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Fertilization affects soil nitrogen cycling and nitrous oxide (N2O) emissions, which are mainly driven by microbes. A 32-year field experiment was conducted to investigate the effects of chemical fertilizers and their combination with organic materials on the abundance of denitrifying functional genes (nirS, nirK, nosZ I and nosZ II) in Ultisol. The treatments comprised no fertilizer (CK), chemical fertilizer, chemical fertilizer+peanut straw, chemical fertilizer+rice straw, chemical fertilizer+radish and chemical fertilizer+pig manure.