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An efficient protocol of plant regeneration through direct and indirect organogenesis in Swertia chirayita was developed. Explants cultured on Murashige and Skoog medium supplemented with 2,4-D (0.5 mg L−1) with combination of Kinetin (0.5 mg L−1) showed the highest frequency (84%) of callusing and 1.0mg L−1 6-benzyladenine (BA) in combination with (100 mg L−1) Adenine sulphate (Ads) + (0.1 mg L−1) Indole acetic acid (IAA) was excellent for maximum adventitious shoot (12.69 ± 1.30) formation in four week of culture. A maximum number of (7.14 ± 0.99) shoots were developed per leaf explants through direct organogenesis. The highest frequency of rooting (11.46 ± 1.56) was observed on MS medium augmented with IAA (1.0 mg L−1). Well-rooted shoots transferred to plastic pots containing a soilrite: sand mix and then moved to the greenhouse for further growth and development. Four major …