Plants are subjected to a variety of abiotic stressors, including drought stress, that are fatal to their growth and ability to produce under natural conditions. Therefore, the present study was intended to investigate the drought tolerance potential of faba bean (Vicia faba L.) plants under the co-application of biochar and rhizobacteria, Cellulomonas pakistanensis (National Culture Collection of Pakistan (NCCP)11) and Sphingobacterium pakistanensis (NCCP246). The experiment was initiated by sowing the inoculated seeds with the aforementioned rhizobacterial strains in earthen pots filled with 3 kg of sand-mixed soil and 5% biochar. The morphology of biochar was observed with highly porous nature, along with the detection of various essential elements. The biochemical and physiological data showed that phenolic compounds and osmolytes were adversely affected by the induction of drought stress. However, the application of biochar and rhizobacteria boosted the level of flavonoids on average by 52.03%, total phenols by 50.67%, soluble sugar by 82.85%, proline by 76.81%, glycine betaine by 107.25%, and total protein contents by 89.18% in all co-treatments of biochar and rhizobacteria. In addition, stress indicator compounds, including malondialdehyde (MDA) contents and H2O2, were remarkably alleviated by 54.21% and 47.03%, respectively. Similarly, the amplitude of antioxidant enzymes including catalase, peroxidase, superoxide dismutase, ascorbate peroxidase, and guaiacol peroxidase was also enhanced by 63.80%, 80.95%, 37.87%, and 58.20%, respectively, in all co-treatments of rhizobacteria and biochar. Conclusively, biochar and rhizobacteria have a magnificent role in enhancing the drought tolerance potential of crop plants by boosting the physio-biochemical traits and enhancing the level of antioxidant enzymes.

• MeSH
• Antioxidants metabolism   MeSH
• Charcoal * chemistry   MeSH
• Phenols metabolism   MeSH
• Flavonoids metabolism   analysis   MeSH
• Stress, Physiological * MeSH
• Plant Roots microbiology   growth & development   MeSH
• Malondialdehyde metabolism   MeSH
• Droughts * MeSH
• Soil Microbiology MeSH
• Vicia faba * microbiology   growth & development   MeSH
• Publication type
• Journal Article MeSH

• Publication type
• Overall MeSH

Trisaccharides bind to their interaction partners-lectins relatively weakly, which makes detection of their complexes challenging. In this work, we show that an osmolyte presence improves the distinguishing complexes of lectin Sambucus nigra with trisialyllactoses with various binding affinities. The addition of osmolyte, non-binding sugar mannose significantly improved the precision of binding experiments performed using chronopotentiometric stripping at the electrode surface and fluorescence analysis in solution. Osmolytes minimized nonspecific interactions between binding sugar and lectin. Obtained findings can be utilized in any in vitro methods studying interactions of carbohydrates, respectively their conjugates with proteins. The study of carbohydrate interactions appears important since they play essential roles in a variety of biological processes including carcinogenesis.

• MeSH
• Sambucus nigra * chemistry   metabolism   MeSH
• Sugars MeSH
• Lectins * metabolism   MeSH
• Trisaccharides metabolism   MeSH
• Publication type
• Journal Article MeSH

Plant survival in response to freezing stress depends on the efficient activation of tolerance mechanisms. Fritillaria imperialis exposure to freezing stress enhanced signalling molecules Ca2+ and H2O2 along with overexpression of Ca2+ signalling proteins (Ca2+ dependent protein kinases, CPK), followed by upregulation of NHX1 (Na+/H+ antiporter), LEA (late embryogenesis abundant proteins) and P5CS (1-pyrroline-5-carboxylate synthetase). Overexpression of OsCNGC6 was responsible for high accumulation Ca2+, Na+ and K+. The NHX1 gene product transported Na+ to vacuoles and increased cytosolic K+ content to re-establish ionic homeostasis under stress conditions. The reduced water potential of leaves was due to high accumulation of osmolytes and ions. No changes were observed in relative water content of leaves, which might be correlated with overexpression of the LEA gene, which protects against dehydration. High accumulation of H2O2 under freezing stress was responsible for activation of antioxidant systems involving SOD, phenols, anthocyanins, catalase and ascorbate peroxidase. Photosynthesis, suppressed in freezing-stressed plants, returned to normal levels after termination of freezing stress. Taken together, our findings suggest that Fritillaria efficiently tolerated freezing stress through induction of signalling mechanisms and overexpression of cold stress-responsive genes, and prevention of cold-induced water stress, oxidative stress and photosynthetic damage.

