wheat vacuolar iron transporter tavit2 transports fe and mn and is effective for biofortification increasing the intrinsic nutritional quality of crops known as biofortification is viewed as a sustainable approach to alleviate micronutrient deficiencies in particular iron deficiency anemia is a major global health issue but the iron content of staple crops such as wheat triticum aestivum is difficult to change because of genetic complexity and homeostasis mechanisms to identify target genes for the biofortification of wheat we functionally characterized homologs of the vacuolar iron transporter vit the wheat genome contains two vit paralogs tavit1 and tavit2 which have different expression patterns but are both low in the endosperm tavit2 but not tavit1 was able to rescue the growth of a yeast saccharomyces cerevisiae mutant defective in vacuolar iron transport tavit2 also complemented a manganese transporter mutant but not a vacuolar zinc transporter mutant by overexpressing tavit2 under the control of an endospermspecific promoter we achieved a greater than 2fold increase in iron in white flour fractions exceeding minimum legal fortification levels in countries such as the united kingdom the antinutrient phytate was not increased and the iron in the white flour fraction was bioavailable in vitro suggesting that food products made from the biofortified flour could contribute to improved iron nutrition the singlegene approach impacted minimally on plant growth and also was effective in barley hordeum vulgare our results show that by enhancing vacuolar iron transport in the endosperm this essential micronutrient accumulated in this tissue bypassing existing homeostatic mechanisms