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

Authors: GURURAJ CHALAGERI, UDDAGIRI VENKANNA BABU

Abstract: Viola patrinii DC. (Violaceae) petiole explants were used for inducing calli. Significant callus proliferation was observed on MS medium supplemented with 16.12 \muM NAA and 13.33 \muM BA. Shoot regeneration was achieved upon transferring the green friable calli to MS medium with a 2-fold dilution of potassium dihydrogen phosphate supplemented with 23.25 \muM Kn and 2.68 \muM NAA. Maximum shoot regeneration was achieved in 4 weeks. Multiple shoots were separated and further cultured in half-strength MS medium supplemented with 9.85 \muM IBA with 2% sucrose. Profuse root development was observed after 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.

Keywords: Viola patrinii, regeneration, micropropagation, leaf explants, RAPD, genetic fidelity

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