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Explant culture of Juniperus virginiana
Předota, Václav ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
1 ABSTRACT Charles University in Prague Faculty of pharmacy in Hradec Králové Department of pharmacognosy Student: Václav Předota Supervisor: PharmDr. Marie Kašparová, PhD. Title of diploma thesis: Plant tissue culture of Juniperus virginiana The derivation of callus cultures from leaves of Juniperus virginiana (varieties Hetzii, Glauca and Grey Owl) and determination of their growth curves were studied in this work. The cultures were cultivated at the temperature of 25 řC and light period of 16 hours light/8 hours dark on the Schenk and Hildebrandt medium with the addition of 3.0 mg.l-1 α-NAA and 0.2 mg. l-1 kinetin. It is clear from the growth curves, that all three varieties reached their maximum in growth on 35th day of the cultivation. The best results were achieved by variety Glauca.
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Secondary metabolites of plant tissue culture of Trifolium pratense L.
Novotná, Hana ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Hana Novotná Supervisor: PharmDr. Marie Kašparová, Ph.D. Title of diploma thesis: Secondary metabolites of plant tissue culture of Trifolium pratense L. Explant cultures are perspective sources of secondary metabolites. Nevertheless production of flavonoids and isoflavonoids by the suspension culture of Trifolium pratense L. is not high. Elicitation is one of the methods used to enhance the biosynthesis of secondary metabolites. Elicitation induces physiological changes, stimulates defensive or stress-induced reactions in plants and subsequently triggers the synthesis of secondary metabolites. The objective of this study was to observe the influence of two elicitors - abscisic acid and ascorbic acid - on the production of flavonoids and isoflavonoids by the Trifolium pratense L. suspension culture (Sprint variety). The culture was cultivated in Gamborg medium to which 2 mg.l-1 of 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine were added, at the temperature of 25 řC and 16 hours light / 8 hours dark period. The best elicitation effect of abscisic acid on the production of flavonoids and isoflavonoids was observed after a 6-hour application of the highest 500 µmol.l-1...
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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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Explant culture of Juniperus virginiana
Předota, Václav ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
1 ABSTRACT Charles University in Prague Faculty of pharmacy in Hradec Králové Department of pharmacognosy Student: Václav Předota Supervisor: PharmDr. Marie Kašparová, PhD. Title of diploma thesis: Plant tissue culture of Juniperus virginiana The derivation of callus cultures from leaves of Juniperus virginiana (varieties Hetzii, Glauca and Grey Owl) and determination of their growth curves were studied in this work. The cultures were cultivated at the temperature of 25 řC and light period of 16 hours light/8 hours dark on the Schenk and Hildebrandt medium with the addition of 3.0 mg.l-1 α-NAA and 0.2 mg. l-1 kinetin. It is clear from the growth curves, that all three varieties reached their maximum in growth on 35th day of the cultivation. The best results were achieved by variety Glauca.
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Secondary metabolites of plant tissue culture of Trifolium pratense L.
Novotná, Hana ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Hana Novotná Supervisor: PharmDr. Marie Kašparová, Ph.D. Title of diploma thesis: Secondary metabolites of plant tissue culture of Trifolium pratense L. Explant cultures are perspective sources of secondary metabolites. Nevertheless production of flavonoids and isoflavonoids by the suspension culture of Trifolium pratense L. is not high. Elicitation is one of the methods used to enhance the biosynthesis of secondary metabolites. Elicitation induces physiological changes, stimulates defensive or stress-induced reactions in plants and subsequently triggers the synthesis of secondary metabolites. The objective of this study was to observe the influence of two elicitors - abscisic acid and ascorbic acid - on the production of flavonoids and isoflavonoids by the Trifolium pratense L. suspension culture (Sprint variety). The culture was cultivated in Gamborg medium to which 2 mg.l-1 of 2,4- dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine were added, at the temperature of 25 řC and 16 hours light / 8 hours dark period. The best elicitation effect of abscisic acid on the production of flavonoids and isoflavonoids was observed after a 6-hour application of the highest 500 µmol.l-1...
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Optimization of the growth of plant tissue culture of Juniperus virginiana
Damborská, Vendula ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Student: Vendula Damborská Supervisor: PharmDr. Marie Kašparová, Ph.D. Title of diploma thesis: The Optimization of the growth of the plant tissue culture of Juniperus virginiana The main goal of this diploma thesis lies in the optimization of cultivation medium composition for the tissue culture of Juniperus virginiana (varieties 'Hetzii', 'Glauca', 'Grey Owl'). The best results of all tested media were achieved by using Schenk and Hildebrandt medium with an addition of α-NAA (3.0 mg.l-1 ), kinetin (0.2 mg.l-1 ) and ascorbic acid (15.0 mg.l-1 ). The cultures were cultivated at the temperature of 25 řC and light period of 16 hours light/8 hours dark. The growth curve for the tissue culture of Juniperus virginiana 'Glauca' was set for 2 different cases: with and without the addition of phenylalanin (biogenetic precursor of phenylpropans biosynthesis). The maximal growth was reached on 35th day of the cultivation in both cases, phenylalanin increases the growth rate by 29.65 %.
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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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