1st part of this study, the optimized protocol for induction of Egyptian henbane hairy roots is to use excised leaf explant had 2 months age and infected with A. rhizogenes strain LBA 9402/12 at intensity corresponding to OD600 = 0.6 by using the infection method normal dipping for 30 min. combined with acetosyringon (500 miM). Leaf explants were incubated on MS medium for 2 days under dark condition. Then, transferred to MS medium containing 500 mg/l of rifampicin and incubated for 16 days under 16/8 h (day/night). Leaf explants infected with LBA 9402/12 were cultured on MS and incubated on 25ºC at day/night conditions for 16 days produced significant higher response for hairy root and it has a longest growth curve in growth kinetic study. 2nd part, the highest callus fresh weight was obtained from H. muticus leaf explants cultured on MS containing 0.5 mg/l BAP combined with 1mg/l 2, 4-D and incubated for 4 weeks at 25ºC under day/night condition. 3rd part, the transformed hairyroot clone induced from the strain LBA 9402/12 showed the highest concentrations of hyoscyamine & scopolamine as compared to the concentration of the other plant parts or induced callus.
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