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Portulaca quadrifida (Portulacaceae) is an annual succulent herb having medicinal value and is consumed as a vegetable or salads in India. In the present study, leaf explants were inoculated on Murashige and Skoog’s (MS) medium fortified with sucrose (3%) and combinations of N6-benzyladenine (6-BA) and kinetin (KIN) individually and in combination with 1-naphtalene acetic acid (NAA). Rapid regeneration was observed in medium fortified with combinations of 6-BA (8 μM) and NAA (1 μM) which formed 19.40 ± 0.64 shoots with 100% response. Variation in sucrose concentrations (4-6%) was tried but it failed to increase the shoot number. When the optimized medium was fortified with different carbon sources viz. dextrose, glucose and maltose, they could not evoked better response and sucrose proved to be more effective for regeneration. Rooting of in vitro shoots was achieved in ½MS + sucrose (1%) + indole-3-butyric acid (IBA, 2 μM).