The present investigation was carried out to find out the effect of L-Glutamine on in vitro callus induction using different explants from 3-4 months old plantlets of S. rebaudiana reported by (Vasudevan et al., 2004). Best response was obtained on MS medium supplemented with L- Glutamine at 15 mg/l in combination with 2, 4 –D (1.5mg/L) and BAP (0.5mg/L). Glutamine at this concentration induced 88 % callus with 37.77 and 104.24 mg callus weight on 14^th and 30^th day, respectively. This combination can only able to produce greenish, healthy nodular calli with more embryogenic potential and less necrotic lesion which further performed better in regeneration medium fortified with 2,4 –D (0.5mg/L), BAP (1.5mg/L) and L Glutamine (15 mg/L) produced large number of shoots.