Many plant species in subalpine regions are under threat of extinction as a result of climate change. In this study, the genetic diversity and geographic differentiation of three regions and six populations of Primula farinosa subsp. modesta (Bisset & Moore) Pax in Korea were assessed using the ISSR (Inter Simple Sequence Repeat) marker. The average genetic diversity (P = 60.62, SI = 0.299, h = 0.190) was relatively lower than that of other long-lived perennials, even though it is a self-incompatible species. AMOVA analysis showed that 50% of the total genetic diversity was partitioned among regions and Bayesian cluster analysis showed some remarkable geographic trends that were structured into 2 or 3 regions, suggesting limited gene flow among regions. Considering the population fragmentation, low level genetic diversity, and high genetic differentiation, it is essential to establish in situ and ex situ conservation strategies for P. farinosa subsp. modesta.