Genetic transformation of olive somatic embryos through Agrobacterium tumefaciens and regeneration of transgenic plants
Transformed olive plants were regenerated from inoculated somatic embryos with Agrobacterium tumefacience strain GV3101, which carries the plasmid pBI-P5CS containing Arabidopsis thaliana P5CS cDNA, kanamycin marker (npt) gene and uidA reporter gene. Initially, repetitively embryogenic cultures were established from radicles and cotyledonary segments of mature olive zygotic embryos. Single somatic embryos at cotyledon stage were used for transformation. Through repetitive somatic embryogenesis, non-chimer secondary embryos were selected and propagated on kanamycin containing medium. Resistant embryos were converted to plantlets by subjecting them to desiccation. Transformation and P5CS gene expression was confirmed by β-glucuronidase (GUS) assay polymerase chain reaction (PCR) and reverse transcription (RT)-PCR analysis.
Key words: Olea europaea, somatic embryogenesis, transformation, β-glucronidase, P5CS gene.