Helichrysum oocephalum is a medicinal plant of the genus Helichrysum that have limited distribution in Iran. Local geographical populations may differ in their genetic content and form different gene pools. Therefore, we carried out population genetic investigation and morphological studies in five geographical populations of Helichrysum oocephalum by using ISSR molecular markers. AMOVA produced the significant genetic differences. The mean Nm value revealed some degree of gene flow among Helichrysum oocephalum 8. Molecular and morphological analysis indicated that we have 2 groups in the studied populations. The present findings may be of use in the conservation of this medicinal plant in Iran.