An in vitro protocol for plant regeneration through organogenesis was established for Picrorhiza kurroa Royle Ex. Benth (Scrophulairaceae). The plant is a principle source of glycoside that is Picroside-I, Picroside-II and Kutkoside. Callus was induced on Murashige and Skoog basal medium supplemented with cytokinin (Kn or BA) and auxin (2,4-D/IBA/NAA) using leaf, petiole and stem explants. Shoots were regenerated on MS media with BA ( to mgl-1 ) combined with auxin (IBA or NAA) in the range of to mgl-1 . Rooting was initiated with full or half media supplemented with to mgl-1 of IBA or NAA. In vitro regenerated plantlets were successfully transferred to the soil. Pots having garden soil with organic compost and regular irrigation of half strength MS solution at initial stages are desirable for higher survival of plantlets. | Standardization of in vitro multiplication protocol for Picrorhiza kurroa (Royle Ex. Benth)