To study impact of climate and fertilization on soil microbial community, a transplant experiment of black soil (Phaeozem) was conducted at Hailun, Fengqiu and Yingtan Experiment Stations located separately in the temperate, warm temperate and midsubtropical zone in East China. Soil microbial communities were measured by Biolog ECOPlate^TM. In different climate conditions, balance application of N, P and K increased soil microbial metabolic activity （expressed as AWCD, average well color development）in black soilsIn the teaseling stage of maize, AWCD of black soil followed a sequence of Yingtan station>Hailun station>Fengqiu station with fertilization (NPK), and in a sequence of: Yingtan station>Fengqiu station>Hailun station without fertilization (CK). The soil bacterial metabolic activity increased with the monthly mean temperature without fertilization, while the interaction of rainfall and fertilization increased soil bacterial metabolic activity and affected the impact of temperature. Principal component analysis of soil microbial communities showed that the metabolic fingerprint of soil microbial communities varied slightly between black soils in Hailun station and Fengqiu station, but significantly from that in Yingtan station under midsubtropical monsoon climate. The effect of climate on variation of the carbon source of soil microbial communities was displayed in αKetobutyric Acid, Putrescine, D,L-α-Glycerol and L-Threonine (R＞0.9), while the effect of fertilization was in β-Methyl-D-Glucoside, Glucose-1- Phosphate and Pyruvic Acid Methyl Ester. To sum up, variation of the hydrothermal conditions in different climate zones and fertilization affects integrally metabolic activity and metabolic fingerprint of microbial communities in black soil.