. majus is produced after 10 weeks of Agrobacterium inoculation.