Nitovska I.O., Shakhovsky A.M., Komarnitsky I.K., Kuchuk M.V.

Production of Brassica oleraceae(+Arabidopsis thaliana) and Brassica napus cell lines resistant to spectinomycin/streptomycin as a result of plastome genetic transformation

Cytology and Genetics 2006, v 40, number 3, 3-10

Plastid genetic transformation has been performed using both the PEG-treatment of protoplasts of somatic hybrids of B. oleracea carrying A. thaliana chloroplasts and the particle bombardment of regenerable calluses of B. napus sv. Westar. The chloroplast transformation vector pCB040 carried resistance (aadA) gene flanked by rapeseed plastid DNA sequences to target its insertion between the trnV-rps7 fragments. Selection of transplastomic cell lines has been performed according to their ability to grow on the medium supplied with spectinomycin and streptomycin in high concentrations. Antibiotic resistant cell lines have been obtained using the both transformation methods. The presence of the aadA gene in the A. thaliana and B. napus plastomes was confirmed by PCR analysis for two cell lines of B. oleracea (+A. thaliana) and three lines of B. napus.