Rubus crataegifolius is a shrub species and is primarily distributed throughout Asia and Europe. We collected 550 representative samples of the eleven populations in Korea to estimate leaf variation within species. The 35 morphological characteristics allowed us to resolve well-supported fixed characteristics and variable characteristics. Sixteen of twenty-three quantitative characteristics and five of twelve qualitative characteristics showed significant differences among populations. We argued that the population differentiation can accounted for the variation in phenetic characteristics such as spine in this species, but were less successful in accounting for variation in other traits. Within a polystatistic leaf structure, their morphological differences could be modulated by ecological pressure such as temperature, altitude, duration of sunshine, and solar radiation. The phenogram showed two distinct clades. One was a group in central Korea and the other was a group in the southern regions of Korea. If morphological characteristics in R. crataegifolius populations reflect their ecological niche, morphology should be regarded with caution as the main criterion for population studies in R. crataegifolius.