Transformation and Regeneration
Successful transformation of A. carvifolia with A. tumefaciens GV3101 harboring the rol B and rol C genes was carried out. Two independent transformation experiments were performed and 300 explants were used per transformation event. Transformation efficiency was found to be 20–30%, but only 4 rol B transformants and 3 rol C transformants survived to maturity on the selection media. Morphological variability was observed between wild type plants and transgenics bearing rol B and rol C genes. Rol B transgenics, which grew faster on the selection media, had wider leaves and more inflorescence, while rol C transgenics, which were recalcitrant to regeneration, showed a narrow leaf blade and decreased internode length (Fig 5).