Ajuga bracteosa Wall ex. Benth is an important medicinal plant growing in Kashmir Himalaya. During the present study an efficient and rapid in vitro protocol has been established viz; Callus induction and multiple shoot regeneration. Callus induction has been achieved from different explants viz., leaf, petiole and internodal cuttings. Maximum callus production was obtained when leaf explants were inoculated on Murashige and Skoog (1962) medium supplemented with BAP (benzylaminopurine 5.0 mg/l) after 19 days of inoculation. In the petiole explant callus was induced on MS (Murashige and Skoog ) medium supplemented with BAP (2.0 mg/l) + IAA ( Indole-3-acetic acid 3.0 mg/l) after 51 days of inoculation and from internodal explant callus was induced on MS medium supplemented with BAP (2.0 mg/l) + NAA(α-Naphthalene acetic acid 5.0 mg/l) after 35 days of inoculation. Callus derived from leaf explants differentiated into multiple shoots on MS medium supplemented with different concentrations of auxins (IAA, NAA) and cytokinins (BAP, Kinetin). Maximum multiple shoots regenerated on MS medium supplemented with BAP (5.0 mg/l) after 28 days of inoculation.