Human deficiencies of iron (Fe) and zinc (Zn) are worldwide problems. Biofortification of wheat could reduce Fe and Zn deficiencies in societies that depend on wheat consumption. This study investigated the effects of foliar application of Fe with or without Zn on the concentrations of Fe and Zn in grain and especially in flour of three wheat cultivars. On average, grain Fe concentration was increased significantly from 29.5 mg kg(-1) in the control to 37.8, 35.9, or 34.9 mg kg(-1) by application of FeSO4, ferric citrate plus ZnSO4, or ferric citrate, respectively. As expected, grain Zn concentration was increased from 29.0 mg kg(-1) in the control to 45.7 or 39.6 mg kg(-1) by application of ferric citrate plus ZnSO4 or a complex of micronutrients. Although the Fe and Zn concentrations in flour were inherently lower than in bran and shorts made by experimental mill, the concentrations in flour were simultaneously increased from 10.4 to 12.4 mg kg(-1) for Fe and from 11.8 to 17.4 mg kg(-1) for Zn by application of ferric citrate plus ZnSO4. Importantly, Fe was peripherally localized within grain fractions and strictly limited to transport to endosperm, making it more difficult to increase the quantity of Fe in flour products by foliar Fe application, but the situation with Zn is promising because Zn is more readily transported to the endosperm than Fe. The current study increases the understanding of agronomic biofortification.