Rapid shoot multiplication of Nyctanthes arbortristis was achieved from axillary meristems on Murashige and Skoog (MS) basal medium supplemented with 1.0−1.5 mg/l 6-benzyladenine (BA), 50 mg/l adenine sulfate (Ads) and 3% (m/v) sucrose. Inclusion of indole-3-acetic acid (IAA) in the culture medium along with BA + Ads promoted a higher rate of shoot multiplication. Maximum mean number of microshoots per explant (6.65) was achieved on the MS medium supplemented with 1.5 mg/l BA, 50 mg/l Ads and 0.1 mg/l IAA after 4 weeks of culture. The elongated shoots rooted within 13 to 14 days on ½ strength MS medium supplemented either with indole-3-butyric acid (IBA), IAA or naphtylacetic acid (NAA) with 2% sucrose. Maximum percentage of rooting was obtained on medium having 0.25 mg/l IBA, 0.1 mg/l IAA and 2% sucrose. About 70% of rooted plantlets survived in the greenhouse. The in vitro raised plants were grown normally in the soil condition. This result will facilitate the conservation and propagation of the important medicinal plant.