The genetic reduction of linolenic acid levels increases the quality and stability of soybean oil. The objective of this study was to determine the inheritance and evaluate the nature and magnitude of gene effects on soybean seed linolenic acid level. Means and variances of F1, F2, and F3 generations were made from the cross between accession BARC-12 (low linolenic acid content) and the commercial Brazilian cultivar CAC-1 (normal linolenic acid content). The results demonstrated that linolenic acid content in soybean is under the genetic control of a small number of genes. The additive model explained the means for the three generations and for the parents. Non-allelic gene interactions had little effect on the determination of genotypic values for the individuals. The generation means and population variation analyses demonstrated that the dominance deviations contribute little to the trait. These results showed that backcross breeding programs can be used to introduce the low linolenic acid content trait into soybean seeds, since it is possible to identify with very high accuracy the desired genotypes in segregating populations.