Predicted increases in atmospheric carbon dioxide (CO2) concentrations could modify crop resistance to insect herbivores by altering plant quality. The short generation times of aphids may allow them to exploit such changes and colonise previously resistant plant genotypes. Lucerne (Medicago sativa) has undergone global selective breeding against aphids, including the pea aphid, Acyrthosiphon pisum. The purpose of this study was to characterise how ambient CO2 (aCO2) and elevated (eCO2) (400 and 600 µmol mol−1, respectively) affected plant physiological traits potentially linked to aphid resistance, focussing on foliar amino acid concentrations, across five M. sativa genotypes with varying resistance to A. pisum. These included susceptible (Hunter River), low (Trifecta), moderate (Aurora and Genesis) and high resistance (Sequel). Under eCO2, root nodulation doubled and essential amino acid concentrations increased by 86% in resistant Sequel, whereas essential amino acid concentrations decreased by 53% in Genesis. Moreover, concentrations of lysine, an amino acid whose deficiency has been linked previously to A. pisum resistance in M. sativa, increased by 127% in Sequel at eCO2. Compared with aCO2, aphid colonisation of Sequel plants rose from 22% to 78% and reproduction rates increased from 1.1 to 4.3 nymphs week−1 under eCO2 conditions. In contrast, Genesis became more resistant at eCO2 compared with plants at aCO2; aphid colonisation rates fell from 78% to 44% of plants and reproductive rates decreased from 4.9 to 1.7 nymphs week−1. In conclusion, predicted changes in atmospheric CO2 concentrations could either reduce (Sequel) or enhance (Genesis) resistance to aphids, which might be linked to quantitative and qualitative changes in foliar amino acids.