Abstract: All the direct or indirect benefits that an ecosystem provides to human beings are called ecosystem services, which are the direct source of human well-being. Due to the multi-functionality of ecosystems and human choice preferences, the value of ecosystem services has different spatiotemporal scale effects and heterogeneity. Lishui City, Zhejiang Province, as a pioneering area for ecological civilization construction in China, is a model of practicing the principle that green mountains and clear waters are as valuable as mountains of gold and silver. Taking Lishui City as the research area, based on the land use data of four periods in 2005, 2010, 2015 and 2021, the spatio-temporal changes of ecosystem service value (ESV) in Lishui City were quantitatively evaluated by using the modified equivalent factor method and GIS spatial statistical analysis method. The research results reveal that: (1) During the period from 2005 to 2021, the ESV in Lishui City as a whole showed a trend of first increasing and then decreasing. The spatial distribution presented a distribution feature of low in the middle and high in the periphery, and the high-value areas of ESV in Lishui City gradually concentrated and the low-value areas gradually spread in the four periods. (2) The value-added hotspots are mainly distributed in the water areas and reservoir areas of Yunhe County, Longquan City and other places, while the loss hotspots are mainly concentrated in the concentrated areas of cultivated land and urban construction land in Songyang County, Liandu District and other places. By comparison, it is found that the ratio of ecosystem service value to GDP in this study is similar to the results of similar studies. This research can provide a reference basis for the spatial planning of Lishui City.