**5. Conclusions**

The present work led to the identification of high number of *VTL* genes from hexaploid wheat. Because of polyploidization, a very high number of genes from this sub-family was identified. The presence of high number of VTL been restricted to only chromosome 2, 4 and 6 of the wheat genomes. The expression of these gene under metal stress including changes in the presence of Fe and Zn concentrations and exposure to heavy metals reinforce the importance of this gene-family during metal homeostasis. Our work will help in better understanding of the Fe transporters significance in metal homeostasis so as to biofortify wheat.

**Supplementary Materials:** The following are available online at http://www.mdpi.com/2223-7747/9/2/229/s1, **Table S1**: List of 31 VIT family genes extracted from ensembl biomart using Pfam ID: PF01988. **Table S2**: Subcellular localization (WolfPsort). **Table S3**: Conserved motifs identified in VTL and VIT proteins using MEME suite. The color code, consensus sequence logo, E-value and the number of proteins in which each motif was found are listed in the table. **Table S4**: Metal-responsive cis-elements found in *VTL* and *VIT* gene promoter regions. **Table S5**: List of gene specific primers used for qRT-PCR for *TaVTL* genes. **Figure S1**: Phylogenetic tree for VIT family genes from *Arabidopsis*, *Brachypodium*, *Oryza sativa*, *Zea mays* and *Triticum aestivum.* Sequences were extracted using Pfam ID followed by alignment by Muscle and construction of NJ tree using MEGA software. **Figure S2**: Transmembrane domains in wheat VIT family proteins. Figure shows potential TM domains in *TaVIT* and *TaVTL* proteins, based on the alignment with *EgVIT1* protein, using MUSCLE. **Figure S3**: Heatmap depicting the expression of VIT family genes (VIT and VTL genes) in Control (FPKM\_Control) and Fe starved (FPKM\_Fe) wheat roots. FPKM values were extracted using Cu fflinks pipeline from SRA projectID SRP189420. Increasing intensity of blue colour shows increase in expression as shown by the colour bar above. **Figure S4**: E ffect of di fferent metals on the phenotype and growth of wheat seedlings. (A) Phenotype of wheat seedlings showing retarded growth of shoots and roots. (B) Impact of di fferent metals on the growth (in cm) of roots and shoots. **Figure S5**: Heatmap depicting the expression of VIT family genes (VIT and VTL genes) in Control (FPKM\_Control) and Fe starved (FPKM\_Fe) wheat roots. FPKM values were extracted using Cu fflinks pipeline from SRA project ID SRP189420. Increasing intensity of blue colour shows increase in expression as shown by the colour bar above.

**Author Contributions:** Conceptualization, S.S., A.K.P.; methodology, S.S., A.K., V.M. and A.K.P.; formal analysis, S.S., A.K.P., G.K. and H.R.; investigation, S.S., V.M.; writing—original draft preparation, A.K.P., A.K., J.K.; writing—review and editing, A.K.P., A.K., G.K., and H.R.; visualization, A.K.P.; funding acquisition, A.K.P. All authors have read and agreed to the published version of the manuscript.

**Funding:** This work was supported by the institutional NABI-CORE gran<sup>t</sup> to AKP.

**Acknowledgments:** All the authors thank Executive Director, NABI for facilities and support. Support from International Wheat Genome Sequencing Consortium for providing the high-quality wheat genome resources is highly appreciated. DBT-eLibrary Consortium (DeLCON) is acknowledged for providing timely support and access to e-resources for this work.

**Conflicts of Interest:** The authors declare no conflict of interest.
