Rice bran, the byproduct of brown rice, with a million tons produced annually, is the major storage organ for lipids. Increasing the α-linolenic acid (ALA) content in rice bran by biotechnology strategies is beneficial for both human health and the oil industries. We introduced two ω3/Δ15 fatty acid desaturase genes cloned from rice and soybean into rice under the control of an embryo-specific promoter, REG. The ALA content (dry weight) in embryos and bran was increased up to 6.09 and 5.86 mg/g, respectively, in transgenic lines, which was 25.4- and 27.9-fold higher than the 0.24 and 0.21 mg/g in the nontransformant control. ALA accounted for 46.7 and 44.3 % of the total fatty acids in embryos and bran, respectively, of transgenic plants, which was comparable to that in linseed and perilla seeds. The trait of high ALA content was stably inherited. The enhanced ALA content was preferentially located at the sn-2 position in triacylglycerols, which are digestible and absorbable for humans. ALA-enriched rice bran may help alleviate human health problems caused by ALA deficiency, with the production of healthy bran oil for humans and feed for animals.