Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture

The influence of cytokinins and culture conditions including medium volume, harvest time and elicitation with abiotic elicitors (SA/MeJ) have been studied for the optimal production of biomass and withanolides in the multiple shoot culture of Withania somnifera. Elicitation of shoot inoculum mass (2 g l(-l) FW) with SA at 100 mu M in the presence of 0.6 mg l(-l) BA and 20 mg l(-l) spermidine for 4 h exposure time at the 4th week in 20 ml liquid medium recorded higher withanolides production (withanolides A [8.48 mg g(-l) DW], withanolides B [15.47 mg g(-l) DW], withaferin A [29.55 mg g(-l) DW] and withanone [23.44 mg g(-l) DW]), which were 1.14 to 1.18-fold higher than elicitation with MeJ at 100 mu M after 5 weeks of culture. SA-elicited cultures did not exhibit much variation in biomass accumulation when compared to control. This cytokinin induces and SA-elicited multiple shoot culture protocol provides a potential alternative for the optimal production of biomass and withanolides utilizing liquid culture.