Аgrobacterium rhizogenes – опосредованная трансформация и особенности регенерантов Digitalis purpurea L.
Aim. The influence of Agrobacterium rhizogenes-mediated transformation on morphological and biochemical features of D.purpurea. The goal was achieved through the study of the properties of the transformed cell cultures and regenerated plants. Methods. Transformation of D. purpureausing soil bacterium strain A.rhizogenes 15834, regeneration, PCR analysis, HPLC chromatography Results. A stable growing «hairy roots» culture and plant regenerates of Digitalis purpurea L. were obtained. Cultural mediums for different types of explants cultivation were chosen and optimized. pRi-transformants were obtained by spontaneous regeneration of transformed roots. HPLC analysis showed that the synthesis of cardiac glycosides in hairy roots exceeds 3.5 times its synthesis in callus, and in Ri-transformants synthesis of cardiac glycosides was amplified by 0.6 compared to intact plants. Conclusions. Hairy roots culture and Ri-transformated plants of D.purpurea which were able to synthese the whole range of secondary compounds typical to intact plants were obtained. The amount of cardiac glycosides in hairy roots was higher than in callus culture, but wasn’t enough for commercial cultivation. In transformed plants, the synthesis of secondary metabolites is increased compared to intact plants. We associate it with the incorporation of the rol-genes A.rhizogenes into the nuclear genome.
Keywords: Digitalis purpurea L., «hairy roots», regenerated plants, secondary metabolites, cardiac glycosides.