Starch is one of the most important quality characteristics of potatoes, which is widely used in the food, medical, petrochemical and other industries. The demand for starchy potato varieties in the market is increasing year by year. To explore the characteristics and key genes of starch accumulation and regulation of potato tuber, in this study, transcriptome profiling of creeping stems (a), pre-tuber expansion (b), mid-tuber expansion (c), late tuber expansion (d), and mature stage (e) of high and low starch potato cultivars DXY and DS1 were conducted by RNA sequencing analysis, and a total of 9494 differentially expressed genes (DEGs) were identified at five stages. Binding function annotation revealed that the differential genes were mainly enriched in molecular functions such as binding and catalytic activity. Metabolic pathway analysis revealed that the differential genes were mainly enriched in carbohydrate-related metabolic pathways, and 137 DEGs were associated with starch and sucrose metabolism. Nine key genes regulating starch synthesis were examined, and the relative expression level of the sucrose synthase gene PGSC0003DMG400013547 was higher during growth period of DXY than that at any stage. The relative expression level of the fructokinase gene PGSC0003DMG400026916 was significantly higher in DXY than in DS1 during the a and b periods. The relative expression level of amylase gene PGSC0003DMG400009891, PGSC0003DMG400001549, and glucan endo-1,3-β-glucosidase gene PGSC0003DMG400024642, PGSC0003DMG400003181 in DS1 was significantly higher than that in DXY with in DXY with starch-rich cultivars during the c and e periods. The described gene might be a key regulatory gene for starch synthesis and accumulation. This study provides a clue for the investigation of the regulatory mechanism of tuber starch metabolism in different potato cultivars. [ABSTRACT FROM AUTHOR]