Introduction of asymbiotically propagated seedlings of Cephalanthera falcata (Orchidaceae) into natural habitat and investigation of colonized mycorrhizal fungi

Asymbiotic seedling propagation and introduction of seedlings into a natural habitat were achieved for Cephalanthera falcata. For immature seeds collected 65 days after pollination, high germination rate (av. 50%) was achieved on Hyponex agar medium plates. Root development occurred in about 10% of the protocorms 5 months after seed sowing. Rooted protocorms were transferred to a culture bottle containing 100 ml of the Hyponex agar medium and incubated continually. In about 30% of the transferred individuals, shoot height reached 1.5-2 cm 8 months after the transfer. After acclimatization in wet vermiculite at 4 degrees C for 6 months, 135 individuals were planted in a natural stand of C. falcata in mid February 2001. Shoot appearance rate was 44.4% at the first year and flowering was noted in some plants. At the fifth year, shoots with an average height of 21.6 cm still appeared in four plants, and flowering was noted in three of them. Colonization of mycorrhizal fungi was examined in two of them as well as one wild plant, in which the mycorrhizal fungi were identified to be in Thelephoraceae or Russulaceae. These fungi are known to form ectomycorrhiza with trees, and thus a tripartnership symbiosis consisting of C. falcata, mycorrhizal fungi and trees was suggested. The involvement of ectomycorrhizal fungi might be the reason for the low survival rate in the field experiment, because the distribution of ectomycorrhizal fungi relevant to this orchid is assumed to be uneven. The possibility of introducing artificially propagated orchids into natural habitats was discussed.