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Alaide Braga De Oliveira - One of the best experts on this subject based on the ideXlab platform.
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effect of digitalis lanata matrix composition on the lanatoside c partition coefficient and its consequence on rotation locular counter current chromatography efficiency
Journal of Chromatography A, 1998Co-Authors: Fernao Castro Braga, Wolfgang Kreis, Alaide Braga De OliveiraAbstract:Lanatoside C standard samples had their partition coefficients (K) determined in dichloromethane-methanol-water (4:4:3), (5:6:4) and (7:13:8), by the shake flask method and the obtained values were similar. On the other hand, K for lanatoside C accomplished by calculation from rotation locular counter-current chromatography (RLCC) traces of Digitalis lanata pre-purified extracts, as well as by the shake flask method, in the same solvent systems, showed a significant variation. Such variation was attributed to differences in the pre-purification procedures for D. lanata extract, which resulted in starting materials for RLCC with very distinct cardenolide profiles, reflecting, thus, in the K values for lanatoside C and, consequently, in the lanatoside C purity at the end of the chromatographic process.
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Avaliação quantitativa de cardenolídeos no cultivar experimental de Digitalis lanata do maciço do itatiaia e perspectivas de seu emprego industrial
Sociedade Brasileira de Química, 1997Co-Authors: Braga,fernão Castro, Récio,rubens Almeida, Alaide Braga De OliveiraAbstract:The content of digoxin and lanatoside C (jointly quantified), lanatoside A, lanatoside B, glucoevatromonoside, odorobioside G, glucogitoroside, glucoverodoxine, glucodigifucoside and digitalinum verum was determined by HPLC in Digitalis lanata harvested in Brazil, as well as in clones industrially employed for the production of cardenolides. The Brazilian plants presented greater variation in the contents of cardenolides than the analyzed clones. Lanatoside C and digoxin concentrations were higher in the clones (6120±640 nmol/g dry leaf) than in the Brazilian plants (1820±900 nmol/g dry leaf). The concentrations of these glycosides were found to be within the range described for native species and also were similar to an European cultivar, what makes possible its industrial use for the cardenolides production
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Avaliação quantitativa de cardenolídeos no cultivar experimental de Digitalis lanata do maciço do itatiaia e perspectivas de seu emprego industrial Quantitative evaluation of cardenolides in a brazilian digitalis lanata cultivar and perspectives of its industrial use
Sociedade Brasileira de Química, 1997Co-Authors: Fernao Castro Braga, Wolfgang Kreis, Rubens Almeida Récio, Alaide Braga De OliveiraAbstract:The content of digoxin and lanatoside C (jointly quantified), lanatoside A, lanatoside B, glucoevatromonoside, odorobioside G, glucogitoroside, glucoverodoxine, glucodigifucoside and digitalinum verum was determined by HPLC in Digitalis lanata harvested in Brazil, as well as in clones industrially employed for the production of cardenolides. The Brazilian plants presented greater variation in the contents of cardenolides than the analyzed clones. Lanatoside C and digoxin concentrations were higher in the clones (6120±640 nmol/g dry leaf) than in the Brazilian plants (1820±900 nmol/g dry leaf). The concentrations of these glycosides were found to be within the range described for native species and also were similar to an European cultivar, what makes possible its industrial use for the cardenolides production
Ekrem Gurel - One of the best experts on this subject based on the ideXlab platform.
