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Iron deficiency is a nutritional problem in plants and reduces crop productivity, quality and yield. With the goal of improving the iron (Fe) storage properties of plants, we have investigated the function of three Arabidopsis proteins with homology to  V acuolar  I ron  T ransporter1 (AtVIT1). Heterologous expression of   V acuolar Iron  T ransporter- L ike1  ( AtVTL1 ; At1g21140),  AtVTL2  (At1g76800) or  AtVTL5  (At3g25190) in the yeast vacuolar Fe transport mutant,  Δccc1 , restored growth in the presence of 4 mM Fe. Isolated vacuoles from yeast expressing either of the  VTL  genes in the  Δccc1  background had a three- to four-fold increase in Fe concentration compared to vacuoles isolated from the untransformed mutant. Transiently expressed GFP-tagged AtVTL1 was localized exclusively and AtVTL2 was localized primarily to the vacuolar membrane of onion epidermis cells. Seedling root growth of the Arabidopsis  nramp3/nramp4  and  vit1-1  mutants was decreased compared to the wild type when seedlings were grown under Fe deficiency. When expressed under the 35S promoter in the  nramp3/nramp4  or  vit1-1  backgrounds,  AtVTL1 ,  AtVTL2  or  AtVTL5  restored root growth in both mutants. The seed Fe concentration in the  nramp3/nramp4  mutant overexpressing  AtVTL1 ,  AtVTL2  or  AtVTL5  was between 50 and 60% higher than in non-transformed double mutants or wild-type plants. We conclude that the VTL proteins catalyze Fe transport into vacuoles and thus contribute to the regulation of Fe homeostasis  in planta .

Vacuolar-Iron-Transporter1-Like Proteins Mediate Iron Homeostasis in Arabidopsis