We studied the effect of polyamines (spermidine, putrescine and spermine) along with benzyladenine, kinetin and napthaleneacetic acid on multiple shoot regeneration of sugarcane. Murashige & Skoog medium containing a combination of benzyladenine (8.3 μM), kinetin (4.6 μM), napthaleneacetic acid (2.6 μM) and spermidine (68 μM) induced the maximum number of shoots (42 shoots/explant) compared to benzyladenine (8.3 μM), kinetin (4.6 μM) or napthaleneacetic acid (2.6 μM) alone or with putrecine (68 μM), spermine (32 μM) or combinations of polyamines. Plantlets raised were successfully transplanted in soil with a 90% survival rate.