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Shoot-tip Cultivation and Cardenolide Content Analysis of Natural Populations of Digitalis lanata Ehrh. subsp. lanata (wooly foxglove) in Thrace Region
Abant İzzet Baysal University, 2018Co-Authors: Buhara Yücesan, Wolfgang Kreis, İsmail Eker, Luiz Eduardo Santos Lazzarini, Noreen Aslam, Aliyu Mohammad, Jose Eduardo Brasil Pereira Pinto, Ekrem GurelAbstract:Digitalis lanata, due to the presence of cardenolides in the leaves, is widely used in drug industry for the treatments of congestive heart diesases. In this present study, natural populations of D. lanata in Thrace region were collected. Thereafter, cardiac glycoside analysis of D. lanata subsp. lanata grown in natural habitats and micropropagation from shoot tips were established as a model work for a consistent and a sustainable plant production. Of the cardenolides, Lan A (ranging between 24.8 and 300.4 mg 100g-1) and Lan C (42.1 and 258 mg 100 g-1 were predominantly found in the samples as compared to lanatoside B, digoxin and digitoxin. After seed germination, shoot-tip cultivation was achieved after a subsequent shoot and root formation on MS medium. Indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) depending on their concentrations were found to be effective in shoot and root formation accordingly. Maximum two shoots from shoot tips were produced after 8 weeks of cultivation, and at the end of 12th weeks of cultivation each shoots produced an average number of 6.4 roots in 8.1 cm length. Acclimatization was also achieved successfully with a maximum survival rate (95 %) for 4 weeks. This study is the first report on cardenolide profiling of Thracian D. lanata at different locations, and shoot tip cultivation of the population selected under in vitro conditions
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an overview of cardenolides in digitalis more than a cardiotonic compound
Current Pharmaceutical Design, 2017Co-Authors: Ekrem Gurel, Buhara Yücesan, İsmail Eker, Serhan Karvar, Muhammad SameeullahAbstract:Abstract The genus Digitalis L. containing species, commonly known as the "foxglove", is the main source of cardenolides, which have various pharmacological properties effective against certain pathological conditions including myocardial infarction, arterial hypertension, cardiac dysfunction, angina, and hypertrophy. Togehter with a prime effect of controlling the heart rhythm, many workers demonstrated that lanatoside C and some other cardiac glycosides are effective in several cancer treatments such as prostate and breast cancers. Due to digoxigenin derivatives of cardenolides, which are mainly used for medicinal purposes, such as digoxigenin, D. lanata as a main source is of great interest for commercial scale production of cardenolides in Europe. Phytochemical studies on cardenolides, naturally occurring plant secondary metabolites, have mainly focused on the species of the genus Digitalis L., as the members of this family have a high level and diverse content of cardenolides. During the last few decades, plant tissue culture techniques have been optimised for many plant species including Digitalis, however, the production capacity of cardenolides somehow failed to reach a commercially desired extent. In this review paper, the genus Digitalis is evaluated in terms of its main botanical and physiological features, traditional uses, molecular genetics and metabolomics, cellular mechanism of action, medicinal uses, clinical pharmacology, drug interactions, therapy in the management of cardiovascular disorders, potential utility of therapy in extracardiac conditions, and toxicity.
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Investigation of Relationship Between Chemical Stress Factors and Certain Metabolites Including Cardenolides in Callus Cultures of Endemic Turkish Digitalis L. Species
International Journal of Secondary Metabolite, 2017Co-Authors: Gunce Sahin, Sandeep Kumar Verma, Ekrem GurelAbstract:The aim of the present research is to obtain relationship between different stress treatments [Cu (copper) and Hg (mercury)] and content of cardiac glycosides (digoxigenin, gitoxigenin, lanatoside C, digoxin and digitoxin) as secondary metabolites of commercial value for the pharmaceutical industry and to determine the antioxidant metabolites against stress conditions in callus cultures of endemic Turkish Digitalis species. The effects of different stress treatments on cardiotonic glycoside accumulation in D. lamarckii Ivanina, D. trojana Ivanina, D. davisiana Heywood and D. cariensis Boiss. ex Jaub. et Spach were investigated using HPLC. HPLC analysis revealed that all stress conditions were significantly effective at 5% significance level according to their control groups. The