Shirui lily (Lilium mackliniae), an endangered Asiatic lily species, hails from the Shirui peak of Siroi hill range in Ukhrul district of Manipur, India. This rare species is now at the verge of its extinction due to uncontrolled trampling and plucking by the human intrusions and extensive slash and burning cultivation in this region, as a result of which, it has been recorded in the ‘Red Data Book’ of ‘Indian Plants: Volume I’. Conservation of this endangered species under changing climatic conditions is a great concern for the researchers. An attempt has been made in the present investigation to establish an efficient in vitro regeneration protocol in L. mackliniae for its conservation. Callus cultures were established in MS medium containing different concentrations of 2,4-D (1-2mg/l). The regeneration ability in friable callus was 12 fold higher as compared to the compact callus. Direct regeneration and somatic embryogenesis was achieved in friable callus at 6-8 weeks of subculture in MS with benzyladenine (BA, 1.0mg/l). Multiple shoot proliferation and rooting were achieved in MS+0.5mg/l BA and 0.5mg/l napthalene acetic acid (NAA), respectively. Hardening under mist house resulted in higher survivability (85%) over poly house (62.2%) and shed net (55.5%). The in vitro plantlets were evaluated under different temperature and moisture regimes. The first time report of in vitro regeneration protocol will be helpful to augment breeding strategies in this rare endangered species for wide adaptation under changing climate conditions.