Abstract:Phosphorus (P) deficiency is a limiting factor for agricultural production in the region covered with lime concretion black soil. Application of chemical P fertilizer is a common strategy to improve P availability. Numerous studies have demonstrated the essential role of P fertilization on soil fertility and crop yield, whereas relatively little is known about the effects of long-term P application on the belowground fungal community. The overall objective of this study was to investigate the effects of long-term P fertilization on fungal community diversity, composition, and intraspecific interactions in a lime concretion black soil, and to assess how the effects varied with application rate. Soils from a 21-year field experiment located in Mengcheng County, Anhui Province, were used, with treatments including three levels of P application rates, i.e. (1) P0:unamended control, (2) P1:chemical P fertilizer was applied at P2O5 45 kg/hm2, and (3) P2:chemical P fertilizer was applied at P2O5 90 kg/hm2. ITS1 gene pyrosequencing was employed to characterize the fungal community diversity and composition. The results showed that long-term P fertilization significantly improved soil fertility. Compared with the P0 treatment, the total P (TP) from the P2 and P1 treatments was 40.00% and 24.00% higher, and the available P (AP) was 384% and 144% higher, respectively. The Shannon index decreased with the P application rates, and was significantly negatively correlated with TP (r=-0.678, P=0.045) and AP (r=-0.677, P=0.045). Ascomycota was the dominant phylum in the studied soil, with its relative abundance being 65.71%-79.86%. Principal coordinate analysis (PCoA) revealed that the fungal community structure differed significantly among the different treatments. Redundancy analysis (RDA) further suggested that the fungal community structure was mainly determined by TP and dissolved organic carbon (DOC). Compared with the P0 treatment, the relative abundance of Ascomycota from P2 and P1 was 9.87% and 11.75% higher, and that of Sordariomycetes was 28.63% and 15.97% higher, respectively. Conversely, the relative abundance of Mortierellomycota from P2 and P1 was 42.63% and 43.81% lower, and that of Mortierellomycetes was 43.82% and 42.63% lower, respectively. Long-term P fertilization resulted in a significant enrichment of saprophytic fungi, namely, the genera of Plenodomus, Penicillium, and Arthrobotrys were enriched in the P1 treatment, and the genera of Cyphellophora, Zopfiella, Guehomyces, Mortierella, and Mucor were enriched in the P2 treatment. However, pathogenic (i.e., Simplicillium and Magnaporthiopsis) and symbiotic (i.e., Sagenomella and Nigrospora) fungi were depleted in the P1 and P2 treatments, relative to that of the P0 treatment. Co-occurrence network analysis revealed that long-term P fertilization increased network complexity, as reflected by the larger edge number and higher average degree in the P1 and P2 treatments than in the P0 treatment, which may enhance the network stability by reducing the spread of indirect effects in the treatments with P application. Moreover, the P1 and P2 treatments had a higher ratio of positive edges, indicating more cooperators in the P1 and P2 treatments, whereas more defectors occurred in the P0 treatment. Our results demonstrated that long-term P application can improve P availability, reduce fungal community diversity, alter fungal community structure, increase saprophytic taxa, eliminate pathogenic and symbiotic taxa, and enhance network complexity and stability in lime concretion black soil.