The annotated chromosome numbers of 25 species from 6 families of monocotyledons, most of them (14) belonging to Poaceae family, are presented here. The data, except three chromosome counts (Allium oleraceum from Hungary and Calamagrostis villosa from Slovakia), are all based on plants collected in the Czech Republic. The karyological data of 21 species represents new information. While the majority of species presented here originated from one or two localities each, the species Calamagrostis villosa has been studied more extensively: all plants, collected altogether at 13 localities (mountain and lower altitudes), are characterized by an invariable decaploid level (2n = 70). The record of triploid Allium oleraceum is only the second reference to this rare ploidy level in this species. All original karyological data are compared with literature references to particular species, preferentially from Europe.