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Abstract This study focused on the idea that the toxic effect of zearalenone (ZEA) and the protective actions of the brassinosteroid - 24-epibrassinolide (EBR) as well as selenium are dependent on its accumulation in chloroplasts to a high degree. These organelles were isolated from the leaves of oxidative stress-sensitive and stress-tolerant wheat cultivars that had been grown from grains that had been incubated in a solution of ZEA (30 μM), Na 2 SeO 4 (Se, 10 μM), EBR (0.1 μM) or in a mixture of ZEA with Se or EBR. Ultra-high performance liquid chromatography techniques indicated that ZEA was adsorbed in higher amounts in the chloroplasts in the sensitive rather than tolerant cultivar. Although the brassinosteroids and Se were also accumulated in the chloroplasts, higher levels were only found in the tolerant cultivar. The application of EBR increased the homocastasterone content, especially in the chloroplasts of the tolerant plant and after the addition of ZEA. The presence of both protectants caused a decrease in the ZEA content in studied organelles and resulted in diminishing of the oxidative stress (i.e. changes in the activity of the antioxidative enzymes). Moreover, a recovery of photosystem II and decrease in the negative impact of ZEN on Hsp 90 transcript accumulation was observed in plants. Highlights • Zearalenone, brassinosteroids and selenium were accumulated in chloroplasts. • 24-Epibrassinolide and Se absorption in chloroplasts was stimulated by zearalenone. • 24-Epibrassinolide and Se absorption in chloroplast decreased of zearalenone content. • Defense mechanism against zearalenone involved protection of PSII. [ABSTRACT FROM AUTHOR]

Steroids constitute one of the most important groups of compounds of regulatory properties both in the animal and plant kingdom. In plants, steroids such as brassinosteroids or progesterone, by binding to protein receptors in cell membranes, regulate growth and initiate processes leading to increased tolerance to stress conditions. Due to their structural similarities to sterols, these steroids may also directly interact with cellular membranes. Our aim was to determine the changes of the structural parameters of lipid membranes under the influence of hydrophobic steroid compounds, i.e., 24-epibrassinolide (EBR) and its precursor—24-epicastasterone (ECS) and progesterone (PRO). Lipids were isolated from wheat seedlings with different tolerances to frost, grown at low temperatures (5 °C) for 1.5 and 3 weeks (acclimation process). Control plants were cultured continuously at 20 °C. From galactolipids and phospholipids, the main polar lipid fractions, the monolayers were formed, using a technique of Langmuir trough. EBR and ECS were introduced into monolayers, together with lipids, whereas the PRO was dissolved in the aqueous sub-phase upon which the monolayers were spread. Measurements performed at 25 °C and 10 °C showed a significant action of the tested compounds on the physicochemical properties of the monolayers. EBR and PRO increased the area per lipid molecule in monolayers, resulting in formation of more flexible surface structures while the presence of the ECS induced the opposite effect. The influence of the polarity of lipids and steroids on the interactions in the monolayer was discussed. Lipids extracted from the membranes of wheat with the most tolerance to frost were characterized by the highest fatty acid unsaturation and steroids had a relatively weak effect on the parameters of the structure of their monolayers. [ABSTRACT FROM AUTHOR]

The aim of this research was to characterize the changes of structural organization of chloroplasts of sensitive (Maresi) and tolerant (Cam/B1) barley genotypes upon soil drought (10 days), which was applied in two stages of plant growth, i.e. seedlings and flag leaves. The electron paramagnetic resonance (EPR) technique was used for the determination of changes in the concentration and nature of long-lived radicals and metal ions (Mn, Fe), measured directly in the structures of fresh leaves, occurring after stress treatment. Stronger variations of EPR parameters were found after drought stress application in the flag-leaf phase and for sensitive genotype. Chloroplasts of Cam/B1 were characterized by a larger surface area and less degradation of their structure during drought stress in comparison to Maresi. The data obtained from Raman spectra showed that better stress tolerance of the genotype was accompanied by greater accumulation of carotenoids in chloroplasts and was correlated with an increase in carotenoid radicals. The increase of the value of the electrokinetic potential (relative to control), which was slightly larger for the chloroplasts of Maresi than of Cam/B1, indicated the chemical reconstruction of the membrane leading to a reduction of their polarity during drought action. [ABSTRACT FROM AUTHOR]

