The aim of this study was to evaluate the genetic diversity among 36 wild strawberry tree (Arbutus unedo L.) genotypes belonging to different altitude and geographical origins, using 16 ISSR (Inter Simple Sequence Repeats) markers. Results revealed that ISSR primers produced a total of 344 bands, of which 94.1% were polymorphic, indicating a high level of genetic variation among the studied populations. The mean values of polymorphism information content (PIC), efective multiplex ratio (EMR), resolving power (Rp) and marker index (MI) were 0.34, 19.13, 23.46 and 6.45, respectively. The total gene diversity (Ht), gene diversity within populations (Hs) and total genetic differentiation coefficient among the populations (Gst) values were 0.33, 0.15 and 0.50, respectively. The genetic distance between all pairwise combinations of the genotypes ranged from 0.113 to 0.512. The cluster analysis revealed three main groups and which are subdivided into eight subgroups and a two independant branches. Therefore, the grouping of strawberry tree genotypes was independent of their geographical origins and altitude with exception for the genotypes belonging to Bin El-ouidane and Laanoucer populations. The results showed that ISSR markers are suitable tools for the evaluation of genetic diversity among strawberry tree genotypes. The data of this work will be useful for strawberry tree breeding programs and conservation strategies.