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In vitro cultures of medicinal of plants - VIII
Špandelová, Veronika ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
In vitro cultures of medicinal of plants - VIII Method of elicitation is one of the possibilities how to increase production of secondary metabolites in cultures in vitro. The effect of abiotic elicitor (3-jodo-4-methylfenyl)amide 5- terc-butyl-6-chloropyrazine-2-karboxylic acid in three concentrations on the flavonolignans and flavonoid taxifolin production in Silybum marianum callus and suspension culture was tested in this study. Elicitation was run in this time intervals: 6, 12, 24, 48, 72 and 168 hours. Cultivation proceeded on Murashige - Skoog medium with the addition of 10 mg/l of α- naphtylacetic acid. The content of flavonolignans and flavonoid taxifolin was determined by HPLC. The maximum content of flavonolignans (0,05%) in callus culture was found after 168 hours of elicitor treatment in concentration of c2 = 2,33 x 10-4 mol/l. The maximum production of silychristin (0,04%) was found after 6 hours of affecting elicitor c1 = 2,33 x 10-3 mol/l. There wasn't statistical significant increasing of flavonolignans and taxifolin production in suspension culture. The maximum production of silybin A (0,01%) in medium of suspension culture was found after 24 hours of elicitor treatment in concentration of c1 = 2,33 x 10-3 mol/l.
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Silybum marianum in vitro-abiotic elicitation.
Adamová, Eva ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Elicitation is the method to increase the production of secondary metabolites in plant tissue. Elicitation by electric field is one of the method of abiotic elicitation to stimulate plants to produce higher amount of secondary metabolites. In vitro cultures of Silybum marianum were cultivated on Murashige-Skoog medium. Suspension cultures were exposed to electric field (50 mA, voltage 5, 7, 8, 10, 15, 20, 24 V) during 10, 30 and 60 minutes. The content of silymarin complex components was measured by (HPLC). The maximal content of taxifolin (0.04 %), silychristin (0.28 %), silydianin (0.06 %) in suspension culture was detected after elicitation by electric field 8V (10 minutes, 6 hours). The maximal content of taxifolin, silychristin, silydianin in suspension culture was detected after elicitation by electric field 8V (60 minutes, 6 hours) and 10V (60 minutes, 24 hours). The significant increase of silymarin complex components were not detectable after the other values of voltage .
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Thesis The effect of flavoinoids and chosen chelators on the oxidation of natural substances
Malá, Marie ; Martin, Jan (advisor) ; Tůmová, Lenka (referee)
Mgr. Marie Malá, Thesis The effect of flavoinoids and chosen chelators on the oxidation of natural substances, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, 2014, 67 pages. Thesis "The effect of flavonoids and chosen chelators on the oxidation of natural substances" deals with Fenton's reaction. For the needs of this work it means the effect of chosen flavonoids and chelators on oxidation of salicylic acid in presence of peroxide and ferrous ions. Chelators were efficient antioxidants at very low concentrations, but inefficient or pro-oxidative at higher concentrations. The most significant antioxidant effect of flavonoid genistein was estabilished at pH 4,15, pH optimum was in range 3,3 - 5,91.
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Explant cultures of Higher plants 27
Kostřiba, Jan ; Dušková, Jiřina (advisor) ; Tůmová, Lenka (referee)
Kostřiba J.: In vitro cultures of higher plants XXVII. Diploma Paper, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, 2007, p. 61 The work aimed at biotransformation of exogenous precursors of arbutin with in vitro cultures of Centella asiatica (L.). The precursors of arbutin were added into the medium (hydroquinone, tyrosine, p-coumaric acid, hydroxybenzoic acid). The precursors were used in a concentration of 200 mg/l and the period of their action was 6, 12, 24, 48 and 168 hours (1 week). The cultivation of calluses was created by two different ways - by suspension culture and by culture cultivated on the filter-paper bridge. The cultivation was prosecuted on the light. Positive results (both TLC and HPLC analysis) in arbutin production were obtained in all suspension cultures after an addition of hydroquinone. The largest amount of arbutin in callus cultures was measured after a 48 hours cultivation (0,043 %). After a 168 hours cultivation was amount of arbutin only 0,038 %. In this experimental variant arbutin was also released into the medium (0,096 %). Biotransformation experiments with the others precursors of arbutin were not successful. The way of cultivation by the filter-paper bridges was also unsuccessful.
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Physical mapping of centromeric and terminal markers of linkage groups in Xenopus tropicalis
Tůmová, Lucie ; Krylov, Vladimír (advisor) ; Nedvídek, Josef (referee)
Xenopus tropicalis belongs together with its related genus Xenopus laevis to important model organisms on the field of study of developmental processes. In comparison to well introduced models (i.e. Melanogaster, Mus etc.) for Xenopus tropicalis there is an absence of linkage and physical maps that have invaluable importance for the identification of developmentally relevant genes, including their syntheny with human. Currently, there exists provisional linkage map constructed by scientific group of Department of Biology and Biochemistry, University of Huston, Texas, USA. It is constituted of ten linkage groups (LG1-LG10) and six linkage clusters (A-F) composed by total of 1713 Simple Sequence Repeat (SSR) polymorphic markers. The laboratory of developmental biology, Charles University in Prague, participates in completing the linkage map by physical mapping of chosen polymorphic markers on chromosomes Xenopus tropicalis, which led to allocation of each linkage group and cluster to the right chromosome and to assessment of the orientation in most of them. The physical mapping of terminal markers will show us, how far chromosomes are covered with linkage groups and which areas still remain unmapped. The cooperative laboratory from National Institute for Medical Research, London, UK, originated a list of...
