In this study about concentration and combined the application of plant growth regulators on the MS media for leaves culture. the alive and callus induction were determining factors affecting induction direct callus induction from leaves of mulberry. in the first experiment the best response of callus induction was observed from leaves culture on the MS medium supplement with 3 mg/l 2,4-D and 1 mg/l BA for 8 weeks found that expanded and created a lot of calluses, showed the most favorable response in terms of combination of plant growth regulators supplement on the MS media for leaves culture (table 3). All dead leaves after culture for 6 weeks Because media inappropriate, experimental results are inconsistent with the experiments of Liu et al, 2004 reported percentage of callus induction in medium with 25 and 50 μM ( 5 and 10 mg/l.) were also high than in medium without 2iP. The second experimental , no callus induction. And the leaves explant eventually dies and different result may be about prepared media error or different kinds of plants make the experimental results were different. Leaves explant high alive when treat in darkroom which it rate more than treat in light room. And has callus induction more than, so it may be said that dark promote alive and callus induction of leaves as final experiment, leaves explant culture on the MS medium supplement with 2 mg/l BA and 1 mg/l NAA treat in dark room. have 100 % alive, 100% callus induction.