Media chemical composition effect on production of embryogenic calli and production of transformed plants from Hyoscyamus muticus L (Egyptian henbane) were assessed using gene gun. Hyoscyamus muticus L., (Egyptian Henbane) is medically and economically important plant as it contains widely used atropine alkaloids, scopolamine, hyoscyamine. Callus induction and regeneration ability from leaf of Hyoscyamus muticus L were examined. Effects of 7 media were evaluated on type II callus production and regeneration. T3 medium showed greater positive response in embryogenic calli formation frequency (84.95%), reflected on shoot formation frequency (7.65/explant) and produced a high root yields after transfer on the rooting medium. Transformed henbane plants were achieved by particle bombardment using plasmid pAB-6 harboring the gus and bar genes with transformation efficiency 13.4%. Total alkaloid contents were reached to 6.05% in transformed plants. It compared with untransformed plants which contained 2.95%.