The temperature of a wine fermentation strongly affects lipid metabolism and thus, aromatic profiles. Most of the metabolic studies are done in well-controlled laboratory conditions, yet wine is produced in less-reproducible industrial conditions. The aim of this study is to analyse the effect of fermentation temperature (13 degrees C and 25 degrees C) and culture media (synthetic media and grape must) on yeast lipid composition and volatile compounds in wine. Our results show that yeast viability was better at 13 degrees C than at 25 degrees C whichever growth medium is used, but that the complexity of the grape must enabled cells to reach higher viable population size. Viability was also related to the incorporation of linoleic acid and beta-sitosterol, which were present in the grape must. A lower temperature modified the cellular lipid composition of yeast, increasing the degree of unsaturation at the beginning of fermentation and decreasing the chain length as fermentation progressed. We also found that medium-chain fatty acids, mainly dodecanoic acid, were present in the cell phospholipids. Wines produced from grape must were more aromatic and had a lower volatile acidity content than those derived from a synthetic medium. Fermentations that were performed at the lower temperature also emphasized this feature.