This data record contains arbuscular mycorrhizal fungi (AMF) operational taxonomic unit (OTU) occurrences in native prairie plant roots collected from remnant prairies and reconstruction of tallgrass prairies in Minnesota and Iowa, from April-December of 2022. The research aims to improve prairie reconstruction methods using AMF to improve prairie plant performance, species diversity, and resistance to invasive cool-season grasses. The data in this release includes 5 data sets. Three describe the AMF: 1) SampleID_and_OTU_readcount_tier2_data.csv provides the number of reads for each OTU identified in each sample; 2) OTU_assigned_taxonomy_FunGuild.csv includes a collated list of all the taxonomic guilds of the identified OTUs; 3) AMF_phylotypes.csv summarizes the 21 distinct phylotypes represented in the data. Competition_experiment_SamplesIds_and_replicates.csv provides a link between the AMF OTUs and the experimental units from which they were extracted. Finally, Greenhousecountdata.csv is the complete growth and weight data for each plant in the experiment with its corresponding ID, which allows these data to be linked to the AMF data.