Establish an in vitro propagation system and produce secondary metabolites in Ashwagandha (Withania somnifera L.)

Ashwagandha (Withania somnifera L.) is an endangered and vital medicinal Indian plant. The present study focused on in vitro micropropagation of Ashwagandha plant material and studied the effects of Chitosan levels as an elicitor on secondary metabolic bioactivity withanolides (withanolide-A, withaferin-A). MS medium fortified by BAP (1.0mg/L) and NAA (0.125mg/L) achieved a maximum of 6 shoots per explant. In vitro shoot elongation and leaf number (12.33cm and 10.67, respectively) were improved on MS medium supplemented with BAP at 2.00mg/L and NAA at 0.25. High frequency of rooting 100% and maximum root length (8.00cm), as well as the number of white roots (1.00), were achieved on an MS medium supplemented by NAA (0.5mg/L). Referring to the impacts of Chitosan levels on in vitro micropropagation, all growth parameters (shoots, roots) were decreased significantly in an inverse relationship with Chitosan levels. HPLC method was used for the quantitative importance of bioactive chemical constituents, withanolide-A, and withaferin-A, to estimate the quantitative importance of bioactive chemical constituents withanolide-A, and withaferin-A affected by Chitosan levels as the elicitor in vitro. Chitosan at 100 ppm achieved the maximum accumulation of the withanolide-A (259.12mg/g), while the highest concentration of withaferin-A (11.96mg/g) was obtained by 200 ppm Chitosan in root tissues. In general, the results indicate the possibility of obtaining an ideal medium for the propagation of Ashwagandha in vitro and the importance of the effect of Chitosan as a stimulant on the vegetative parameters and increasing accumulation of active alkaloids in the shoot and root tissues of Ashwagandha.