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Osmunda

Osmunda

Osmunda

The publication of a detailed phylogeny of the family by Metzgar et al. in 2008 showed that Osmunda as circumscribed was paraphyletic and that Osmunda cinnamomea, despite its morphological similarity to Osmunda claytoniana, was sister to the rest of the family, and resurrected the segregate genus Osmundastrum, by elevating it from subgenus, to contain it and render Osmunda monophyletic. Their phylogeny of Osmundaceae genera is shown in the following cladogramOsmunda regalis, commonly called royal fern, is a tall, deciduous, native fern which usually occurs on moist bluffs and ledges and along streams (sometimes growing in the water). Typically grows in clumps to 5-6' tall, but with constant moisture can reach 6' in height. Broad fronds have large, well-separated pinnae (leaflets) which give this fern an almost pea-family appearance. Fronds typically turn yellow to brown in autumn. Spores are located in brown, tassel-like, fertile clusters at the tips of the fronds, thus giving rise to the additional common name of flowering fern for this plant. Osmunda fiber used in the potting of orchids comes from the fibrous roots of these ferns.Osmunda plants originated from the Botanical Garden of the University of Kaiserslautern (Germany). They were identified by Mr. Bernd Simon, who is a known expert for plant taxonomy. A voucher specimen was deposited at the Herbarium of the University of Wuerzburg; (Index Herbariorum Code: WB) under the number 2017_HNO001. The black roots (Fig. 1) were cleaned, dried and minced. The ethanolic extract was prepared as follows: 18.5 g of minced roots were homogenized in 30 ml 70% ethanol with a power homogenizer and subsequently agitated overnight at 37 °C.

After 14 days incubation with daily agitation, the supernatant was cleared by centrifugation and sterile filtration. The yield after centrifugation and sterile filtration was 20 ml. A 1 ml aliquot of the extract was dried by centrifugal evaporation. According to the weight of the dried substance the concentration of the extract was adjusted to 6 mg/ml with 70% ethanol. Aliquots of the stock solution were stored at −80 °C. For experiments the stock solution was diluted 1:10 with culture medium without supplements (0.6 mg/ml). This working solution was finally diluted to 6, 15, 30, 60 and 90 μg/ml for MTT assays. One batch was used for all experiments.Cells were seeded at 5000 cells/well in 96 well plates. O. regalis ethanolic extract (6 mg/ml) was diluted 1:10 with cell culture medium without supplements and subsequently further diluted. Cells were treated with increasing concentrations (0, 6, 15, 30, 60, 90 μg/ml) of Osmunda regalis extract for 48 h. After 48 h cell culture medium was removed and replaced by medium supplemented with MTT (1 mg/ml) [12]. Following 4 h incubation, isopropanol replaced medium for 45 h at 37 °C. Colour conversion of the MTT reaction was measured at a wavelength of 570 nm. Relative toxicity was calculated as % surviving cells by setting control cells treated with vehicle as 100% surviving cells. Viability was calculated to the following formula: absorption Control cells (untreated)/100 * absorption treated cells.Due to historical sources it wasn’t clear, which parts of the plant had been used formerly. Williams et al. [6] used the “bulb” for production of an alcoholic extract. The bulb of Osmunda fern plants is composed of a hard wooden splintery part, surrounded by black roots (Fig. 1). In a first attempt (data not shown) we tried to extract active ingredients from the wooden parts by shredding and extracting them in 70% ethanol for prolonged periods. The preliminary MTT results, however, were disappointing. Unlike the wooden bulb, the black roots proved to be much more promising. (Source: www.ncbi.nlm.nih.gov)

 

 

The publication of a detailed phylogeny of the family by Metzgar et al. in 2008 showed that Osmunda as circumscribed was paraphyletic and that Osmunda cinnamomea, despite its morphological similarity to Osmunda claytoniana, was sister to the rest of the family, and resurrected the segregate genus Osmundastrum, by elevating it from subgenus, to contain it and render Osmunda monophyletic. Their phylogeny of Osmundaceae genera is shown in the following cladogramOsmunda regalis, commonly called royal fern, is a tall, deciduous, native fern which usually occurs on moist bluffs and ledges and along streams (sometimes growing in the water). Typically grows in clumps to 5-6' tall, but with constant moisture can reach 6' in height. Broad fronds have large, well-separated pinnae (leaflets) which give this fern an almost pea-family appearance. Fronds typically turn yellow to brown in autumn. Spores are located in brown, tassel-like, fertile clusters at the tips of the fronds, thus giving rise to the additional common name of flowering fern for this plant. Osmunda fiber used in the potting of orchids comes from the fibrous roots of these ferns.Osmunda plants originated from the Botanical Garden of the University of Kaiserslautern (Germany). They were identified by Mr. Bernd Simon, who is a known expert for plant taxonomy. A voucher specimen was deposited at the Herbarium of the University of Wuerzburg; (Index Herbariorum Code: WB) under the number 2017_HNO001. The black roots (Fig. 1) were cleaned, dried and minced. The ethanolic extract was prepared as follows: 18.5 g of minced roots were homogenized in 30 ml 70% ethanol with a power homogenizer and subsequently agitated overnight at 37 °C.

After 14 days incubation with daily agitation, the supernatant was cleared by centrifugation and sterile filtration. The yield after centrifugation and sterile filtration was 20 ml. A 1 ml aliquot of the extract was dried by centrifugal evaporation. According to the weight of the dried substance the concentration of the extract was adjusted to 6 mg/ml with 70% ethanol. Aliquots of the stock solution were stored at −80 °C. For experiments the stock solution was diluted 1:10 with culture medium without supplements (0.6 mg/ml). This working solution was finally diluted to 6, 15, 30, 60 and 90 μg/ml for MTT assays. One batch was used for all experiments.Cells were seeded at 5000 cells/well in 96 well plates. O. regalis ethanolic extract (6 mg/ml) was diluted 1:10 with cell culture medium without supplements and subsequently further diluted. Cells were treated with increasing concentrations (0, 6, 15, 30, 60, 90 μg/ml) of Osmunda regalis extract for 48 h. After 48 h cell culture medium was removed and replaced by medium supplemented with MTT (1 mg/ml) [12]. Following 4 h incubation, isopropanol replaced medium for 45 h at 37 °C. Colour conversion of the MTT reaction was measured at a wavelength of 570 nm. Relative toxicity was calculated as % surviving cells by setting control cells treated with vehicle as 100% surviving cells. Viability was calculated to the following formula: absorption Control cells (untreated)/100 * absorption treated cells.Due to historical sources it wasn’t clear, which parts of the plant had been used formerly. Williams et al. [6] used the “bulb” for production of an alcoholic extract. The bulb of Osmunda fern plants is composed of a hard wooden splintery part, surrounded by black roots (Fig. 1). In a first attempt (data not shown) we tried to extract active ingredients from the wooden parts by shredding and extracting them in 70% ethanol for prolonged periods. The preliminary MTT results, however, were disappointing. Unlike the wooden bulb, the black roots proved to be much more promising. (Source: www.ncbi.nlm.nih.gov)

 

 

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