Asymbiotic seed germination of Dactylorhiza fuchsii as a tool for the introduction of a legally protected species into the natural environment

Dactylorhiza fuchsii (Druce) Soó is a legally protected species in some countries with considerable commercial potential. The main objective of this study was to develop an efficient protocol for seed propagation using in vitro cultures that could serve as a basis for the successful reintroduction of regenerated plants into their natural habitats. The most effective sterilisation procedure involved immersion in a 0.8% aqueous detergent solution for 18 h, followed by treatment with a 5% aqueous sodium hypochlorite solution for 60 min. The highest germination rate was observed on Orchimax medium supplemented with 10 mg/L-1 kinetin. The resulting protocorms were regenerated on several media, whereas plant development was achieved on media lacking phytohormones. The greatest number of regenerated shoots was obtained on half-strength Murashige and Skoog medium. The results demonstrate the feasibility of propagating this species under in vitro conditions and provide a basis for its conservation and potential commercial use.