A study of the distribution of orchid species in Puglia, based on an analysis of 2084 bibliographic reports from 2000 to 2022, was carried out with the aim of revising and updating the information on the consistency of the Orchidaceae family in Puglia, with a special focus on assessing threatened species occurring inside and outside protected areas. The work presents a checklist of the Orchidaceae taxa (genera, species, and subspecies) found in the region, including observations on genera and species that present taxonomic challenges. A total of 113 taxa (i.e., species and subspecies), distributed across 16 genera, are listed in alphabetical order. The most representative genera wereOphrys(51 taxa),Serapias(15 taxa), andEpipactis(11 taxa). Additionally, 49 taxa (43.4%) were found to be endemic to Italy, with 21 of these, mostly belonging to theOphrysgenus, being exclusive to Puglia. Our study notes two different trends of distribution: a predominantly coastal distribution for orchid records located in southern Puglia (the Salento peninsula) and a more widespread distribution for the other provinces. Our study also shows that the greatest number of records locate orchids in protected areas with a positive correlation between their presence and habitats cited in Directive 92/43/EEC.