The aim of the present study was to investigate the effect of different concentrations of plant growth regulators on in vitro induction of callus and plantlets regeneration from anthers culture of  Datura metel L. The activity of catalase (CAT) as one of the important antioxidant enzymes was measured during different stages of callus and organogenesis under all combinations of hormones. Anthers were cultured under aseptic conditions on Murashige and Skoog (MS) medium supplemented with benzyladenine (BA) (0.0, 3.0, 5.0 and 7.0 mg/l) in combination with 2,4-D (2,4-dichlorophenoxy acetic acid) (0.0, 0.01 or 0.05 mg/l) for callus induction and organogenesis. Results showed that MS containing different combinations of BA and 2,4-D induced calli with percentage ranging between 20-100%. High concentrations of growth regulators enhanced the highest callus percentage and  promoted plantlets regeneration by indirect organogenesis  reached to higher numbers of regenerated plantlets (4.5 and 7.4) at 5.0 mg/l BA + 0.05mg/l 2,4-D and 7.0 mg/l BA + 0.05 mg/l 2,4-D, respectively. Results revealed that catalase  activity increased significantly in organogenic callus with regenerated plantlets (23.5 and 47.3 unit) at higher concentrations of BA and  2,4-D (5.0 mg/l BA + 0.05 mg/l 2,4-D and 7.0 mg/l BA + 0.05 mg/l 2,4-D, respectively).