The genus Gladiolus L. (Iridaceae) includes about 250–280 species of perennial herbaceous corms, distributed in Africa, Madagascar and Eurasia. The taxonomy of the genus is not simple, since the characteristics of the species are quite confusing. Our article presents the results of a comparative palyno-morphological analysis of all 9 species and 2 subspecies of the genus Gladiolus of the Armenian flora. The purpose of the work is to identify diagnostic characteristics of pollen used to distinguish individual taxa. Six morphological characters, namely the length of the polar axis (P), as well as the length of the large and small equatorial diameters (E1 and E2, respectively), exine thickness and the number of echinae and perforations per unit area of the pollen surface were measured using light microscopy (LM) and scanning electron microscopy (SEM). Literature data, as well as the results of our previous studies, showed that the genus is characterized by stenopalynous anasulcate pollen grains with a two-lane operculum. In the presence of significant uniformity in pollen characteristics, our studies revealed that among the species studied, G. hajastanicus and G. kotschyanus have slightly larger pollen grain sizes, and the pollen of the species G. tenuis is characterized by a minimal number of both echinae and perforations per unit surface area of the pollen grain (using SEM). A certain difference has also been established between the subspecies of the species G. kotschyanus. In particular, the sizes of pollen grains of G. kotschyanus subsp. kotschyanus (based on E1) is slightly larger than in G. kotschyanus subsp. distichus. At the same time, the number of echinae and perforations per unit surface of a pollen grain (at the SEM level) in the subspecies G. kotschyanus subsp. distichus is approximately twice as large as G. kotschyanus subsp. kotschianus.