Micro propagation and tissue culture of the endangered medicinal plant withania somnifera by the direct shoot and root initiation method.

ABSTRACT

Leaf and Cotyledon explants of Withania somnifera (L). Dunal were used to evaluate the effect of different growth regulators on the in vitro direct shoot and root initiation methods. Four different explants were used to establish callus shoot and root direct regeneration. In the first experiment leaf segments were cultured on MS basal supplemented with 2,4 – Dichlorophenoxyacetic acid (2,4 – D, 0.1-20.0 mg/L), with combination of Naphthalene acetic acid (NAA 0.1-20 mg/L) and Benzylaminopurin (0.1-20 mg/L). This new protocol was standardized for easy mass propagation of W. somnifera medicinal plant. Callus initiation was observed best in MS media with (2,4- D 1.0-5.0 mg/L) after 16-20 days (93%). Highest maximum number of multiple shoots was obtained on MS medium (BAP 3.0 – 5.0 mg/L). The shoots were seaperated from the multiple-shoots, transferred to MS medium supplemented with 1.5 – 20 mg/L NAA favored roots formation occurred in most of the shoot let 88% were successfully achieved in the MS media. The rooted plantlets were transferred to polythene bags which was containing vermi compost, sand and red soil in the ratio of 122 and kept in a mist house. After acclimatization in the mist house for 2-months, it transferred to greenhouse. The plantlets were successfully planted in the field.