Callusing in Stevia rebaudiana (Natural Sweetener) for Steviol Glycoside Production

Abstract—Stevia rebaudiana Bertoni (natural sweetener) belongs to Asteraceae family and can be used as substitute of artificial sweeteners for diabetic patients. Conventionally, it is cultivated by seeds or stem cutting, but seed viability rate is poor. A protocol for callus induction and multiplication was developed to produce large no. of calli in short period. Surface sterilized nodal, leaf and root explants were cultured on Murashige and Skoog (MS) medium with different concentrations of plant hormone like, IBA, kinetin, NAA, 2,4-D, and NAA in combination with 2,4-D. 100% callusing was observed from leaf explants cultured on combination of NAA and 2,4-D after three weeks while with 2,4-D, only 10% callusing was observed. Calli obtained from leaf and root explants were shiny green while with nodal explants it was hard and brown. The present findings deal with induction of callusing in Stevia to achieve the rapid callus multiplication for study of steviol glycosides in callus culture.