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Induction of hairy roots in Arnica montana L. by Agrobacterium rhizogenes
- Petrova, Mariya, Zayova, Ely, Vlahova, Mariana
- Open life sciences 2013 v.8 no.5 pp. 470-479
- Arnica montana, Rhizobium rhizogenes, biomass production, cefotaxime, coculture, culture media, genes, genetic transformation, polymerase chain reaction, roots, secondary metabolites
- The induction of hairy roots in Arnica montana L. by Agrobacterium rhizogenes mediated system was established. The frequency of genetic transformation varied from 4.8 to 12% depended on method of infection. The cefotaxime at concentration of 200 mg/l proved to suppress effectively the growth of A. rhizogenes after co-cultivation. Among the three tested nutrient media: Murashige and Skoog (MS), Gamborg’s (B5) and Schenk and Hildebrandt (SH), MS medium was superior for growth and high biomass production of transformed roots compared to other culture media. After culturing for 40 days the fresh weight of clone T4 increased 7.6 fold over the non-transformed roots. The transfer of rol A, rol B and rol C genes into Arnica genome was confirmed by PCR analysis. Established genetic transformation techniques in A. montana efficiently provided and generated a large number of transformed roots — an excellent system for studying gene function and could be used for the production of secondary metabolites synthesized in roots.