Long-term fertilization promotes soil organic nitrogen accumulation by increasing the abundance of keystone microbial cluster across aggregates

Nitrogen (N) in soil mainly occurs in organic forms, with soil organic nitrogen (SON) playing an essential role in supplying N to crops. However, it is unclear how the content and distribution of SON vary among different aggregate sizes, and how SON content is related to soil microorganisms. In this study, we investigated the associations between SON content and various microbial community characteristics across different aggregate sizes using data from a long-term field fertilization experiment in paddy soils. We also performed functional prediction to explore potential microbial mechanisms underlying SON accumulation. The results showed that SON content decreased as aggregate size decreased, with macroaggregates having the highest SON content and the silt-clay fraction having the lowest. Furthermore, our linear regression and structural equation modeling analysis revealed that keystone microbial clusters (highly interconnected taxa within Firmicutes, Proteobacteria, Chloroflexi, Actinobacteriota, and Ascomycota) were significantly correlated with SON content. Finally, functional prediction analysis suggested that this relationship may be due, at least in part, to the large number of metabolic pathways involved in biosynthesis of N-containing compounds present in keystone microbial clusters. Overall, our study highlights the importance of keystone microbial clusters in influencing the accumulation of SON and contributes to our understanding of soil N biogeochemical cycling.