Based on satellite remote sensing image, GIS and Fragstats, this study modeled and calculated the dynamic changes of land use, land cover and landscape patterns in Guizhou Province, China, and calculated the changes of ecosystem service values (ESVs). The impacts of the evolution of landscape patterns on the ESVs were analyzed, and reasonable policy recommendations were made. The findings are as follows: (1) In the past two decades, the area of cropland and grassland has decreased; the area of water bodies, urban and rural, industrial and mining, and residential areas has increased; the area of forestland has increased first and then decreased. (2) The two major types of landscapes, cropland and grassland, are clearly being replaced by two land types, forest land and water bodies. (3) Overall, the degree of landscape aggregation and adjacency has decreased, and the landscape heterogeneity has increased. (4) The total amount of ESV in 2000, 2008, 2013 and 2017 was 2574 × 10 Yuan RMB, 2605 × 10 Yuan RMB, 2618 × 10 Yuan RMB and 2612 × 10 Yuan RMB, respectively. The changes of landscape patterns had important impacts on the ESVs. In order to solve the problems caused by the increasingly prominent changes in the landscape patterns and improve the ESVs, it is necessary to rationally plan and allocate land resources, optimize the industrial structures, and develop effective regulatory policies.