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Sugarcane (Saccharum officinarum L.) is a plant that is used as raw material for sugar production. Sugarcane productivity is influenced by many factors, one of which is water supply. Therefore, it is necessary to carry out plant breeding efforts through the induction of colchicine mutagens to produce superior sugarcane varieties with multiple chromosome numbers and resistance to drought stress. The purpose of this study was to determine the dose and duration of immersion of colchicine on the anatomical character of the stomata. This research was conducted on 24 varieties of GMP6 mutant with 3 repetitions. Observations were made on 4 stomata characters, namely length, width, number, and density of stomata. The average value of the tested characters is then used to perform principal component analysis and cluster analysis using MVSP software. Cluster analysis was carried out using the Unweighted Pair Group Method with Arithmetic Average (UPGMA) to classify the asesions based on the similarity of anatomical characters. The results showed that the administration of colchicine was able to affect the stomatal character which was different from the control. The character of the length and width of the stomata decreased compared to the control along with the addition of concentration and duration of immersion. The most influential treatment was a concentration of 0.1 ppm with an immersion time of 1 day and a concentration of 1 ppm with an immersion time of 2 days. Based on the results of PCA analysis, 24 GMP6 mutant varieties showed different stomata characteristics from the GMP6 variety. [ABSTRACT FROM AUTHOR]