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The selenium effect on secondary metabolites production in in vitro cultures of medicinal plants - II
Ošťádalová, Tereza ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Tereza Ošťádalová Supervisor: Doc. PharmDr. Lenka Tůmová, CSc. Title of diploma thesis: The selenium effect on secondary metabolites production in in vitro cultures of medicinal plants - II. Key words: callus, suspension culture, abiotic elicitor, selenium, Fagopyrum In vitro plant cultures usually produce only a small amount of secondary metabolites. The method of elicitation is one of the options how to increase the production of these substances. The effect of selenium as abiotic elicitor on rutin production in callus and suspension culture of Fagopyrum esculentum Moench., variety Pyra was observed in this study. The release of rutin into the nutrient medium was studied as well. The cultivation was performed in Murashige and Skoog (MS) nutrient medium which was enriched with 1 mg/l of 2,4-dichlorophenoxyacetic acid. The solution of selenium in three different concentrations (c1 = 9.012.10-3 mol/l, c2 = 9.012.10-4 mol/l, c3 = 9.012.10-5 mol/l) was used. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of elicitor treatment. The rutin content was determined by high-performance liquid chromatography (HPLC). The elicitation led to increasing of rutin amount in callus and also in suspension...
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The selenium effect on secondary metabolites production in in vitro cultures of medicinal plants - II
Ošťádalová, Tereza ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy Candidate: Tereza Ošťádalová Supervisor: Doc. PharmDr. Lenka Tůmová, CSc. Title of diploma thesis: The selenium effect on secondary metabolites production in in vitro cultures of medicinal plants - II. Key words: callus, suspension culture, abiotic elicitor, selenium, Fagopyrum In vitro plant cultures usually produce only a small amount of secondary metabolites. The method of elicitation is one of the options how to increase the production of these substances. The effect of selenium as abiotic elicitor on rutin production in callus and suspension culture of Fagopyrum esculentum Moench., variety Pyra was observed in this study. The release of rutin into the nutrient medium was studied as well. The cultivation was performed in Murashige and Skoog (MS) nutrient medium which was enriched with 1 mg/l of 2,4-dichlorophenoxyacetic acid. The solution of selenium in three different concentrations (c1 = 9.012.10-3 mol/l, c2 = 9.012.10-4 mol/l, c3 = 9.012.10-5 mol/l) was used. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of elicitor treatment. The rutin content was determined by high-performance liquid chromatography (HPLC). The elicitation led to increasing of rutin amount in callus and also in suspension...
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The selenium effect on secondary metabolites production in in vitro cultures of medicinal plants - I
Černá, Pavlína ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Higher plants represent an important source of valuable substances, so called secondary metabolites, which can be obtained through explant cultures of plants. Elicitation is a method of increasing the secondary metabolites production. This study aims to evaluate the secondary metabolites production in Fagopyrum esculentum variety Spacinska cultures in vitro after abiotic elicitor treatment. The experiment was focused on alteration of rutin production in callus and suspension cultures of F. esculentum var. Spacinska after selenium application. Murashige and Skoog nutrient medium supplemented with 1 mg l-1 2,4-dichlorophenoxyacetic acid was used for the cultivation. Selenium solutions of various concentrations (c1 = 9.012×10-3 mol l-1 , c2 = 9.012×10-4 mol l-1 , c3 = 9.012×10-5 mol l-1 ) were affecting the cultures for 6, 12, 24, 48, 72 and 168 hours. The content of rutin was determined by HPLC. The release of secondary metabolites into the nutrient medium was studied as well. After elicitor application, the rutin production increased in both callus and suspension cultures. Higher levels of rutin content were detected in callus culture. The maximum rutin content (0.6 mg g-1 DW) was reached after 12 h of elicitor treatment of c2 concentration in callus culture. Concerning suspension culture, the...
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In vitro cultures of medicinal plants IXX
Seidlová, Markéta ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
In vitro cultures of medicinal plants - IXX The subject of this thesis is the evaluation of secondary metabolites production in in vitro cultures of Silybum marianum L. after elicitor treatment. In this study selenium dioxide as elicitor in concentrations of 9,012.10-3 mol/l; 9,012.10-4 mol/l; 9,012.10-5 mol/l was used. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of elicitor treatment. The effect of elicitor was compared with control samples, which were cultured without elicitation. The content of taxifolin and flavonolignans was determined by the method of HPLC. The results showed, that almost all observed metabolites were released into a nutrient medium. Cells of callus and suspension cultures produced only small amounts of taxifolin (0.01 mg/g DW). Taxifolin and silymarin complex releasing into nutrient media was observed as in control and also in the elicitated samples of callus and suspension cultures. Selenium dioxide elicitation caused statistically significant increases in releasing taxifolin and silymarin complex into the nutrient medium. The statistically significant releasing of flavonolignans (2.2 mg/100 ml) to the medium of suspension culture was reached after 72 hours of treatment with selenium dioxide in concentration of c1 (9,012.10-3 mol/l). The statistically...
