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The Influence on Production of Secondary Metabolits in Tissue Culture Hypericum perforatum (L.)
Machačková, Jana ; Martin, Jan (advisor) ; Tůmová, Lenka (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Farmacognosy Candidate Mgr. Jana Machačková Consultant PharmDr. Jan Martin, Ph.D. Title of Thesis The influence on production of secondary metabolits in tissue culture Hypericum perforatum (L.) Using of precursors is one of the biotechnological methods, where precursors are added to the culture medium. Callus and suspensions cultures of Hypericum perforatum L. were used. Fenylalanin and cinnamoyl acid in concentrations 20,0 g/l, 2,0 g/l and 0,2 g/l, resp. 25,0 g/l, 2,5 g/l and 0,25 g/l were used as precursors. The effect of precursors on the production of flavonoids was evaluated after 4, 24 and 168 hours. Cultures were cultivated on Murashige and Skoog medium with addition 2 mg/ml growthing hormon BAP. HPLC method for analysis was used. Generally, the highest increase of flavonoids was made by precursor fenylalanin in concentration B in callus and suspensions cultures. The top in production of quercitrin was detected in suspension cultures after application fenylalanin in concentration C after 168 hours cultivation (from 0,16 % to 0,22 %). Hyperosid was the most increased by fenylalanin in concentration C after 4 hours cultivation (from 0,24 % to 0,35 %). Cinnamoyl acid as precursor is not suitable for increase...
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Transport mechanisms of secondary metabolites across membranes of plant cells
Kubeš, Jan ; Tůmová, Lenka (advisor) ; Babula, Petr (referee) ; Obložinský, Marek (referee)
Faculty of Pharmacy in Hradec Králové, Charles University Department of pharmacognosy Candidate: Mgr. Jan Kubeš Supervisor: PharmDr. Lenka Tůmová, CSc. Title of Doctoral thesis: Transport mechanisms of secondary metabolites across membranes of plant cells An attention was focused on the metabolism of isoflavones in recent years, including their transport within plants and cultures in vitro. A research is focused on the transport mechanism identification across cell or vacuolar membrane. There are localized different transporters as or the transport of some metabolites can take place through membrane vesicules. The release of five isoflavones (genistin, genistein, biochanin A, daidzein and formononetin) into nutrient medium was observed in suspension culture of Genista tinctoria L and Trifolium pratense L. These cultures were treated with NH4VO3 solutions (1 a 10 M concentration) when isoflavon content in medium was higher after 24 hours. This increased volume of isoflavones was supressed by different transport mechanism inhibitors. The transport of isoflavones in T. pratense was influenced by ABC inhibitors from MRP subfamily, but genistein concentration in medium was lower after treatment with MDR subfamily inhibitors. Brefeldin A, which blocks vesicular transport, also decreased concentration of...
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Explant cultures of Higher plants 32
Macháčková, Hedvika ; Dušková, Jiřina (advisor) ; Tůmová, Lenka (referee)
Machackova H.: Explant cultures of Higher plants 32. Thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Kralove, Department of Botanic and Environmental Science, Pharmacy (2010) This paper focused on the study of biotransformation activity of suspension culture Datura meteloides DC. ex Dunal after addition of arbutin exogenous precursor hydroquinone into nutrient medium. The percentage of arbutin content was tested depending on the type and concentration of growth regulator in nutrient medium (IAA, IBA - concentrations of 0,1; 1,0; 10,0 mg/l), concentration of arbutin precursor hydroquinone (100 and 200 mg/l) and intervals between collection of samples for analysis, i.e. cultivation time with precursor (24, 48, 168 hours). Culture transformed hydroquinone into arbutin regardless of the type and concentration of growth regulator, precursor concentration and the length of the experiment. The highest percentage of arbutin content in culture extract (8,11 %) was detected after 24 hour cultivation using hydroquinone in concentration of 200 mg/l and growth regulator IAA in concentration of 0,1 mg/l. The presence of arbutin was also detected in nutrient medium after 48 and 168 hours (0,68 %). In most cases, cultures using hydroquinone in concentration of 200 mg/l had higher production of...