Abstract: Two kinds of wheat genotypes with different tolerance to osmotic stress (NaCl and PEG-treatment) were investigated with biochemical analyses, including the measurements of total antioxidant capacity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, reducing power and starch content. The results were compared with electron paramagnetic resonance (EPR) data concerning the nature and amounts of stable long lived radicals present in the control and stressed plants. In addition, the changes in manganese content upon stress conditions were monitored. Different mechanisms of protection against PEG stress in sensitive and tolerant wheat genotypes were postulated. In sensitive genotypes, electrons were created in excess in stress conditions, and were stabilized by polysaccharide molecules, whereas in tolerant genotypes, protection by antioxidants dominated. Moreover, the quinone–semiquinone balance shifted towards semiquinone, which became the place of electron trapping. NaCl-treatment yielded significant effects mainly in sensitive genotypes and was connected with the changes of water structure, leading to inactivation of reactive oxygen species by water molecules. [Copyright &y& Elsevier]

Abstract: Grains of five genotypes of wheat (four Polish and one Finnish), differing in their tolerance to drought stress were chosen for this investigation. Electron paramagnetic resonance spectroscopy allowed observation of transition metal ions (Mn, Fe, Cu) and different types of stable radicals, including semiquinone centers, present in seed coats, as well as several types of carbohydrate radicals found mainly in the inner parts of grains. The content of paramagnetic metal centers was higher in sensitive genotypes (Radunia, Raweta) than in tolerant ones (Parabola, Nawra), whereas the Finnish genotype (Manu) exhibited intermediate amounts. Similarly, the concentrations of both types of radicals, carbohydrates and semiquinone were significantly higher in the grains originating from more sensitive wheat genotypes. The nature of carbohydrate radicals and their concentrations were confronted with the kinds and amounts of sugars found by the biochemical analyses and microscopy observations. It is suggested that some long lived radicals (semiquinone and starch radicals) occurring in grains could be indicators of stress resistance of wheat plants. [Copyright &y& Elsevier]

Abstract: The putative role of cytokinins in processes leading to reproductive development of plants was investigated by analysing the shoot apical parts of a winter cultivar of oilseed rape (Brassica napus L. var. oleifera, cv. Górczański). The endogenous cytokinin levels were measured by liquid chromatography–tandem mass spectrometry (LC–MS) in the shoot apices of vegetative plants (grown at 20/17°C with a 16/8h day/night regime) and vernalized plants (56 days at 5/2°C with a 16/8h photoperiod) at different times during floral transition. During vernalization, the content of all isoprenoid cytokinins increased significantly, coinciding well with the onset of the early stages of reproductive development. Cytokinin levels reached their maxima when most of the plants became irreversibly reproductive (after 42 days of cold treatment). cis-Zeatin riboside (unequivocally identified by quadrupole-time-of-flight MS) accounted for ca. 87–89% of the total isoprenoid cytokinin content in control and vernalized plants, whilst N6-isopentenyladenosine (∼6% in control and ∼8% in vernalized plants) and cis-zeatin (∼2% in control and ∼1% in vernalized plants) were the next most abundant cytokinins. In the post-vernalization period, endogenous cytokinin levels decreased, but remained significantly higher in the reproductive plants than in the vegetative controls. These results suggest that cytokinins, especially those of the cis-zeatin type, are involved in vernalization-induced reproductive development of B. napus. [Copyright &y& Elsevier]

