Iron is an essential micronutrient for Citrus, playing an important role in photosynthesis and yield. The aim of this paper is to evaluate the tolerance to Fe deficiency of five Citrus rootstocks: sour orange (S), Carrizo citrange (C), Citrus macrophylla (M), Troyer citrange (T) and Volkamer lemon (V). Plants were grown for 5 weeks in nutrient solution that contained the following Fe concentrations (in µM): 0, 5, 10, 15 and 20. At the end of the experiment, biomass (dry weight), leaf area, total leaf chlorophyll (CHL) and the activity of root chelate activity (FCR) were registered. Additionally, the mineral composition of roots (R) and shoots (S) was evaluated. Principal Component Analysis was used to study the relationships between all parameters and, subsequently, the relations between rootstocks. In the first component, N-S, P-S, Ca-S, Cu-S, Zn-S, Mn-S and Zn-R and Mn-R concentrations were related to leaf CHL and FCR. Increases in leaf CHL, Mg-R, DW (shoots and roots) were inversely related to Cu-R, which was shown in the second component. The values obtained were consistent for V10, C15, and C20, but in contrast for S0 and S5. Overall, the Fe/Cu ratio was significantly related to CHL, which may be used to assist rootstock performance.