ABSTRACT The present work was conducted in order to investigate the effect of explant type (leaf and stem internode) and auxin type (2,4-Dichlorophenoxyacetic acid, 2,4-D and Naphthalene Acetic Acid ,NAA) at different concentrations (0.00, 0.25, 0.50, 1.00 and 2.00 mg/l) as well as  different concentrations (0, 5, 10, 20 and 40 mg/l)  of L-phenylalanine(Phe) or tryptophan (Trp) on Verbesina encelioides callus cultures growth and production of secondary metabolites. Obtained results demonstrated that using leaf explant and supplementation MS medium with 0.5 mg/l NAA could enhanced and recorded the ultimate values of callus fresh weight, antioxidant activity (%), total flavonoids, total phenolic compounds contents and yields of Verbesina encelioides callus. Also, when the highest concentration (40 mg/l) of either L-phenylalanine or tryptophan was combined with 0.5 mg/l NAA, the maximum values of the abovementioned parameters were obtained. L-phenylalanine proved to be more effective than tryptophan in this regard.