Abstract: A protocol has been developed for plant regeneration from protoplast culture of Crocus pallasii subsp. haussknechtii using regenerable embryogenic calli obtained from shoot meristem culture on MS+9.28 μM kinetin+4.52 μM 2,4-D. Protoplasts were isolated directly from embryogenic calli, embedded in Ca-alginate beads and cultured with nurse cells in MS+4.64 μM kinetin+4.52 μM 2,4-D+5.68 μM ascorbic acid+0.3 M mannitol at 20±2°C in darkness. After appearing of microcalli on the surface of the beads, they were transferred onto 1/2MS+2.32 μM kinetin+2.26 μM 2,4-D+5.68 μM ascorbic acid for growth of embryogenic calli. Somatic embryos matured on MS medium growth regulator free and germinated on 1/2MS+14.45 μM GA3 +4.43 μM BA at 20±2°C in a 16/8 h light/dark cycle.