Kultura korenova Gentiana punctata L. transformisanih pomoću Agrobacterium tumefaciens C58Cl(pArA4b)
High biomass producing root cultures of Gentiana punctata L. transformed with Agrobacterium tumefaciens C58Cl(pArA4b)
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Gentiana punctata L. shoot cultures were transformed with Agrobacterium tumefaciens clone C58C1 harbouring A. rhizogenes A4 Ri plasmid (pRiA4b) to produce hairy roots capable of autonomous growth on growth regulator-free medium. Optimal medium comprised WPM macro-nutrient salts, MS micro-salts and iron, LS vitamins and 2% sucrose. Successful transformation was confirmed by PCR amplification of awe 1 and rol C sequences. Excised, transgenic root cultures characterized by intensive lateral root branching enabled high biomass production. Fast growth required use of explants with small initial weight and frequent subculturing. Cultures were maintained for 10 years (more than 100 subcultures) without changes in morphology or physiological responses.