Direct Shoot Bud Proliferation Protocol from Stevia rebaudiana Leaf Culture for Healthy Biomass Production

Abstract

AN EFFICIENT and simple regeneration protocol using leaf explants was described on MS medium in the presence of various plant growth regulators. After four weeks of culture, the highest induction of adventitious shoot buds from leaf culture (90%) was achieved on MS medium with 1.0mg/L BA. The largest number of shoot buds (3.02) per leaf explant with no callus formation was achieved on MS medium with 2.0mg/L BA. The best combination for the induction of numerous shoot buds was found to be TDZ + BA + IBA (0.5, 1.0, and 0.5mg/L, respectively), which presented 10.4 shoots per explant with 3.15cm in length. For in vitro root formation stage, Healthy shoots were collected and cultivated on a half-strength MS with various concentrations of NAA and IAA alone or in combination with 2,4-D or IBA. Furthermore, the highest roots (8.17) per shoot with root formation (100 %) was observed using a half-strength MS medium in the presence of 2.0mg/L IAA. Therefore, healthy in vitro rooted plantlets were acclimatized successfully after four weeks in the greenhouse.