Protocol Optimization for the Acclimatization of Stevia rebaudiana Bertoni Under Controlled Conditions

Stevia rebaudiana explants (like nodal region, leaf, shoot apex) has been cultured in Ms. media with different supplements of plant growth hormones like (BAP), BA,2,4-D,NAA (0.25 to 1.5mg/lt) and has been developed invitro with slight variations to the mother plant. When these plants are shifted to natural environmental conditions needs proper acclimatization procedure to be followed because the survival percentage of the plants has been reduced which leads to the reduction in the multiplication of the plants. Acclimatization continues to be a major bottle neck in the micro propagation of stevia too with lower success reported. In the present investigation stevia plants had undergone two phases of hardening procedure like primary and secondary hardening in the green house. The plants are washed thoroughly with fungicides before starting the hardening procedure. The temperature maintenance and the variation in the potting mixture composition have a major role to play in the sustenance of the invitro stevia plant. The total procedure takes a time period of five weeks to enhance the survival percentage of plants.