Ficus genus is widely distributed in all the climatic stages and great diversity. The exploration of genetic diversity is a pre-requisite for genome organization in the wild species and the related domesticated ones. Inter-Simple Sequence Repeat (ISSR) markers were used to assess the identification of 23 important Ficus accessions and determination of the genetic relationships among these accessions. Out of 21 ISSR primers tested, five primers produced 116 detectable fragments, of which 106 (91.3%) were polymorphic across the accessions. Each of the five primers produced a fingerprint profile unique to each of the accessions studied, and thus could be solely used for their identification. Thirteen unique bands specific to nine species were detected. These may be converted into species-specific probes for identification purposes. Genetic relationships among these accessions were evaluated by generating a similarity matrix based on the Dice coefficient and the Unweighted Pair Group Method with Arithmetic Average (UPGMA) dendogram. The results showed a clear cut separation of the 23 Ficus accessions and were in broad agreement with the morphology. Both molecular and morphological markers will be useful for preservation of the Ficus germplasm.