The biological concept of species is founded on the presence of barriers that limit the gene flow and species delimitation represents a vivid topic in orchids. Studies on the reproductive isolation in mediterranean orchids often examine the gene flow between co-occuring species providing a relevant understanding of how species boundaries are maintained. In Ophrys, the role of post-mating boundaries are always been tought to be weak and, despite the relevant morphological and ethological differences, low phylogenetic diversity has been recorded. In contrast, not many studies are focused on the relevance of each barriers in close related species in island condition. Using a morphological approach and experimental crosses to evaluate post-mating barriers, here, we clarify the taxonomic position of two sardinian endemic orchid sister-species, Ophrys annae and Ophrys chestermanii, currently ascribed in the bornmuelleri group. In Ophrys, due their iper specialized pollination syndrome, premating barriers are often considered more relevant than post mating barriers. We found that the two endemisms differs sustantially in 16 functional traits adopted. We further detected asymmetric results in manual crosses suggesting that post-mating barriers may act as reinforcement of the pre-mating barriers. Our results suggest that in close related orchid species of the genus Ophrys, gene flow is limited also by post-mating barriers which have a relevant act in keeping the two reproductively isolated.