Soil sustainability is based on soil microbial communities’ abundance and composition. Straw returning (SR) and nitrogen (N) fertilization influence soil fertility, enzyme activities, and the soil microbial community and structure. However, it remains unclear due to heterogeneous composition and varying decomposition rates of added straw. Therefore, the current study aimed to determine the effect of SR and N fertilizer application on soil organic carbon (SOC), total nitrogen (TN), urease (S-UE) activity, sucrase (S-SC) activity, cellulose (S-CL) activity, and bacterial, fungal, and nematode community composition from March to December 2020 at Guangxi University, China. Treatments included two planting patterns, that is, SR and traditional planting (TP) and six N fertilizer with 0, 100, 150, 200, 250, and 300 kg N ha –1. Straw returning significantly increased soil fertility, enzymatic activities, community diversity, and composition of bacterial and fungal communities compared to TP. Nitrogen fertilizer application increased soil fertility and enzymes and decreased the richness of bacterial and fungal communities. In SR added plots, the dominated bacterial phyla were Proteobacteria, Acidobacterioia, Nitrospirae, Chloroflexi, and Actinobacteriota; whereas fungal phyla were Ascomycota and Mortierellomycota and nematode genera were Pratylenchus and Acrobeloides. Co-occurrence network and redundancy analysis (RDA) showed that TN, SOC, and S-SC were closely correlated with bacterial community composition. It was concluded that the continuous SR and N fertilizer improved soil fertility and improved soil bacterial, fungal, and nematode community composition.