It has been noted previously that nutrient-stressed plants generally release more soluble carbohydrate in root exudates and consequently support more mycorrhizae than plants supplied with ample nutrients. Fertilization may select strains of vesicular-arbuscular mycorrhizal (VAM) fungi that are inferior mutualists if the same characteristics that make a VAM fungus successful in roots with a lowered carbohydrate content also reduce the benefits that the fungus provides a host plant. This two-phase study experimentally tests the hypothesis that fertilizing low-nutrient soil selects VAM fungi that are inferior mutualists. The first phase examines the effects of chemical fertilizers on the species composition of VAM fungal communities in long-term field plots. The second phase measures the effects of VAM fungal assemblages from fertilized and unfertilized plots on big bluestem grass grown in a greenhouse. The field results indicate that 8 yr of fertilization altered the species composition of VAM fungal communities. Relative abundance of Gigaspora gigantea, Gigaspora margarita, Scutellispora calospora, and Glomus occultum decreased while Glomus intraradix increased in response to fertilization. Results from the greenhouse experiment show that big bluestem colonized with VAM fungi from fertilized soil were smaller after 1 mo and produced fewer inflorescences at 3 mo than big bluestem colonized with VAM fungi from unfertilized soil. Fungal structures within big bluestem roots suggest that VAM fungi from fertilized soil exerted a higher net carbon cost on their host than VAM fungi from unfertilized soil. VAM fungi from fertilized soil produced fewer hyphae and arbuscules (and consequently provided their host with less inorganic nutrients from the soil) and produced as many vesicles (and thus provisioned their own storage structures at the same level) as fungi from unfertilized soil. These results support the hypothesis that fertilization selects VAM fungi that are inferior mutualists.