This study provides the results of the developmental biology of the highly rare and endangered moss species Bruchia vogesiaca (recorded in less than 30 localities in the Northern Hemisphere, mainly western, central, and southwestern Europe). The aim of the study was to achieve the fully developed gametophyte and to propagate it for the purpose of conservation, reintroduction, and introduction to potential habitats free from xenic contamination. These gametophytes will be used for the study of genetics and genomics of this species. The micropropagation of B. vogesiaca was successfully applied on BCD medium supplemented with 0.1 μM BA and on BCD supplemented with 0.3 μM IBA and 0.3 μM BA for numerous gametophore production. The highest production of secondary protonema was achieved on MS/2 S/2 medium enriched with 0.1 or 0.3 μM IBA and 0.3 μM BA. Rather successfully applied micropropagation of this threatened moss species enables better knowledge of its biology and is of great value for its conservation biology and developmental research. Chemical names used: indole-3-butyric acid (IBA), N6-benzyladenine (BA), Murashige and Skoog medium (MS).
Rindera umbellata (Waldst. & Kit.) Bunge is a rare, critically endangered and horticulturally appealing plant with unexplored pharmaceutical potential. Its distribution is restricted to sandy habitats, whereas propagation in nature is limited by fungal infections of the seeds. To initiate its ex situ conservation and provide material for metabolomic studies, we have introduced R. umbellata into in vitro culture using immature embryos as primary explants. A 72% of the embryos germinated on growth regulator-free medium. The optimization of growth conditions was based on varying carbohydrates (sucrose, glucose, and fructose) in the medium. In vitro growth and development of R. umbellata plants were significantly affected by both the type and concentration of the applied sugars. For most recorded parameters, including leaf elongation, biomass production, rooting percentage, and the number and length of roots, 0.1 m sucrose was optimal. The highest percentage of explants with developed buds was achieved on 0.06 m sucrose (38.77%) or 0.3 m glucose (27.43%). The plantlets obtained on 0.1 m sucrose were successfully acclimatized to greenhouse and field conditions with survival rates of 71.43% and 42.86%, respectively. To our best knowledge, this is the first publication dedicated to this species.
A rapid, one-step method for direct leafy and rooted bulblets regeneration of Lilium martagon var. cattaniae Vis. was established using seeds as the starting explants for in vitro culture initiation. Adventitious bulblets were regenerated from one scale explants on MS (Murashige and Skoog, 1962) medium supplied with various concentrations of plant growth regulators. The most efficient medium for multiplication was MS supplemented with 0.2 mg/l 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA) ranging from 0.25 to 2 mg/l. On average, five bulblets were obtained per explant after six weeks in culture. Upon rooting, about 200 plantlets were successfully hardened in the green house with a 95% survival rate. Preliminary experiments indicated that the plantlets from the greenhouse could be successfully used for field cultivation. In vitro propagation of this endemic and protected plant species is of great importance for germplasm conservation.
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