The development of biochemical from plant cells marks the beginning of a new phase in biotechnological research and increasing global demand for scopolamine. From this point, somatic calli of Egyptian henbane (Hyoscyamus muticus L.) were produced from leaf segments cultured on Murashige and Skoog (MS) medium containing various concentrations of Kin (0.5, 1, and 2 mg/l) or 2,4-D (1 and 2 mg/l). Immobilisation callus was conducted using various alginate treatments at 1.0, 1.5 and 2.0% combined with glycerol at 2, 3 and 4%. To complete gleization, two sources of calcium ions (CaCl2 and Ca (NO3)2.4H2O) were used at three concentrations 10, 20 and 30% or 1.5, 3 and 5%, respectively. The encapsulated somatic cell aggregates were cultured in liquid MS medium with 100, 200 and 300 mg/l of each tryptophan, glutamine and jasmine oil at 0.1, 0.2 and 0.3 % (v/v). The dried and fresh weights of immobilised cells were obtained after every two weeks of incubation for one month.
The highest callus production (100%) was obtained on a medium containing 0.5 mg/l of Kin. While the medium containing 0.5 mg/l Kin plus 1 mg/l 2,4-D produced the highest percentage of somatic embryos (91.67%) with 147 embryos/g, this count produced a regeneration percentage of 39.27% (equivalent to 57.67 embryos/g).
HPLC analysis revealed that the highest values of scopolamine and hyoscyamine (the major alkaloids) were obtained from applied tryptophan. When cells were treated with 100 mg/l of tryptophan, the medium contained 0.008 mg/ml of scopolamine and 0.09 mg/ml of hyoscyamine, respectively.