EFFECT OF L-PHENYLALANINE ON THE PRODUCTION OF SOME ALKALOIDS AND STEROIDAL SAPONINS OF FENUGREEK COTYLEDONS DERIVED CALLUS

Abstract

The aim at this study is to increase some alkaloids and steroidal saponins in Trigonella foenum-graecum L. Callus initiation was carried out culturing cotyledons of the fenugreek on MS medium achieved by 1.0 mg/lˉ¹ 2, 4-D and 0.4 mglˉ¹ kinetin. The best medium to maintain callus was MS medium accomplished by 0.5 mg/lˉ¹ 2, 4-D and 0.5 mg/ lˉ¹ kinetin. Callus formation was allowed to take place for thirty days in the dark at 25±2 °C. Callus was analyzed using high performance liquid chromatography (HPLC). Methanol callus extract showed high concentrations of some alkaloids and steroidal saponins compared to the methanol extract of cotyledons. For increasing, the concentration of secondary metabolites, L-phenylalanine was added at concentrations 0, 100, 200 or 300 mg/lˉ¹. L-phenylalanine at 200 mg/lˉ¹ led to significant increase in diosgenin, choline and trigonelline reaching177.82, 415.12 and 1815.53 μg/ mlˉ¹ per 100 mg fresh weight of callus respectively, L-phenylalanine at 100 mg/lˉ¹ increased smilagenin, tigogenin and carpaine reached 259.52, 122.87 and756.72 μg/ mlˉ¹ per 100 mg respectively.