Int. J. Biosci.14( 4), 292-304, April 2019
The present study was under taken to establish an efficient protocol considering various cultural aspects for plant regeneration, multiplication and rooting in vitro of Withania somnifera L. because of its medicinal value and to its survivability. Shoot tip and nodal segment were cultured aseptically on MS medium containing different concentrations and combinations of cytokinins (BAP and KIN) and auxins (NAA, IBA, and 2, 4-D) for direct and indirect regeneration. The explants were inoculated on MS medium supplemented with 8g/l Agar and 30g/l sucrose, different combination and concentration of plant growth regulators (auxin and cytokinins) was used for plant regeneration. The optimum pH was adjacent to 5.8 and autoclaved at 121±2°C for 20 minutes and incubated at 25±2°C under 16/8 hours photoperiod. In this study, the best shoot multiplication for direct regeneration was observed in 2.0mg/l BAP+0.2mg/l NAA, the longest shoot was observed 5.20cm and 5.00cm respectively. In case of indirect regeneration, highest response of callus was obtain by shoot tip (80.2%) and nodal segment (84.0%) culture on MS medium supplemented with 2mg/l 2, 4-D+0.2mg/l KIN. The best proliferation (6.5%) of adventitious bud was observed in BAP 2mg/l maximum rooting of the micro cuttings of regenerated shoots were successfully achieved with 2.0mg/l IBA and 1/2 MS+IBA2mg/l, the result was obtained 85% and 80% respectively. The plantlets were successfully transferred to soil and the percentage of survivability in such condition was obtains 87%. Micropropagation technique is the most effective way to rescue this medicinally endangers plant.
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