Sprekelia formosissima (L.) Herbert is a bulbous ornamental species of the Amaryllidaceae family, distributed from northern Mexico to Guatemala. Different reports on cytogenetic research on this species show differences in chromosome numbers (2n = 60, 120, 150, 180) with a basic number of x = 30. Since the chromosome number of plants can provide useful information for various fields of research including karyotaxonomy, genetics, and cytogenetics, it is important to determine the chromosome number for this species. Bulbs of S. formosissima were collected in three different locations in the state of Jalisco, Mexico. Meristematic root cells were analyzed cytogenetically using a modified steam-drop method. The three populations showed different chromosome numbers and ploidy levels, 2n = 2x = 60, 2n = 4x = 120, and 2n = 5x = 150. The ploidy level was compared with morphological traits showing higher values related to higher ploidy levels. The populations analyzed correspond to new polyploid cytotypes of S. formosissima.