Rice is a tropical plant, so cold temperature may be detrimental to its development, depending on the genotype and environmental conditions. Degree of lipid unsaturation has been related to cold tolerance due to its effect on membrane stability. So, the aim of this study was to characterize the fatty acid composition and its alterations due to cold temperature in rice genotypes of diversified origin. Forty-four rice genotypes at the V4 stage were submitted to two temperature conditions: 10°C and 28°C for two days and after this they had their leaves collected for lipid extraction and quantification. Control plants were allowed to regrow until presenting four leaves fully expanded and then were subjected to 10°C for ten days for cold tolerance evaluation. Plant survival was measured seven days after recovery at 28°C and the genotypes were grouped in three cold tolerance classes: tolerant, intermediate and sensitive. These classes differed for total saturated and unsaturated fatty acids only under the cold temperature treatment. Further analysis of the more abundant fatty acids: linoleic, linolenic and palmitic, showed that the two last ones differed between tolerant and sensitive genotypes. Linolenic acid increased after cold exposure in cold tolerant genotypes while palmitic acid decreased, and an opposite behavior was found in the cold sensitive genotypes. These evidences indicate that these fatty acids are potential molecular markers useful for breeding programs as well as for future basic studies on cold tolerance in rice.