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Micropropagation of Stevia rebaudiana Using Shoot Tip Explants – A Magical Sweetener and Medicinal Plant

Centre for Biotechnology, Maharshi Dayanand University, Rohtak-124 001 (Haryana), India
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(Received : August 21, 2017; Accepted : October 13, 2017)


Stevia rebaudiana is emerging as a new natural sweetener crop world-wide and expected to be a potential alternative crop for diversification in agriculture. Diterpene glycosides produced from the leaves of this plant are safe, non-nutritive as well as high-potency sweeteners. It may be used as a better alternative to sucrose and other artificial sweeteners, being 250-300 times as sweet as sucrose. Stevia produces sterile seeds and the availability of cuttings is seasonal and limited. The availability of quality planting material is emerging as a major constraint in the commercial exploitation of this magical sugar free sweetener crop. The present study was carried out to develop a cost effective in vitro technique for large scale multiplication to sustain the supply of quality planting material of stevia using shoot tip explants. Murashige and Skoog medium was used for the in vitro propagation of stevia through multiple shoot regeneration from the culture of shoot tip explants. The MS media was supplemented with different concentrations of naphthalene acetic acid, indole acetic acid, indole butyric acid, 6- benzyleadenine, kinetin, ascorbic acid and citric acid. Shoot tip explants varied in shoot induction response to different media supplementations used. Maximum shoot regeneration (97.0 and 95.0%) was observed on MS basal+BA 1.5 mg/ l+Kin 1.5 mg/l+IAA 0.5 mg/l with and without additives (AA 20 mg/l+CA 40 mg/l) using shoot tip explants, respectively. Maximum shoot proliferation (32.7 shoots /culture) was obtained on the medium MS+BA 2.0 mg/l+Kin 2.0 mg/l after 30 days of culturing. Maximum rooting (92%) was observed on half MS basal+IBA 0.5 mg/l. The rooted plantlets were acclimatized and transplanted under open field conditions.

Key words : Shoot tip, stevia, diversification, additives