Optimized production and characterization of in vitro shoots of Withania somnifera as alternate to field grown tissues

Abstract

Withania somnifera is one among the 32 prioritized medicinal plants by newlinethe National Medicinal Plant Board(NMPB). Its global demand has increased newlinerecently owing to several pharmacological activities, but faces difficulty in newlinereaching the market due to inconsistency of quality standard. This study aims newlineto increase biomass production of in vitro shoots of W. somnifera thereby newlinedeveloping a source for continuous supply of raw materials with uniform newlinemetabolite content as an alternate to field grown materials for pharmaceutical newlinepurposes. In the present study, two biotic elicitors, fungal mycelial extract newline(Aspergillus niger) and a seaweed extract (Kappaphycus alverizii) were newlinestudied at different concentrations and exposure times individually and in newlinecombinations for their effect on Growth index (GI) and metabolite content. The newlineinfluence of these elicitors was then studied under three culture conditions newline(Solid, Suspension and Bioreactor). Among all the treatments, the seaweed newlineelicited shoots in bioreactor(TG3C3) recorded GI increase of 2.58 and 3.75 newlinefold over control shoots(TG1) in suspension(C2) and solid(C1) respectively. newlineThe antioxidant potential of the shoots also increased on seaweed elicitation newlineas exhibited by DPPH radical scavenging activity. The HPTLC quantification newlineof the withanolides (withanolide A, withaferin A and withanone) and GC-MS newlineprofiling of metabolites showed an overall decrease in bioreactor cultures newlinecompared to the solid, suspension cultures and the field grown shoots. This newlinedecreased metabolite content was comparable to the metabolite content of newlinethe field grown roots and thus was closely related to the field roots as newlinerevealed in the clustering analysis. This biomass enhanced in vitro shoots of newlineW. somnifera showed suppression of ROS levels, DNA damage and newlineapoptosis comparable to that of field roots in LPS induced raw 264.7 cells. newlineFurther the in vitro validation of two main hub genes of oxidative stress newlineobtained through in silico analysis showed that field root and in vitro shoot newlineextracts of W. somnifera