GC Analysis of Invitro Developed Shoots of Stevia Rebaudiana Through Rapid Tissue Culture

Abstract

Stevia rebudiana, commonly known as stevia, is an economically important sweetening medicinal herb species from the family Asteraceae. Micropropagation of Stevia is needed for multiplication in large amount to meet the industrial demand along with supplementation for high content of medically important secondary metabolites in shortest possible timing.So this study is carried out to develop rapid micro propagation of Stevia rebudiana by using indirect organogenesis where leafs and nodal segment explants were inoculated on Eleven MS media for callus induction with different concentration of auxin and cytokines. MS media supplemented with IBA (0 to 3.5mg), IAA (0 to 3mg/l), Kinetin (0 to 3mg/l) and TDZ (0 to 3mg/l) in different concentration and combination were tested. MS medium with 1mg/l Kinetin and 3mg/l IAA showed 100% of callus induction from leaf explants in 17 days of culturing. Calli were cultured on regeneration media and 4-5 micro shoots were regenerated on MS medium supplemented with Kinetin (1mg/L) IBA (3mg/L) (1.5mg/l) BAP in 15-20 days of incubation. These shoots were further subculture to develop into plantlet. One year old shoot from the field,which is the most desired commercial state and two months old tissue cultured shoots which is in well developed form were analyzed for phytochemicals by GC analysis and we found that the two month old tissue cultured shoots has 56 compounds and 86 compounds detected in 1 year field grown shoots of Stevia rebaudiana. So we found that in the present study the optimized tissue culture conditions are producing quality rich material which could be used by pharma or herbal industry for developing any products.