In vitro protocorm development and mass multiplication of an endangered orchid, Dactylorhiza hatagirea.

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. [ABSTRACT FROM AUTHOR]