DETERMINATION OF TROPANE ALKALOIDS IN HAIRY ROOT CULTURE OF HYOSCYAMUSMUTICUS USING DIFFERENT STRAINS OF AGROBACTERIUM RHIZOGENES

Hairy Root Culture is the new route for large scale secondary metabolite production because of their fast and plagiotropic growth, genetic and biochemicalstability. In this study, Hyoscyamusmuticus hairy root clones were established following infection with Agrobacterium rhizogenes strains A4, LBA 9402/12 and NRC 5149. The transformation frequency obtained with the LBA 9402/12 strain was found to be highest one (98.33%), then that of the A4 (76.00%), followed by NRC 5149 strain (31.33%). Subsequently, the culture condition for the hairy root were established, however, different hairy root lines showed differentbiomass at 4 weeks of culture under 16 hlight/8 h dark photoperiod in liquid MS medium. The LBA 9402/12 hairy roots showed the longest log phase and prolonged for three weeks at stationary phase in comparison with A4 and NRC 5149. Significant differences appeared between the tropanalkaloid concentrations of the different hairy root clones andall explants of the intact plant. The maximum hyoscyamineand scopolamine concentration was found with Transformed roots with LBA 9402/12 (5.913 and 1.530 mg/gd.w.t), respectively.