Somatic embryogenesis and root regeneration in Hyoscyamus niger L. for the production of hyoscyamine

Embryogenic calli could be induced from the leaf, petiole and root pieces of Hyoscyamus niger using different induction medium. The embryogenic calli produced were different in term of colour and morphology. Small globular somatic embryos could only be produced from the leaf explants cultured on Murashige and Skoog (MS) medium supplemented with 6 mg/L picloram. The globular embryos increased in size and developed into torpedo, heart shape or bipolar embryos accompanied by root formation after cultured onto MS basal medium for one week then followed by two weeks of culture in the maturation medium (MS + 1.0 mg/L BA). More roots could be regenerated from the leaf-derived somatic embryos when they were transferred into liquid MS medium without any plant growth regulator. Our result indicated that the leaf-derived embryogenic callus induced on solid MS supplemented with 6.0 mg/L of picloram did not show the presence of hyoscyamine. However, the H. niger roots regenerated from the somatic embryos showed the presence of hyoscyamine. Although the somatic embryos failed to germinate to complete plantlets, the regenerated roots from the somatic embryos could be mass propagated to obtain hyoscyamine, the main tropane alkaloid produced in H. niger.