Cadmium (Cd) is a major environmental pollutant and one of the most toxic metals in the environment. Arbuscular mycorrhizal fungi (AMF) assisted phytoremediation can be used to remove Cd from polluted soils but the role of AMF, which mediate in Cd accumulation and tolerance, remains poorly understood. Here we inoculated Lolium perenne with two different AMF species (Glomus etunicatum and Glomus mosseae). Mycorrhizal L. perenne and non-mycorrhizal controls were exposed to Cd stress and we tested the effects of AMF mycorrhization on Cd uptake and subsequent tolerance, as well as the underlying mechanisms. Mycorrhizal infection increased root Cd2+ uptake and we observed that net Cd2+ influx was coupled with net Ca2+ influx. The inactivation of Ca2+ transporter channels decreased Cd2+ uptake in non-inoculated roots to a greater extent than in inoculated roots, indicating that AMF activates additional ion transport channels. In consequence, inoculated plants exhibited higher Cd accumulation in both roots and shoots than non-inoculated controls. However, AMF-inoculated plants showed higher chlorophyll concentrations, photosynthesis, and growth under Cd, indicating lower Cd toxicity in AMF-inoculated plants, despite the increase in Cd uptake. We observed that AMF-inoculated favored the isolation of Cd within cell wall and vacuole, and had higher concentrations of superoxide dismutase activity and glutathione concentration in roots than non-inoculated plants, consequently experienced less stress upon Cd exposure, relative to non-AMF controls. Our results highlight the potential and mechanism of AMF for enhancing phytoremediation of L. perenne in heavy metal contaminated environments.