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In vitro cultures of medicinal plants XX
Chreňová, Kateřina ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
The study is aimed to enhance in vitro production of secondary metabolites in Genista tinctoria L. via elicitor treatment. The different levels of elicitor concentration - selenium dioxide were utilized to affect the quantity of isoflavonoids occurred in cultures. Experiment was perfomed in callus and suspension cultures on MS nutrient media supplemented with 10 g l-1 of NAA (α-naphtylacetic acid) as growth regulator. The elicitor was added in the form of solution in concentrations of 9,012.10-3 mol l-1 ; 9,012.10-4 mol l-1 and 9,012.10-5 mol l-1 . It was exposed for 6, 12, 24, 48, 72 and 168 hours. The content of isoflavonoids was determined by HPLC in dry weight (DW) and medium. The most effective production of genistin (6,20 mg. g-1 DW, 8,30 mg. g-1 DW) in callus culture was measured. It was reached in concentrations of 9,012.10-4 mol l-1 and 9,012.10-5 mol l-1 after 168 h elicitor treatment. The second most satisfactory genistin level 5,20 mg g-1 DW was detected after elicitor application in concentration of 9,012.10-4 mol l-1 after 6 h. The content of genistein, daidzein and formononetin in callus culture was low and in the most cases equal zero compared to control samples. The content of biochanin A was equal zero compared to control samples. The most efficient daidzein production (37,10 mg...
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In vitro cultures of medicinal plants XVIII
Bremertová, Iva ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
Genista tinctoria, family Fabaceae, is a potent source of isoflavonoids (genistin, genistein, daidzein, formononetin, biochanin A) with a wide spectrum of potential medical impact. Genista also contains quinolizidin alkaloids (cytisin, anagyrin, lupanin, spartein, etc.), which are toxic. The reason why in vitro cultures are used is an absence of toxic alkaloids production and higher yield of isoflavonoids in comparison with intact plant. For an increase of isoflavonoid production method of elicitation is beeing used. Isoflavonoids are studied for their phytoestrogenic effects, for which they could be used in treatment of postmenopausal symptoms and even in treatment of hormon-dependent tumours. The elicitor ethephon in concentration of 7000 µM, 700 µM and 70 µM was used in this work. The effect of ethephon inhibitor (AgNO3) in concentration of 120 µM was investigated too. Samples were examined after 24, 48, 72, 96 and 168 hours and then analysed by HPLC method. An effect of ethephon and its combination with AgNO3 was observed in callus and suspension cultures. Release of isoflavonoides into culture media was studied too. Daidzein production was the highest of all isoflavonoids in the callus culture after the treatment of ethephon in concentration of 700 µM after 96 hours (45,10 mg/g DW). The most...
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In vitro cultures of medicinal plants XVII
Cinková, Lucie ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Milk thistle, Silybum marianum L. Gaertn., is a source of flavonoid taxifolin and flavonolignans - silymarin complex (silybin, silydianin, silycristin and isosilybin). Milk thistle is usually obtained by field cultivation. Alternative way for getting the active components, is the use of in vitro cultures. But the production of secondary metabolites by the in vitro cultures is low in comparison with plant. One of the possibilites how to increase this produciton is the method of elicitation. In this study, ethephon as the elicitor, in the concentrations of 500 µmol/l, 400 µmol/l, 200 µmol/l, 100 µmol/l and 50 µmol/l was used with the aim to increase secondary metabolite production in suspension and callus cultures. The effect of ethephon was compared to its inhibitor (AgNO3, 120 µmol/l). The levels of flavonolignans and taxifolin were measured by the method of HPLC. The samples were taken 24, 48, 72, 96 and 168 hours after the ethephon application and inhibitor treatment. The nutrient medium of suspension culture was also tested for the possibity of secondary metabolites releasing into medium. The highest content of flavonoid taxifolin was found in the suspension culture medium after 48 h treatment with ethephon in conc. of 400 µmol/l. The level of taxifolin was increased by 197-fold to 1,97 mg/100...
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