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Fig. S1 Element elution profiles of the standard reference material National Institute of Standards and Technology (NIST) 1573a (tomato leaf) from the anion exchange resin column used to separate and purify Cu for isotope analysis.

Fig. S2 Standard compound Cu K-edge X-ray absorption near-edge structure (XANES) spectra used for principal component analysis.

Fig. S3 Measured and fitted X-ray absorption near-edge structure (XANES) spectra of root samples of tomato and oat plants.

Fig. S4 Measured and fitted X-ray absorption near-edge structure (XANES) spectra of leaf samples of tomato and oat plants.

Fig. S5 Extended X-ray absorption fine structure (EXAFS) spectra and Fourier transform of oat roots grown in an Fe-sufficient nutrient solution.

Fig. S6 Extended X-ray absorption fine structure (EXAFS) spectra and Fourier transform of oat roots grown in an Fe-deficient nutrient solution.

Fig. S7 The δ65Cu value in the root and shoot of tomatoes as a function of the fraction of Cu translocated to the shoot.

Table S1 The GEOCHEM predicted species distribution in the three different nutrient solutions used for plant growth: Fe-N,N′-bis(2-hydroxybenzyl)ethylenediamine-N,N′-dipropionic acid (HBED) solution used for tomato plants, Fe-ethylenediamine-N,N′-bis(2-hydroxyphenylacetic acid) (EDDHA) solution used for oat plants, and a no Fe solution, used on both tomato and oat plants to induce Fe deficiency

Table S2 Copper recoveries following digestion and column purification of National Institute of Standards and Technology (NIST) 1573a standard reference material

Table S3 Summary of the anion exchange column purification procedure for Cu using AG-MP-1 resin

Table S4 Macro- and micronutrient concentrations (mg kg−1) for tomato and oat plant tissues grown in Fe-sufficient (+Fe) or Fe-deficient (−Fe) conditions

Table S5 Linear combination fitting of X-ray absorption near-edge structure (XANES) Cu K-edge spectra for tomato and oat plants grown in Fe-sufficient (+Fe) and Fe-deficient (−Fe) nutrient solutions

Table S6 Results for extended X-ray absorption fine structure (EXAFS) curve fitting of tomato and oat plant roots

Methods S1 A more detailed overview of the methods used to grow and digest plants, as additional information on the Cu purification procedure and isotope measurement conditions.

Methods S2 A description of how X-ray absorption spectroscopy (XAS) model compounds were prepared, analysed and interpreted.

Notes S1 A Rayleigh fractionation model is presented and used to explain in more detail the root to shoot copper fractionation observed and discussed in the main text.