The lipid contents and fatty acid profiles of Chlorella zofingiensis cultured in the dark with various carbon sources were investigated. Glucose was found to be the best carbon source for the growth and lipid production. When cultivated with 50gL(-1) glucose, C. zofingiensis accumulated lipids up to 52% of the dry biomass, with triacylglycerols (TAGs) accounting for 72.1% of the total lipids. Fatty acid profiles revealed that glucose contributed to the highest yield of total fatty acids (TFAs) and proportion of oleic acid (35.7% of TFAs), which corresponded to the strongest up-regulation of biotin carboxylase (BC) and stearoyl ACP desaturase (SAD) genes. In fed-batch cultivation based on glucose, the lipid yield and productivity of C. zofingiensis were further increased to 20.7gL(-1) and 1.38gd(-1)L(-1) respectively, representing 3.9-fold of those achieved in batch culture. We conclude that C. zofingiensis has great potential for biodiesel production.