Soybean has around 20% oil in total seed compound. Fatty acid concentration of soybean oil is about 12% palmitic acid, 4% stearic acid, 23% oleic acid (ω-9), 54% linoleic acid(ω-6) 54% and 8% linolenic acid(ω-3). To improve oxidative stability and quality of oil, the breeding programs mainly focused on reducing saturated fatty acids, increasing oleic acid and reducing linolenic acid in soybean oil. In plant oil, an essential fatty acid omega-3 fatty acid is in the form of α-linolenic acid (ALA) therefore, increasing ALA in soybean oil became one of the breeding goals for human health. In our research group, we have two breeding programs for concentration of ALA in soybean oil. Wild soybeans have almost twice ALA than that in cultivated soybeans. Introgression of alleles from wild soybean to cultivated soybean may lead to the increase of ALA in soybean seed oil for various applications. We developed several backcross populations by elite cultivars x wild soybean to select high ALA with good agronomic traits. In the case of low linolenic acid program, we developed an EMS (ethyl methane sulfonate) mutation population to select low ALA concentration line and found a mutant line with low ALA in seed oil. The scheme for developing high ALA concentration from wild soybean and molecular characterization for low ALA line will be discussed.