An efficient protocol for high frequency in vitro regeneration of Meizotropis pellita an endangered and endemic plant was developed. In vitro germination of the seeds of Patwa was achieved successfully after incubation for 10 -15 days and plantlets up to a height of 5 -7 cm with profuse rooting were observed after incubation for 7 weeks in hormone free MS medium. This in vitro germination totally reduced contamination that used to occur in the explant taken from the field. Callus induction and proliferation were observed in leaf explants after 15 -20 days of incubation in MS medium containing 2 -4, D (9.06 μM) alone or in combination with 2 -4, D (9.06 μM) + 2-iP (7.38 μM). Shoot regeneration was achieved from 2 months old callus explant in MS medium supplemented with BA (17.6 μM) + GA3 (1.0 μM). Shoot multiplication was also achieved from cotyledonary node of M. pellita in MS medium supplemented with Kinetin + GA3 (4.6 μM + 1.0 μM) or BA (13.2, 17.6 μM) + GA3 (1.0 μM) after 30 -45 days of incubation. IBA (4.9 μM) was more effective in root regeneration from micro shoots. The plantlets after acclimatization over a period of 1 month were further hardened in a polyhouse for two months. On the basis of available literature this is the first and significant study regarding the comparative effect of different PGRs on in-vitro propagation of Meizotropis pellita by using different explants and their subsequent effect on rooting. This significant study could be useful for large scale production of successfully hardened plants and conservation of this shrub.