RELATIVE PRODUCTION AND QUANTIFICATION OF STEVIOSIDE FROM In-vitro GENERATED SHOOTS, CALLUS, SUSPENSION CULTURE AND SYNSEEDS OF Stevia rebaudiana (Bertoni) Bertoni

The well known antidiabetic medicinal plant Stevia rebaudiana (Bertoni) Bertoni shows very poor seed germination in both in vitro and in vivo culture conditions. Various tissue culture techniques have been hypothesized to enhance its production as well as production of medicinally important secondary metabolite. The present endeavor is a collective attempt to analyse 4 techniques of tissue culture for production of stevioside. For this in vivo grown plants of S. rebaudiana were subjected to grow in the laboratory. Kinetin (0.1 mgL-1) with BAP (0.05 mgL-1) was found the best hormonal combination for direct shoot regeneration from nodal segment while leaves responded its best for production of callus at BAP (3 mgL-1) and NAA (4 mgL-1). Single cells of S. rebaudiana grow rapidly in the liquid MS medium fortified with BAP (3 mgL-1), NAA (4 mgL-1) and ascorbic acid (1mg L-1). Like this, synthetic seeds of S. rebaudiana were proliferated in the medium containing 0.1 mg L-1 Kinetin and 0.1 mg L-1 BAP. The stevioside quantification study was conducted on 15th and 30th day old samples, and found single cell suspension culture medium best to achieve the higher amount of stevioside while plantlets grown from synthetic seeds shown steady amount of stevioside by increasing the time.