The second version of the checklist and Red List of bryophytes of the Czech Republic is provided. Generally accepted infraspecific taxa have been incorporated into the checklist for the first time. With respect to the Red List, IUCN criteria version 3.1 has been adopted for evaluation of taxa, and the criteria used for listing in the respective categories are listed under each red-listed taxon. Taxa without recent localities and those where extinction has not been proven are listed as a subset of DD taxa. Little known and rare non-threatened taxa with incomplete knowledge of distribution which are worthy of further investigation are listed on the so-called attention list. In total, 849 species plus 5 subspecies and 19 varieties have been accepted. 23 other historically reported species and one variety were evaluated as doubtful with respect to unproven but possible occurrence in the territory, and 6 other species with proven occurrence require taxonomic clarification. 43 taxa have been excluded from our flora compared to the last checklist version. 48.6 % of evaluated taxa have been listed in either of the Red List categories (EX (RE), CR, EN, VU, LR or DD), which is comparable to other industrialized regions of Central Europe.