A pot experiment was conducted in an open greenhouse to explore the use of citrate-coated cobalt ferrite nanoparticles (CoFe2O4 NPs) as a source for Fe fortification of three wheat lines (Triticum aestivum L.). Two of the three wheat lines tested differ in their efficiency concerning Zn storage in their grains (efficient and inefficient), and one had inefficient P-absorption. The NPs were supplied by foliar or soil application of Fe at 330 mg L-1, and 46 or 68 mg kg-1 soil, respectively. A positive control (Fe-EDTA salt, a conventional iron fertilizer) and a negative control (no fertilization) were also included to compare the efficiency of NP fertilization. Soil fertilization with NPs improved the grain yield and Fe concentration in the grains compared with the foliar application of NPs and conventional Fe fertilizer. Application of soil NPs at 68 mg kg-1 increased the grain yield by 52% and 21% compared with the control and soil Fe-EDTA treatments, respectively. Likewise, grain Fe concentration increased by 96% and 72% compared with the control and soil Fe-EDTA treatments, respectively. The phytic acid concentration in grains and the phytic acid:Fe ratio decreased by 6% and 62%, respectively, due to the soil application of NPs (68 mg Fe per kg). The Fe grain concentration of lines inefficient for Zn storage and P-uptake in plants from soil fertilized with NPs (68 mg Fe per kg) was 1.37 and 0.26 fold above the target biofortification concentration (60 mg Fe per kg). Cobalt concentration in grains ranged from 9 to 16 mg kg-1. These concentrations were below the maximum allowable limit of Co in grains (50 mg kg-1) recommended by FAO and the WHO. Our results showed that Fe supplied as NPs may improve the nutritional quality of wheat grains, and the economic yield. However, there remains a long way to go to achieve effective and economic use of nanotechnology for the nutritional development of wheat.