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Those microbial responses to biochar addition were higher in 60-year-old soil relative to 20-year-old soil, leading to a higher enhancement of N retention and mitigation of N leaching. Soil pH was the prime factor that influenced soil microbes, and it strongly correlated with microbial biomass, enzyme activity, the relative abundance of dominant phyla and α-diversity indices. Therefore, the enhancement of microbial biomass, activity and shifts of bacterial community composition related to N cycling in response to biochar additions that incr