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Integration programmes for immigrant children in the Czech Republic and Great Britain
Tůmová, Lenka ; Ženíšek, Jakub (advisor) ; Farrell, Mark Robert (referee)
This bachelor thesis deals with immigration integration politics and programmes of the Czech Republic and the Great Britain, with the main focus on the integration of immigrant children into the society and education system. The work is divided into two parts: each one gives information about Czech and British immigration history and development, integration policies, and the main issues that each of the country is dealing with. All the information and data used in the text come from the latest research and from updated source documents.
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Hypericum perforatum in vitro- abiotická elicitace
Majerová, Jitka ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Farmacognosy Candidate Mgr. Jitka Majerová Consultant Doc. PharmDr. Lenka Tůmová, CSc. Title of Thesis Hypericum perforatum in vitro- abiotická elicitace The effect of cerium chloride at 3 different levels of concentration (4,0571x 10-3 mol/l, 4,0571x 10-4 mol/l, 4,0571x 10-5mol/l) on production of hyperoside in cell and suspension culture of Hypericum perforatum L. was studied. The sample were taken after elicitation in fixed time period 0, 6, 12, 24, 48, 72 and 168 hours after elicitation. The colntrol sample (without elicitor treatment) ware taken after 24 and 168 hours. The sample ware analysed by HPLC method. Cerium chloride influenced hyperoside production in suspension and callus cultures. The highes content of hyperosid (0,27 mg/g DW) ware reached in suspension culture after cerium chloride treatment at concentration of c2 (4,0571x 10-4 mol/l) after 48 hours. Callus and suspension cultures did not release any hyperoside into nutrient medium.
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The Effect of Elicitation on Production of Secondary Substances in Cultures in vitro
Špandelová, Veronika ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
The effect of elicitation on production of secondary substances in cultures in vitro Plant secondary metabolites are important sources for pharmaceutical, chemical and food industry. Elicitation is one of the possibilities how to increase the production of secondary subtances in cultures in vitro. The effect of abiotic elicitor 2- (3-brombenzylsulfanyl)pyridine-4-karbothioamide with antituberculotic activity on the isoflavonoids production Genista tinctoria callus and suspension culture was tested in this study. The elicitor was applied in three concentrations. Elicitation was run in this time intervals: 6, 12, 24, 48, 72 and 168 hours. Cultures were cultivated on nutrient medium, that described Shenk and Hildebrant with the addition of 0,5 mg/l of 2,4- dichlorfenoxyacetic acid and 0,1 mg/l of kinetin. The content of isoflavonoids was determined by HPLC. The highest content of isoflavonoids (0,03 %) in callus culture was found after the application of elicitor's solution in concentration of c3 = 2,96 x 10-5 mol/l after 24 hours of treatment. The maximum production of daidzein (0,02 %) in callus culture was detected after 48 hours of elicitor treatment in concentartion of c1 = 2,96 x 10-3 mol/l and after 24 hours of elicitor treatment in concentration of c3 = 2,96 x 10-5 mol/l. Suspension culture...
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In vitro cultures of medicinal plants - XI.
Sojková, Kristýna ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
In vitro cultures of medicinal plants - XI. Elicitation is one of the few strategies that can be used in enhancement of secondary metabolites production from explant cultures. The effect of abiotic elicitor (silver nitrate) on flavonolignan and flavonoid taxifolin production in suspension culture of Silybum marianum L. (Gaertn.) and on isoflavones production in suspension culture of Genista tinctoria L. was tested. Silver nitrate in various concentrations (5.887.10-3 mol/l; 5.887.10-4 mol/l; 5.887.10-5 mol/l) was used as elicitor. Content of secondary metabolites in suspension cultures was determinated by high performance liquid chromatography (HPLC). The samples were taken after 6, 12, 24, 48, 72 and 168 hours after elicitor treatment. The highest content of taxifolin production (0.02 %) in suspension culture of Silybum marianum L. (Gaertn.) after silver nitrate (5.887.10-4 mol/l) treatment and 72 h sampling was detected. The highest content of genistin (0.05 %) in suspension culture of Genista tinctoria L. was found after silver nitrate (5.887.10-4 mol/l) treatment and 48 h sampling. The highest content of daidzein (0.09 %) was detected after elicitor (5.887.10-4 mol/l) treatment and 168 h sampling.