predominant cardiac glycoside was lanatoside C (Lan C) followed by digitoxin, digoxigenin, gitoxigenin and digoxin. No digoxin was detected in all treatments as well as in control groups. For the calibration curves, concentrations of 5, 10, 20, 30 and 40 mg/l digoxigenin, gitoxigenin, lanatoside C, digoxin and digitoxin were used (R2= 0.99). Cardenolides were eluted with acetonitrile (A) and water (B) gradients as follows: 0 to 20 min 20% (A), 80% (B); 20 to 23.40 min 30% (A), 70% (B); 23.40 to 30 min 25% (A), 75% (B) and 30 to 40 min 40% (A), 60%(B). Average peak area of the glycoside in samples was automatically calculated and monitored by ChemStation LC/MS software against that of standards. Enhanced production of cardenolides was achieved from callus cultures elicited with 50 μm CuSO4 and HgCl2. Higher amounts of cardenolides were obtained when callus of four Digitalis species were elicited with CuSO4. Results demonstrated that catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) activities, the total contents of phenolics and proline were markedly stimulated under stress conditions. All these results indicated that treatments have induced changes in the redox state of callus cells and suggest that this alteration change cardenolides accumulation and antioxidative status in Digitalis L. callus cultures
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hydrogen peroxide induced antioxidant activities and cardiotonic glycoside accumulation in callus cultures of endemic digitalis species
Plant Physiology and Biochemistry, 2014Co-Authors: Gunce Sahin Cingoz, Sandeep Kumar Verma, Ekrem GurelAbstract:Abstract The effect of hydrogen peroxide (H2O2) on callus cultures of four Digitalis species (Digitalis lamarckii, Digitalis trojana, Digitalis davisiana and Digitalis cariensis) increased catalase (CAT), superoxide dismutase (SOD), total phenolic, proline activity and cardiotonic glycoside production. Callus derived from hypocotyl explants was cultured on Murashige and Skoog medium supplemented with 0.25 mg L−1 indole-3-acetic acid (IAA) and 0.5 mg L−1 thidiazuron (TDZ). After a month of culture, callus was transferred to MS medium containing 10 mM H2O2 and then incubated for 6 h. The amount of five cardenolides (Lanatoside C, Digitoxin, Digoxigenin, Gitoxigenin and Digoxin) as well as CAT, SOD, total phenolic, proline activity from Digitalis species were compared. No digoxin was detected in all treatments and control groups. The total cardenolides estimated were in the order of D. lamarckii (586.65 μg g−1 dw), D. davisiana (506.79 μg g−1 dw), D. cariensis (376.60 μg g−1 dw) and D. trojana (282.39 μg g−1 dw). It was clear that H2O2 pre-treatment resulted in an increase in enzymatic and nonenzymatic antioxidants. However, a significant negative relationship between cardenolides production and overall activities of CAT, SOD, total phenolic and proline was evident. The described protocol here will be useful for the development of new strategies for a large-scale production of cardenolides.
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embryogenesis plant regeneration and cardiac glycoside determination in digitalis ferruginea subsp ferruginea l
Plant Cell Tissue and Organ Culture, 2014Co-Authors: Gunce Sahin, Sandeep Kumar Verma, Buhara Yücesan, Ekrem GurelAbstract:The present study reports, for the first time, an efficient in vitro plant regeneration protocol for Digitalis ferruginea subsp. ferruginea L. (rusty foxglove). We have used different concentrations of gibberellic acid (GA3) on Murashige and Skoog (MS) medium to assess the germination frequency of seeds. High frequency of germination was achieved on MS medium with 1.0 mg l−1 GA3. 6-Benzylaminopurine (BAP) combined with α-naphtaleneacetic acid (NAA) or 2, 4-dichlorophenoxy acetic acid (2, 4-D) in the induction MS medium induced both somatic embryogensis and shoot organogenesis. The highest percentage of callus growth (85 %) was obtained when hypocotyl explants were cultured on MS medium containing 0.5 mg l−1 2, 4-D plus 1.0 mg l−1 BAP. The maximum mean number of somatic embryos (7.3 ± 1.3 embryos) or shoots (12.0 ± 1.1 shoots) per callus was obtained when medium contained 0.25 mg l−1 NAA plus 1.0 mg l−1 BAP or 0.5 mg l−1 NAA plus 2.0 mg l−1 BAP. The regenerated shoots easily rooted on MS medium. Higher amounts of lanatoside C [13.2 ± 0.5 mg 100 g−1 dry weight (dw)] and digoxin (2.93 ± 0.31 mg 100 g−1 dw) accumulation were obtained when shoots were obtained by indirect regeneration. We also investigated derivatives of cardenolides, i.e., digitoxigenin (730 ± 180 mg 100 g−1 dw), gitoxigenin (50 ± 20 mg 100 g−1 dw) and digoxigenin (490 ± 170 mg 100 g−1 dw) from natural samples.
Elio Jiménez - One of the best experts on this subject based on the ideXlab platform.
