Tropane Alkaloids Production of Immobilized Cells of Egyptian Henbane (Hyoscyamus muticus L.)

Abstract

T HE 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 (CaCl 2 and Ca (NO 3 ) 2 .4H 2 O) 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.