A most effective protocol for axillary shoot proliferation was established on MS medium supplemented with different concentrations of cytokinin and auxin using nodal and shoot tip explants from field grown mature plant of Gynura porcambens. In vitro response of the explants to multiple shoot regeneration varied greatly with the position of the explanting branch on the donor plant. Highest frequency 98.21% of shoot formation, maximum number 6.2 of shoots per explant and average 4.5 cm length of shoot were obtained on MS medium supplemented with 4.0 µM 6-Benzylaminopurine (BAP). Shoot multiplication and growth were significantly affected by level of concentration of sucrose. Optimum sucrose concentration was also estimated for in vitro shoot regeneration from nodal explant in MS medium with different concentrations of sucrose. The highest rate (97.31%) of shoot production was achieved in a shootregenerating medium with 30 gm/l sucrose. For rooting, the in vitro proliferated and elongated shoots were excised into 2-4cm long microcuttings, which were planted individually on a root-induction MS medium containing 4.0 µM Indole-3-butyric acid (IBA). Within 4 weeks of transfer to the rooting medium, all the cultured microcuttings produced 2-6 roots. The in vitro regenerated plantlets were transferred to soil, and the survival rate ex vitro was 95.00%.