Efficient in vitro plant regeneration from leaf-derived callus and genetic fidelity assessment of an endemic medicinal plant Ranunculus wallichianus Wight & Arnn by using RAPD and ISSR markers

Ranunculus wallichianus is a medicinally important plant and an endemic species to the Western Ghats of South India. An efficient reliable indirect regeneration protocol system for R. was developed from leaf explants in present investigation. Leaf were cultured on both full-strength half-strength MS (Murashige & Skoog) medium supplemented with different concentrations (0.5 mg L− 1 3.0 1) 2,4-D NAA. Among tested, highest percentage yellowish-green compact nodular callus formation observed 2.0 2, 4-D. Then, vitro raised organogenic containing various BA, KIN, TDZ 0.5 NAA 10 % CW shoot regeneration. The response (97 %) maximum number shoots (11.1 ± 0.13 shoots/culture 9.2 0.21 cm mean length) obtained 2.5 BA CW. well elongated effectively rooted IBA + 250 activated charcoal. well-rooted plantlets successfully hardened acclimatized survival rate 94 %. Clonal fidelity assessed by using DNA-based RAPD ISSR molecular markers. A total 56 47 monomorphic bands markers respectively. This propagation could be effective conservation their genetic purity its further investigations. study develops mass multiplication Ranunuculus through organogenesis evaluates clonal regenerants