Transport ability of Mx IRT1 and Mx DsIRT1 in Arabidopsis irt1 and yeast fet3fet4 mutants. (A) Diagrams of the Mx IRT1-eGFP and Mx DsIRT1-eGFP constructs; (B) Phenotype analysis of WT (wild type) and irt1 transformed lines; WT, irt1:vector, irt1:Mx IRT1, and irt1:Mx DsIRT1 lines were grown under +Fe (CK) and −Fe conditions; (C) The germination and early growth comparisons for irt1:vector, irt1:Mx IRT1, and irt1:Mx DsIRT1 lines. Seeds from each of the three lines were germinated and grown in 1/2 MS without iron (−Fe) for 5 days; (D) Immunoblot analysis showing the presence of At IRT1 in WT, but not in Arabidopsis
irt1 mutant plants. Proteins were harvested from plants after 3 days iron deficiency. The blots were probes with At IRT1 antibody; (E) Immunoblot analysis showing the presence of full length and truncated proteins in Mx IRT1-eGFP and Mx DsIRT1-eGFP transgenic lines, respectively, prepared by transforming Arabidopsis IRT1 mutant plants. The blots were probed using GFP antibody; (F) Complementation and Immunoblotting analysis of Mx IRT1 and Mx DsIRT1 in DEY1453 (left); Blots probed with At IRT1 antisera show the presence of full length Mx IRT1 and truncated Mx DsIRT1 in the transformed yeast strains; the iron content was measured using inductively coupled plasma mass spectrometry (ICP-MS) after yeast cells had been cultured for 4 h in YPD (yeast extract peptone dextrose) liquid supplemented with 100 μM Fe2+-EDTA (ethylene diamine tetraacetic acid) (right); and (G) Cadmium sensitivity and Immunoblotting experiments in DY1457 (left); Blots probed with At IRT1 antisera show the presence of full length Mx IRT1 and truncated Mx DsIRT1 in the transformed yeast strains; and the cadmium content (right) of 3 transformed DY1457 was measured using ICP-MS after 100 μM treatment for 4 h.