ABSTRACT Arbuscular mycorrhizal fungi (AMF) have been reported to have a wide distribution in terrestrial ecosystems and to play a vital role in ecosystem functioning and symbiosis with Stipa grasses. However, exactly how AMF communities in the rhizosphere change and are distributed along different Stipa population with substituted distribution and their relationships remain unclear. Here, the changes and distribution of the rhizosphere AMF communities and their associations between hosts and the dynamic differences in the glomalin-related soil protein (GRSP) in the rhizosphere soil of seven Stipa species with spatial substitution distribution characteristics in arid and semiarid grasslands were investigated. Along with the substituted distribution of the Stipa populations, the community structures, taxa, species numbers, and alpha diversity index values of AMF in the rhizosphere changed. Some AMF taxa appeared only in certain Stipa species, but there was no obvious AMF taxon turnover. When the Stipa baicalensis population was replaced by the Stipa gobica population, the GRSP tended to decline, whereas the carbon contribution of the GRSP tended to increase. Stipa grandis and Stipa krylovii had a great degree of network modularity of the rhizosphere AMF community and exhibited a simple and unstable network structure, while the networks of Stipa breviflora were complex, compact, and highly stable. Furthermore, with the succession of zonal populations, the plant species, vegetation coverage, and climate gradient facilitated the differentiation of AMF community structures and quantities in the rhizospheres of different Stipa species. These findings present novel insights into ecosystem functioning and dynamics correlated with changing environments. IMPORTANCE This study fills a gap in our understanding of the soil arbuscular mycorrhizal fungal community distribution, community composition changes, and diversity of Stipa species along different Stipa population substitution distributions and of their adaptive relationships; furthermore, the differences in the glomalin-related soil protein (GRSP) contents in the rhizospheres of different Stipa species and GRSP’s contribution to the grassland organic carbon pool were investigated. These findings provide a theoretical basis for the protection and utilization of regional biodiversity resources and sustainable ecosystem development.