Ettlia oleoabundans, a freshwater green microalga, was grown under different environmental conditions to study its growth, lipid yield and quality for a better understanding of the fundamental physiology of this oleaginous species. E. oleoabundans showed steady increase in biomass under low temperature and low light intensity, and at high temperature lipid cell content significantly increased independent of nitrate depletion. Studies on light quality showed that red light treatment did not change the biomass concentration, but stimulated lipid yield especially oleic acid, the most desirable biodiesel precursor. Moreover, no photoreversibility in lipid production was observed when applying alternating short-term red and far-red lights, which left the phytochrome effect still an open question. In addition, carbon dioxide enrichment via an air sparging system significantly boosted exponential growth and increased carbon conversion efficiency. Finally, a practical study demonstrated the feasibility of growing E. oleoabundans for high lipid production using a diluted agricultural anaerobic waste effluent as the medium. Together, these studies showed the potential of E. oleoabundans as a promising high yield feedstock for the production of high quality biodiesel.