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Phytochemical study of individual plant species of Bergenia genus
Hendrychová, Helena ; Tůmová, Lenka (advisor) ; Nagy, Milan (referee) ; Šmejkal, Karel (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Mgr. Helena Hendrychová Supervisor: doc. PharmDr. Lenka Tůmová, CSc. Title of Doctoral Thesis: Phytochemical study of individual plant species of Bergenia genus Bergenia, native to central Asia, is promising source of medicinal compounds mainly in the system of Chinese and Indian traditional medicine. These species have been used for centuries in therapy of various diseases. They are valued for their ability to dissolve kidney or bladder stones. Bergenia plants are used for therapy of pulmonary diseases and cold. They have demonstrated antioxidant, antibacterial, antiviral, immunostimulant, anti-inflammatory, anticancer, antidiabetic, antitussive, hepatoprotective and diuretic effects. The main aim of this thesis was to determine the content of secondary metabolites (arbutin, total polyphenols, bergenin and anthocyanins) in three taxons of Bergenia genus - Bergenia crassifolia (L.) Fritsch, Bergenia ciliata (Haw.) Sternb. a Bergenia x ornata Stein., to compare their biological activities and to identify the most appropriate kind of Bergenia plants with the greatest potential for use in therapy. Plant hybrid B. x ornata was evaluated on the content of phenolic compounds and biological activity...
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The effect of pyrazine derivatives on secondary metabolites in in vitro cultures
Kovalíková, Zuzana ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
Vplyv derivátov pyrazínu na tvorbu sekundárnych metabolitov v rastlinných kultúrach in vitro - Zuzama Kovalíková - 71 - 11 Abstract Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Mgr. Zuzana Kovalíková Consultant: doc. PharmDr. Lenka Tůmová, CSc. Title of Thesis: The effect of pyrazine derivatives on secondary metabolites in in vitro cultures. This thesis investigates the effect of new synthetic pyrazine derivatives as potential elicitors on rutin production in callus and suspension cultures of Fagopyrum esculentum, var. Bambi. All tested chemical elicitors increased the production of rutin in both callus and suspension cultures. Callus and suspension culture was cultivated in Murashige and Skoog nutrient medium containing growth regulator (2,4-D) under the specified temperature and light conditions. The samples were taken in 6, 12, 24, 48, 72 and 168 hours after elicitor exposition, control samples were taken in 24 and 168 hours after exposition. The determination of the rutin content was performed by high performance liquid chromatography (HPLC). Elicitor P.G.R.1, propyl 5-(3-(4.chlorophenyl)ureido)pyrazine-2-carboxylate showed the highest increase in rutin production after 12 hours treatment in the callus culture, whereas the highest increase in...
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Possibilities of affecting of secondary metabolites production in culture Silybum marianum in vitro
Kunzová, Lucie ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
1 ABSTRAKT Mgr. Lucie Kunzová Possibilities of affecting of secondary metabolites production in culture Silybum marianum in vitro Elicitation is the method making use defensive mechanism of plants to increasing poduction of secondary metabolites in plants and cultures in vitro. The effect of 6, 12, 24, 48, 72 and 168 hours influence by three concentrations of the abiotic elicitor (3-jodo-4-methylfenyl)amide 5-methylpyrazine-2-karboxylic acid on the flavonolignans production in Silybum marianum callus and suspension culture was monitored in this study. The in vitro culture was cultivated on Murashige-Skoog medium with the addition of 10 mg/l of α-naphtylacetic acid as a growth regulator. The content of flavonolignans was determined by HPLC. The maximum content of flavonolignans (0,03 %) in callus culture was demostrated after 24 hours of elicitation (c1 = 2,83 · 10-3 mol/l). The maximum content of flavonolignans (0,04 %) in suspension culture was demonstrated after 48 hours of elicitation (c3 = 2,83 · 10-5 mol/l).
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