Genetic variation at 10 allozyme loci was analyzed in 14 populations of Polygala reinii (Polygalaceae), a perennial herb endemic to central Honshu, Japan, with a fragmented geographical distribution. The levels of genetic variation within species ( P=80.0, A=3.10, H(E)=0.303) and within populations ( P=42.1, A=1.61, H(E)=0.163) were considerably higher than the mean for other endemic plants or short-lived perennial herbs. Genetic differentiation among populations was also high ( G(ST)=0.404). The genetic distance phenogram tended to show a clustering of the populations reflecting the fragmentation of the species range. A principal component analysis revealed the same tendency, as well as three groupings of populations in the Tokai district, on the Kii Peninsula and in the northern Kinki district. A negative correlation was obtained between the levels of gene flow and geographical distance among the populations ( r=-0.745, P<0.0001). These results indicated limited gene flow among populations in P. reinii, presumably due to the geographical isolation accompanying the fragmented distribution. On the other hand, the geographical differentiation between the Japan Sea and Pacific Ocean sides was found in P. reinii, suggesting the influence of postglacial migration on the establishment of the genetic structure of this species.