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Study of secondary metabolites in explantat culture of Trifolium pratense L.
Jindřišková, Zuzana ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Zuzana Jindřišková The Study of Secondary Metabolites in Explant Culture of Trifolium pratense L. The basic prerequisite for a successful elicitation that is used to increase the production of secondary metabolites is, among others, finding a suitable elicitor, its concentration and optimal duration of effect of the elicitor on the plant in vitro culture, which was the main subject of this diploma thesis. The focus of our observations was the influence of 6-, 24- , 48- and 168-hour effect of nickel chloride solution (in the concentrations of 0.1 mmol, 1 mmol, 10 mmol and 100 mmol) and zinc sulphate (in the concentrations of 0.1 µmol, 1 µmol, 10 µmol a 100 µmol) on the production of flavonoids and isoflavonoids in the suspension culture of Trifolium pratense L. variety Tempus. The culture was cultivated on the Gamborg nutrien medium with the addition of 2 mg.l-1 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 6- benzylaminopurine at 25řC and the light period of 16 hours light/ 8 hours dark. The maximum content of flavonoids, which was found out by the photometric determination of the Czech Pharmacopoeia 2009, was proved in the suspension culture of Trifolium pratense L. variety Tempus (0.406%) after 48-hour elicitation of nickel chloride solution in the concentration of 0.1 mmol, when there was a...
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Effect of 2-benzylthiopyridine-4-carbothioamide derivatives on isoflavonoids and flavonoids accumulation in suspension culture of Trifolium pratense L.
Machová, Veronika ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
1 Abstract Veronika Machová Effect of 2-benzylthiopyridine-4-carbothioamide derivatives on isoflavonoids and flavonoids accumulation in suspension culture of Trifolium pratense L. The paper examinate the effect of three concentration of 2-(2-fluoro-6- nitrobenzylsulfanyl) pyridine-4-carbothioamide on the production of flavonoids and isoflavonoids by the Trifolium patense L. (variety DO-8, variety Tempus). The culture were cultivated in the Gamborg nutrient media with addition of 2 mg·l-1 2,4- dichlorophenoxyacetic acid a 2 mg·l-1 6-benzylaminopurine, at the temperature of 25řC, 16-hr light/8-hr dark period. The elicited and the inspection samples underwent the photometric determination of flavonoids in accordance with the Czech Pharmacopoeia 2009 ant the determination of isoflavonoids via the HPLC method. The results show that the maximum content of flavonoids in suspension cultures of Trifolium pratense L. (variety DO-8) was caused by 48-hour elicitation by 100 µmol·l-1 solution of 2-(2-fluoro-6-nitrobenzylsulfanyl) pyridine-4-carbothioamide, when the a statistically significant increase in production was 93% compared to control cultures. The maximum content of flavonoids in variety Tempus was caused by 48-hour elicitation by 10 µmol·l-1 solution of 2-(2-fluoro-6-nitrobenzylsulfanyl) pyridine-4-...
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Influencing of production of plant tissue culture of Trifolium pratense L. II
Částková, Lucie ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Lucie Částková Influencing of production of plant tissue culture of Trifolium pratense L. II The aim of this thesis was to observe the effect of the abiotic elicitor vanadyl sulfate on the production of flavonoids in the suspension culture of Trifolium pratense L., variety DO-8 and variety Sprint. The effects on the production of isoflavonoids caused by the elicitor have been also studied. The cultures were cultivated at the temperature of 25 žC, 16 hour light/8 hour dark period, in the Gamborg's nutriet medium with the addition of 2 mg.l-1 of 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine. Vanadyl sulfate was added in four concentrations and its influence was observed after 6, 24, 48 and 168 hours. The quantity of flavonoids was determined spectrophotometrically according to the Pharmacopoeia Bohemica 2009. The quantity of isoflavonoids was determined using HPLC. The maximal content of flavonoids, 0,459 %, was measured in the suspension culture of Trifolium pratense L., variety DO-8, taken after 48 hours of action of elicitor vanadyl sulfate, which concentration was 1 µmol.l-1 . The statistically significant increase in production was 106 % compared to the control. The production of flavonoids in variety Sprint was most influenced by the action of vanadyl sulfate, which...