Abstract: Biochemical analyses of antioxidant content were compared with measurements of fluorescence and electron paramagnetic resonance (EPR) to examine the alteration of radicals in wheat seedlings exposed to 2 days of selenium stress. Two genotypes of Polish and one of Finnish wheat, differing in their tolerance to long-term stress treatment, were cultured under hydroponic conditions to achieve the phase of 3-leave seedlings. Afterwards, selenium (sodium selenate, 100μM concentration) was added to the media. After Se-treatment, all varieties showed an increase in carbohydrates (soluble and starch), ascorbate and glutathione content in comparison to non-stressed plants. These changes were more visible in Finnish wheat. On the basis of lipid peroxidation measurements, Finnish wheat was recognized as the genotype more sensitive to short-term Se-stress than the Polish varieties. The antioxidant enzyme activities (superoxide dismutase, ascorbate peroxidase and glutathione reductase) increased in Polish genotypes, whereas they decreased in Finnish wheat plants cultured on Se media. The action of reactive oxygen species in short-term action of Se stress was confirmed by the reduction of PSII and PSI system activities (measured by fluorescence parameters and EPR, respectively). EPR studies showed changes in redox status (especially connected with Mn(II)/Mn(III), and semiquinone/quinone ratios) in wheat cell after Se treatment. The involvement of the carbohydrate molecules as electron traps in production of long-lived radicals is postulated. [Copyright &y& Elsevier]

Abstract: Currents flowing through slow vacuolar SV channels of rape (Brassica napus) growing on media supplemented with Cd2+ (400μM), and/or SeO4 2− (2μM) were examined. The aim of the study was to investigate the role of Cd2+ in modulation of SV channel activity and to determine whether Se reverses the effect of cadmium. Vacuoles were isolated using a quick surgical method to avoid application of any cell wall-degrading enzymes. Vacuoles of rape exhibited typical SV channel activity with slow activation at positive potentials and strong rectification into the vacuolar lumen. Single-channel conductance in cytoplasm-side-out tonoplast patches ranged between 68.8±1.9pS in the control, 80.1±2.5pS, in Cd2+-, 74.2±2.4pS in Cd2+/selenate-, and 80.1±1.8pS in selenate-pretreated plants. The lack of a clear tendency was likely an effect of equilibration of the pipette solution (without Cd2+/SeO4 2−) with that of the luminal side of the vesicles. In the vacuole-attached configuration, in which natural vacuolar solution was not exchanged, there was a significant reduction in single-channel conductance in the Cd2+- (40.3±2.8pS), Cd2+/selenate- (47.1±2.8pS) and selenate-pretreated (42.3±1.4pS) plants, compared to the control (60.2±1.7pS). The reduction in single-channel conductance only partially explained the significant decline in the densities of ion current flowing through the vacuolar membrane in the whole-vacuole configuration in the plants growing on Cd2+ and Cd2+/selenate media. Thus, Cd2+ accumulation in the vacuole reduced the number of active SV channels from 0.28±0.05μm−2 in the control to 0.021±0.005 and 0.039±0.004μm−2 in Cd2+ and Cd2+/selenate-pretreated plants, respectively. [Copyright &y& Elsevier]

Abstract: Selenium appears to be an important protective agent that decreases cadmium-induced toxic effects in animals and plants. The aim of these studies was to investigate the changes of properties of chloroplast membranes obtained from Cd-treated rape seedlings caused by Se additions. Chloroplasts were isolated from leaves of 3-week-old rape plants cultured on Murashige-Skoog media supplied with 2μM Na2SeO4 and/or 400μM CdCl2 under in vitro conditions. The following physicochemical characteristics of chloroplasts were chosen as indicators of Se-effects: average size, zeta potential, ultrastructure, lipid and fatty acid composition and fluidity of envelope membrane. The results suggest that Se can partly counterbalance the destructive effects of Cd. This protective action led to an increase of chloroplast size reduced by Cd treatment and rebuilt, to some extent, the chloroplast ultrastructure. Lipid and fatty acid composition of chloroplast envelopes modified by Cd showed a decrease in digalactosyl-diacylglycerol content and an increase of content of monogalactosyl-diacylglycerol and phospholipid fractions, as well as an increase of fatty acid saturation of all lipids studied. The change in fatty acid saturation correlated well with a decrease of membrane fluidity and with a diminishing of absolute values of zeta potential. The presence of selenium in cultured media caused a partial reversal of the detected changes, which was especially visible in properties related to the hydrophobic part of an envelope, i.e. fatty acid saturation and fluidity. [Copyright &y& Elsevier]

