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The effect of elicitors on the secondary metabolites production in vitro cultures -I.
Damaskinos, Antonios ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Active compounds have been always originated from plants. Plants though, were able to produce only very low amounts of them and that was the reason for trying many alternative ways of production, one of them being plant tissue culture cultivation. This method is any fragment of living tissue or organ taken from an intact plant or an already existing explant culture, with the intention of growing an artificial growth medium. Even this method though, is not able to produce large amounts compared to extraction from field plants. Elicitation is considered a possible way to increase the production of secondary metabolites. This method used the plant's own defense system, in order to increase the production of secondary metabolites in vitro. The compound which is used to produce the effect is called elicitor. During our experimental work I used as an elicitor the compound Ethephon (2-Chloroethylphosphonic acid) upon callus and suspension cultures of Hypericum perforatum, with intention to observe its effect on flavonoid production. This experiment was based on three different concentrations and six different withdrawal times, being 6, 12, 24, 72, 168 hours. The maximum effect of elicitor was reached with concentration c1 (1mg/100ml) after 12 hours and with concentration c3 (100mg/100ml) after 72 hours.
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The effect of abiotic elicitors on secondary metabolites content in plant cultures in vitro - II.
Tomaidesová, Barbora ; Tůmová, Lenka (advisor) ; Karlíčková, Jana (referee)
The Effect of Abiotic Elicitors on the Content of Secondary Metabolites in In vitro Plant Cultures - II. The method of elicitation is used for the increased production of secondary metabolites in in vitro plant cultures. In this study, there the effect of pyridine derivate N-(5-chlorpyridin-2-yl)-4-ethylbenzamide in three concentration - 3.845.10-3 mol/l; 3.845.10-4 mol/l and 3.845.10-5 mol/l for the production of flavonolignans in callus and suspension cultures of Silybum marianum was tested. The evaluation of elicitation effect proceeded after 6, 24, 48, 72 a 168 hours against control samples without the elicitor treatment after 24 and 168 hours. In the same intervals the samples of culture medium were taken and assessed. The cells were cultivated on Murashige and Skoog medium with 10 mg/ml of α-naphtylacetic acid as growth regulator. The obtained and adjusted samples were analysed by the HPLC method. From the parts of silymarin complex were detected silychristin, silybin A, isosylibin A, isosilybin B. Flavonoid taxifolin wasn̕ t detected in any case. In callus culture, there was achieved a maximal production of all detected components of silymarin complex (30.508 μg/g DW) after 168 hours of elicitor treatment in concentration of 3.845.10-4 mol/l, it was about only substance - silychristin. The...
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The effect of abiotic elicitors on secondary metabolites content in plant cultures in vitro - I.
Teplá, Klára ; Tůmová, Lenka (advisor) ; Siatka, Tomáš (referee)
Plants are a source of a wide range of secondary substances, which due to their effects find use in many areas of focus. By a method called elicitation, we can achieve their higher and thus more efficient production. This diploma thesis aimed to determine whether the abiotic elicitor 2-(4-chlorophenyl)-N-(5-chloropyridin-2-yl)acetamide can positively affect the production of the flavonoid hyperoside in callus and suspension culture of Hypericum perforatum L. The elicitor was added to the in vitro cultures in three concentrations: C1 = 3,571.10-3 mol/l; C2 = 3,571.10-4 mol/l and C3 = 3,571.10-5 mol/l. A sample was taken at regular intervals after 6, 24, 48, 72 and 168 hours of elicitor treatment. Control samples were taken after 24 and 168 hours. The content of hyperoside produced was subsequently determined using High Performance Liquid Chromatography. Simultaneously, the amount of hyperoside released into the nutrient media of both plant cultures was also monitored. Maximum hyperoside production was recorded in suspension culture after 6 (17,7 µg/g DW) and 48 hours (3,69 µg/g DW) of elicitor treatment with the lowest concentration of C3 (3,571.10-5 mol/l). The content of hyperoside in the first case was 1770 % higher compared to the control sample. There was a significant release of hyperoside...
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The effect of elicitors on the secondary metabolites production in vitro cultures -I.
Damaskinos, Antonios ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
Active compounds have been always originated from plants. Plants though, were able to produce only very low amounts of them and that was the reason for trying many alternative ways of production, one of them being plant tissue culture cultivation. This method is any fragment of living tissue or organ taken from an intact plant or an already existing explant culture, with the intention of growing an artificial growth medium. Even this method though, is not able to produce large amounts compared to extraction from field plants. Elicitation is considered a possible way to increase the production of secondary metabolites. This method used the plant's own defense system, in order to increase the production of secondary metabolites in vitro. The compound which is used to produce the effect is called elicitor. During our experimental work I used as an elicitor the compound Ethephon (2-Chloroethylphosphonic acid) upon callus and suspension cultures of Hypericum perforatum, with intention to observe its effect on flavonoid production. This experiment was based on three different concentrations and six different withdrawal times, being 6, 12, 24, 72, 168 hours. The maximum effect of elicitor was reached with concentration c1 (1mg/100ml) after 12 hours and with concentration c3 (100mg/100ml) after 72 hours.
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