The nodal segment explants of Glycyrrhiza glabra L. were utilized to develop an efficient in vitro callus induction protocol. Full strength MS medium was tested with NAA and 2, 4-D alone and BAP with NAA and 2, 4-D respectively. Concentrations 1.0-5.0 mgL^-1 of NAA and 2, 4-D were used alone and BAP (3.0 mgL^-1) with 1.0-2.0 mgL^-1 of NAA and 2, 4-D concentrations were used for the incubation of the nodal segments respectively. Amongst all, the combined concentration of BAP (3.0 mgL^-1) with NAA (3.0 mgL-¹) was given superior results as 100 % of callus formation with 13.07 ± 0.39 average days taken to initiate callus. Today, Glycyrrhiza species became endangered because of overexploitation and demolition of a home as it contains many valuable remedial properties. Hence the present investigation was undertaken to establish an in vitro shoot formation protocol to preserve the important medicinal plant Glycyrrhiza glabra L.