To reveal the genetic diversity and genetic relationships of China’s Bergenia germplasm, 28 Bergenia accessions from different regions in China were analyzed by 24 intersimple sequence repeat (ISSR) markers. The results showed that 318 sites were amplified in all germplasm, including 307 polymorphic sites, and the percentage of polymorphic sites was 96.54%. Cluster analysis showed that the 28 accessions were divided into three categories, with a similarity coefficient of 0.5475. Bergenia purpurascens was clustered into one category; B. scopulosa was clustered into one category; and B. tianquaninsis, B. emeiensis, B. stracheyi, and B. crassifolia were clustered into one category. The results of the cluster analysis indicated that the 28 accessions were not completely classified by origin. Using the ISSR marker technique to analyze the phylogenetic relationship of Bergenia germplasm is helpful for identifying valuable resources and providing a theoretical basis for the selection of breeding parents.