N-induced root exudates mediate the rhizosphere fungal assembly and affect species coexistence

Root exudates play essential roles in shaping root-associated microbial communities in plant-soil systems. However, knowledge regarding the influence of root exudates on soil communities, particularly concerning their assembly processes and species coexistence patterns, remains limited. In this study, we performed a 20-month pot experiment using a nitrogen (N) addition gradient (0, 2.5, 5, 7.5, 10, and 15 g N m^-2 yr^-1), amplicon sequencing, and metabolomics to investigate the effect of short-term N addition on the assembly process and species coexistence of fungal communities, as well as their association with root exudates in the rhizosphere and bulk soils around Bothriochloa ischaemum. The results demonstrated that short-term N addition led to distinct differences in the diversity, composition, assembly process, and co-occurrence networks of fungal communities in the rhizosphere and bulk soils. The diversity of fungal communities in the rhizosphere soil increased with the rate of N input and peaked at N10 treatment; this could be correlated with the increased abundance in long-chain organic acids (LCOAs). However, above the threshold N rate of 10 g N m^-2 yr^-1, diversity decreased probably because of the high N-induced inhibitory effect on root exudates (i.e., LCOAs). N addition increased the relative abundance of Sordariomycetes in the rhizosphere and decreased the relative abundance of Mortierellomycetes in the bulk soil, while enhancing the abundance of pathotrophs in both bulk and rhizosphere soils. The rhizosphere fungal community was dominated by a stochastic process at a low N input (N0 and N2.5) and by deterministic processes at a high N input (N10 and N15), which is opposite to the trends in the bulk soil. These fungal assembly processes determine the coexistence of fungal species; deterministic processes lead to less interconnected networks in rhizosphere soils that harbor a more complex network than the bulk soil. Associations between the assembly process and species coexistence in the rhizosphere of B. ischaemum were closely related to the changes in root exudates, such as amino acids, short-chain organic acids, and phenols, which were stimulated by N addition. Collectively, our study emphasizes the key roles of root exudates in the establishment of fungal communities in the plant-soil system and furthers our understanding of plant-microbe interactions.