[Objective] The spatio-temporal variation pattern and change law of ecological service value (ESV) in the Lijiang River basin during 2000—2020 were determined in order to provide a spatial governance method for optimizing land use structure and achieving the “dual carbon” goal. [Methods] Based on remote sensing information mining and a geographic information analysis platform, ecological service values were evaluated by value accounting and its dynamic and sensitivity analysis methods. The spatio-temporal pattern of ESV was studied by spatial autocorrelation methods. The leading factors and drivers of ESV differentiation were diagnosed by geographic detection methods. [Results] ① The spatial heterogeneity of land use/cover in the Lijiang River basin was significant. The area of forest land and cultivated land was the largest, and change was stable. The area of construction land grew rapidly. ② The ecological service value for the Lijiang River basin was about 3.75×10⁷ yuan. The ESV of forest land and grassland was the highest, accounting for 52.44% of the total value. The overall change initially increased rapidly and then decreased slowly. The value of raw material production and soil and water conservation increased significantly, while the value of food supply and water conservation decreased significantly. ③ In general, ESV showed a pattern of low values in the midstream region and high values in the upstream and downstream regions, exhibiting a positive spatial agglomeration feature. [Conclusion] Urban expansion is the core driving force of the spatio-temporal differentiation of ESV and its dynamic changes in the Lijiang River basin. Disturbance of ESV by human activities is increasing over time.