The present study was aimed to develop an efficient reproducible protocol for the large scale propagation of medicinally important Boerhaavia diffusa L using the shoot-tip, nodal, internodal and leaves as explants. Highest percentage (97.2 ± 0.68 for nodal segments and 95.6 ± 0.82 for Shoot tip) of multiple shoot proliferation (14.4 ± 0.64 for nodal segments and 7.83 ± 0.38) was observed on MS medium augmented with 1.5 mg/l of BAP. The maximum number of shootlets regeneration (11.6 ± 0.38) was observed from MS medium supplemented with 3.0 mg/l of BAP. Maximum percentage (73.2 ± 0.47) of shootlets was observed in MS medium supplemented with 3.0 mg/l of BAP. Highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in ½ MS medium supplemented with 1.0 mg/l of IBA.