Plant height is among the most important agronomic traits influencing crop yield. Wheat lines carrying Rht genes are important in plant breeding due to their both higher yield capacity and better tolerance to certain environmental stresses. However, the effects of dwarf-inducing genes on stress acclimation mechanisms are still poorly understood. Under the present conditions, cadmium stress induced different stress responses and defence mechanisms in the wild-type and dwarf mutant, and the mutant with the Rht-B1c allele exhibited higher tolerance. In the wild type after cadmium treatment, the abscisic acid synthesis increased in the leaves, which in turn might have induced the polyamine and proline metabolisms in the roots. However, in the mutant line, the slight increment in the leaf abscisic acid content accompanied by relatively high salicylic acid accumulation was not sufficient to induce such a great accumulation of proline and putrescine. Although changes in proline and polyamines, especially putrescine, showed similar patterns, the accumulation of these compounds was antagonistically related to the phytochelatin synthesis in the roots of the wild type after cadmium stress. In the dwarf genotype, a favourable metabolic shift from the synthesis of polyamine and proline to that of phytochelatin was responsible for the higher cadmium tolerance observed.
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
Short-term (24 h) responses of Cladonia arbuscula subsp. mitis and Cladonia furcata to copper (CuII) or chromium (CrIII) excess (10 or 100 μM) were compared. C. arbuscula accumulated more Cu and Cr at higher metal doses but both species revealed depletion of K and/or Ca amount. Not only Cu but also Cr typically elevated reactive oxygen species (ROS) formation (fluorescence microscopy detection of total ROS and hydrogen peroxide) and depleted nitric oxide (NO) signal, with Cu showing more negative impact on lipid peroxidation (BODIPY 581/591 C11 staining reagent). Metals and staining reagents also affected anatomical responses and photobiont/mycobiont visibility. Principally different impact of Cu and Cr was observed at antioxidative metabolites level, indicating various ways of metal-induced ROS removal and/or metal chelation: Cu strongly depleted glutathione (GSH) and stimulated phytochelatin 2 (PC2) content while ascorbic acid accumulation was depleted by Cu and stimulated by Cr. Subsequent experiment with GSH biosynthetic inhibitor (buthionine sulfoximine, BSO) revealed that 48 h of exposure is needed to deplete GSH and BSO-induced depletion of GSH and PC2 amounts under Cu or Cr excess elevated ROS but depleted NO. These data suggest close relations between thiols, NO and appearance of oxidative stress (ROS generation) under metallic stress also in lichens.
- MeSH
- antioxidancia metabolismus MeSH
- buthionin sulfoximin MeSH
- chrom toxicita MeSH
- fytochelatiny metabolismus MeSH
- glutathion metabolismus MeSH
- kovy toxicita MeSH
- kyselina askorbová metabolismus MeSH
- látky znečišťující životní prostředí toxicita MeSH
- lišejníky účinky léků metabolismus fyziologie MeSH
- měď metabolismus MeSH
- oxidační stres fyziologie MeSH
- peroxid vodíku metabolismus MeSH
- peroxidace lipidů MeSH
- reaktivní formy kyslíku metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Responses of Scenedesmus quadricauda grown in vitro and differing in age (old culture-13 months, young culture-1 month) to short-term cadmium (Cd) or nickel (Ni) excess (24h) were compared. Higher age of the culture led to lower amount of chlorophylls, ascorbic acid and glutathione but higher signal of ROS. Surprisingly, sucrose was detected using DART-Orbitrap MS in both old and young culture and subsequent quantification confirmed its higher amount (ca. 3-times) in the old culture. Cd affected viability and ROS amount more negatively than Ni that could arise from excessive Cd uptake which was also higher in all treatments than in respective Ni counterparts. Surprisingly, nitric oxide was not extensively different in response to age or metals. Strong induction of phytochelatin 2 is certainly Cd-specific response while Ni also elevated ascorbate content. Krebs cycle acids were more accumulated in the young culture but they were rather elevated in the old culture (citric acid under Ni excess). We conclude that organic solid 'Milieu Bristol' medium we tested is suitable for long-term storage of unicellular green algae (also successfully tested for Coccomyxa sp. and Parachlorella sp.) and the impact of age on metal uptake may be useful for bioremediation purposes.
- MeSH
- fytochelatiny metabolismus MeSH
- kadmium farmakokinetika toxicita MeSH
- kyseliny karboxylové metabolismus MeSH
- metabolom účinky léků MeSH
- nikl farmakokinetika toxicita MeSH
- oxid dusnatý metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- Scenedesmus účinky léků růst a vývoj metabolismus MeSH
- sulfhydrylové sloučeniny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Increasing emissions of heavy metals such as cadmium, mercury, and arsenic into the environment pose an acute problem for all organisms. As a mass of protection, many of them, develop mechanisms of full resistance or at least exhibit partially resisting toward these effects. In this way, based on the chemical similarity of the involved metallic species, they are able, to replace them with viable metals necessary for the effective functioning of the cell. These heavy metals may be bound to the functional groups of proteins and modify their structure and through this also affect their physiological function 1, 2. Higher plants, algae, certain yeasts and animals are able to respond to heavy metals by synthesizing phytochelatins (PCs) and related cysteine-rich polypeptides. Phytochelatin synthases are γ-glutamylcysteine (γ-Glu-Cys) dipeptidyl transpeptidases that catalyze the synthesis of heavy metal-binding PCs 3, 4. PCs, cysteine-rich peptides, are produced from glutamine, cysteine and glycine. Unlike commonmetal-binding structures, MT and GSH, PCs are not gene-encoded, but enzymatically synthesized peptides 5. PCs have been identified in a wide variety of plant species, microorganisms and some invertebrates 6-10. They are structurally related to glutathione (GSH) and were presumed to be the products of a biosynthetic pathway. Numerous physiological, biochemical and genetic studies have confirmed GSH as the substrate for PCs biosynthesis 11, 12. The general structure of PCs is (c-Glu-Cys)n-Gly, with increasing repetitions of the dipeptide Glu-Cys linked through a c-carboxylamide bond (Fig 1), where n varies from 2 to 11, but typically reaching not further than five 13. Except glycine, also other amino acid residues can be found on C-terminal end of (γ-Glu-Cys)n peptides. Examples of which, like Ser, Glu, Gln and Ala are often found at this position in some plant species, and they are assumed to be functionally analogous and synthesised via essentially similar biochemical pathways 14, 15. In in vitro studies of PC synthase expressed in E. coli or in S. cerevisiae, the enzyme was activated to varying extents by Cd, Cu, Ag, Hg, Zn and Pb ions 16-18. PC synthase genes were also isolated in A.thaliana 16 and T.aestivum 18. Genes homologous to those from A.thaliana and T.aestivum were also found in S.pombe and C.elegans, suggesting the existence of PC synthase genes in more species 19.