• MeSH
• Antioxidants metabolism   MeSH
• Photosynthesis physiology   MeSH
• Fritillaria genetics   metabolism   MeSH
• Stress, Physiological genetics   MeSH
• Cold Temperature * MeSH
• Oxidative Stress physiology   MeSH
• Hydrogen Peroxide metabolism   MeSH
• Reactive Oxygen Species metabolism   MeSH
• Gene Expression Regulation, Plant * MeSH
• Plant Proteins genetics   metabolism   MeSH
• Publication type
• Journal Article MeSH

Being a chilling-sensitive staple crop, rice (Oryza sativa L.) is vulnerable to climate change. The competence of rice to withstand chilling stress should, therefore, be enhanced through technological tools. The present study employed chemical intervention like application of sodium nitroprusside (SNP) as nitric oxide (NO) donor and elucidated the underlying morpho-physiological and biochemical mechanisms of NO-mediated chilling tolerance in rice plants. At germination stage, germination indicators were interrupted by chilling stress (5.0 ± 1.0 °C for 8 h day-1), while pretreatment with 100 μM SNP markedly improved all the indicators. At seedling stage (14-day-old), chilling stress caused stunted growth with visible toxicity along with alteration of biochemical markers, for example, increase in oxidative stress markers (superoxide, hydrogen peroxide, and malondialdehyde) and osmolytes (total soluble sugar; proline and soluble protein content, SPC), and decrease in chlorophyll (Chl), relative water content (RWC), and antioxidants. However, NO application attenuated toxicity symptoms with improving growth attributes which might be related to enhance activities of antioxidants, mineral contents, Chl, RWC and SPC. Furthermore, principal component analysis indicated that water imbalance and increased oxidative damage were the main contributors to chilling injury, whereas NO-mediated mineral homeostasis and antioxidant defense were the critical determinants for chilling tolerance in rice. Collectively, our findings revealed that NO protects against chilling stress through valorizing cellular defense mechanisms, suggesting that exogenous application of NO could be a potential tool to evolve cold tolerance as well as climate resilience in rice.

• MeSH
• Nitric Oxide Donors pharmacology   MeSH
• Homeostasis drug effects   MeSH
• Germination drug effects   MeSH
• Malondialdehyde metabolism   MeSH
• Nitroprusside pharmacology   MeSH
• Cold Temperature MeSH
• Crop Protection methods   MeSH
• Nitric Oxide metabolism   MeSH
• Hydrogen Peroxide metabolism   MeSH
• Lipid Peroxidation drug effects   MeSH
• Peroxidases metabolism   MeSH
• Cold-Shock Response physiology   MeSH
• Oryza drug effects   MeSH
• Seedlings drug effects   MeSH
• Superoxides metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Aldehyde dehydrogenases (ALDHs) are responsible for oxidation of biogenic aldehyde intermediates as well as for cell detoxification of aldehydes generated during lipid peroxidation. So far, 13 ALDH families have been described in plants. In the present study, we provide a detailed biochemical characterization of plant ALDH2 and ALDH7 families by analysing maize and pea ALDH7 (ZmALDH7 and PsALDH7) and four maize cytosolic ALDH(cALDH)2 isoforms RF2C, RF2D, RF2E and RF2F [the first maize ALDH2 was discovered as a fertility restorer (RF2A)]. We report the crystal structures of ZmALDH7, RF2C and RF2F at high resolution. The ZmALDH7 structure shows that the three conserved residues Glu(120), Arg(300) and Thr(302) in the ALDH7 family are located in the substrate-binding site and are specific to this family. Our kinetic analysis demonstrates that α-aminoadipic semialdehyde, a lysine catabolism intermediate, is the preferred substrate for plant ALDH7. In contrast, aromatic aldehydes including benzaldehyde, anisaldehyde, cinnamaldehyde, coniferaldehyde and sinapaldehyde are the best substrates for cALDH2. In line with these results, the crystal structures of RF2C and RF2F reveal that their substrate-binding sites are similar and are formed by an aromatic cluster mainly composed of phenylalanine residues and several nonpolar residues. Gene expression studies indicate that the RF2C gene, which is strongly expressed in all organs, appears essential, suggesting that the crucial role of the enzyme would certainly be linked to the cell wall formation using aldehydes from phenylpropanoid pathway as substrates. Finally, plant ALDH7 may significantly contribute to osmoprotection because it oxidizes several aminoaldehydes leading to products known as osmolytes.

