Abstract
Most cultivated orchids, contributing to a worldwide highly profitable industry, are originated from tropic regions. Conversely, a considerable number of spontaneous orchids, belonging to the terrestrial orchids and widely diffused throughout the European continent, are not considered for trading due to their less gorgeous appearance and for technical difficulties in seed propagation. However, a breeding programme was undertaken aimed at developing a new hybrid between Himantoglossum adriaticum H. Baumann and H. robertianum (Loisel.) P. Delforge [syn. Barlia robertiana (Loisel.) Greuter] by applying techniques of anther conservation, manual pollination and in vitro asymbiotic germination of the obtained seeds. The plantlets that originated from the protocorms after seed germination were successfully acclimatised after potting in a proper medium. The parentage of the progenies of the hybridisation experiment was assessed by sequencing the Internal Transcribed Spacer assembly (ITS) and plastid barcoding markers of the parental lines and of the hybrids. The method proved to be effective in revealing the origin of the hybrids and to validate the maternal inheritance of the plastid DNA.
Highlights
Orchidaceae is the largest family of flowering plants comprising of approximately32,000 species in around 800 genera
All H. adriaticum flowers hand cross-pollinated with H. robertianum pollinaria were successfully fertilized, showing a rapid development and an evident swelling of the capsules (Figure 3A)
Being H. adriaticum a strictly entomophilous species, the inflorescence was covered with an anti-insect tulle bag to avoid the female being pollinated by stray pollen carried by insects
Summary
Orchidaceae is the largest family of flowering plants comprising of approximately32,000 species in around 800 genera. Orchids account for a large share of global floriculture trade both as cut flowers and potted plants [5] With their uniqueness of shape, breath-taking colours and exceptionally long shelf life provide a source of great aesthetic value. Several mycorrhizal fungi, in particular Basidiomycetes, of the Tulasnellaceae family, and some species of Ascomyc etes, such as Helotiales, Sordariomycetes and Exophiala, have been found to be involved in the network with Himantoglossum adriaticum [16] These symbiotic associations are somewhat problematic to recreate and maintain in co-cultivation during the in vitro propagation phases, there is a need to develop protocols for asymbiotic in vitro seed culture on culture media provided with fundamental nutrients for germination. There has been a considerable increase in the number of studies on tissue culture over the past few years to support the goal of restoring endangered and threatened species and to promote commercial production and economic growth
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