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The effect of pyrazine derivatives on secondary metabolites content in plant cultures in vitro - III.
Blahnová, Kristýna ; Tůmová, Lenka (advisor) ; Karlíčková, Jana (referee)
One of the possibilities how to increase the production of secondary metabolites (SM) in plant in vitro cultures is the method of elicitation. The aim of this work was to determine the effect of an abiotic elicitor from the class of pyrazine derivatives 1-benzyl-3-(pyrazin-2-yl)urea on the production of flavonolignans of silymarin complex and flavonoid taxifolin in the plant culture Silybum marianum (L.) Gaertn. Pyrazine derivatives are investigated for their herbicidal properties, so elicitation proceeds by the mechanism of stress inducing effect. Tissue cultures were grown on Murashige and Skoog growth medium with the addition of the growth regulator α-naphthylacetic acid. Elicitation was performed on both callus and suspension cultures. The elicitor was used in three different concentrations: c1 = 100.0 mg/100 ml; c2 = 10.0 mg/100 ml; c3 = 1.00 mg/100 ml. Particular samples were taken after 6, 24, 48, 72 and 168 hours of elicitor effect, control samples after 6, 48 and 168 hours. After drying, the callus and suspension tissues were extracted with methanol and the content of the monitored secondary metabolites was determined by HPLC. It was also tested if SM are released into the growth medium. Flavonolignans silybinin A and silybinin B were not detected in any of the analyzed samples. The...
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Constituents of Rosmarinus officinalis and their biological effects
Klížová, Aneta ; Karlíčková, Jana (advisor) ; Tůmová, Lenka (referee)
CHARLES UVIVERSITY THE FACULTY OF PHARMACY IN HRADEC KRÁLOVÉ DEPARTMENT OF PHARMACOGNOSY AND PHARMACEUTICAL BOTANY Title of the diploma thesis: CONSTITUENTS OF ROSEMARY AND THEIR BIOLOGICAL EFFECTS Candidate: Aneta Klížová Supervisor: PharmDr. Jana Karlíčková, Ph.D. Diploma thesis 2021/2022, pp. 89 Rosemary (Rosmarinus officinalis L.) belongs to the Lamiaceae family. It is a bushy, evergreen perennial plant coming from the Mediterranean. Two pharmacopoeia herbal substances are used from this plant - leaves and essential oil. Rosemary has a wide range of uses. It is mainly used as a spice in the kitchen. Due to its antioxidant effects, it is used as a food preservative. It is also added to various cosmetic products such as soaps, creams, shampoos and perfumes. Rosemary and its components (carnosol, carnosic acid and rosmarinic acid and others) show many effects, which include analgesic, antioxidant, antiinflammatory, anticancer, antirheumatic, spasmolytic, antihepatotoxic, carminative, atherosclerotic, choleretic, antidepressant effects and others. It also works against bacteria and viruses. Rosemary is widely used as a traditional medicine, so it is included in the EMA monograph. Extract of rosemary or substances isolated from it have been tested in many in vitro and in vivo studies. Essential oil...
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Evaluation of flavonoids content in various species of Bergenia genus
Marková, Almíra ; Tůmová, Lenka (advisor) ; Karlíčková, Jana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany Author: Almíra Marková Supervisor: Assoc. Prof. PharmDr. Lenka Tůmová, CSc. Title of diploma thesis: Evaluation of flavonoids content in various species of Bergenia genus Originally an Asian genus Bergenia from the family Saxifragaceae includes 32 species described so far and many hybrids that were created by crossing them. The leaves and rhizomes of bergenia used widely in folk medicine of China, India, Mongolia, and Russia are rich in a wide range of chemical constituents. The most important secondary metabolites of this plant are tannins, arbutin, bergenin, but also flavonoids. The drug is valued for its litholytic, antioxidant, anti-inflammatory, antipyretic, or antitussive properties. This work aimed to determine flavonoid content in red and green leaves of Bergenia crassifolia (L.) Fritsch, Bergenia ciliata (Haw.) Sternb. and Bergenia x ornata Guill. and their comparison with each other depending on the harvest period and climatic conditions. The samples were obtained from the botanical garden of the Faculty of Horticulture of Mendel University in Lednice. Flavonoid content was determined spectrophotometrically. The highest flavonoid content was measured in B. ciliata (1.1829 ±...
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Isolation and biological activity of the pigment myxoxanthophyll from microalgæ.
Nováková, Michaela ; Tůmová, Lenka (advisor) ; Kubáč, David (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate Mgr. Michaela Nováková Supervisor doc. PharmDr. Lenka Tůmová, Ph.D. Co-supervisor Dr. José Carlos Cheel Horna, Ph.D. Title of PharmDr. Thesis: Isolation and biological activity of the pigment myxoxanthophyll from microalgae Global demand for natural pigments has increased in the past few years. Myxoxanthophyll, a glycosylated monocyclic carotenoid, is a pigment that occurs naturally in cyanobacteria but any scalable isolation process has not yet been developed to obtain it from its natural source. In this study, myxoxanthophyll was isolated from unicellular cyanobacterium Synechocystis salina (S. salina) using high performance countercurrent chromatography (HPCCC), where the lower phase of the biphasic solvent system composed of n-heptane-ethanol-water (2:4:4, v/v/v) was used as a mobile phase, whereas its upper phase was the stationary phase. For the HPCCC isolation, a multi-injection method was developed, and four consecutive sample injections (70 mg per each) were performed using 20 mg of myxoxanthophyll, which was finally purified with high performance liquid chromatography (HPLC). Overall, a final myxoxanthophyll yield of 15 mg (98% purity) was obtained. The target pigment showed weak antioxidant and...
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In vitro-cultures of medicinal plants - V
Sedlářová, Marcela ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
The elicitation of the therapeutically important cultures in vitro is a metod used for increasing the production of secondary metabolites. The effectiveness of elicitation depends on a complex interaction between the elicitor and the plant cell. The elicitation is influenced by many factors: the elicitors specificity, their concentration, the treatment interval, the growth phase of cell culture, the composotion of nutritional medium and the light. A wide array of external stimuli are capable of triggering changes in the plant cell which leads to a cascade of reactions, ultimately resulting in the formation and accumulation of secondary metabolites which helps the plant to overcome the stress factors. This work the influence of UV radiation on the flavonoids production in the callus cultures Genista tinctoria in vitro investigated. The highest production of flavonids was obtained at genistin under the exposure of callus culture to UV radiation of 254 nm wave lenght for the time 300 s with taking of sample after 48 hours. This callus culture grew on SH medium. The content of genistin was 3,03 %. Keywords: elicitation, elicitor, secondary metabolites, UV radiation, Genista tinctoria
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In vitro cultures of medicinal plants - VI
Čvejdlíková, Zuzana ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
Zuzana Čvejdlíková Cultures of medicinal plants in vitro - VI Abstract This thesis deals with the plant cultivation in vitro. I studied the influence of abiotic elicitor N-(2- brom-3-methylphenyl)-5-terc-butylpyrazin-2-carboxamid (in concentrations: c1 = 8.36.10-4 mol/l, c2 = 8.36.10-5 mol/l, c3 = 8.36.10-6 mol/l) on the production of flavonoids in the callus and suspension cultures of Ononis arvensis L. The content of flavonoids was determined by photometric method. The highest content of flavonoids (about 5800 %) was observed after elicitor treatment at concentration c3 (8.36.10-6 mol/l) after 48 hours.
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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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