The aim of the present study was to determine the unique molecular markers among, three species of the genus Anthemis and the construction of phylogenetic tree using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) technique. Genetic diversity was analyzed among 15 populations of three species of Anthemis (Anthemis melampodina, Anthemis, pseudocotula and Anthemis, bornmuelleri), collected from different locations at Saudi Arabia by using RAPD primers. Pairwise genetic distance was calculated based on Nei and Li coefficient. Unweighted pair group method with an average (UPGMA) was used for construction of dendrogram, based on the similarity matrix data. Results showed wide variations among A. bornmuelleri and other two species. A wide close genetic relation was observed between A. melampodina and A. pseudocotula. RAPD-PCR technique was shown to be an accurate tool, in other to ascertain plant relationships among species of genus Anthemis.