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Increased cardenolides production by elicitation of Digitalis lanata shoots cultured in temporary immersion systems
Plant Cell Tissue and Organ Culture (PCTOC), 2012Co-Authors: Naivy Pérez-alonso, Alina Capote, André Gerth, Elio JiménezAbstract:Digitalis lanata is an important source of cardenolides such as digoxin and lanatoside C, which have been widely applied in the treatment of cardiac insufficiencies. Elicitation is one of the most effective methods to enhance the biosynthesis of several secondary metabolites in medicinal plants. We studied the effect of elicitation with Chitoplant^®, Silioplant^® and methyl jasmonate on biomass and cardenolides accumulation in shoots of D. lanata cultivated in temporary immersion systems. Morphological response of the shoots was influenced by elicitors. A reduction in length and number of shoots was evident with all MJ concentrations. Regarding biomass production, Chitoplant^® (0.1 g l^−1) was found to impact significantly on fresh and dry weight of the shoots. HPLC analysis revealed a higher content of lanatoside C compared to digoxin in all treatments. The highest accumulation of lanatoside C was achieved with Chitoplant^® (0.1 g l^−1), which resulted in 316 μg g-DW^−1 and with Silioplant^® (0.01 g l^−1; 310 μg g-DW^−1), which accounted for a 2.2-fold increase in lanatoside C content compared to non-elicited shoot cultures. Additionally, elicitation of D. lanata shoots in temporary immersion systems resulted in an oxidative stress characterized by hydrogen peroxide and malondialdehyde accumulation. These observations point to a connection between hydrogen peroxide generation, lipid peroxidation and cardenolide accumulation. The optimization of elicitor treatment and culture conditions for cardenolide production as well as the advantages of TIS for this purpose are discussed.
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cardiotonic glycosides from biomass of digitalis purpurea l cultured in temporary immersion systems
Plant Cell Tissue and Organ Culture, 2009Co-Authors: Naivy Perezalonso, André Gerth, Dirk Wilken, Annett Jahn, Horstmichael Nitzsche, Gerhard Kerns, Alina Capoteperez, Elio JiménezAbstract:Cardiotonic glycosides are extracted mostly from leaves of Digitalis plants. Commercial production of bioactive secondary metabolites by traditional agriculture is an inefficient process and can be affected by climatic and soil conditions. Strategies, based on in vitro culture methods, have been extensively studied to improve the production of specific plant derived chemicals. The aim of the present research was to obtain biomass of D. purpurea using the temporary immersion system (TIS) and to determine the content of cardiotonic glycosides (digitoxin, digoxin and lanatoside C) as secondary metabolites of commercial value for the pharmaceutical industry. Shoots were cultured in 1,000 ml TIS during 28 days. The effect of four immersion frequencies (once every 2, 4, 6, and 12 h) was studied. Biomass accumulation was influenced by immersion frequency. The maximum biomass accumulation (values in respect of fresh and dry weight) was obtained with immersions every 4 h (six immersions per day). HPLC analysis revealed the presence of digoxin and digitoxin for all immersion frequencies. No lanatoside C was detected in the biomass cultured in TIS. Digoxin concentrations varied depending on the frequencies tested. In contrast, the digitoxin content showed no dependency on the immersion frequency. Net production of digoxin and digitoxin per TIS were found to be higher with immersions every 4 h. The best net production of digitoxin and digoxin per TIS were 167.6 and 119.9 μg, respectively. The development of organ culture based on temporary immersion system can be a reliable method for the steady production of biomass for cardiotonic glycosides production, which is reported for the first time for Digitalis genus in this communication.
Wolfgang Kreis - One of the best experts on this subject based on the ideXlab platform.
