In this study, 2 subspecies; Dactylorhiza romana (Seb.) Soo subsp. romana and Dactylorhiza romana (Seb.) Soo subsp. georgica (Klinge) Soo ex Renz & Taub. mainly distributed in Turkey were investigated in terms of morphological, anatomical characters and habitat properties. Plant and soil samples of D. romana subsp. romana were taken from 9 localities and D. romana subsp. georgica from 7 localities in Turkey. We observed that some of morphological characters such as; underground part length, tuber length, the longest leaf width, and bract length had different properties between both subspecies. Cross sections of the leaves, stems, tubers and roots were taken by free-hand and stained with Sartur solution and Safranin. The same procedure was performed for the surface sections of leaves and stern. Stomatal and epidermal cells, starch granules dimensions, lengths of raphides, stomata index, and cuticular thickness were measured. Anatomical characters of leaves such as cuticle thickness and stomata index displayed between two subspecies. Although D. romana subsp. georgica grows in high altitudes, D. romana subsp. romana prefers lower altitudes. The most common habitat of D. romana is meadow, macchie and forest lands. We obtained that both of them had similar soil properties, while the differencies came out by habitat properties. Morphological and habitat characters were compared with independent-T test.