CALLUS VIA SHOOT ORGANOGENESIS AND PLANT REGENERATION OF STEVIA REBAUDIANA BERTONI

Stevia rebaudiana Bertoni is important medicinal plant which is native between southern Brazil and northern Paraguay. A reliable protocol for shoot organogenesis and regeneration was developed for an antidiabetic medicinal plant species. Regeneration via indirect shoot organogenesis was established from two types of explant (leaf and stem) on MS medium [1] with different concentrations and combinations of plant growth regulators (PGR) such as 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP) and gibberellic acid (GA(3)) for inducing organogenic callus of S. rebaudiana. In the present investigation the highest percentage of response was shown by 90% and 80% callogenesis from leaf and stem explants on MS medium supplemented with 2.0 mg/l 2,4-D + 0.5 mg/l BAP + 0.5 mg/l GA(3) (MSC3) was observed. The high frequency of regenerants was observed from S. rebaudiana leaves at 0.5 mg/l 2,4-D + 2.0 mg/l BAP + 0.5 mg/l GA(3) (MSR3). Optimal shoot multiplication and rooting were obtained on 2.0 mg/l BAP and 0.1 mg/l indole-3-butyric acid (IBA), respectively, followed by successful acclimatization of regenerants (90% survival) to greenhouse condition. The purpose of this study was to establish a reliable S. rebaudiana regeneration protocol that can be used to facilitate the cultivation, propagation and planting of this plant species.