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Biological nitrogen (N) fixation (BNF) via diazotrophs is an important ecological process for the conversion of atmospheric N to biologically available N. Although soil diazotrophs play a dominant role in BNF and arbuscular mycorrhizal fungi (AMF) serve as helpers to favor BNF, the response of soil BNF and diazotrophic communities to different long-term fertilizations and the role of AMF in diazotrophs-driven BNF are poorly understood. Herein, a 33-year fertilization experiment in a wheat-maize intercropping system was conducted to investigate the changes in soil BNF rates, diazotrophic and AMF communities, and their interactions after long-term representative fertilization (chemical fertilizer, cow manure, wheat straw, and green manure). We found a remarkable increase in soil BNF rates after more than three decades of fertilization compared with nonfertilized soil, and the green manure treatment rendered the highest enhancement. The functionality strengthening was mainly associated with the increase in the absolute abundance of diazotrophs and AMF and the relative abundance of the key ecological cluster of Module #0 (gained from the co-occurrence network of diazotrophic and AMF species) with dominant diazotrophs such as Skermanella and Azospirillum. Furthermore, although the positive correlations between diazotrophs and AMF were reduced under long-term organic fertilization regimes, green manuring could reverse the decline within Module #0, and this had a positive relationship with the BNF rate. This study suggests that long-term fertilization could promote N fixation and select specific groups of N fixers and their helpers in certain areas. Our work provides solid evidence that N fixation and certain groups of diazotrophic and AMF taxa and their interspecies relationship will be largely favored after the fertilized strategy of green manure.

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Bacterial community is a key factor affecting aerobic composting, and understanding bacterial community succession is important to revealing the mechanism of organic matter degradation. In this study, the succession and metabolic characteristics of bacterial communities were explored in 45 days composting of sheep manure and wheat straw by using high-throughput sequencing technology and bioinformatics tools, respectively. Results showed that the alpha diversity of bacterial community significantly decreased in the thermophilic (T2) phase and then recovered gradually in the bio-oxidative (T3) and the maturation (T4) phases. Bacterial communities varied at different stages, but there were 158 genera in common bacterial species. Unclassified_f_Bacillaceae, Oceanobacillus, Bacillus, Pseudogracilibacillus, and Nocardiopsis were identified as keystone bacterial genera. Eleven genera were significantly correlated (p < 0.05), or even extremely significantly correlated (p < 0.001), with the physicochemical factors. Redundancy analysis (RDA) showed that changes of bacterial community diversity correlated with physicochemical factors. The highest relative abundances were amino acid and carbohydrate metabolism among the metabolic groups in the compost. These results will provide theoretical support for further optimizing sheep manure composting conditions and improving the quality of organic fertilizers.