AGROBACTERIUM-MEDIATED TRANSFORMATION AND REGENERATION OF THE DIPLOID STRAWBERRY
Transformed calli and plants of the diploid strawberry (Fragaria vesca FRA 197) were obtained using Agrobacterium tumefaciens carrying a binary vector plasmid pBI121 which contains a nopaline synthase (NOS) promoter-driven neomycin phosphotransferase (NPT II) gene and a CAMV35S promoter-driven B-glucuronidase (GUS) marker gene. Following cocultivation, petioles and leaf discs were placed on non-selective shoot induction medium for 5 days, and then on shoot selection medium (SSM) containing 50 mg/L kanamycin and 250 mg/L mefoxin. Leaf discs and petioles were transferred every 6 weeks to fresh SSM, and after the third subculture the kanamycin was reduced to 25 mg/L. Calli from two petioles gave rise to shoots which were easily rooted. GUS activity, assayed histochemically, was detected in putative transformants but not in control plants. Presence of the NPT II and the GUS marker genes in leaves of transformed plants was fonfirmed by the Polymerase Chain Reaction (PCR), using primers specific to each gene. Amplification of an alcohol dehydrogenase sequence was utilized as an internal positive control in the PCR.
Haymes, K. M. and Davis, Thomas M. (1993). AGROBACTERIUM-MEDIATED TRANSFORMATION AND REGENERATION OF THE DIPLOID STRAWBERRY. Acta Hortic. 348, 441-441