Effect of Different Growth Hormones and In-vitro Propagation of Stevia rebaudiana and Quantification of Secondary Metabolites

Abstract

Stevia rebaudiana being a natural sweetener, being utilized worldwide in food entities like green teas, syrups, ready to eat meals, etc. The traditional sugar we use in our daily lives can be substituted by Stevia as well. In this study, we have utilized MS media of varied concentrations and growth hormones, which have enhanced the effect of Stevia, with respect to the composition of growth hormones for callus induction with KN and IBA their concentration being 1 :3 (mg/l) and BAP and NAA with a concentration of 0.5 : 2 (mg/l) ; these conditions have been optimized for shoot regeneration and multiplicity of In-vitro plantlets. GC analysis was carried for both field grown leaves and in-vitro leaves which reported the presence Diterpene which is an alkaloid. Both sample types were detected with compounds containing Diterpene and Sesquiterpene properties. In one year old field grown shoots, 86 molecules are identified (Table 6), whereas in two month old tissue cultured shoots, 59 molecules were identified (Table 7) in which n alkenes, n-alkanes, fatty acids, terpenes, diterpene, monoterpenes sesquiterpenes were identified. HPLC was carried out for the estimation of steviol glycoside concentration in field grown tissue and in-vitro grown tissue. The concentration reported in field grown tissues was 0.5287% and in in-vitro grown tissues was 3.5932% for the first time. We can conclude that these optimized conditions conducted in the laboratory for a period of 1 year, have high significance for commercial purposes.