A study was conducted in the Department of Floriculture, Medicinal and Aromatic Plants, Uttar Banga Krishi Viswavidyalaya, Cooch Behar, West Bengal, India in the year 2022 and 2023 to evaluate thirty gladiolus genotypes and classifying them into clusters as a pre-breeding exercise to select genotypes with maximum dissimilarity. The genotypes included both Indian and exotic varieties which were collected from different parts of India. Considering sixteen qualitative traits, as described in DUS testing guidelines, Shannon’s and Simpson’s diversity analyses were performed to indicate the characters that are primarily responsible for the dissimilarity between the genotypes and also to identify the qualitative traits worthy to be considered while selection of parents for breeding programmes. Shannon’s and Simpson’s indices, deducted for all the sixteen qualitative characters, were found to be maximum in Flower main colour (H) and Inner tepal: colour of stripe (D) – the characters primarily used to identify genotypes phenotypically. The germplasm was further classified following UPGMA cluster analysis and five major clusters were formed. The genetic distance between the thirty genotypes ranged from 29% to 75% where cv. Arka Kesar, cv. Arka Gold, cv. Pusa Dhanvantri and cv.Punjab Lemon Delight were found to have maximum genetic distance with most other genotypes under study.