Based on remote sensing image interpretation data of land use/cover change (LUCC) for six periods from 1995 to 2020, we used the equivalent factor method to assess the value of ecosystem services (ESV) in Jiangsu Province and investigated its spatial and temporal evolution characteristics on the grid scale in combination with spatial autocorrelation analysis. The results showed that total value of ecosystem services in Jiangsu Province is on the rise, with an increase of 3.15 billion USD compared with that in 1995. Water area has the largest proportion of ESV and hydrological regulation makes the greatest contribution among ecosystem services, reaching 32.99 billion USD in 2020. The value of ecosystem services measured by the coefficient of sensitivity index of various land use types is as follows: water area>cropland>forestland>grassland>unused Land. Ecosystem service value is positively correlated to spatial distribution, but the correlation is gradually weakening, and the clustering relationship is dominated by high–high and low–low aggregation. The study’s findings will serve as useful guidelines for creating policies in Jiangsu Province pertaining to land use planning and eco-environmental protection.