Different long-term fertilization regimes affect soil protists and their top-down control on bacterial and fungal communities in Mollisols

Soil protists represent a vastly diverse component of soil microbial communities and significantly contribute to biogeochemical cycling. However, how different fertilization regimes impact the protistan communities and their top-down control on bacteria and fungi remain largely unknown. Here, using high-throughput sequencing, we investigated the differences in protist communities and their relationships with bacterial and fungal communities in Mollisols of Northeast China that were subjected to chemical and organic fertilization over 30 years. The results showed that manure addition rather than chemical fertilization significantly increased protistan alpha diversity and changed protistan community structure. Manure amendments markedly increased the relative abundances of protistan consumers (such as Cercozoa) and reduced the proportion of phototrophic protists (such as Chlorophyta). Soil pH was the most influential factor driving microbial communities, and protists were less sensitive to environmental disturbances than bacteria and fungi. Protistan communities exhibited more stronger relationships with bacterial communities than fungal communities, and Chlorococcum was the most important contributor in regulation of microbial taxa and functional genes. Furthermore, manure addition slightly simplified the microbial network, and chemical plus manure fertilization improved network stability with the highest robustness. Manure addition specifically mitigated the negative interactions between protists and bacteria while reinforced the positive interactions between protists and fungi. This study advanced our knowledge about the roles of protistan groups in regulating microbial communities and ecosystem functions associated with