An experiment with seven N, P, K-fertilizer treatments, i.e., control (no fertilizer), NP, NK, PK, NPK, NP2K, and NPK2 where P2 and K2 indicate double amounts of P and K fertilizers respectively, was conducted to examine the effect of long-term continuous application of chemical fertilizers on microbial biomass and functional diversity of a black soil (Udoll in the USDA Soil Taxonomy) in Northeast China. The soil microbial biomass C ranged between 94 and 145 mg kg-1, with the NK treatment showing a lower biomass; the functional diversity of soil microbial community ranged from 4.13 to 4.25, with an increasing tendency from control to double-fertilizer treatments, and to triple-fertilizer treatments. The soil microbial biomass, and the microbial functional diversity and evenness did not show any significant differences among the different fertilizer treatments including control, suggesting that the long-term application of chemical fertilization would not result in significant changes in the microbial characteristics of the black soil.