Guizhou Province is a typical karst mountainous area where strong human activities are concentrated in the basin region; therefore, it is of great value to study the evolution of land use function to enhance sustainable usage of the basin area. Based on land use change, this paper uses the theory of Ecological-production-living function to construct a classification system of land use function using remote sensing image data of Guizhou Province in 1990, 2000, 2010, and 2016 generated through ArcGIS10.2 software to construct a land use function transfer matrix and basin functional hot spot analysis distribution map. The results show that:(1) The land use function in Guizhou basins has shifted from the single-function dominance of traditional agricultural production to multi-functional dominance of traditional agricultural production and rural living functions. (2) From 1990 to 2016, basin function increased 10 types, 8 of which are composite types, diversifying the function of the basin; (3)The basins ratio change curve for single function and composite functions is "X" type, and by 2016 the proportion of composite-function basins was greater than that of single-function basins; (4) The basin's land use function changed slowly between 1990 to 2000; after 2000, the basin changed rapidly; (5) Functional shifts were the strongest from traditional agricultural production, which mainly change to urban living, rural living, industrial production, associate compound, and natural ecological functions; (6) The spatial distribution of hot spots in various functional types of the basin are changing in a complicated manner; as a whole, the spatial distribution of each function has changed from agglomeration to dispersion.