Nitrogen-limited conditions have been routinely prepared for efficient lipid production by oleaginous microorganisms. However, it is difficult to attain high cellular lipid contents with natural nitrogen-rich substrates. In the present study, we showed that lipid accumulation by Rhodosporidium toruloides Y4 was directly linked to the carbon to phosphorus (C/P) molar ratios of the culture media. Moreover, such lipid accumulation phenomena were effective regardless of the presence of high amounts of nitrogen sources. Thus, cellular lipid content and lipid yield were 62.2% and 0.205 g/g glucose, respectively, using a medium with a carbon to nitrogen (C/N) molar ratio of 6.1 and a C/P molar ratio of 9552. This work suggested that phosphorus limitation can be equally effective and efficient to mediate lipid accumulation, which in turn, provides opportunities to produce microbial lipid more economically using natural or waste materials with high nitrogen content. (C) 2010 Elsevier Ltd. All rights reserved.