A Simple and Efficient In Vitro Mass Multiplication Procedure for Stevia rebaudiana Bertoni and Analysis of Genetic Fidelity of In Vitro Raised Plants Through RAPD

An investigation was carried out to develop an efficient micropropagation protocol for Stevia rebaudiana Bertoni. Experiments were conducted to optimize suitable media for in vitro shoot multiplication and root induction and to study the effect of culture vessel on shoot multiplication. Out of the different media compared for in vitro shoot multiplication, hormone-free liquid medium was found the most suitable. The results also revealed that strength of Murashige and Skoog (MS) salt, culture medium consistency (solid or liquid) and type of culture vessel significantly influenced the in vitro shoot multiplication rate. The shoots when placed on half strength hormone free half MS medium and 100 ppm charcoal showed cent percent root induction with maximum number of roots per shoot (21.2) as well as maximum root length (4.22 cm). Hardening of rooted plants on biopeat showed more than 80 % of survival rate. Further, clonal fidelity of the in vitro raised plants was carried out using RAPD markers and results indicated that all the tissue cultured derived plants are true-to-type and there are no somaclonal variations among these plants.