Bupleurum latissimum is a narrowly endemic and endangered plant, restricted to only two small populations on steep cliffs of a small island, Ulleung Island, in Korea. The genetic diversity and population differentiation in the two remnant populations of the species were investigated using RAPD (random amplified polymorphic DNA) analysis. The Neis gene diversities were 0.146 in the smaller population of 45 individuals, and 0.151 in the larger population of 61 individuals. The genetic variation was not significantly different between these two populations. Genetic diversity within populations was not low considering the very small size of populations. Analysis of molecular variance (AMOVA) revealed higher variation within populations (65.9%) than genetic differentiation between them (34.1%). B. latissimum revealed higher population differentiation than other outbreeding species. The differentiation of the populations corresponded to low gene flow (Nem = 0.482). The cluster and principal coordination analyses provide strong support for high population differentiation, showing that all individuals of the two populations have built up population-specific clusters. Although gene flow between the two populations of B. latissimum was limited, they have preserved relatively high levels of genetic variation.