Studying stress pathways on the level of secondary metabolites that are found in very small concentration in the cells is complicated. In the algae, the role of individual metabolites (such as carotenoids, phenolic compounds, organic acids, and vitamins) and miRNAs that participate in plant's defence are very poorly understood during stressful conditions. Therefore, in the present experiment, the model organism Chlamydomonas reinhardtii was exposed to stress conditions (Lyc and UV-C irradiation) to detect these substances, even at very low concentrations. The purpose was to monitored changes at each response level with a future view to identifying their specific roles under different stress factors. In stress-treated cultures, numerous transcriptomic and metabolomic pathways were triggered in C. reinhardtii. Although Lyc significantly decreased the concentration of AA, suggesting that Lyc has a similar function in C. reinhardtii as in plants. The negative effect of UV-C radiation was based on the production of ROS and enhancement of antioxidant responses, resulting in increased levels of polyphenols and simple phenolic compounds. Both treatments did lead to extensive changes in transcript levels and miRNA expression patterns.
- MeSH
- alkaloidy amarylkovitých farmakologie MeSH
- Chlamydomonas reinhardtii účinky léků genetika metabolismus účinky záření MeSH
- fenantridiny farmakologie MeSH
- mikro RNA * MeSH
- polyfenoly metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- regulace genové exprese u rostlin účinky léků účinky záření MeSH
- RNA rostlin * MeSH
- ultrafialové záření * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nanotechnology, new fascinating field of science, is bringing many application's options. However, it is necessary to understand their potential environmental risk and toxicity. Zinc selenide quantum dots (ZnSe QDs) are getting valuable due to wide industrial usage, mainly as cadmium free diodes or stabilizing ligand. Thanks to unique properties, they could also open the possibilities of application in the agriculture. Their effects on living organisms, including plants, are still unknown. Therefore, the attention of this work was given to antioxidant response of Arabidopsis thaliana to 100 and 250 μM ZnSe QDs foliar feeding. ZnSe QDs treatment had no statistically significant differences in morphology but led to increased antioxidant response in the leaves at the level of gene expression and production secondary antioxidant metabolites. Concurrently, analysis of growth properties of Agrobacterium tumefaciens was done. 250 μM ZnSe solution inhibited the Agrobacterium tumefaciens viability by 60%. This is the first mention about effect ZnSe QDs on the plants. Although QDs induced oxidative stress, the apply treatment dose of ZnSe QDs did not have significant toxic effect on the plants and even no morphological changes were observed. However, the same amount of ZnSe QD induced an inhibitory effect on Agrobacterium tumefaciens.
- MeSH
- Agrobacterium tumefaciens účinky léků růst a vývoj MeSH
- antioxidancia metabolismus MeSH
- Arabidopsis genetika růst a vývoj metabolismus MeSH
- exprese genu * MeSH
- kvantové tečky toxicita MeSH
- metabolomika * MeSH
- sloučeniny selenu aplikace a dávkování toxicita MeSH
- sloučeniny zinku aplikace a dávkování toxicita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This work has analysed the influence of CdCl2, NiCl2 and CuCl2 on simple phenolic acids, such as the 3PPP (phenylpyruvic, phenylacetic, and 4-hydroxyphenylpyruvic) and 2DR (3,4-dihydroxyphenylacetic and rosmarinic) phenolic acids for the first time and studied their interactions with antioxidant systems and the glutathione-ascorbate cycle in the freshwater green microalga Scenedesmus quadricauda. The compounds investigated are related to both the catabolic and anabolic pathways of l-phenylalanine and l-tyrosine, the main molecules in the biosynthesis of polyphenols. The concentrations of the simple phenolic acids responded significantly to NiCl2, CdCl2 and CuCl2 at 5, 20 and 40 μM concentrations. The 3PPP phenolic acid concentrations after 24 h were always higher in metal-treated cells than in controls, while the 2DR concentrations were significantly lower in the metal-treated cells than the controls. The GSH/GSSG ratio was lower in all experimental groups treated with the selected metals (especially so at 40 μM). Pearson correlation analysis indicated a strong negative correlation between ascorbate and rosmarinic acid content (-0.670; p < 0.05) in NiCl2-treated samples and reduced glutathione and 3,4-dihydroxyphenylacetic content (-0.700; p < 0.05) in CdCl2-treated samples. The GSSG content in samples exposed to CuCl2 was correlated with the concentrations of all of the investigated phenolic acids (4 negative, 1 positive).
