Abstract: The landscape pattern in rural areas is constantly changing with urbanization and new rural development, resulting in corresponding changes in ecosystem services. Taking Lishui District of Nanjing as the research area, this study used landscape pattern index, InVEST model and geographical detector model to analyze the temporal and spatial changes of landscape pattern and ecosystem services in 2000, 2010 and 2018. The relationship between landscape pattern index and ecosystem services was identified. The following conclusions were drawn: (1) During the past 18 years, the aggregation degree of all land types in Lishui District were high. The fragmentation degree of cultivated land, forest land and construction land changed greatly, while that of other land types changed little. The overall landscape fragmentation was low and changed slightly. The patch Statistical dispersion and landscape spatial connectivity between different land use types changed inversely. (2) During the past 18 years, the level of water and soil conservation in Lishui District was stable, but the level of carbon sequestration, water production and habitat quality showed a downward trend. In terms of spatial distribution, the areas that demonstrated a high carbon sequestration, water production and habitat quality decreased, while those that had low values increased. (3) The mean fractional dimensionality index and the aggregation index were the main landscape pattern factors affecting ecosystem services in Lishui District. The correlation between the interaction of any two of the four landscape pattern indices and the three ecosystem services were higher than that of the single index effect. The combined effects of multiple factors of landscape pattern led to changes in ecosystem services.