Orchids are unique and highly evolved group of flowering plants with a remarkable diversity and exotic beauty. They are rich in alkaloids and other phytochemicals thereby are used in indigenous medicines to cure a variety of ailments. Dactylorhiza hatagirea (D. Don) Soő tubers are used to prepare ‘salep’ a local drug having aphrodisiac properties. It is a medicinally important orchid with threatened status and used as cut flower in many countries including India, so in the present investigation micropopagation techniques have been employed. The technique of asymbiotic seed germination has added new domains to orchid propagation for cultivation and conservation. The ex-situ conservation strategies involved propagation using different explants like inflorescence, petioles, apical tips, stems, leaves, tubers and seeds. Green and mature pods containing seeds germinated on medium supplemented with cytokinin and auxin leading to seedling regeneration via PLB’s regeneration. In another treatment auxins particularly IBA have been found to induce germination and plantlet regeneration in tuber sections. Therefore an efficient method of propagation was developed so that this critically endangered medicinal orchid can be easily propagated to achieve its mass multiplication.