- MeSH
- antioxidancia metabolismus MeSH
- chlorid kademnatý toxicita MeSH
- fytochelatiny metabolismus MeSH
- glutathion metabolismus MeSH
- hydroxybenzoáty metabolismus MeSH
- kyselina askorbová metabolismus MeSH
- měď toxicita MeSH
- mikrořasy účinky léků růst a vývoj metabolismus MeSH
- nikl toxicita MeSH
- oxidační stres účinky léků MeSH
- polyfenoly metabolismus MeSH
- Scenedesmus účinky léků růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Epigenetic changes are important mechanisms in the regulation of chromatin structure and gene expression. Cytosine methylation is one of the major epigenetic modifications, mediated by DNA methyltransferases, which transfer methyl groups from S-adenosyl-L-methionine (SAM) to the fifth carbon of cytosine. Various external environmental conditions can change the global hypo/hypermethylation pattern of DNA. These alterations may affect the organism's response to stress conditions. In this study, for the first time, we investigated the effects of 5-azacytidine, a DNA methyltransferase inhibitor, and cadmium, a toxic metal and environmental pollutant, on the growth, biosynthesis of secondary metabolites (phenols, flavonoids, carotenoids), SAM, S-adenosylhomocysteine, 5'-methylthioadenosine and global 5-methylcytosine (5-mC) in the green microalgae Chlamydomonas reinhardtii and Scenedesmus quadricauda. The studied species showed major differences in 5-mC content, secondary metabolite content, and antioxidant activity. Cadmium increased GSH (glutathione) content in C. reinhardtii by 60% whereas 5-azacytidine did not affect GSH. The biosynthesis of GSH in S. quadricauda in response to the stressors was the opposite. Global 5-mC content of C. reinhardtii was 1%-1.5%, and the content in S. quadricauda was 3.5%. Amount of some investigated methionine cycle metabolites (SAM, S-adenosyl homocysteine [SAH], methionine) in S. quadricauda distinctly exceeded C. reinhardtii as well. However, chlorophylls a and b, carotenoids, total phenolic content, total flavonoid content and, antioxidant activity were significantly higher in C. reinhardtii than S. quadricauda. Therefore, in further studies it would be advisable to verify whether methylation of cytosine affects the expression of genes encoding certain secondary metabolites.
In this study, 14 commercial products (dietary supplements) containing alga Chlorella vulgaris and cyanobacteria Spirulina platensis, originated from China and Japan, were analysed. UV-vis spectrophotometric method was applied for rapid determination of chlorophylls, carotenoids and pheophytins; as degradation products of chlorophylls. High Performance Thin-Layer Chromatography (HPTLC) was used for effective separation of these compounds, and also Atomic Absorption Spectrometry for determination of heavy metals as indicator of environmental pollution. Based on the results obtained from UV-vis spectrophotometric determination of photosynthetic pigments (chlorophylls and carotenoids), it was confirmed that Chlorella vulgaris contains more of all these pigments compared to the cyanobacteria Spirulina platensis. The fastest mobility compound identified in Chlorella vulgaris and Spirulina platensis using HPTLC method was β-carotene. Spectral analysis and standard calibration curve method were used for identification and quantification of separated substances on Thin-Layer Chromatographic plate. Quantification of copper (Cu2+, at 324.7 nm) and zinc (Zn2+, at 213.9nm) was performed using Flame Atomic Absorption Spectrometry with air-acetylene flame atomization. Quantification of cadmium (Cd2+, at 228.8 nm), nickel (Ni2+, at 232.0nm) and lead (Pb2+, at 283.3nm) by Electrothermal Graphite Furnace Atomic Absorption Spectrometry; and quantification of mercury (Hg2+, at 254nm) by Cold Vapour Atomic Absorption Spectrometry.
- MeSH
- Chlorella vulgaris chemie MeSH
- chlorofyl chemie MeSH
- chromatografie na tenké vrstvě metody MeSH
- karotenoidy chemie MeSH
- potravní doplňky analýza MeSH
- spektrofotometrie atomová metody MeSH
- Spirulina chemie MeSH
- těžké kovy chemie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Čína MeSH
- Japonsko MeSH
Zinc belongs to the mineral elements, the so-called micronutrients, which are essential for all types of plants. Embedding itself into the enzymes associated with proteosynthesis and energy processes, zinc is necessary for maintaining the integrity of biomembranes and also plays an important role in the development of seeds and generative organs. This review focuses on summarising the findings on the interaction of zinc and plants and translates into the knowledge of the effect of zinc nanoparticles on plants. The findings include an overview of both positive and negative effects on plants. In conclusion there is a great interest in nano-zinc as improving the knowledge about individual forms of zinc and their uptake and assimilation within higher plants may be the first step towards a wider involvement of zinc nanoparticles into agriculture.
