tailieunhanh - In vitro plant regeneration via petiole callus of Viola patrinii and genetic fidelity assessment using 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. | Research Article Turk J Bot 36 (2012): 358-368 © TÜBİTAK doi: In vitro plant regeneration via petiole callus of Viola patrinii and genetic fidelity assessment using RAPD markers Gururaj CHALAGERI*, Uddagiri Venkanna BABU Plant Tissue Culture Lab, Research & Development Centre, The Himalaya Drug Company, Makali, Bangalore 562 123 - INDIA Received: Accepted: Abstract: Viola patrinii DC. (Violaceae) petiole explants were used for inducing calli. Significant callus proliferation was observed on MS medium supplemented with μM NAA and μM 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 μM Kn and μM NAA. Maximum shoot regeneration was achieved in 4 weeks. Multiple shoots were separated and further cultured in half-strength MS medium supplemented with μM 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. Key words: Viola patrinii, regeneration, micropropagation, leaf explants, RAPD, genetic fidelity Introduction Viola patrinii DC. (Violaceae), commonly known as China violet, is an important medicinal herb, mentioned in traditional medicine for a variety of therapeutic applications including the purification of blood and the treatment of bruises and ulcers (Kirtikar & Basu, 1995); in the Chinese system of medicine it is recommended for use against .