Urginea is a very important and rare medicinal plant, found growing in India, Africa and Mediterranean regions. Due to its medicinal properties the bulb of Urginea has found its place in British and European pharmacopeia. In the present study, genetic fingerprints of seven accessions of Urginea from different parts of Karnataka, South India were developed using PCR-based markers-ISSR. 3 ISSR primers were used to elucidate genetic diversity important for management and conservation. A total of 935 clear, distinct and reproducible bands were observed corresponding to an average of 311 bands per primer of which 899 bands were polymorphic in nature and percentage of polymorphism obtained was 96%. The binary matrix was used to calculate the distance matrix by Neighbour joining using free tree software. The matrix has a genetic distance of 0.52715 to 0.68282 with a mean of 0.5823. The smallest distance value of 0.52715 was observed between Ranganathittu and Bukkapatna indicating that these ecotypes are similar. The maximum distance value 0.68282 suggesting high divergence detected between Gulbarga and Channamallipura accessions. Dendrogram was constructed according to the data recorded. The accession B. R. Hills falls to cluster I and Ranganathittu, Magadi, C M Pura, Karighatta, Gulbarga and Bukkapatna falls to cluster II with three subclusters. High genetic diversity documented in the present study provides a baseline data for optimization of conservation of the Urginea species.