• MeSH
• Aldehyde Dehydrogenase chemistry   genetics   metabolism   MeSH
• Phylogeny MeSH
• Pisum sativum enzymology   genetics   MeSH
• Isoenzymes chemistry   genetics   metabolism   MeSH
• Catalytic Domain genetics   MeSH
• Kinetics MeSH
• Crystallography, X-Ray MeSH
• Zea mays enzymology   genetics   MeSH
• Models, Genetic MeSH
• Models, Molecular MeSH
• Molecular Sequence Data MeSH
• NAD metabolism   MeSH
• Plant Proteins chemistry   genetics   metabolism   MeSH
• Plants enzymology   genetics   MeSH
• Amino Acid Sequence MeSH
• Gene Expression Profiling MeSH
• Substrate Specificity MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Springtails are closely related to insects, but they differ from these with respect to water balance, in particular because springtails are small and have high integumental permeability to water. Here we report a series of experiments addressing the dynamics of osmoregulation, water content and accumulation of free amino acids (FAAs) in three springtail species during exposure to a gradually increasing environmental desiccation simulating conditions in drought exposed soil. Folsomia candida and Protaphorura fimata (both living in the deeper soil layers; euedaphic species) were active throughout the 3week exposure, with the developing drought regime ending at -3.56MPa (the soil water activity at the permanent wilting point of plants is -1.5MPa) and remained hyperosmotic (having an body fluid osmolality higher than the corresponding environment) to their surrounding air. Sinella curviseta (living in upper soil/litter layers; hemiedaphic species) also survived this exposure, but remained hypoosmotic throughout (i.e. with lower osmolality than the environment). The body content of most FAAs increased in response to drought in all three species. Alanine, proline and arginine were the most significantly upregulated FAAs. By combining our results with data in the literature, we could account for 82% of the observed osmolality at -3.56MPa in F. candida and 92% in P. fimata. The osmolality of S. curviseta was only slightly increased under drought, but here FAAs were considerably more important as osmolytes than in the two other species. We propose that FAAs probably have general importance in drought tolerance of springtails.

• MeSH
• Amino Acids metabolism   MeSH
• Adaptation, Biological physiology   MeSH
• Arthropods physiology   MeSH
• Droughts * MeSH
• Osmoregulation MeSH
• Soil chemistry   MeSH
• Water MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

I přes zvyšující se úspěšnost léčby neplodnosti metodami asistované reprodukce zůstává nadále problém vybrat nejkvalitnější embryo, které má v sobě potenciál dalšího vývoje a implantace. V současnosti je výběr embrya založen především na morfologických znacích. Tento přístup je subjektivní, proto je snahou nalézt jinou, objektivnější a robustnější metodu, vhodnou k výběru embrya. Metabolismus embrya představuje jednu z možností. Tato neinvazivní metoda umožňuje pozorovat změny v hladinách různých metabolitů v kultivačním médiu před inkubací a po inkubaci jednoho embrya. Nejčastěji diskutovanými látkami jsou sacharidy a aminokyseliny jako hlavní komponenty kultivačních médií. Sacharidy slouží převážně jako zdroj energie, zatímco aminokyseliny jsou využívány k syntéze proteinů a nukleotidů, působí jako antioxidanty, osmolyty a pufry. Bylo navrženo několik metod, které hodnotí metabolický profil embrya a je mnoho teorií, jak podle metabolismu embrya vybrat to nejvhodnější k transferu. Klíčová slova: aminokyseliny, metabolismus embrya, výběr embrya po IVF, sacharidy

Despite the increasing success of infertility treatment methods of assisted reproduction, it still remains a problem how to select the best embryo that has the potential for further development and implantation. At the present time, embryo selection is based especially on morphological criteria. This approach is subjective; therefore there is a trend to find another more objective and robust method for embryo selection. Embryo metabolism can be used as an indicator of viability. This non-invasive method allows observing changes in the levels of different metabolites in culture medium before and after incubation of the only one embryo. The most mentioned substances are carbohydrates and amino acids as important components of culture medium. Carbohydrates serve predominantly as energy sources, whereas amino acids are precursors of protein and nucleotides, antioxidants, osmolytes, pH regulators etc. Several methods have been proposed for evaluating of embryo metabolic profile of embryo. There are many hypotheses for embryo selection according its metabolic profile. Keywords: amino acids, embryo metabolism, embryo selection in IVF, carbohydrates

