Abstract: In order to improve the soybean fatty acid content, improve the quality of soybean, through the use of sulfur, reveal the influence of different genotypes of soybean fatty acid content and search for of the best sulfur level of the 3 genotypes. 3 soybean varieties (‘Heinong48’, high-protein cultivar; ‘Heinong37’, middle cultivar; and ‘Heinong44’, high-oil cultivar) grown more in Heilongjiang Province were investigated. The soybean seeds were planted in pots. 4 levels of sulfur (S) treatment were designed, which were S1, S2, S3 and S4 (0, 0.02, 0.04, 0.06 g of sulfur per kilogram of soil). Used gas chromatography, the content of the mature soybean fatty acid was determined. It showed that: the fatty acid content of three genotypes of soybean seed were all increased. The same treatment for different varieties of NH44 content increased obviously, with the same variety S3 under different conditions of fatty acid content increased obviously. Under S3 condition, a small amount of myristic acid appeared in HN37 and HN48 grain; linoleic acid and stearic acid content of HN44 grain was much higher than other varieties; the content of oleic acid showed the highest in the HN48 grain; there was no significant difference between α-linolenic acid and palmitic acid content of HN44, HN48 and HN37 grain. Application of sulfur on 3 varieties of soybean fatty acid content had important influence, the suitable treatment of sulfur could improve soybean fatty acid content.