Environmental conditions, specifically heat stress, are important factors in asparagus crop production. Arbuscular mycorrhizal fungi (AMF) have been shown to increase plant growth. Effects of heat stress on nutrient uptake have rarely been examined in intact plants, but the limited results indicate that heat stress will decrease uptake; no studies have examined heat stress effects on asparagus nutrient uptake. We examined the effects of AMF, Glomus intraradices, on the growth, nutrient uptake, heat stress responses, and antioxidative activity in asparagus (Asparagus officinalis L.). We grew AMF-inoculated or non–AMF-inoculated asparagus plants in sand culture at 20 to 25 °C for 14 weeks in a greenhouse and subsequently subjected to three temperature conditions (control = 20 °C/25 °C night/day, mild heating = 30 °C/35 °C night/day, and severe heating = 37 °C/42 °C night/day) in growth chambers. Morphological and physiological growth parameters were compared between AMF-inoculated and non–AMF-inoculated plants. The mycorrhizal symbiosis markedly enhanced biomass production and heat stress responses negatively in plants compared with that in the non–AMF-inoculated plants. Plants grown under non–AMF-inoculated treatment had severe rate of leaf browning (80% to 100%), whereas the mycorrhizal plants showed a minimum rate of leaf browning under heat stress conditions. The results indicated mycorrhizal-inoculated plants showed an increase activity of antioxidative enzymes, such as superoxide dismutase and ascorbate peroxidase. The 2,2-diphenyl-1picrylhydrazyl radical scavenging activity also showed a greater response in mycorrhizal plants than in the control plants under each temperature treatment. Application of AMF enhanced plant growth and mineral nutrients and alleviated heat stress damage through an increased antioxidative activity and the mycorrhizal symbiosis significantly enhanced heat stress tolerance of asparagus.