Abstract: As the largest tributary of the Yellow River, the Weihe River occupies an important position in the governance and development of the Yellow River. The ecological footprint and ecological carrying capacity of the mainstream of Weihe River in Gansu Province from 2009 to 2019 were calculated by applying the improved three-dimensional ecological footprint method, the impact of social and economic activities in the basin on the ecological environment was studied, and the measures aimed at promoting the high-quality development of the river basin was explored. The results showed that from 2009 to 2019, the fossil energy land and cultivated land were the largest components of the ecological footprint in the basin of Weihe River in Gansu, which reached 36 498.64 and 12 869.88 hm² taking provincial hectare as the area equivalent, respectively. The ecological footprint and ecological carrying capacity showed a trend of expanding initially and then shrinking, the proportion of ecological natural resource supply in the regional consumption capacity increased from 13.92% to 15.16%, and the ecological deficit per capita constantly declined at a rate of 2.95% per year. The breadth and depth of ecological footprint increased notably, showing that the unreasonable consumption structure of natural resources has been alleviated, but the mitigation trend of the pressure on the ecological environment in the basin was not obvious. To alleviate the regional ecological pressures, measures such as increasing ecological land, readjusting industrial structure, and optimizing resource consumption structure should be taken in the future.