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Simmondsia chinensis (Link) Schneider (jojoba) is a potential multipurpose plant of arid and semiarid regions of India. An efficient method for in vitro direct plant regeneration from nodal segments of mature male and female S. chinensis has been developed. The nodal explants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). The higher percentage (78.4±2.45) of shoot bud induction was observed at 2.5 mg/l BAP in male explants, while, percentage (86.2±2.14) of shoot bud induction obtained at 2.0 mg/l BAP in female explants. The influence of adenine sulphate (AdS) at 40 mg/l concentration with BAP was found best for vigorous shoot proliferation. For shoot multiplication in vitro derived shoots cultured on MS medium containing 2.0 mg/l BAP+1.0 mg/l NAA+40 mg/l AdS were produced the highest number (10.79± 2.81) of shoots with maximum (3.75±1.34 cm) shoot length in male cultures, while, the best combination of 2.0 mg/l BAP+0.5 mg/l NAA+40 mg/l AdS were yielded the highest number (9.64±1.23) of shoots with maximum (4.10±1.23 cm) shoot length in female cultures. Half strength of MS medium containing 3.0 mg/l IBA with 300 mg/l activated charcoal (AC) was found best for in vitro root induction in both male (98.7±1.64%) and female (97.3±1.04%) micropropagated shoots. The micropropagated plantlets were hardened under green house conditions and transplanted to field conditions with 92% survival rates. This standardized protocol of direct plant regeneration could be effective for large scale propagation of male and female S. chinensis and helps for successful commercialization of this plant.