In order to compare the uptake, utilization and distribution of iron (Fe) as nano and bulk Fe complex (Fe(III)-EDTA), an experiment was conducted using hydroponically-grown Fe-deficient tobacco (Nicotiana rustica L.) plants. Plants were treated with Fe either through roots (root application, RA) or leaves (foliar application, FA).Leaf chlorophyll concentration and plants biomass responded to the Fe re-supply; this response was quicker for the nano Fe complex than the bulk Fe complex, in particular, in the RA plants. Plants re-supplied with the nano Fe complex had lower Fe content but higher Fe use efficiency than plants re-supplied with the bulk Fe complex.Analysis of different plant fractions at two subsequent weeks revealed that the nano Fe complex had higher mobility than the bulk Fe complex, both in the xylem and the phloem. Comparison of RA and FA revealed that the re-supply of Fe through roots was more efficient in retrieving the whole plant growth than the foliar Fe spray.RA, in contrast to FA plants, profited from an enhanced Fe uptake capacity induced under Fe starvation in the roots. Alternatively, FA was effective in extending the green leaf area duration. Our data suggested that nano Fe complex is advantageous as both leaf spray and a long-term feeding of plant through roots.