Vitex negundo is a commonly used medicinal plant in Indian subcontinent. It has broad range of uses including antimicrobial, analgesic, antifertility, antioxidants, antihyperglycemic to anti-inflammatory and antitumor effects. Considering its medicinal importance, an attempt was made with an aim to protect, conserved the germplasm and to improve genotype of V.negundo by in-vitro technique of tissue culture through callus induction. Explants collected from field grown plants were cultured on MS medium with different concentration and combinations of Auxin (2, 4D, NAA) and cytokinin (BAP, Kin) with or without additive like coconut water for callogenesis. Among 12 different hormonal combinations of MS media, media supplemented with 2, 4D, NAA BAP alone or in combination with C12 showed better response for callussing and C18 for multiple shooting and R14 for rooting showed high efficacy to induce callus and organogenesis. Callus induction was observed within 6 days and the callus was subcultured once in 2 week in subculturing media. The multiple shoot regeneration was observed. Maximum number of shoot regeneration was observed (8-12) in S18 media, supplemented with BAP-10-28 ml and coconut water 5 ml/l. The regenerated shoots having size of 10-28 mm were excised and transferred to liquid medium supplemented with low concentration of IBA, NAA for rooting. Among the different concentration of Auxin tested was more effective for inducing roots within 2 weeks of culture with maximum number of 8-16/explants measuring size of 16-20 mm. 85- 90% of the microshoots produced long healthy root system.
