The black soil region of Northeast China is the largest commodity grain base, and the degradation of the black soil has become increasingly severe, seriously affecting the coordinated development of production-ecological functions. This paper used CASA, precipitation storage method, RUSLE and RWEQ to quantify four key ecosystem services in the black soil region of Northeast China: provision, water conservation, soil conservation, and windbreak and sand fixation, and analyzed their spatio-temporal evolution characteristics. The contribution rates of climate change and human activity factors to the provision service were determined using the residual analysis method. The trade-off and synergy characteristics of ecosystem services under different regions and different dominant driving factors were depicted at the grid and county scales. The results show that: (1) In the past 20 years, the provision, water conservation, and windbreak and sand fixation services in the black soil region of Northeast China slightly increased overall, with the number of counties accounting for 96.6%, 76.9% and 75.4%, with the provision service in the eastern Inner Mongolia and Heilongjiang, the water conservation service in Heilongjiang and Jilin, and the windbreak and sand fixation service in Liaoning showing obvious increasing trends. The soil conservation service slightly decreased overall, with the number of counties accounting for 60.7%, especially in Jilin with the most obviously decreasing trend. (2) The change of provision service in the black soil region of Northeast China was mainly driven by the joint promotion of climate change and human activities. Human activity was the dominant driving factor, with a total contribution rate of 62.65%, and the number of counties accounting for 76.6%. (3) The provision service in the black soil region of Northeast China mainly showed a synergistic relationship with water conservation (the number of counties accounting for 88.67%) and windbreak and sand fixation services (the number of counties accounting for 77.14%), while it mainly showed a trade-off relationship with soil conservation service, with the number of counties accounting for 68.25%. Human activity factors aggravated the trade-off effect among the services. This paper helps to clarify the main contradictions of ecosystem services in the black soil region of Northeast China, and to formulate scientific, reasonable, and the targeted policies for ecological protection and restoration, which has important significance for improving the ecological function of Northeast China and ensuring national food security.