|
|
The effect of Hsp70 protein on the infection caused by Potato virus Y
Doričová, Vlasta ; Hýsková, Veronika (advisor) ; Hoffmeisterová, Hana (referee)
Whithin their natural environment, plants are subjected to a combination of stress conditions. Since potential interactions between signal pathways, plants respond to multiple stresses differently from how they do to individual stresses, activating a specific programme. Heat shock proteins (HSP70) overexpressed after heat shock influence the viral infection. On one side HSP70 can participate on refolding of aggregated or partially denaturated proteins, on the other side HSP70 can interact with viral proteins and facilitate propagation of viral replication complexes. In this work the effect of heat shock (42řC, 2. hours) applied before or after the inoculation of plants Nicotiana tabacum L. cv. Petit Havana SR1 with Potato virus Y on viral infection was detected. This effect was studied in two biological experiments. The amount of coat protein of PVYNTN and protein HSP70 were detected simultaneously with the activity assays of Hatch-Slack cycle enzymes, glycosidases and peroxidase. Both experimental approaches (heat shock applied before or after the inoculation by PVYNTN ) enhanced amount of the virus and in the 2nd experiment it accelerated infection development. Immediately after application of heat shock the amount of HSP70 was increased. The enhancement of HSP70 by viral infection occurred...
|
|
|
Study of plant stress responces in presence of pharmaceuticals in cultivation medium
Bystroňová, Jana ; Soudek, Petr (advisor) ; Podlipná, Radka (referee)
The aim of this study was to verify the possibility of ibuprofen degradation by selected plant cultures and determination of activities of antioxidant enzymes (peroxidase, catalase, ascorbate peroxidase and glutathione-S-transferase) as markers of oxidative stress caused by ibuprofen. Nicotiana tabaccum (cv. La Burley 21, cv. SR 1 and their GMOs) and Nicotiana glauca were used as experimental plants. The rate of removal of ibuprofen tested by tobacco was decreasing in the following order: N. tabaccum SR1 > N. tabaccum Zm-P60-1-T4 > N. tabaccum TRI 2T2 > N. glauca > N. tabaccum TRI 2T1 > N. tabaccum cv. La Burley > N. tabaccum Zm-P60-1-T5. As the most suitable tobacco for the removal of ibuprofen seemed untransformed N. tabaccum SR1. The long-term experiment showed that plant stress is being manifested even after longtime. N. tabaccum cv. La Burley 21 seemed to be the most tolerant to ibuprofen in compare with the total enzyme activities in cultures with the presence of ibuprofen and controls. N.glauca was the least tolerant cultivar. Keywords: phytoremediation, ibuprofen, Nicotiana tabaccum, Nicotiana glauca, HPLC, peroxidase, catalase, ascorbate peroxidase, glutathion-S-transferase
|
|
|
Polymeric framework materials as noble metal free heterogeneous catalysts
Piatková, Mária ; Bojdys, Michael Janus (advisor) ; Tarábek, Ján (referee)
4 Abstract A functional triazine-based polymer framework with embedded copper (Cu) nanoparticles (Cu@TzP) is obtained from a one-pot, "wet" chemistry process that is easily scaled up to industrial demand. The polymer framework has permanent, guest-accessible microporosity and can be obtained as a membrane with 124 m2 g-1 , or as a bulk powder with 660 m2 g-1 (by Ar sorption). The Cu nanoparticles are generated in situ during the formation process of the polymer framework, and they serve as ideal, heterogenised active sites for C-N bond formation and enzyme-mimetic peroxidase catalysis. Further, we tune the porosity of the polymeric support matrix by thermal tempering (carbonisation). It turns out, that Cu@TzP performs best as a peroxidase-mimic in the form of a thin, accessible flake with 2 nm-sized Cu nanoparticles (NPs). Although no records were broken in terms of catalytic activity, we establish an attractive design principle of a scalable membrane support for noble-metal free catalysis.
