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<p><strong>Background:</strong><em> Tigridia pavonia</em> (L.f.) DC is a wild species with great ornamental value, of which nine plant varieties are known. Within the evolutionary process of this species, Penélope has been considered a natural hybrid, product of the cross between the varieties Trinidad and Dulce.<strong> Objective:</strong> In the present study, the cytogenetic relationships among Trinidad, Dulce and Penélope were analyzed. <strong>Methodology:</strong> The karyotype of the varieties Trinidad and Penélope was determined through classic cytogenetics and the physical mapping of the genes 5s and 45s rDNA through Fluorescent In Situ Hybridization. <strong>Results:</strong>  The results showed for the first time the karyotype and the physical mapping of the genes 5s and 45s rDNA in the varieties Trinidad and Penélope. <strong>Implications:</strong> The information generated can be the basis for future evolutionary analyzes, and / or breeding programs in the species. <strong>Conclusion:</strong> A higher cytogenetic similarity of Penélope with Trinidad and Dulce has been revealed, suggesting that the latter may be the parents.</p>