Colchicum luteum L. is an economically important and endangered medicinal plant of the Kashmir Himalaya. The corm extract is used for the treatment of rheumatism, gout, Behcet’s syndrome, and Alzheimer disease. It is also used extensively in plant breeding programs for the doubling of chromosomes. The present study was carried out for two years (2017–2019) to study the genetic diversity of C. luteum, an economically important and endangered medicinal plant of Kashmir Himalaya. The mapping of genetic diversity of C. luteum was estimated using Mahalanobis D2 analysis in the Aharbal (Kulgam), Dhara (Theed), and Baera Baal Hills (Harwan) of Kashmir Valley. The results showed the presence of 5 clusters for 30 populations. There were 17 populations in cluster-1, 1 in cluster-2, 2 in cluster-3, 3 in cluster-4, and 7 in cluster-5. The majority of the population was a group in cluster-1 followed by cluster-5. The maximum intracluster distance (D2 values) was observed in cluster-5 (46.55588) followed by cluster-3 (41.61871), and the maximum inter-cluster distance (D2 values) was observed in cluster-3 (46.55588) followed by cluster-5 (41.61871). Our study revealed that plant species possessed sufficient genetic diversity among the populations. Cluster-5 showed superiority in plant−1 respect of the maximum mean plant height (28.46 cm), leaf area (47.0 cm2), number of seeds plant−1 (26.85), corm length (5.15 cm), corm width (3.17 cm), fresh weight of corm plant (6.87 g), and dry weight of corm plant (4.81 g) as compared to other clusters. Out of five clusters, cluster-5 is a promising one for better yield and yield attributing traits. The present study revealed that plant species possessed sufficient genetic diversity among the populations as 30 populations were arranged into 5 clusters. Therefore, cluster-5, consisting of seven populations from the undisturbed area of Harwan, and consequently the populations from the same cluster can be multiplied for initiating a conservation and breeding program and can serve as a tool for the scientific community to evolve better contemporary varieties of C. luteum with profitable characters such as more yield of corms, etc. This will assist farmers, particularly the marginal farmers, to alleviate their income.