Summary: Currents passing through slowly activating vacuolar channels (SV) in isolated vacuoles from winter (Górczański) and spring (Młochowski) varieties of rape (Brassica napus) were examined using the patch-clamp technique. Eight-week-long vernalization at 5/2°C (day/night) was applied to obtain the generative stage of winter rape. SV channels of vacuoles isolated from vegetative (rosette) and generative leaves of both varieties were examined in order to investigate a possible role of these ion channels in rape flowering. Single SV channel conductance measured in a vacuole-attached configuration (natural cell sap) ranged from 60 to 83pS. Lower values were observed in the generative leaves of both varieties. Unitary conductance measured in excised cytoplasm-out membrane patches did not differ significantly among the experimental variants, with the exception of spring generative vacuoles, where it was significantly lower. There was also no difference in SV current densities measured in the whole-vacuole configuration. Gibberellic acid (GA3) (2mg/l) caused lowering of macroscopic SV currents by 20%, and had no significant effect on the single channel conductance. We conclude that SV channels play a role in rape vernalization and flowering owing to their multifactor regulation abilities rather than structural changes. [Copyright &y& Elsevier]

Abstract: The aim of presented researches was to investigate the physicochemical properties of Langmuir monolayer of galactolipids extracted from two different kinds of plastids: immature embryos and inflorescences. Differences between the physicochemical properties of the plastid membranes may help to explain different physiological processes, such as plant regeneration. Surface pressure (π) vs. molecular area (A) isotherms of the monogalactosyldiacylglycerol (MGDG)/digalactosyldiacylglycerol (DGDG) monolayers of various molar ratios were measured at 15°C. Galactolipids were extracted from two different types of tissue: inflorescences and embryos. Based on the analysis of the π–A isotherms, the properties of monolayers, such as collapse pressure (π coll), limiting area (A lim), compressibility modulus (), excess free energy of mixing (ΔG EXC) and free energy of mixing (ΔG MIX), were calculated. The results show that pure MGDG and DGDG and their mixtures form liquid-expanded monolayers, independently on the kind of tissue. Galactolipids originating from inflorescences produce more compressible films at the air/water interface, with larger limiting area per molecule and lower stability against the collapse process. MGDG and DGDG are miscible and form non-ideal mixed monolayers at the air/water interface. Negative values of ΔG EXC were calculated for the mixture of galactolipids originating from inflorescences, with the content of MGDG, x MGDG >0.6. In the case of embryos, the negative values of ΔG EXC were found for x MGDG ≈0.5. Therefore, the attractive interactions between MGDG and DGDG exist in the mixtures of these compositions. As it is shown by negative values of ΔG MIX, mixed monolayers are more stable compared with unmixed ones. [Copyright &y& Elsevier]

Summary: The effect of selenium (Se) on rape (Brassica napus) seedlings subjected to cadmium (Cd) stress was studied in vitro by investigating plant growth and changes in fatty acid composition, activity of antioxidative enzymes and DNA methylation pattern. Physiological experiments were carried out on seedlings cultured for 2 weeks on Murashige-Scoog (MS) media with Cd concentrations of 0, 400 and 600μM, and on corresponding media supplied with Se (2μM). Exposure to increasing Cd concentrations reduced the fresh weight of the upper part (hypocotyls+cotyledons) of the seedlings more strongly than that of the root system, which was accompanied by higher Cd accumulation in these tissues. In the upper part, Cd exposure led to significant changes in the biochemical parameters: fatty acid unsaturation of plasmalemma decreased, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPOX) diminished and that of ascorbate peroxidase (APX) increased. In contrast, the roots showed an increase in fatty acid unsaturation and in the activity of antioxidative enzymes. In both parts of rape seedlings H2O2 level and lipid peroxidation increased. Se addition to medium considerably reversed the Cd-induced decrease in fresh mass as well as the changes in lipid unsaturation and peroxidation. Se applied separately or in combination with Cd did not significantly affect the activity of antioxidative enzymes in the roots, but diminished it in the upper part. Moreover, the presence of Se in medium prevented changes in the DNA methylation pattern triggered in rape seedlings by high Cd concentrations. Two possible mechanisms for the action of Se were considered: (1) removal of Cd from metabolically active cellular sites, and (2) reduction of oxygen radicals. [Copyright &y& Elsevier]

