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Stanovení biologicky aktivních metabolitů jednobuněčných řas za různých kultivačních podmínek
Malyugina, Svetlana
This thesis studies effect of cultivation conditions on the production of secondary me-tabolites in the green microalgae Scenedesmus quadricauda. During the experiment was studied influence of pH, UVC radiation, light intensity and various temperature conditions on the amount of biologically active compounds (polyphenols, flavonoids, antioxidants). Metabolites were determined using biochemical analytical methods. The total content of polyphenols was determined using the Folin-Ciocalteu method, which is based on chemical reaction of phenols with Folin-Ciocalteu reagent. The total con-tent of flavonoids was determined spectrophotometrically using AlCl3.6H2O with the addition of NaNO2 and 1 M NaOH. The total antioxidant capacity (TAC) was deter-mined using phosphomolybdenun antioxidant assay. Antioxidant activity was stimu-lated by UVC radiation. The highest value being recorded when samples were exposed to UVC for 5 minutes. Light intensity and the temperature changes did not have significant effect on antioxidant activity of studied samples. There was an increase in TAC in samples cultured in acid pH medium (4 and 5). The content of polyphenolic and flavonoid compounds with high light intensity (3500, 4000 lx) and high tempera-tures (35 °C, 40 °C) decreased. Conversely, changes in pH and UV-C stimulate biosyn-thesis of both polyphenols and flavonoids. The greatest increase of these compounds was observed in the medium with pH = 4 and the time of exposure to UV-C for 5 minutes a day.
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Vliv inhibitorů BSO, Lycorine a AIP na biosyntézu sekundárních metabolitů jednobuněčné řasy Scenedesmus quadricauda
Rankić, Ivan
The theoretical overview summarize the characteristics of algae and their primary and secondary metabolism. It also deals with inhibitors of ascorbic acid, glutathione and polyphenolic compounds synthesis. The practical part studies the influence of selected inhibitors (BSO, Lyc, AIP) and CdCl2 on the secondary metabolites production and biomass growth in freshwater microalgae Scenedesmus quadricauda. The content of secondary metabolites (eg. caffeic acid, chlorogenic acid, salicylic acid, cinnamic acid, p-coumaric acid) was detected by HPLC-MS. Furthermore, the total antioxidant capacity, the total polyphenol content and the total flavonoid content was determined spectrophotometrically. All the inhibitors used, reduced S. quadricauda growth in comparison with control samples. The most pronounced inhibitory effect was observed in the samples treated with Lyc + Cd after 7 days of experiment. All the treated samples responded to the stress factors by altering metabolic pathways and inhibiting growth. As a result, there were changes in the composition and the amount of selected secondary metabolites versus the control samples. At the same time, in most cases, the biosynthesis of polyphenolic and flavonoid substances has been stimulated. The oxidative reduction equilibrium and the induction of oxidative stress were probably impaired.
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Study of photosynthetic response of green algae on presence of selected heavy metals
NOVÁK, Jan
With the world population growth, more and more agricultural areas fall into regions affected by waste products. Therefore it is important to determine their impact on environment toxicity. The aim of this thesis is to evaluate the effect of selected ions of heavy metals (CoSO4, CuSO4, CdSO4, ZnSO4 and K2Cr2O7) on the process of photosynthesis in a selected green algae species (Scenedesmus quadricauda) by means of monitoring of dynamics of oxygen release into solution of nutrient medium. The experiments with individual compounds proceeded by pH 7, temperature 25°C and light 450 ?mol m-2 s-1. Algareact device developed by the company BVT Technologies was used for measuring. Comparison with growth curves Scenedesmus quadricauda by different pH was used for the validation of the method. The method of the measurement of dissolved oxygen in medium is suitable for the measurement of immediate impact of chemical compounds on photosynthetic activity of examined organism. The method is restricted by the fluidity of measured solution. The lowest quantity of oxygen disolved inmedium was measured by CdSO4 in concentration 0,1 M.
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Bioaccumulation and toxicity of selenium compounds in the green alga Scenedesmus quadricauda
UMYSOVÁ, Dáša
Three strains of the green alga Scenedesmus quadricauda were selected to be specifically resistant to the presence of high concentrations of inorganic Se. The strain SeIV was found to be resistant to high doses of selenite, the strain SeVI was resistant to selenate, and the strain Se(IV+VI) to the presence of both Se compounds. It has been shown that both the dose and chemical form of Se are the critical factors in the cellular response. In this PhD. thesis are results from experiments with the resistant strains (activity of thioredoxin reductase, content of selenium and selenomethionine, effect of sulphur deficiency).
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DNA damage response in green algae Chlamydomonas reinhardtii and Scenedesmus quadricauda
HLAVOVÁ, Monika
The effect of FdUrd, zeocin, caffeine and their combination on the cell cycle of green algae Chlamydomonas reinhardtii and Scenedesmus quadricauda and response of these model organisms to DNA damage were studied. Both, FdUrd and zeocin, caused DNA damage that led to cell cycle arrest in these algae. In contrast, caffeine partially abolished G2 phase block imposed by zeocin. Protein levels of three crucial cell cycle regulators - CdkA, CdkB and Wee1 kinases were measured to identify mechanisms controlling reaction to DNA damage.
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