In this study, in order to know the variability for a rootstock breeding program genetic diversity and relationships among 55 Iranian almond genotypes and seven related <i>Prunus</i> species were investigated. Morphological and molecular analyses were used. Principal component analysis showed that three components explained 67.6% of the total morphological variation for the first year and 68.06% for the second year of the study. Leaf traits were predominant in the first component and contributed most of the total variation. Leaf length and width, as well as, leaf area were highly correlated with each other and correlated to vigor. Also a negative correlation was found between leaf length/width ratio and vigor. Ward's method was used to construct cluster from morphological data which allocated individuals into their respective species. Out of 100 pre-screened RAPD primers, 16 with reproducible bands and maximum polymorphism were selected. Two-hundred and sixty bands were scored of which 250 of them were polymorphic. Average value of polymorphism per primer was 95.81% and maximum value for polymorphism (100%) was obtained from TIBMBA-14, TIBMBA-17, TIBMBB-05, TIBMBB-08, TIBMBD-09, and TIBMBD-10. On the other hand, the minimum value was obtained from TIBMBB-16 (86%). Primer TIBMBB-5 gave the maximum number of bands (25 fragments) and the minimum obtained from TIBMBE-18 (11 fragments). Genetic similarity based on Jaccard's coefficient ranged from 0.28 to 0.79 with an average of 0.53. Molecular analysis revealed a high degree of separation among samples regarding their geographical origin. Correlation between two approaches was low (R =−0.38). High molecular and morphological variability indicated that this collection includes rich and valuable plant materials for almond rootstock breeding.