Planta Med 2012; 78 - P_2
DOI: 10.1055/s-0032-1307510

In Vitro Propagation and Metabolic Profiling of Elite Stevia rebaudiana Bert. Plants

H Lata 1, S Chandra 1, YH Wang 1, RM Moraes 1, 2, MA ElSohly 1, 3, IA Khan 1, 2, 4
  • 1National Center for Natural Products Research, School of Pharmacy, The University of Mississippi, University, MS 38677, USA
  • 2Center of Water and Wetland Resources, The University of Mississippi Field Station, MS, Abbeville, USA
  • 3Department of Pharmaceutics, School of Pharmacy, The University of Mississippi, University, MS 38677, USA
  • 4Department of Pharmacognosy, School of Pharmacy, The University of Mississippi, University, MS 38677, USA

Stevia rebaudiana Bertoni (Asteraceae) is a sweet, perennial herb and an important medicinal plant. The leaves of Stevia contain a non-caloric natural sugar (diterpenoid steviol-glycosides, Fig.1), which is alternative to artificially produced sugar substitutes and 300 time sweeter than sugar (sucrose). Due to a very low seed germination percentage and a poor vegetative propagation, tissue culture is the most suitable and reliable process for the mass propagation of Stevia. The present study was carried out to standardize an efficient and improved protocol for the in vitro propagation of S. rebaudiana Bertoni. In vitro shoot multiplication from the nodal segments was best obtained on MS medium supplemented with 0.2mg/L TDZ, whereas rooting was found highest in half strength MS media without any growth regulators. These plants were successfully acclimatized and hardened in ex-vitro conditions. Furthermore, metabolic profiling using HPLC analysis was done to determine what chemical differences, if any, exist between mother plant and in vitro propagated and hardened plants. Chemical profile of the major constituents (rebaudioside A, stevioside and steviolbioside) were identified and quantified. Our results reveal no significant differences in the chemical profile and content, among the mother plant and in vitro propagated plants of Stevia rebaudiana (Fig.2).

Fig.1: (1) Rebaudioside A: MF- C44H70O23 and MW- 967.01, (2) Stevioside: MF- C38H60O18, MW- 804.87 and (3) Steviolbioside: MF- C32H50O13, MW- 642.734.

Fig.2: Metabolic profiling of mother plant and a randomly selected tissue culture raised plant of Stevia rebaudiana. SM: standard of rebaudioside A (peak 1), stevioside (peak 2) and steviolbioside (peak 3), MP: mother plant and P1: a randomly selected tissue culture raised plant derived from mother plant.

Acknowledgement: This research was partially funded by the USDA, Agricultural Research Service Specific Cooperative Agreement No. 58–6408–7-012.