Summary: The electric and structural properties of envelope membranes of chloroplasts obtained from vegetative and generative plants of rape and the effect of hormone (IAA, GA3 and zearalenone) treatment were determined by zeta potential and small-angle X-ray scattering (SAXS) methods. Chloroplasts were isolated from leaves cut off from the vegetative (before cooling) and generative apical parts of plants. The lipid composition of chloroplast envelope membranes were analyzed by chromatographic techniques. Envelopes from generative plants contained higher levels of digalactosyldiacylglycerol (DGDG) and smaller amounts of phospholipids (PLs) in comparison to those obtained from vegetative ones. Moreover, envelopes of generative plants were characterized by higher fractions of unsaturated fatty acids. The zeta potential changes caused by hormone treatment were higher for chloroplasts isolated from vegetative plants in comparison to chloroplasts isolated from generative ones. An especially strong effect was observed for chloroplasts treated with IAA. The thickness of bilayers of untreated chloroplasts from vegetative plants were larger by 0.4nm when comparing to the thickness of layers obtained from generative ones. The effect of hormones (GA3 and zearalenone) was detected only for vegetative chloroplasts. Both applied methods indicated differences in the properties of untreated and hormone-treated chloroplasts obtained from vegetative and generative plants. [Copyright &y& Elsevier]

Summary: Surface parameters of polar lipids extracted from winter wheat plastids were investigated by the Langmuir and X-ray differentiation scattering methods. Highly purified plastids were isolated from non-embryogenic (NE) and embryogenic (E) calli initiated from inflorescences. NE plastids contained more monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) and less phospholipids (PL) fraction than E plastids. Moreover, in E calli, unsaturated fatty acids were detected in a higher proportion than in NE for both MGDG and DGDG. No significant differences in fatty acids saturation of PL between NE and E objects were detected. Aqueous surface monolayers were prepared from separate lipids and from mixtures of glycolipids and PL. In the case of MGDG, isotherms showed specific shoulders, contrary to continuous isotherms obtained for other investigated lipids. On the base of isotherms, the surface parameters: limiting area (A lim) and collapse pressure () were calculated. Indole-3-acetic acid (IAA) increased the A lim of all separated lipids about 4–10Å2/mol. However, for NE lipid mixture, the effect of IAA was much smaller (about 2Å2/mol) than for other objects (usually about 5Å2/mol). X-ray experiments for liposomes, obtained from mixtures of glycolipids and PL of NE and E plastids, showed continuous scattering curves with maxima characteristic for lipid bilayer membranes. Calculations of distance distribution functions indicated that bilayer thickness was 41 and 38Å for NE and E, respectively. IAA influence on membrane structures was detected especially in E liposomes and increased the distance between head groups by about 2Å. It is suggested that changes occur during embryogenesis in specific structure of plastid membranes determined also the formation of domains, similar to that suggested for plasmalemma (Plant Sci. 165 (2003) 265). IAA treatment influenced the membrane structure, especially E plastids increasing distances between polar groups. [Copyright &y& Elsevier]

The structure of phospholipid bilayers in the presence of sterols was investigated using the X-ray technique. Both phospholipids and sterols were isolated from the plasmalemma of winter wheat calli initiated from immature inflorescences. Calli were cultured to non-embryogenic (NE) and embryogenic (E) types by changing the proportion of 2,4-D in culture medium. Sterols presence in phospholipid mixture did not markedly influence the P–P distance in bilayers in comparison to sterols-free bilayers. Thus reduced thickness of membranes constructed from E lipids in comparison to NE was detected. Additionally, phytohormone (IAA, 2,4-D, kinetin) influence on sterols/phytohormone bilayers was visible only in the case of lipids extracted from E cells. These results confirm our earlier suggestion about higher flexibility and existence of specific phospholipid domains for phytohormone adsorption. Sterols could precisely fix the conformational ordering of the phospholipid. Lower levels of sterols in NE membranes may favour looser packing of bilayer, and consequently lead to higher permeability to phytohormones. On the other hand, the differences in phospholipid domains composition could result in different phytohormone actions on NE and E membranes. [Copyright &y& Elsevier]

