Abstract: The unambiguous identification of plant varieties is important in many areas of plant biology research. Eremophila (E) is a genus of plant characterized by a great economic aspects and distribution throughout the dry and warm climates. Saudi Arabia was succeeded to cultivate seven species of Eremophila (E. bignoniiflora, E. divaricata, E. glabra, E. laanii, E. maculata, E. oppositifolia and E. pterocarpa). This study employed Amplified Fragment Length Polymorphism (AFLP) to investigate the genetic variation in the selected Eremophila species introduced to Saudi Arabia. Eremophila DNA was isolated, purified and restricted using EcoRI and MseI enzymes. Restricted DNA fragments were amplified using different sets of primer combination in order to illustrated differences between the amplified DNA fragments of each species. Our results verified 195 polymorphic DNA fragments including 14 unique markers between the seven cultivars. Phylogenetic tree was showed highest genetic interrelation (81%) between E. bignoniiflora and E. oppositifolia while the lowest interrelation (15%) was illustrated between E. glabra and E. pterocarpa in comparing to the other selected species. This study succeeded to verify unique DNA markers that can easily use to distinguish between the seven species of Eremophila genus cultivated in Makah Al-Mukaramah region and identified the interrelation between them.