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Shoot-tip Cultivation and Cardenolide Content Analysis of Natural Populations of Digitalis lanata Ehrh. subsp. lanata (wooly foxglove) in Thrace Region
Abant İzzet Baysal University, 2018Co-Authors: Buhara Yücesan, Wolfgang Kreis, İsmail Eker, Luiz Eduardo Santos Lazzarini, Noreen Aslam, Aliyu Mohammad, Jose Eduardo Brasil Pereira Pinto, Ekrem GurelAbstract:Digitalis lanata, due to the presence of cardenolides in the leaves, is widely used in drug industry for the treatments of congestive heart diesases. In this present study, natural populations of D. lanata in Thrace region were collected. Thereafter, cardiac glycoside analysis of D. lanata subsp. lanata grown in natural habitats and micropropagation from shoot tips were established as a model work for a consistent and a sustainable plant production. Of the cardenolides, Lan A (ranging between 24.8 and 300.4 mg 100g-1) and Lan C (42.1 and 258 mg 100 g-1 were predominantly found in the samples as compared to lanatoside B, digoxin and digitoxin. After seed germination, shoot-tip cultivation was achieved after a subsequent shoot and root formation on MS medium. Indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) depending on their concentrations were found to be effective in shoot and root formation accordingly. Maximum two shoots from shoot tips were produced after 8 weeks of cultivation, and at the end of 12th weeks of cultivation each shoots produced an average number of 6.4 roots in 8.1 cm length. Acclimatization was also achieved successfully with a maximum survival rate (95 %) for 4 weeks. This study is the first report on cardenolide profiling of Thracian D. lanata at different locations, and shoot tip cultivation of the population selected under in vitro conditions
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cardenolide biosynthesis in light and dark grown digitalis lanata shoot cultures
Plant Physiology and Biochemistry, 1999Co-Authors: Marina Eisenbeis, Wolfgang Kreis, Ernst ReinhardAbstract:Abstract Shoot cultures of the cardenolide-producing species Digitalis lanata Ehrh. accumulated up to 0.6 μmol cardenolides per g dry mass when cultivated under continuous white light. After transfer to permanent dark, the cardenolide content of cultured shoots gradually decreased and reached non-detectable levels after 12 weeks. After transfer back to light conditions, cardenolides started to accumulate and reached the levels of light-grown controls after 4 weeks. Radiolabelled pregnenolone and progesterone were incorporated into cardenolides in both green light-grown and white dark-grown shoots. It was thus established that cardenolides are synthesised de novo in chloroplast-free tissues without apparent cardenolide accumulation, indicating that these compounds are efficiently turned over in the dark and that tissue differentiation, but not intact chloroplasts, is essential for cardenolide formation. The time course of two late anabolic enzymes of cardenolide metabolism, acetyl-CoA:digitoxin 15′-O-acetyltransferase (DAT, EC 2.3.1.-) and UDP-glucose:digitoxin 16′-glucosyltransferase (DGT, EC 2.4.1.-) was established during transfer of shoots from light to dark and vice versa. Only DAT was affected and was not measurable any more under dark conditions. The DGT may not be down-regulated because of its important, maybe even vital, role as an enzyme providing the vacuolar storage forms of cardenolides. Two catabolic cardenolide-specific enzymes, lanatoside 15′-O-acetylesterase (LAE, EC 3.1.1.6.) and cardenolide 16′-O-glucohydrolase I (CGH I, EC 3.2.1.21), were also investigated and it was demonstrated that CGH I is inactive in dark-grown shoots. These observations indicate that CGH I is not involved in cardenolide degradation in situ, but may instead play a role in cardenolide remetabolisation and activation after wounding or in developmental programs.
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effect of digitalis lanata matrix composition on the lanatoside c partition coefficient and its consequence on rotation locular counter current chromatography efficiency
Journal of Chromatography A, 1998Co-Authors: Fernao Castro Braga, Wolfgang Kreis, Alaide Braga De OliveiraAbstract:Lanatoside C standard samples had their partition coefficients (K) determined in dichloromethane-methanol-water (4:4:3), (5:6:4) and (7:13:8), by the shake flask method and the obtained values were similar. On the other hand, K for lanatoside C accomplished by calculation from rotation locular counter-current chromatography (RLCC) traces of Digitalis lanata pre-purified extracts, as well as by the shake flask method, in the same solvent systems, showed a significant variation. Such variation was attributed to differences in the pre-purification procedures for D. lanata extract, which resulted in starting materials for RLCC with very distinct cardenolide profiles, reflecting, thus, in the K values for lanatoside C and, consequently, in the lanatoside C purity at the end of the chromatographic process.
