THE AIM of the current investigation was to study biodiesel production byFusarium oxysporum AUMC 3224 cultured on two types of agricultural wastes , the first one called el-ghasheem and the second was wheat straw. In the case of el-ghasheem medium, the maximum lipid production was attained with 125 gl-1 ghasheem concentration and 0.1 gl-1 NaNO3 for 7 days, at pH 5.5 and temperature 28°C. Maximum lipid yield was 1.558±0.003 gl-1 lipid weight and 32.44±0.035% lipid percentage. In the case of wheat straw, the maximum lipid production was achieved with 5 g wheat straw, 50 ml of 2 gl-1 NaNO3, for 14 days, at 28°C. Maximum lipid yield was 0.45±0.005 g/L lipid weight and 28.81±0.305 % lipid percentage. The profile of extracted lipids from F.oxysporum was studied using Fourier transform infrared spectrometer (FTIR) indicating the presence of triglycerides and after transesterification of lipids showed the presence of fatty acid methyl esters. The fatty acids profiles were also determined by gas chromatography (GC) coupled with flame ionization detector and GC-mass spectroscopy. Data revealed the presence of significant amounts of palmitic, oleic, stearic, linoleic and other methyl esters.The results showed that lipids from F.oxysporum was a potential alternative resource for biodiesel production.