Biochar application has the potential for mitigating N2O emissions from agricultural soils and has been suggested as a management practice to ameliorate soil fertility and increase crop productivity. Nevertheless, the influence of biochar addition on N2O emissions from soils with different pH levels is not yet clear, which results in a poor understanding of the mechanisms regarding biochar application to soil N2O mitigation. A 40-day incubation experiment was carried out in the present study to investigate the impact of biochar on N2O emissions from soils with different natural pH. Four treatments (control, nitrogen fertilizer application, biochar amendment, and N plus biochar amendment) were set up separately in soils with three different natural pH levels (acidic vegetable soil, neutral rice soil, and alkaline soil). Our results showed that adding biochar significantly decreased N2O emissions by 20.8% and 47.6% in acidic vegetable soil for both N and no N addition treatments, respectively. For neutral and alkaline soils, the reduction of N2O emissions by biochar amendment was only significant for N addition treatments in alkaline soil. Soil pH and NO3–N concentration were significantly affected by biochar amendment (soil pH increased by 1.43-1.56, 0.57-0.70, and 0.29-0.37 units for acidic vegetable soil, neutral rice soil, and alkaline soil, respectively). Thus, biochar amendment could be used as an effective management practice for mitigating N2O emissions from acidic and alkaline soils.