In karst regions (KRs), unique surface morphology and irrational human exploitation have led to increasingly prominent issues such as land fragmentation and rocky desertification. Understanding the spatiotemporal evolution of cultivated land (CL) in these areas is of great significance for supporting regional socioeconomic development, food security, and ecological sustainability. This study focuses on Guilin, combining GIS spatial analysis with methods including kernel density analysis, dynamic degree, spatial transfer matrix, and a Geodetector to examine the spatiotemporal distribution characteristics, evolution trends, and driving factors of land use based on five-phase of land use data from 2000 to 2020. The results show that: (1) over the past two decades, land use in Guilin has been dominated by CL and forest land, with CL exhibiting a spatial pattern of more in the east and south, and less in the west and north; (2) the CL transfer-out rate exceeded the transfer-in rate, mainly shifting to construction land and forest land; (3) the overall density of CL showed a declining trend, with a relatively stable spatial pattern; and (4) driving factor analysis indicates that the spatiotemporal changes in CL are jointly influenced by multiple factors, with natural factors exerting a stronger influence than socio-economic factors. Among them, the interaction between elevation and temperature had the greatest impact and served as the dominant factor. Although GDP and population were not dominant individually, their explanatory power and sensitivity increased significantly when interacting with other factors, making them key sensitive factors. The results can provide a scientific reference for the protection and rational utilization of CL resources in KR.