Turkish Journal of Botany




Immature seeds were cultured on 10 different media for germination. Maximum germination was achieved on Lindeman orchid medium (37.12%) within 17 days of culturing. Protocorms with leaf primordia were cultured on BM-2 and 7 different modifications of Murashige and Skoog (MS) medium with various hormone combinations [0-3 mg/L indole butyric acid (IBA) and 0-3 mg/L kinetin (Kin)] for plantlet regeneration and mass multiplication. Maximum number of shoots (18.12 ± 0.3), highest shoot length (17.80 cm ± 2.16), maximum root number (8.25 ± 0.69), and highest root length (8.02 cm ± 1.45) were found on MS medium with 3 mg/L IBA and 1 mg/L Kin. Plantlets with 2-3 shoots were transferred to different potting mixtures for acclimatization to field conditions and further multiplication. One hundred percent survival was obtained in C-8 potting mixture consisting of cocopeat + vermiculite + perlite (1:1:1), which produced 75 shoots (25 plantlets) after 1 month of transplantation in the greenhouse. The current study describes for the first time a rapid in vitro protocorm development and mass multiplication protocol for Dactylorhiza hatagirea (D.Don) Soo that holds robust potential for large-scale propagation and metabolite production.


Conservation, development, mass multiplication, potting mixture, protocorm formation

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