Implementation of the recently developed Cyprus National Register of Commercial Varieties mandates proper cataloguing of the material conserved in the ex situ Olive Collection at the Agricultural Research Institute of Cyprus. A total of 125 trees belonging to 32 accessions were analyzed using 11 morphological endocarp traits and 14 microsatellite [simple sequence repeat (SSR)] markers. The SSR variability allowed segregation of 16 genotypes among accessions, which were clustered into three main groups based on genetic similarity. Similarity indexes among groups ranged from 0.63 to 0.65 and within groups were ≥0.9. Lower morphological variability was detected among accessions, which conformed to three morphological profiles; moreover, the three morphological profiles corresponded to the three groups of genetic similarity obtained by SSR markers. The identification, based on the unique combination of SSR genotypes and endocarp morphologies, revealed the presence of three cultivars and 15 molecular variants that presented limited molecular differences but morphological profiles identical to their catalogued cultivars. Two cultivars, ‘Ladoelia’ and ‘Kato Drys’, demonstrated molecular variation. The current study underpins the efficient management of olive germplasm collections based on combined molecular and phenotypic characterization of their accessions. The complete cataloguing of olive germplasm ensures the correct distribution of identified and authenticated material across national or international institutions.