The goal of biofortification is enhancing and developing the nutrients in the edible parts of the plants. In order to investigate the effect of Iron, Zinc and mycorrhiza on barley yield and yield components, a field study was conducted during 2013-14 years in agricultural experimental station of Islamic Azad University Arak branch. A factorial experiment was arranged in randomized complete block desing with three replication. Three Levels of Zn, Z1 (no application), Z2 (25 kg/ha-1) and Z3 (50 kg/ha-1) were used. Mycorrhiza fungi including no inoculation and Glomus intraradices and three levels of Iron (no application, 2/5 kg/ha-1 and 5 kg/ha-1) were used. Simple effect of Mycorrhiza on yield showed highest grain yield with (3.90 ton/ha-1) was obtained from Glomus intraradices. Highest levels of straw yield with 3.43 were obtained in treatment Glomus intraradices. The simple effect of mycorrhiza, Iron and Zinc showed the highest biologic yield with 6.91, 6.70, 6.49 was obtained in treatment Glomus intraradices, 5 kg/ha Iron and 50 kg/ha Zn. The highest P content with 1.43 ppm was obtained in treatment Mycorrhiza fungi. Simple effect of mycorrhiza, Zn and Fe on Zn content showed, the highest amount of this traits with 41.32, 39.46 and 40.90 conducted in Glomus intraradices, 2.5 kg/ha-1 iron and 50 kg/ha-1 Zn application alternatively.