Complex responses of soil prokaryotes, fungi and protists to prairie restoration on retired agricultural lands

Restoring native ecosystems on marginal croplands has many benefits but the impacts on belowground biodiversity are less clear, in part because the limiting factors regulating soil biota are complex and poorly described. Here, we studied how grassland prairie restoration of marginal croplands affected the diversity and composition of soil microbiota on 5 conventional farms from Ontario, Canada. Soil samples (0-15 cm) were collected from annually cultivated fields and adjacent planted perennial grassland where cultivation and chemical inputs had ceased several years previously. Following DNA extraction, we estimated bacterial and fungal abundance using quantitative PCR, and microbial diversity of prokaryotes, fungi and protists using amplicon high-throughput sequencing. Under both land uses, prokaryotic communities were dominated by Proteobacteria, Actinobacteria and Acidobacteria, fungal communities by Ascomycota, and protist communities by Rhizaria (TSAR), Evosea (Amoebozoa) and Chlorophyta (Archaeplastida). Prairie restoration did not have a consistent effect on soil microbial abundance, richness or evenness, which responses varied across farms. Microbial genetic and taxonomic community composition (i.e., sequence variant and genus level) were affected by land use, farm and the interaction between these two factors. Generally, prairie soils had higher relative abundance of Latescibacterota, Desulfobacterota, Acidobacteriota and Glomeromycota, and lower of Deinococcota, Chytridiomycota and Amoebozoa_X. In terms of differentially abundant fungal genera, prairies promoted more fungal plant symbionts, less saprotrophs and no plant pathogens. Interkingdom networks revealed changes in potential microbe-microbe associations with prairie restoration, with only 8 associations in common between land uses. The relationship between soil microbial diversity and physicochemical properties varied across microbial groups, diversity metrics and land uses. Our results evidence the complexity associated with restoring soils from agricultural land to natural ecosystems, with unspecified farm-specific factors (e.g., soil type, prairie species, management history) strongly modulating the response of different microbial groups and variables.