The Main aim of this study was determine the effect of different croton(Codiaeum variegatum) genotypes (Gold Dust and Petra) and growth hormones on callus induction, micro propagation and antibacterial activities. Murashige and skoog (MS) medium with four compositions of growth hormones for callus induction, murshM1 (2 mg/l 2, 4 Dichlorophenoxy acetic acid (2, 4-D), M2 (3 mg/l 2, 4-D), M3 (2mg/l 1-Naphthalen acetic acid (NAA), M4 (1mg/l 2, 4-D + 1 mg/l NAA) were used. For micro propagation, four different hormone balance were used, i.e., M1 (1 mg/l Benzyl adenine (BA) + 25 mg/l peptone), M2 (1mg/l BA + 50mg/l peptone), M3 (3mg/l BA + 25mg/l peptone), M4 (3mg/l BA + 50mg/l peptone).For rooting induction the combinations of growth regulators were used as follows, Mr1 (1mg/lIndolbuteric acid(IBA), Mr2 (2mg/l IBA), and Mr3 (1mg/l IBA+ 1mg/l NAA). Results showed that the highest genetic response for callus induction frequencies were seen for Petra genotype (91.25%) followed by Gold Dust. Medium M3 showed the highest response for callus induction frequencies (82.4%) followed by M1.Interaction between genotypes and media each alone were highly significant in callus fresh weight character. Regarding the micro propagation, Petra genotype gave highly genetic response followed by Gold Dust, also medium (M1) gave highly response for micro propagation followed by M2. In vitro roots were successfully induced by (1 or 2) mg/l of IBA which gave longest and few roots, while 1mg/l NAA gave shorter and more root number. Plants with roots were moved to the green house for acclimatize in pots contain sand /farmyard manure (8:2VN). Alkaloids extracted from callus for both genotypes were detected for antibacterial activities against Gram positive and negative bacteria. Alkaloids extracted from calli for media M1 and M2 were gave a highest antibacterial activity. Finally the present results gives scientific evidence on the ideal composition of the media to product the alkaloid extracts from C. variegatum (Gold Dust and Petra) as medicinal plants to be used as antibacterial agents against different pathogens.