The present study is the first report of the establishment of embryogenic callus cultures from seedling
tissue, the regeneration of plants via somatic embryogenesis and the development of a regeneration system
from protoplast to plant, using three wild species of Cyclamen, Cyclamen graecum Link, Cyclamen
mirabile Hildebrand, Cyclamen trochopteranthum Schwarz (syn. Cyclamen alpinum hort. Dammann ex
Sprenger). The ability to form embryogenic callus and to regenerate via somatic embryogenesis was
strongly genotype-dependent for each species. From 0.5 g callus, up to 1461 somatic embryos were
formed in the case of C. mirabile. Culture media with different concentrations of plant growth regulators,
CaCl2 and activated charcoal significantly influenced embryo formation in this species. Up to
1.4×106 protoplasts were isolated from 1 g of C. graecum cell suspension. Diverse growth responses of
the protoplasts in two embedding agents, agarose and alginate, were observed for the different Cyclamen
species. These specific growth characteristics could be used as a selection marker for future fusion
experiments. From both protoplast culture systems, somatic embryos were regenerated, grown to
plantlets and acclimatised to greenhouse conditions.
The present study is the first report of the establishment of embryogenic callus cultures from seedling
tissue, the regeneration of plants via somatic embryogenesis and the development of a regeneration system
from protoplast to plant, using three wild species of Cyclamen, Cyclamen graecum Link, Cyclamen
mirabile Hildebrand, Cyclamen trochopteranthum Schwarz (syn. Cyclamen alpinum hort. Dammann ex
Sprenger). The ability to form embryogenic callus and to regenerate via somatic embryogenesis was
strongly genotype-dependent for each species. From 0.5 g callus, up to 1461 somatic embryos were
formed in the case of C. mirabile. Culture media with different concentrations of plant growth regulators,
CaCl2 and activated charcoal significantly influenced embryo formation in this species. Up to
1.4×106 protoplasts were isolated from 1 g of C. graecum cell suspension. Diverse growth responses of
the protoplasts in two embedding agents, agarose and alginate, were observed for the different Cyclamen
species. These specific growth characteristics could be used as a selection marker for future fusion
experiments. From both protoplast culture systems, somatic embryos were regenerated, grown to
plantlets and acclimatised to greenhouse conditions.
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