In vitro plant regeneration via petiole callus of Viola patrinii and genetic fidelity assessment using RAPD markers

Abstract

Viola patrinii DC. (Violaceae) petiole explants were used for inducing calli. Significant callus proliferation was observed on MS medium supplemented with16.12 μM NAA and 13.33 μM BA. Shoot regeneration was achieved upon transferring the green friable calli to MS medium witha 2-fold dilution of potassium dihydrogen phosphate supplemented with23.25 μM Kn and 2.68 μM NAA. Maximum shoot regeneration was achieved in 4 weeks. Multiple shoots were separated and further cultured in half-strengthMS medium supplemented with9.85 μM IBA with2% sucrose. Profuse root development was observed aft er 20-25 days of culturing. Regenerated plants were successfully transferred to soil, showing an 85% survival rate. The genetic fidelity of the micropropagated plants was analysed using random amplified polymorphic DNA (RAPD) markers. The RAPD profile of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrating the homogeneity of the micropropagated plants. This protocol could be successfully used for the mass multiplication and germplasm conservation of this medicinal plant. © TÜBİTAK.