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Avaliação quantitativa de cardenolídeos no cultivar experimental de Digitalis lanata do maciço do itatiaia e perspectivas de seu emprego industrial Quantitative evaluation of cardenolides in a brazilian digitalis lanata cultivar and perspectives of its industrial use
Sociedade Brasileira de Química, 1997Co-Authors: Fernao Castro Braga, Wolfgang Kreis, Rubens Almeida Récio, Alaide Braga De OliveiraAbstract:The content of digoxin and lanatoside C (jointly quantified), lanatoside A, lanatoside B, glucoevatromonoside, odorobioside G, glucogitoroside, glucoverodoxine, glucodigifucoside and digitalinum verum was determined by HPLC in Digitalis lanata harvested in Brazil, as well as in clones industrially employed for the production of cardenolides. The Brazilian plants presented greater variation in the contents of cardenolides than the analyzed clones. Lanatoside C and digoxin concentrations were higher in the clones (6120±640 nmol/g dry leaf) than in the Brazilian plants (1820±900 nmol/g dry leaf). The concentrations of these glycosides were found to be within the range described for native species and also were similar to an European cultivar, what makes possible its industrial use for the cardenolides production
Sandeep Kumar Verma - One of the best experts on this subject based on the ideXlab platform.
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Investigation of Relationship Between Chemical Stress Factors and Certain Metabolites Including Cardenolides in Callus Cultures of Endemic Turkish Digitalis L. Species
International Journal of Secondary Metabolite, 2017Co-Authors: Gunce Sahin, Sandeep Kumar Verma, Ekrem GurelAbstract:The aim of the present research is to obtain relationship between different stress treatments [Cu (copper) and Hg (mercury)] and content of cardiac glycosides (digoxigenin, gitoxigenin, lanatoside C, digoxin and digitoxin) as secondary metabolites of commercial value for the pharmaceutical industry and to determine the antioxidant metabolites against stress conditions in callus cultures of endemic Turkish Digitalis species. The effects of different stress treatments on cardiotonic glycoside accumulation in D. lamarckii Ivanina, D. trojana Ivanina, D. davisiana Heywood and D. cariensis Boiss. ex Jaub. et Spach were investigated using HPLC. HPLC analysis revealed that all stress conditions were significantly effective at 5% significance level according to their control groups. The predominant cardiac glycoside was lanatoside C (Lan C) followed by digitoxin, digoxigenin, gitoxigenin and digoxin. No digoxin was detected in all treatments as well as in control groups. For the calibration curves, concentrations of 5, 10, 20, 30 and 40 mg/l digoxigenin, gitoxigenin, lanatoside C, digoxin and digitoxin were used (R2= 0.99). Cardenolides were eluted with acetonitrile (A) and water (B) gradients as follows: 0 to 20 min 20% (A), 80% (B); 20 to 23.40 min 30% (A), 70% (B); 23.40 to 30 min 25% (A), 75% (B) and 30 to 40 min 40% (A), 60%(B). Average peak area of the glycoside in samples was automatically calculated and monitored by ChemStation LC/MS software against that of standards. Enhanced production of cardenolides was achieved from callus cultures elicited with 50 μm CuSO4 and HgCl2. Higher amounts of cardenolides were obtained when callus of four Digitalis species were elicited with CuSO4. Results demonstrated that catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) activities, the total contents of phenolics and proline were markedly stimulated under stress conditions. All these results indicated that treatments have induced changes in the redox state of callus cells and suggest that this alteration change cardenolides accumulation and antioxidative status in Digitalis L. callus cultures
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hydrogen peroxide induced antioxidant activities and cardiotonic glycoside accumulation in callus cultures of endemic digitalis species
Plant Physiology and Biochemistry, 2014Co-Authors: Gunce Sahin Cingoz, Sandeep Kumar Verma, Ekrem GurelAbstract:Abstract The effect of hydrogen peroxide (H2O2) on callus cultures of four Digitalis species (Digitalis lamarckii, Digitalis trojana, Digitalis davisiana and Digitalis cariensis) increased catalase (CAT), superoxide dismutase (SOD), total phenolic, proline activity and cardiotonic glycoside production. Callus derived from hypocotyl explants was cultured on Murashige and Skoog medium supplemented with 0.25 mg L−1 indole-3-acetic acid (IAA) and 0.5 mg L−1 thidiazuron (TDZ). After a month of culture, callus was transferred to MS medium containing 10 mM H2O2 and then incubated for 6 h. The amount of five cardenolides (Lanatoside C, Digitoxin, Digoxigenin, Gitoxigenin and Digoxin) as well as CAT, SOD, total phenolic, proline activity from Digitalis species were compared. No digoxin was detected in all treatments and control groups. The total cardenolides estimated were in the order of D. lamarckii (586.65 μg g−1 dw), D. davisiana (506.79 μg g−1 dw), D. cariensis (376.60 μg g−1 dw) and D. trojana (282.39 μg g−1 dw). It was clear that H2O2 pre-treatment resulted in an increase in enzymatic and nonenzymatic antioxidants. However, a significant negative relationship between cardenolides production and overall activities of CAT, SOD, total phenolic and proline was evident. The described protocol here will be useful for the development of new strategies for a large-scale production of cardenolides.
