In the recent years phytomedicines are employed for curing many diseases as alternative source of synthetic drugs due to many reasons; affordability, low price, no side effects and ecofriendly. Higher demand and hence over exploitation have depleted the plant sources severely. There is an urgent need to conserve the natural stalk by developing conservation strategies. Hypochaeris radicata, one such species needed in large quantity is severely exploited for its medicinal uses in high hills of Nilgiris, the Western Ghats, India. Further, no major scientific reports are available with respect to its bioactive compounds and therapeutic properties. To address this lacuna, the study was carried out to develop protocol for in vitro propagation, to know the phytochemicals by preliminary investigation and quantification and to evaluate the antioxidant properties. The results of the study report that higher number of shoots were obtained from cotyledonary leaf explants in the MS medium fortified with BAP at 2mg/L (100%) followed by BAP and GA3 at 2mg/L and 0.5mg/L (95.63%) respectively. Rooting percentage (98.48%) was better which subcultured onto the MS medium contained NAA at 1mg/L. Rooted plantlets were hardened (garden soil: sand: vermicompost=1:1:1) and established in pots with 75.22% survival rate. The determination of phytochemical constituents by qualitative and quantitative analysis revealed the presence of rich variety of secondary metabolites. The in vitro antioxidant activities of in vitro derived methanolic leaf extract of H. radicata proved this species as potential source for pharmaceutical drug preparations.