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Secondary metabolites of plant cultures in vitro II
Pakánová, Alica ; Kašparová, Marie (advisor) ; Tůmová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Student: Alica Pakánová Supervisor: PharmDr. Marie Kašparová, Ph.D. Title of diploma thesis: Secondary metabolites of plant cultures in vitro II Diploma thesis is focused on callus and suspension plant cultures of Juniperus virginiana 'Glauca' and their production of secondary metabolite. Produced content of podophyllotoxin was observed during 15 subcultivations for both types of cultures. The highest content of podophyllotoxin (0.060 %) was established in the 18th subcultivation of suspension culture derivated from the three-years-old callus culture and then subcultivated in the period of 21 days. The maximal content of podophyllotoxin produced by callus culture (0.0515 %) was found out in the 46th subcultivation. Simultaneously it represented the last observed subcultivation subcultivated in the period of 28 days. This research shows that nor the production of Juniperus virginiana callus culture neither the production of suspension culture is stopped by increased number of subcultivation. Keywords: Suspension cultures, callus cultures, production of secondary metabolites, Juniperus virginiana, podophyllotoxin.
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The Quality of the Dietary Suplements with Content Skullcap
Dvořáková, Lenka ; Martin, Jan (advisor) ; Tůmová, Lenka (referee)
Name of the work: The quality of the dietary suplements with content skullcap Author: Lenka Dvořáková The work was targered on the determination of main flavonoids from skullcap and most often incident flavonoids in the other herbs, which have been by part of polykomponents dietary supmements analyzed in this work. Three extracts from each preparation were prepared under different conditions. The highest amount of flavonoids was achieved in most of cases by extraction in ultrasonic bath and for using lower temperature. These extracts were prepared from capsulae durae. By contraries by extract prepared from capsulae moles, were measured levels (values) higher by extract prepared by using of water bath. Between companies were large differences in measured values and so informations featured on covers were too differently large.
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Explant cultures of Higher plants 29
Faltýnková, Jana ; Dušková, Jiřina (advisor) ; Tůmová, Lenka (referee)
Faltýnková, J.: Explant cultures of higher plants 29. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, 2008 The goal ofmythesis was to verify biotransformation posibility of theDatura meteloides culture. Hydroquine - the precursor of arbutin was added into these cultures. Precursor used in this study was prepared in concentration of 100 mg/l and 200 mg/l. Exposition of precursor was 24, 48, 72, 96, 120, 144 and 168 hours under constant conditions. By the qualitative and quantitative analysis I found the presence of studyied arbutin metabolite in all observed samples. The biggest arbutin volume was obtained after 168 hours of cultivation with precursor of concentration at 200 mg/l ( 3,05%). The in_uence of precursor's concentration and exposition time to the _nal product was thereby con_rmed.
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In vitro cultures of medicinal of plants - X
Maliňáková, Lenka ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
In vitro cultures of medicinal plants - X Aim of this study was to study the effect of abiotic elicitation on production of rutin in callus and suspension culture of Fagopyrum esculentum Moench. Cerium chloride (CeCl3) was used as elicitor at three different levels of concentration. The content of rutin in cell cultures was determined by HPLC. Cell cultures were cultivated on Murashige - Skoog medium with the addition of 10 mg/ml of 2,4-D as a growth regulator. No rutin was produced in both types of culture without the presence of elicitor. The maximal content of rutin in callus culture was determined after 6 hours of treatment with elicitor solution with concentration c1 (4,057.10-3 mol/l.). No production of rutin was observed in suspension culture.
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Antioxidant activity of some cultivars of Sambucus nigra L. fruits.
Nehybová, Monika ; Spilková, Jiřina (advisor) ; Tůmová, Lenka (referee)
8. Abstract The fruit of the European elder, Sambucus nigra L., is used in therapy as a drug Sambuci fructus for its diaphoretic, diuretic and laxative effects. Cultivars of European elder are being studied for the use in the food industry and pharmacy. The fruit contains especially large amount of anthocyanins and flavonoids that are known for their antioxidant activity. The aim of this work is to measure the antioxidant activity of the drug Sambuci fructus and of the fruit of the European elder cultivars Samdal, Mammut, Sambo, Sambu, Sampo, Bohatka, Haschberg, Weihenstephan, Allesö, and Samyl. An extract containing anthocyanins was obtained from fruit by a process based on extraction of frozen material with acidified water at high temperature, followed by separation with polymer non-ionogenic polystyrene-divinylbenzene sorbent. The content of anthocyanins in the extracts was determined spectrophotometrically using a modified pH-differential method. The absorbance was measured along the range of 350 - 600 nm. The content of the anthocyanins was quantified in units of mg of cyanidin 3-glucoside in 1 ml of extract. The content ranged from 0.33 - 2.76 mg cyanidin 3-glucoside/ml of extract. The highest content was in the extract from the fruit of the cultivar Samdal and the lowest was of the cultivar Mammut....
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