|
|
|
The effect of Hsp70 protein on the infection caused by Potato virus Y
Doričová, Vlasta ; Hýsková, Veronika (advisor) ; Hoffmeisterová, Hana (referee)
Whithin their natural environment, plants are subjected to a combination of stress conditions. Since potential interactions between signal pathways, plants respond to multiple stresses differently from how they do to individual stresses, activating a specific programme. Heat shock proteins (HSP70) overexpressed after heat shock influence the viral infection. On one side HSP70 can participate on refolding of aggregated or partially denaturated proteins, on the other side HSP70 can interact with viral proteins and facilitate propagation of viral replication complexes. In this work the effect of heat shock (42řC, 2. hours) applied before or after the inoculation of plants Nicotiana tabacum L. cv. Petit Havana SR1 with Potato virus Y on viral infection was detected. This effect was studied in two biological experiments. The amount of coat protein of PVYNTN and protein HSP70 were detected simultaneously with the activity assays of Hatch-Slack cycle enzymes, glycosidases and peroxidase. Both experimental approaches (heat shock applied before or after the inoculation by PVYNTN ) enhanced amount of the virus and in the 2nd experiment it accelerated infection development. Immediately after application of heat shock the amount of HSP70 was increased. The enhancement of HSP70 by viral infection occurred...
|
|
|
Study of plant stress responces in presence of pharmaceuticals in cultivation medium
Bystroňová, Jana ; Soudek, Petr (advisor) ; Podlipná, Radka (referee)
The aim of this study was to verify the possibility of ibuprofen degradation by selected plant cultures and determination of activities of antioxidant enzymes (peroxidase, catalase, ascorbate peroxidase and glutathione-S-transferase) as markers of oxidative stress caused by ibuprofen. Nicotiana tabaccum (cv. La Burley 21, cv. SR 1 and their GMOs) and Nicotiana glauca were used as experimental plants. The rate of removal of ibuprofen tested by tobacco was decreasing in the following order: N. tabaccum SR1 > N. tabaccum Zm-P60-1-T4 > N. tabaccum TRI 2T2 > N. glauca > N. tabaccum TRI 2T1 > N. tabaccum cv. La Burley > N. tabaccum Zm-P60-1-T5. As the most suitable tobacco for the removal of ibuprofen seemed untransformed N. tabaccum SR1. The long-term experiment showed that plant stress is being manifested even after longtime. N. tabaccum cv. La Burley 21 seemed to be the most tolerant to ibuprofen in compare with the total enzyme activities in cultures with the presence of ibuprofen and controls. N.glauca was the least tolerant cultivar. Keywords: phytoremediation, ibuprofen, Nicotiana tabaccum, Nicotiana glauca, HPLC, peroxidase, catalase, ascorbate peroxidase, glutathion-S-transferase
|
|
|
The effect of heat shock stress on enzyme activities in plants.
Doričová, Vlasta ; Hýsková, Veronika (advisor) ; Liberda, Jiří (referee)
Heat stress is one of abiotic stress factors, which fundamentally influences the growth and development of plants. Plants response to heat stress by series of cell and metabolic changes, the specificity of heat stress is synthesis of molecular chaperons, called heat- shock proteins (HSP). The influence of heat shock (in the form of 1 hour application of 40řC from 20řC) on the activity of NADP-dependent enzymes, enzymes of Hatch-Slack cycle, glycosidases and the activity of peroxidase in tobacco plants (Nicotiana tabacum L. cv. Petit Havana SR1) was studied in this work. Since HSP are involved in proper folding of immature, misfolded or partly denaturated proteins, in degradation of denaturated proteins and in induction of thermotolerance of plants, the aim of this work was to find out, if the activity of studied enzymes will be maintained or modulated within the plant defense response. The highest amount of HSP70 detected immunochemically together with higher activities of NADP-malic enzyme, phosphoenolpyruvate carboxylase, β-hexosaminidase and α- mannosidase compared to control plants was found 1 hour after application of heat shock. Peroxidase activity was most increased 1 day after HS compared to controls. Using native electrophoresis the differences in isoform content between control and...