• Keywords
• metabolismus embrya, výběr embrya,
• MeSH
• Amino Acids metabolism   MeSH
• Reproductive Techniques, Assisted MeSH
• Embryo, Mammalian * metabolism   MeSH
• Embryonic Development MeSH
• Embryo Culture Techniques MeSH
• Culture Media * chemistry   metabolism   MeSH
• Humans MeSH
• Lipid Metabolism MeSH
• Carbohydrate Metabolism MeSH
• Embryo Disposition * MeSH
• Embryo Transfer MeSH
• Check Tag
• Humans MeSH

The review is focused on plant proteome response to salinity with respect to physiological aspects of plant salt stress response. The attention is paid to both osmotic and ionic effects of salinity stress on plants with respect to several protein functional groups. Therefore, the role of individual proteins involved in signalling, changes in gene expression, protein biosynthesis and degradation and the resulting changes in protein relative abundance in proteins involved in energy metabolism, redox metabolism, stress- and defence-related proteins, osmolyte metabolism, phytohormone, lipid and secondary metabolism, mechanical stress-related proteins as well as protein posttranslational modifications are discussed. Differences between salt-sensitive (glycophytes) and salt-tolerant (halophytes) plants are analysed with respect to differential salinity tolerance. In conclusion, contribution of proteomic studies to understanding plant salinity tolerance is summarised and discussed.

• Publication type
• Journal Article MeSH

Misfolding and aggregation of mutant enzymes have been proposed to play role in the pathogenesis of homocystinuria due to cystathionine β-synthase (CBS) deficiency. Chemical chaperones have been recently shown to facilitate proper assembly of several CBS mutants. To asses the number of patients that may respond to chaperone therapy, we examined the effect of selected CBS ligands and osmolytes on assembly and activity of 27 CBS mutants that represent 70% of known CBS alleles. The mutant enzymes were expressed in a bacterial system, and their properties were assessed by native Western blotting and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) assay, respectively. We studied the chaperoning activity of δ-aminolevulinic acid (δ-ALA)-a heme precursor-and of three osmolytes betaine, 2-aminoethanesulfonic acid (taurine), and glycerol. Fourteen mutants responded by at least 30% increase in the amount of correctly assembled tetramers and enzymatic activity to the coexpressional presence of either 0.5 mM δ-ALA, 100 mM betaine, and/or 750 mM glycerol. Eight of these mutants (p.R266K, p.P49L, p.R125Q, p.K102N, p.R369C, p.V180A, p.P78R, p.S466L) were rescuable by all of these three substances. Four mutants showed increased formation of tetramers that was not accompanied by changes in activity. Topology of mutations appeared to determine the chaperone responsiveness, as 11 of 14 solvent-exposed mutations were substantially more responsive than three of 13 buried mutations. This study identified chaperone-responsive mutants that represent 56 of 713 known patient-derived CBS alleles and may serve as a basis for exploring pharmacological approaches aimed at correcting misfolding in homocystinuria.

• MeSH
• Alleles MeSH
• Betaine pharmacology   therapeutic use   MeSH
• Cystathionine beta-Synthase chemistry   drug effects   genetics   metabolism   MeSH
• Escherichia coli metabolism   MeSH
• Glycerol pharmacology   MeSH
• Homocystinuria drug therapy   genetics   metabolism   MeSH
• Polymorphism, Single Nucleotide physiology   MeSH
• Protein Conformation drug effects   MeSH
• Aminolevulinic Acid pharmacology   therapeutic use   MeSH
• Humans MeSH
• Ligands MeSH
• Molecular Chaperones pharmacology   therapeutic use   MeSH
• Protein Multimerization drug effects   MeSH
• Mutant Proteins chemistry   drug effects   metabolism   MeSH
• Protein Folding drug effects   MeSH
• Taurine pharmacology   therapeutic use   MeSH
• Protein Binding MeSH
• Structure-Activity Relationship MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH