European orchid cultivation – from seed to mature plant

We describe a method for growing orchids of the genera Dactylorhiza and Ophrys, two European members of the subfamily Orchidoideae, from seeds to mature plants using asymbiotic in vitro cultures and glasshouse pot cultures. Four media were used: two new media 1/4-2 and Mo2 and two modifications of Michl medium (Michl 1988). We also describe a highly efficient technique for seed disinfection using a syringe. We tested the effects of ethanol treatment on Anacmaptis morio (L) R.M. Bateman, Pridgeon & M.W. seeds, sugar media composition on Dactylorhiza majalis (Rchb.) P.F. Hunt & Summerh., Oeceoclades decaryana (H. Perrier ex Guillaumin & Manguin) Garay & Taylor and Ophrys lojaconoi P. Delforge and the effect of kinetin on Dactylorhiza majalis protocorm growth. Sucrose was the best carbon source, while hexose resulted in the inhibition of protocorm development at early stages. The addition of kinetin at 10 mg/l resulted in the formation of the largest protocorms. Ethanol can have positive effect on seed germination when applied for a short time (2 min), while long-time ethanol exposure (60 min) can kill the seeds.