Clarifying the relationship between human activities and the provision of ecosystem services has received significant interest in recent years because of a growing need for sustainable socio-ecological system development. Using multi-source remote sensing data, we assessed the spatial and temporal distribution of the human footprint index and five ecosystem services under four human activity gradients from 2010 to 2020 in the Xiangjiang River Basin. The five ecosystem services include water supply, soil conservation, food production, habitat quality, and carbon sequestration. The relationship between human footprint and ecosystem services was analyzed from quantitative and spatial perspectives. The results showed that over the past 10 years, water supply and habitat quality decreased by 4.59% and 16.49%, respectively. The other three services increased, and the upstream area of the basin had a higher level of ecosystem services provision. The human footprint index increased by 28.83% over the 10 years and was characterized by point and patchy clustering in the middle and lower reaches. In terms of quantitative characteristics, the relationship between human footprint and ecosystem services was primarily negative. The ecosystem services were sensitive to the human footprint index within the 0−0.4 range. In terms of spatial characteristics, the relationship was dominated by trade-offs. The risky “high–low” trade-offs were mainly distributed in the middle and lower reaches. As the gradients of human activity increased, the maximum fluctuation in ESs was 43%, and the maximum fluctuation in human footprint was 28%, making their relationship more complex. Our results identified response thresholds of ecosystem services to human activities, providing a guide for ecological management and sustainable development of basins.
