SHOOT REGENERATION AND AGROBACTERIUM-MEDIATED TRANSFORMATION OF FRAGARIA-VESCA L

An efficient and reliable method for shoot regeneration from leaf disk s of Fiagaria vesca L. has been developed. This protocol has been succ essfully employed to obtain transformed plants using Agrobacterium tum efaciens as gene vector. Murashige and Skoog basal medium supplemented with benzyladenine (4 mg/l) and indole-3-butyric acid (0.25 mg/l) ind uced the maximum percentage of shoot regeneration (98%) and the highes t number of shoot colonies per explant (4.6) after 8 weeks of culture. Isolated shoots would elongate and proliferate when the benzyladenine concentration was lowered to 0.5 mg/l. The established protocol for s hoot regeneration was employed to transform leaf disks using Agrobacte rium tumefaciens carrying the plasmid pBI121. A 7.7% of the inoculated explants showed kanamycin resistance after 10 weeks of selection in a medium containing 25 mg/l of this antibiotic. The transgenic shoots o btained were rooted in the presence of 25 mg/l kanamycin and successfu lly acclimatized. The final percentage of transformation obtained base d on beta-glucuronidase expression was 6.9%.