|
|
|
Plant alkaloids and their effects on enzymes metabolizing xenobiotics
Višněvská, Kateřina ; Stiborová, Marie (advisor) ; Černá, Věra (referee)
Sanguinarine and chelerythrine are quaternary benzo[c]phenanthridine alkaloids. The first step in sanguinarine metabolism is its reduction to dihydrosanguinarin. Antimicrobial and anti-inflammatory activities of these alkaloids are used in dentistry and as feed additives. Sanguinarine and chelerythrine induce apoptosis of cells. Fluorescence of these alkaloids and intercalation into DNA could be utilized to use the alkaloids as supravital DNA probe. Negative effect of sanguinarine and chelerythrine is their genotoxicity. Cytochrome P450 and peroxidase oxidize ellipticine to detoxication and activation metabolites. Ellipticine is a potent antineoplastic agent exhibiting the multimodal mechanism of its action. Ellipticine intercalates into DNA and inhibits topoisomerase II. Covalent DNA aducts are mediated by CYP or peroxidase oxidation of ellipticine. The anti-tumor activity of ellipticine and its derivatives is caused by a combination mechanism of cell cycle arrest and induction of the apoptotic pathway. Pharmacological efficiencies and geneotoxic side effects of ellipticine is dependent on levels and activities of cytochrome P450 or peroxidase in target tissues. Aristolactams are the major metabolites of biotransformation of aristolochic acid. Nitroreduction is the crucial step in formation of an...
|
|
|
The effect of histone deacetylase inhibitor vaplroate on activity and expression of cytochromes P450 and peroxidases oxidizing ellipticine
Göttlicherová, Markéta ; Souček, Pavel (referee) ; Stiborová, Marie (advisor)
Ellipticine is a potent antineoplastic agent, whose mode of action is considered to be based mainly on DNA intercalation and inhibition of topoisomerase II. Ellipticine was also found to form covalent DNA adducts mediated by its enzymatic activation with cytochromes P450 (CYP) and peroxidases. The next study demonstrated increasing formation of these ellipticine-DNA adducts by histone deacetylase inhibitor valproate (VPA) in neuroblastoma cells. This phenomenon correlates with increasing cytotoxicity of ellipticine induced by this histone deacetylase inhibitor. This observation can be explained by several mechanisms. One of them can be loosening the structure of chromatine, which leads to accessing DNA for modification. Another one is the effect of VPA on activities and expression of enzymes metabolizing ellipticine. This study was aimed to test the second hypothesis. Since VPA has been shown to be metabolized by similar enzymes as ellipticine is, we have studied the effect of VPA (i) on oxidation of ellipticine by cytochromes P450 and peroxidases, (ii) on activities of the CYP enzymes, which significantly participate in oxidation of ellipticine (CYP1A, CYP3A) and (iii) on expression of enzymes oxidizing ellipticine (CYP1A1, CYP3A4, lactoperoxidase). Oxidation of ellipticine in vitro by model...
|
|
|
The technology of cultivation and fertilization of barley (Hordeum) for the manufacture of food supplements' green barley ".
DARDOVÁ, Magdaléna
The aim of this thesis was to study the technology of cultivation and fertilization of barley (Hordeum) for the manufacture of food supplements' green barley ". The main active ingredients in green barley enzymes are superoxide dismutase, catalase, peroxidase, cytochrome oxidase and transhydrogenase. Other ingredients are flavonoids saponarin, glykosylisovitexin and lutonarin. In producing research was determined the best technology for cultivation and fertilization of green barley. Most substances contains barley grown in organic farming. On the contrary, this barley has a higher content of nitrates due to the exclusive use of manure, which are nitrogen-potassium, and in such a system is a factor in low phosphorus. According to the Liebig law nitrogen which is not in balance with the element in the minimum, it is not able to transform plant nitrogen organic substances. From a nutritional point of view seems to be the best ecological cultivation of barley on dry freshwater lakes in the USA. The soil in these areas contain enough nutrients for barley, which is a prerequisite for good quality products. If green barley grown in the Czech Republic, it would be most appropriate cultivation conventional manner. The soils are poor in nutrients, and thus would only manure was insufficient. The increase of the active substances it is appropriate to use any of several kinds of elicitors. The most suitable varieties would malting barley varieties Sebastian, which includes most of the active substances.
|
guest ::
