Recent reports indicate that manganese (Mn), applied as a foliar fertilizer in tank mixtures with glyphosate, has the potential to antagonize glyphosate efficacy and reduce weed control. It was hypothesized that Mn2+ complexed with glyphosate in a similar manner to Ca2+, forming salts that were not readily absorbed and, thereby, reducing glyphosate efficacy. This study was conducted to confirm the interaction of Mn2+ and glyphosate and to measure the effect of Mn on glyphosate absorption and translocation in velvetleaf. In aqueous solutions, Mn2+ binds with solvent molecules and with chelating agents to form hexacoordinate complexes. The distribution of paramagnetic species, both the free manganous ion ([Mn{H2O}6]2+) and the Mn2+–glyphosate complex, in Mn–glyphosate solutions at various pH values were analyzed using electron paramagnetic resonance (EPR) spectroscopy. Glyphosate interaction with Mn appeared to increase as the pH was increased from spray solution levels (2.8 to 4.5) to levels common in the plant symplast (7.5). Growth chamber bioassays were conducted to measure absorption and translocation of 14C-labeled glyphosate in solution with four Mn fertilizers: Mn-ethylaminoacetate (Mn-EAA), Mn-ethylenediaminetetraacetate (Mn-EDTA), Mn-lignin sulfonate (Mn-LS), and Mn-sulfate (MnSO4). Mn-EDTA did not interfere with glyphosate efficacy, absorption, or translocation. However, both MnSO4 and Mn-LS reduced glyphosate efficacy, absorption, and translocation. Mn-EAA severely antagonized glyphosate efficacy, and although glyphosate in tank mixtures with Mn-EAA was absorbed rapidly, little was translocated from the treated leaf. The Mn-EAA fertilizer contained approximately 0.5% iron (Fe) not reported on the fertilizer label. Iron is presumed to be partially responsible for the very limited translocation of glyphosate from the treated leaf in Mn-EAA tank mixtures. Adding ammonium sulfate increased the efficacy, absorption, and translocation of glyphosate for each Mn fertilizer tank mixture.