Doum palm (Hyphaene thebaica L.) has been reported as one of the most beneficial species worldwide. Its fruits are a good source of fibers, antioxidants, B-complex vitamins, essential minerals, monosaccharides, essential oil and flavonoids. Fruit extract shows anti-inflammatory, antioxidant, antimicrobial, anticancer and pharmacological potential. However, there is a lack of investigations dealing with its genetic diversity. Therefore, a collection of twelve mature female palm landraces grown in the botanical garden of Aswan, “Egypt” were used for genetic diversity assessment based on phenotypic and molecular levels. 18 vegetal and chemical traits were evaluated. Fifteen of them showed highly significant variation among the tested landraces. Euclidian-based-dendrogram discriminated the tested landraces based on their phenotypic similarities to three main clusters showing a high level of variability. Molecular analysis using inter simple sequence repeats (ISSR) vis-à-vis start codon targeted (SCoT) polymorphism markers displayed the relationships among the tested landraces molecularly. Both markers showed a reasonable degree of polymorphism, however SCoT was more effective than the ISSR by showing a higher percentage of polymorphism, polymorphism information content, resolving power and diversity index. Cluster analysis based on molecular data showed a clear relationship among the doum landraces. However, ISSR- and SCoT-based dendrograms were slightly different, and completely unlike that based on phenotypic data. Nevertheless, phenotypic and molecular evaluations might complement each other, and by data of both an excellent overview was obtained from principle coordinate analysis. The information herein is valuable and considered as a keystone for further proximate studies on doum genetic diversity to help in improvement and breeding approaches.