In Vitro Propagation And Determination Of Genetic Stability Of Micropropagated Plants Of Stevia Rebaudiana

Biotechnology

In present study, an efficient and economic medium was designed for sprouting of axilliary/apical buds and for shoot multiplication of Stevia rebaudiana. We used thick, vegetative shoot tips (apical buds) and auxiliary buds as explants for inoculation. Auxiliary buds showed better sprouting than apical buds. Such buds gave better response to the medium designed for spouting. Semisolid MS medium with 3% sucrose supplemented with BAP and Kinetin in combinations are used. It was observed that BAP (2mg/L) + Kinetin (0.5mg/L) give best results for shoot induction within a week and shoot grown upto height of about 3 to 4 cm in 2 to 3 weeks and within 30 days on an average highest number of total shoots (20-22) and highest average length of longest shoot (4±0.5 cm) were recorded. Inter-simple sequence repeat (ISSR) markers were used to evaluate the genetic stability of the micropropagated plants and it was found out that most of the ISSR profiles from micropropagated plants were monomorphic and comparable to mother plants. These results suggested that the micropropagation protocol developed by us for rapid in vitro multiplication is appropriate and applicable for clonal mass propagation of Stevia rebaudiana.

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