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embryogenesis plant regeneration and cardiac glycoside determination in digitalis ferruginea subsp ferruginea l
Plant Cell Tissue and Organ Culture, 2014Co-Authors: Gunce Sahin, Sandeep Kumar Verma, Buhara Yücesan, Ekrem GurelAbstract:The present study reports, for the first time, an efficient in vitro plant regeneration protocol for Digitalis ferruginea subsp. ferruginea L. (rusty foxglove). We have used different concentrations of gibberellic acid (GA3) on Murashige and Skoog (MS) medium to assess the germination frequency of seeds. High frequency of germination was achieved on MS medium with 1.0 mg l−1 GA3. 6-Benzylaminopurine (BAP) combined with α-naphtaleneacetic acid (NAA) or 2, 4-dichlorophenoxy acetic acid (2, 4-D) in the induction MS medium induced both somatic embryogensis and shoot organogenesis. The highest percentage of callus growth (85 %) was obtained when hypocotyl explants were cultured on MS medium containing 0.5 mg l−1 2, 4-D plus 1.0 mg l−1 BAP. The maximum mean number of somatic embryos (7.3 ± 1.3 embryos) or shoots (12.0 ± 1.1 shoots) per callus was obtained when medium contained 0.25 mg l−1 NAA plus 1.0 mg l−1 BAP or 0.5 mg l−1 NAA plus 2.0 mg l−1 BAP. The regenerated shoots easily rooted on MS medium. Higher amounts of lanatoside C [13.2 ± 0.5 mg 100 g−1 dry weight (dw)] and digoxin (2.93 ± 0.31 mg 100 g−1 dw) accumulation were obtained when shoots were obtained by indirect regeneration. We also investigated derivatives of cardenolides, i.e., digitoxigenin (730 ± 180 mg 100 g−1 dw), gitoxigenin (50 ± 20 mg 100 g−1 dw) and digoxigenin (490 ± 170 mg 100 g−1 dw) from natural samples.
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regeneration and cardiotonic glycoside production in digitalis davisiana heywood alanya foxglove
Plant Cell Tissue and Organ Culture, 2011Co-Authors: Ekrem Gurel, Sandeep Kumar Verma, Buhara Yücesan, Esra Aglic, Songul Gurel, Munevver Sokmen, Atalay SokmenAbstract:A high-frequency in vitro regeneration of Digitalis davisiana Heywood (Alanya foxglove) and cardiotonic glycoside production from both in vitro produced materials (regenerated plantlets or germinated seedlings) and leaves of natural populations were obtained. Cardiac glycosides regulate heart rhythm and are effective in cancer chemotherapy, in particular for prostate and breast cancer treatments. Testing six different types of culture media revealed that Linsmaier and Skoog (LS) was the most effective for shoot production. Shoot regeneration efficiency was higher when flamingo-bill or hypocotyl explants were cultured on LS medium containing 0.5 mg/l thidiazuron (TDZ) and 0.25 mg/l indoleacetic acid (IAA). Rooting of all shoots that regenerated was readily achieved, even in the absence of plant growth regulators (PGRs). Production of cardenolides (lanatoside C and digoxin) in the materials grown in vitro seemed to correlate with several parameters, such as nutritional and hormonal compositions of the culture medium as well as the duration of culture on the initial regeneration and/or final growth medium. Higher amounts of digoxin accumulation were obtained when shoots were regenerated on LS or Gamborg’s B5 medium containing 0.5 mg/l TDZ and 0.25 mg/l IAA, producing 12.59 and 11.93 mg/kg dry weight (dw) digoxin, respectively. For natural populations, seasonal variations seemed to affect the production of digoxin in the leaves. The highest amount of digoxin (246.58 mg/kg dw) was in leaf samples collected in July, which coincides with the flowering stage of the plant in the region of collection.
