Integration of rye (Secale cereale L.) cover crops into the corn (Zea mays L.) soybean [(Glycine max (L.) Merr.] rotation of the upper Midwest USA can provide many agronomic and agroecological benefits. Integration is made difficult by short growing seasons, but may be facilitated by management of key agroecological interactions such as those between rye and soil microbiota. Rye growth was measured and colonization by arbuscular-mycorrhizal fungi (AMF) was determined in greenhouse experiments using soils from seven different management systems from a long-term cropping-systems experiment in southwest Minnesota. Microbial effects on rye growth were not evident before vernalization, but at final harvest (4 weeks after vernalization) soil microbial populations reduced rye shoot and root growth, relative to a pasteurized control inoculum. At final harvest, shoot biomass in 2-year rotations was 17% greater than 4-year rotations, indicating that microbial populations selected for by 4-year rotations may be more deleterious or pathogenic than those selected for by 2-year rotations. Growth of three rye cultivars was examined in all inocula; cultivars differed in their mean response to soil microbiota and their ability to host AMF. These findings suggest that management factors affect interactions between rye and soil microbiota resulting in altered rye growth.