The early developmental stages of orchids are heterotrophic and obtain energy and carbon from mycorrhizal fungi. Under laboratory in vitro conditions, the mycorrhizal fungi can be substituted with nutrient solution.
However, whole procedure significantly differs between orchid genera and its optimization is very challenging. This methodology describes in vitro propagation optimized for genera Anacamptis, Ophrys and Serapias, which have been difficult to propagate.
It describes seed collection, storage, disinfection and sowing; followed by in vitro cultivation of seedlings and their transfer into greenhouse pot culture. This methodology describes, for the first time, composition of two cultivation media, ER1/4-2A and BM-2b, which were optimized for culture of abovementioned genera.
In addition to this, it uses cultivation in low temperatures and short day, which promotes faster growth and more healthy seedlings comparing with conventional cultivation under laboratory temperatures.