Abstract
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 secondary metabolites were analyzed and quantified using high performance liquid chromatography. Amount of secondary metabolites was found significantly higher, in in vitro plantlets compared to in vivo plantlets and callus raised from S. chirayita. Higher heavy metal accumulation in in vitro as compared to in vivo plantlets correlates higher secondary metabolite production supporting that they play regulatory role in influencing the plant secondary metabolism.
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