- MeSH
- kovové nanočástice * chemie toxicita MeSH
- látky znečišťující životní prostředí chemie metabolismus toxicita MeSH
- nanotechnologie MeSH
- půda chemie MeSH
- rostliny metabolismus MeSH
- zemědělství MeSH
- zinek * chemie metabolismus toxicita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Uniparental silencing of 35S rRNA genes (rDNA), known as nucleolar dominance (ND), is common in interspecific hybrids. Allotetraploid Tragopogon mirus composed of Tragopogon dubius (d) and Tragopogon porrifolius (p) genomes shows highly variable ND. To examine the molecular basis of such variation, we studied the genetic and epigenetic features of rDNA homeologs in several lines derived from recently and independently formed natural populations. Inbred lines derived from T. mirus with a dominant d-rDNA homeolog transmitted this expression pattern over generations, which may explain why it is prevalent among natural populations. In contrast, lines derived from the p-rDNA dominant progenitor were meiotically unstable, frequently switching to co-dominance. Interpopulation crosses between progenitors displaying reciprocal ND resulted in d-rDNA dominance, indicating immediate suppression of p-homeologs in F1 hybrids. Original p-rDNA dominance was not restored in later generations, even in those segregants that inherited the corresponding parental rDNA genotype, thus indicating the generation of additional p-rDNA and d-rDNA epigenetic variants. Despite preserved intergenic spacer (IGS) structure, they showed altered cytosine methylation and chromatin condensation patterns, and a correlation between expression, hypomethylation of RNA Pol I promoters and chromatin decondensation was apparent. Reversion of such epigenetic variants occurred rarely, resulting in co-dominance maintained in individuals with distinct genotypes. Generally, interpopulation crosses may generate epialleles that are not present in natural populations, underlying epigenetic dynamics in young allopolyploids. We hypothesize that highly expressed variants with distinct IGS features may induce heritable epigenetic reprogramming of the partner rDNA arrays, harmonizing the expression of thousands of genes in allopolyploids.
- MeSH
- DNA rostlinná genetika MeSH
- epigenomika * MeSH
- fenotyp MeSH
- genom rostlinný genetika MeSH
- genotyp MeSH
- hybridizace genetická MeSH
- metylace DNA MeSH
- molekulární evoluce * MeSH
- polyploidie MeSH
- regulace genové exprese u rostlin * MeSH
- ribozomální DNA genetika MeSH
- Tragopogon genetika MeSH
- umlčování genů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Low-molecular mass proteins rich in cysteines called metallothioneins (MT) can be considered as markers for the pollution of the environment by metals. Here, we report on suggestion for an automated procedure for the isolation of MT followed by voltammetric analysis. Primarily, we optimized the automated detection of MT using an electrochemical analyser. It was found that the most sensitive and repeatable analyses are obtained at a temperature of 4 °C for the supporting electrolyte. Further, we optimized experimental conditions for the isolation of MT by using antibody-linked paramagnetic microparticles. Under the optimal conditions (4 h long interaction between the microparticles and MT), the microparticles were tested on isolation of various amounts of MT. The lowest isolated amount of MT by antibody-linked paramagnetic microparticles was 5 μg ml(-1) of MT (50 ng). The automated procedure of MT isolation was further tested on isolation of MT from guppy fish (Poecilia reticulata) treated with silver(i) ions (50 μM AgNO(3)). The whole process lasted less than five hours and was fully automated. We attempted to correlate these results with the standard method for MT isolation. The correlation coefficient is 0.9901, which confirms that results are in good agreement. Moreover, the concentration of silver ions in tissues of fish treated with Ag(i) ions was determined by high performance liquid chromatography with electrochemical detection.
- MeSH
- chemické látky znečišťující vodu toxicita MeSH
- magnetismus MeSH
- metalothionein chemie izolace a purifikace metabolismus MeSH
- monitorování životního prostředí metody MeSH
- živorodka metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Matrix metalloproteinases (MMPs) play numerous roles in physiological and pathological processes including cancer. Interactions of MMPs with other zinc-binding proteins are of great interest mainly from the point of view of the participation of these proteins in carcinogenesis. The aim of this study was to utilize chip gel electrophoresis for investigation of matrix metalloproteinase 9 (gelatinase B, MMP-9) interactions with collagen. The interaction was observed and the effect of temperature was tested. We further focused on the study of activation of MMP-9 by the presence of zinc-binding proteins called metallothioneins (MT). We confirmed the basic presumption of the activation of MMP-9 by the presence of MT similar to the temperature effect. Moreover, we chose the method of differential pulse voltammetry Brdicka reaction to verify our results. It can be concluded that the activity of MMP-9 was higher in the presence of MT in comparison with the experimental collagen(type I)-MMP-9 mixture in vitro.
- MeSH
- elektroforéza mikročipová metody MeSH
- kolagen typu I chemie MeSH
- lidé MeSH
- matrixová metaloproteinasa 9 chemie metabolismus MeSH
- metalothionein chemie metabolismus MeSH
- molekulární modely MeSH
- teplota MeSH
- zinek chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