- MeSH
- chemické techniky analytické metody MeSH
- fytochelatiny * biosyntéza chemie metabolismus MeSH
- rostliny metabolismus MeSH
- těžké kovy * metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
In this study, in vitro formed Cd-phytochelatin (PC2) complexes were characterized using ion exchange chromatography (IEC) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The ratio of both studied compounds as well as experimental conditions were optimized. The highest yield of the complex was observed under an applied concentration of 100 µg·mL(-1) PC2 and 100 µg·mL(-1) of CdCl2. The data obtained show that IEC in combination with MALDI-TOF is a reliable and fast method for the determination of these complexes.
- MeSH
- chromatografie iontoměničová metody MeSH
- fytochelatiny chemie MeSH
- kadmium chemie MeSH
- látky znečišťující životní prostředí chemie MeSH
- monitorování životního prostředí metody MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nanomaterials are structures whose exceptionality is based on their large surface, which is closely connected with reactivity and modification possibilities. Due to these properties nanomaterials are used in textile industry (antibacterial textiles with silver nanoparticles), electronics (high-resolution imaging, logical circuits on the molecular level) and medicine. Medicine represents one of the most important fields of application of nanomaterials. They are investigated in connection with targeted therapy (infectious diseases, malignant diseases) or imaging (contrast agents). Nanomaterials including nanoparticles have a great application potential in the targeted transport of pharmaceuticals. However, there are some negative properties of nanoparticles, which must be carefully solved, as hydrophobic properties leading to instability in aqueous environment, and especially their possible toxicity. Data about toxicity of nanomaterials are still scarce. Due to this fact, in this work we focused on studying of the effect of magnetic nanoparticles (NPs) and modified magnetic nanoparticles (MNPs) on tobacco BY-2 plant cell suspension culture. We aimed at examining the effect of NPs and MNPs on growth, proteosynthesis - total protein content, thiols - reduced (GSH) and oxidized (GSSG) glutathione, phytochelatins PC2-5, glutathione S-transferase (GST) activity and antioxidant activity of BY-2 cells. Whereas the effect of NPs and MNPs on growth of cell suspension culture was only moderate, significant changes were detected in all other biochemical parameters. Significant changes in protein content, phytochelatins levels and GST activity were observed in BY-2 cells treated with MNPs nanoparticles treatment. Changes were also clearly evident in the case of application of NPs. Our results demonstrate the ability of MNPs to negatively affect metabolism and induce biosynthesis of protective compounds in a plant cell model represented by BY-2 cell suspension culture. The obtained results are discussed, especially in connection with already published data. Possible mechanisms of NPs' and MNPs' toxicity are introduced.
- MeSH
- antioxidancia metabolismus MeSH
- buněčné kultury MeSH
- chromatografie MeSH
- fluorescenční mikroskopie MeSH
- fytochelatiny metabolismus MeSH
- kovové nanočástice toxicita MeSH
- proteosyntéza účinky léků MeSH
- spektrofotometrie MeSH
- sulfhydrylové sloučeniny metabolismus MeSH
- tabák účinky léků růst a vývoj metabolismus MeSH
- viabilita buněk účinky léků MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- železité sloučeniny toxicita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Silver solid amalgam electrode (AgSAE) was used for differential pulse voltammetric (DPV) measurements of cysteine and cysteine-containing peptides, glutathione, gamma-Glu-Cys-Gly and phytochelatin (gamma-Glu-Cys)(3)-Gly (PC3), in the presence of Co(II) ions. It had been established earlier that cysteine-containing peptides and proteins catalyze hydrogen evolution at mercury electrodes in presence of cobalt salts; these processes are known as the Brdicka reaction. DPV signals measured with the AgSAE, the surfaces of which had been modified by mercury meniscus or mercury film, were qualitatively the same as those obtained with the hanging mercury drop electrode (HMDE). With these electrodes the number and the intensity of Brdicka signals of cysteine, glutathione and PC3 differed, making a distinction among them possible. On the other hand, with the polished silver solid amalgam electrode (the surface of which was completely free of liquid mercury) all three compounds produced only one but strikingly intense peak in the region of Brdicka reaction. Using this signal, cysteine, glutathione as well as PC3 could be determined at 10(-8)M level, representing sensitivity up to 2 orders of magnitude better than attained with the mercury-modified AgSAEs or HMDE.