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Influencing of production of plant tissue culture of Trifolium pratense L. I
Muráriková, Kateřina ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
1 ABSTRACT Kateřina Muráriková Influencing of production of plant tissue culture of Trifolium pratense L. I The aim of this thesis was to check the influence of cinnamic acid, as a precursor of the phenylpropanoids biosynthesis, on the flavonoids production of Trifolium pratense L. suspension culture, to compare two varieties of this plant, the DO-8 one and the Tempus one, in terms of the effect of this precursor, and to help achieve the successful increase of the production of secondary metabolites by the Trifolium pratense L. explant cultures. Concerning the DO-8 variety, the changes of the isoflavonoids production as a result of the cinnamic acid influence have been also tested. The suspension cultures were being cultivated on the Gamborg's nutrient medium with the addition of 2 mg.l-1 2,4-dichlorphenoxyacetic acid and 2 mg.l-1 6-benzylaminopurine. The cultivation was being realised on a slow moving roller at temperature of 25 řC and in the period of 16 hours of light and 8 hours of dark. The cinnamic acid was added in the four concentrations and its influence was observed after 6, 24, 48 and 168 hours. The flavonoids determination was being applied on the check samples and the samples with the addition of cinnamic acid by the spectrophotometric method in agreement with the Český lékopis 2009 and the...
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Influencing of production of plant tissue culture of Trifolium pratense L. II
Částková, Lucie ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Lucie Částková Influencing of production of plant tissue culture of Trifolium pratense L. II The aim of this thesis was to observe the effect of the abiotic elicitor vanadyl sulfate on the production of flavonoids in the suspension culture of Trifolium pratense L., variety DO-8 and variety Sprint. The effects on the production of isoflavonoids caused by the elicitor have been also studied. The cultures were cultivated at the temperature of 25 žC, 16 hour light/8 hour dark period, in the Gamborg's nutriet medium with the addition of 2 mg.l-1 of 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine. Vanadyl sulfate was added in four concentrations and its influence was observed after 6, 24, 48 and 168 hours. The quantity of flavonoids was determined spectrophotometrically according to the Pharmacopoeia Bohemica 2009. The quantity of isoflavonoids was determined using HPLC. The maximal content of flavonoids, 0,459 %, was measured in the suspension culture of Trifolium pratense L., variety DO-8, taken after 48 hours of action of elicitor vanadyl sulfate, which concentration was 1 µmol.l-1 . The statistically significant increase in production was 106 % compared to the control. The production of flavonoids in variety Sprint was most influenced by the action of vanadyl sulfate, which...
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Effect of 2-benzylthiopyridine-4-carbothioamide derivatives on isoflavonoids and flavonoids accumulation in suspension culture of Trifolium pratense L.
Machová, Veronika ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
1 Abstract Veronika Machová Effect of 2-benzylthiopyridine-4-carbothioamide derivatives on isoflavonoids and flavonoids accumulation in suspension culture of Trifolium pratense L. The paper examinate the effect of three concentration of 2-(2-fluoro-6- nitrobenzylsulfanyl) pyridine-4-carbothioamide on the production of flavonoids and isoflavonoids by the Trifolium patense L. (variety DO-8, variety Tempus). The culture were cultivated in the Gamborg nutrient media with addition of 2 mg·l-1 2,4- dichlorophenoxyacetic acid a 2 mg·l-1 6-benzylaminopurine, at the temperature of 25řC, 16-hr light/8-hr dark period. The elicited and the inspection samples underwent the photometric determination of flavonoids in accordance with the Czech Pharmacopoeia 2009 ant the determination of isoflavonoids via the HPLC method. The results show that the maximum content of flavonoids in suspension cultures of Trifolium pratense L. (variety DO-8) was caused by 48-hour elicitation by 100 µmol·l-1 solution of 2-(2-fluoro-6-nitrobenzylsulfanyl) pyridine-4-carbothioamide, when the a statistically significant increase in production was 93% compared to control cultures. The maximum content of flavonoids in variety Tempus was caused by 48-hour elicitation by 10 µmol·l-1 solution of 2-(2-fluoro-6-nitrobenzylsulfanyl) pyridine-4-...
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Study of secondary metabolites in explantat culture of Trifolium pratense L.
Jindřišková, Zuzana ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Zuzana Jindřišková The Study of Secondary Metabolites in Explant Culture of Trifolium pratense L. The basic prerequisite for a successful elicitation that is used to increase the production of secondary metabolites is, among others, finding a suitable elicitor, its concentration and optimal duration of effect of the elicitor on the plant in vitro culture, which was the main subject of this diploma thesis. The focus of our observations was the influence of 6-, 24- , 48- and 168-hour effect of nickel chloride solution (in the concentrations of 0.1 mmol, 1 mmol, 10 mmol and 100 mmol) and zinc sulphate (in the concentrations of 0.1 µmol, 1 µmol, 10 µmol a 100 µmol) on the production of flavonoids and isoflavonoids in the suspension culture of Trifolium pratense L. variety Tempus. The culture was cultivated on the Gamborg nutrien medium with the addition of 2 mg.l-1 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 6- benzylaminopurine at 25řC and the light period of 16 hours light/ 8 hours dark. The maximum content of flavonoids, which was found out by the photometric determination of the Czech Pharmacopoeia 2009, was proved in the suspension culture of Trifolium pratense L. variety Tempus (0.406%) after 48-hour elicitation of nickel chloride solution in the concentration of 0.1 mmol, when there was a...
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