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Wheat grain Zn concentration has important implications for Zn dietary intake in humans. Zn biofortification of wheat by Zn foliar application has been recognized as an efficient tool to enhance grain Zn level. In Serbia, wheat is a staple crop, and no attention is paid to wheat grain as a source of Zn. For the first time, we investigated the effect of Zn foliar application on grain Zn concentration and yield of high-yielding local bread wheat cultivars in Serbia as potential to improve Zn intake in local population. Field experiments were conducted over two growing seasons at two sites located in Vojvodina Province. Zn was applied by spraying of 0.5% water solution of ZnSO4at the end of flowering stage. Zn foliar application had no significant effect on grain yield. Significant increases of Zn grain concentration resulted from Zn treatment in all of five cultivars tested, ranging from 15 to 49%. No significant difference between the two sites was found in the mean increase in grain Zn concentration by Zn foliar application compared with the control. No correlation was found between grain yield and grain Zn concentration, implying that high grain yields common in intensive wheat production in Serbia do not limit Zn biofortification of grain by Zn foliar application. Zn biofortification of wheat grain by a single foliar spray at the end of the flowering stage can be recommended as an efficient tool to increase Zn intake in human population and prevent low dietary Zn intake.