Optimization of root proliferation medium for Hyoscyamus niger L.

Untransformed root culture of Hyoscyamus niger could be induced from petiole explants on solid MS medium supplemented with 2.0 mg l(-1) IBA after four weeks of incubation under continuous light of 32.5 mu mol m(-2) s(-1). However, liquid MS medium supplemented with 0.5 mg l(-1) IBA was found to be better for root proliferation. Cultures at the third subculture cycle during root proliferation showed maximum root biomass production either under continuous light (3.69 +/- 1.00 g) or in total darkness (4.02 +/- 0.94 g). Step-wise optimization of culture medium components was carried out by manipulating the MS macronutrients, myo-inositol, Fe-EDTA and sucrose. The optimum root biomass producing medium was established as modified MS medium containing altered concentrations of macronutrients (15.0 mM NH4NO3, 33.1 mM KNO3, 3.19 mM CaCl center dot 2H(2)O, 1.9 mM MgSO4 center dot 7H(2)O and 1.4 mM KH2PO4), 9.6 mM myoinositol, 0.1 mM Fe-EDTA and 3% sucrose supplemented with 0.5 l(-1) mg IBA.