Strong chelating agents are reported to enhance Cu translocation in plants; however, the mechanisms responsible have not yet been fully established. In this study, both ethylenediaminetetraacetic acid (EDTA) and diethylenetriamine pentaacetic acid (DTPA) were found to increase Cu translocation to shoot tissue, while citric acid did not. Although all three amendments decreased Cu sorption to roots, which should cause greater Cu mobility within plants, this did not correspond with translocation. Energy-dispersive X-ray analysis of root cell walls showed that the endodermis presented a barrier (albeit partial) to the movement of free Cu ions, but this effect was negated by amendment addition. With EDTA, Cu levels in the stele were higher than those in the cortex after 1 week of exposure. Using Si deposition as an indicator, the presence of free Cu increased endodermal development, while amendments prevented this effect. Confocal microscopy and lipid peroxidation observations show that Cu and citric acid increased membrane damage, while EDTA and DTPA had transient effects. Strong chelating agents are less damaging alone than when present in conjunction with elevated Cu levels. Chelating amendments are proposed to enhance Cu phytoextraction by facilitating transport across the endodermis, ostensibly by influencing both membrane integrity and endodermal development.