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Tamarix is the largest genus in the family Tamaricaceae with about 54 species. T. tetragyna var. meyeri and T. tetragyna var. deserti were previously reported from Iran. Due to the presence of overlapping features in the previously designed identification keys for the separation of these two varieties, it could be helpful to use other characters and character states derived from other sources, such as anatomical and molecular studies, in addition to find more effective morphological features for the separation of the two varieties. Therefore, the present study aimed to differentiate these two presumed varieties by multiple approaches using morphological, anatomical and molecular data. For morphological study, 6 quantitative and qualitative characters were examined in 12 samples of the two varieties. ANOVA analysis showed a significant difference between the studied characters. For anatomical study, 10 traits were examined in 4 samples of the two varieties, which also showed a significant difference. AMOVA analysis based on molecular studies using Scot marker showed a significant difference between the two varieties. The PCA biplot show the most variable traits in morphological and anatomical studies. For varieties delimitation, different clustering methods were drawn in all three studies. The results of this study showed that the use of appropriate diagnostic traits in morphological and anatomical studies as well as the use of molecular markers can be effective in showing the separated boundaries of the two varieties studied.