Articles
IN VITRO CULTIVATION OF ALCHEMILLA MOLLIS (ROSACEAE) IN BULGARIA
Article number
955_34
Pages
237 – 242
Language
English
Abstract
Alchemilla mollis is a high-mountain medicinal plant that is critically endangered in Bulgaria.
Recent studies have shown that the plant grows well in ex situ collections under different environmental conditions, making possible its cultivation.
Biotechnological methods can be used for its rapid micropropagation.
In the present study, in vitro cultures of the species were initiated from seeds.
Half of them were stimulated with gibberellic acid and were maintained on solid and liquid MS-based nutrient media in a temporary immersion system.
The media were supplemented with 3 mg/L benzylaminopurine and 1 mg/L α-naphthaleneacetic acid.
The plantlets were subcultured every two or three months by separating shoots from shoot clusters and cutting the big ones into segments.
The removal of the plantlets roots stimulated the formation of shoot clusters.
The propagation coefficient increased progressively with every subcultivation, starting at averages of 1.9, 2.5 and 6.7 shoots per explant during the first passages, and reaching over 850 in vitro plantlets per initial seed over a period of 14 months.
Additional increase of the multiplication effectiveness was achieved in liquid culture with the same composition, which led to the formation of enormous clusters of about 32 shoots within 6 weeks; however, submerged inferior shoots vitrified.
The use of the temporary immersion system RITA® led to the improvement of the plantlets quality rather than of the propagation rate: formation of rhizome was observed, rooting during the next passage on agar medium occurred faster, and roots developed trichomes, which facilitated the ex vitro adaptation in a mixture of soil, sand and coconut fiber.
In vitro rooting took place spontaneously after ca. 2-3 months of cultivation on agar medium.
The first plants to be successfully adapted ex vitro were planted in Vitosha Mt. and are developing well; however, ex vitro adaptation to room conditions is still problematic and needs improvement.
Recent studies have shown that the plant grows well in ex situ collections under different environmental conditions, making possible its cultivation.
Biotechnological methods can be used for its rapid micropropagation.
In the present study, in vitro cultures of the species were initiated from seeds.
Half of them were stimulated with gibberellic acid and were maintained on solid and liquid MS-based nutrient media in a temporary immersion system.
The media were supplemented with 3 mg/L benzylaminopurine and 1 mg/L α-naphthaleneacetic acid.
The plantlets were subcultured every two or three months by separating shoots from shoot clusters and cutting the big ones into segments.
The removal of the plantlets roots stimulated the formation of shoot clusters.
The propagation coefficient increased progressively with every subcultivation, starting at averages of 1.9, 2.5 and 6.7 shoots per explant during the first passages, and reaching over 850 in vitro plantlets per initial seed over a period of 14 months.
Additional increase of the multiplication effectiveness was achieved in liquid culture with the same composition, which led to the formation of enormous clusters of about 32 shoots within 6 weeks; however, submerged inferior shoots vitrified.
The use of the temporary immersion system RITA® led to the improvement of the plantlets quality rather than of the propagation rate: formation of rhizome was observed, rooting during the next passage on agar medium occurred faster, and roots developed trichomes, which facilitated the ex vitro adaptation in a mixture of soil, sand and coconut fiber.
In vitro rooting took place spontaneously after ca. 2-3 months of cultivation on agar medium.
The first plants to be successfully adapted ex vitro were planted in Vitosha Mt. and are developing well; however, ex vitro adaptation to room conditions is still problematic and needs improvement.
Authors
M. Stanilova, R. Gorgorov , A. Vitkova
Keywords
Lady’s mantle, propagation coefficient, liquid culture, TIS
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