Changes in surface structure of liposomes in aqueous solutions obtained from phospholipids of plasmalemma of winter wheat cells were analysed by small-angle X-ray scattering (SAXS) method. Winter wheat cells were obtained from immature inflorescences and cultured in Murashige and Skoog medium supplemented with auxin (non-embryogenic calli, NE) or without phytohormones (embryogenic calli, E). Measurements were performed in the fully hydrated liposomal dispersions in pure water:methanol (100:1 v/v) buffer or in buffer supplemented by auxins (IAA and its chemical analogue melatonine) or cytokinins (kinetin and its analogue mercaptopurine). Both types of liposomes (NE and E) were different in the hydrophilic and hydrophobic phase. The higher amount of PE in comparison to PC as well as the higher content of unsaturated fatty acids, especially 18:2 and 18:3, were observed in E membranes. The SAXS curves from NE and E liposomes indicate the possibility of bilayer formation by these phospholipids. Changes in distance distribution functions derived for NE and E phospholipid liposomes could be explained by changes of bilayer thickness. All investigated hormones were affected only E liposomes and did not have an influence on NE liposomes. These changes of the bilayer size of E membranes after phytohormone treatment were in the range of those detected for NE membranes. This was observed especially for IAA and its chemical analogue. Kinetin influenced bilayer thickness in a lesser degree and inconsiderable differences were noticed for its chemical analogue—mercaptopurine. Thus we suggest that removal of auxins from winter wheat culture media is necessary to maintain structure of membranes characteristic for embryogenic cells. [Copyright &y& Elsevier]

The aim of this work was to investigate the relationships between the chemical composition of oat grains and the tolerance to oxidative stress of oat genotypes. The studies were based on the results of biochemical analyses and both EPR and Raman spectroscopies on whole grains and their parts (embryo, endosperm, seed coat) originating from oat genotypes with different sensitivities to stress. We found that the amounts of fats and especially unsaturated fatty acids, proteins rich in glutamic acid and glycine, as well as phenolics and tocopherols were higher in grains of the tolerant genotype. Moreover, fats and proteins were distributed not only in embryos, but also in endosperms. The grains of tolerant genotypes exhibited high antioxidant activity and contained greater amounts of β-glucan. EPR data pointed to higher concentrations of various kinds of stable organic radicals (semiquinone, tyrosyl and carbon-centered radicals) in whole grains (and their parts) of sensitive genotypes. EPR spectra revealed the character of interactions of paramagnetic transition metal ions Fe(III) and Mn(II) with organic and inorganic structures of grains. The quantitative EPR measurements showed the dependence between the amount of radical species and the content of transition metal ions, mainly Fe(III) bonded to inorganic structures. [ABSTRACT FROM AUTHOR]

The increase of the concentration of ozone in the atmosphere, being the direct source of reactive oxygen species, results in the yield loss of agronomic crops. On the other hand, ozone is also used as a protector against microorganisms, living in plants and present in materials obtained from them, dangerous for human and animal health. In this work it has been studied if ozone in doses similar to those used for removal of microorganisms can have significant influence on the generation of stable organic radicals and changes in the character of transition metal ions and in the antioxidative biochemical parameters of cereal grains. The aim of this work was to find if the response of grains of three cereals (wheat, oat and barley) to ozone depended on their oxidative stress tolerance. The influence of direct short-term ozone application on grains of these cereals, each represented by two genotypes with different oxidative stress tolerance, was studied by biochemical analyses and by electron paramagnetic resonance (EPR) technique. Whole grains as well as their parts: embryo, endosperm and seed coat were subjected to ozone treatment for 30 min. Biochemical investigation of control samples showed that their antioxidant activity increased in order: wheat < oat < barley. EPR method revealed that character and the number of paramagnetic species (transition metal ions: Fe(III), Cu(II), Mn(II) and stable organic radicals) changed upon ozone exposure, depending on the kind of cereal, stress tolerance of particular genotype and the part of grain. The control samples of whole grains and their parts originating from sensitive genotypes contained higher amounts of stable organic radicals (semiquinone, phenoxyl and carbohydrate types) than those from tolerant ones. In embryos of grains from sensitive genotypes their amount increased upon ozone treatment stronger than in embryos from grains of tolerant cultivars. In seed coats and endosperms such relation was not found and the changes in the content of the radicals during ozone application were correlated with the amount of transition metal ions and were more intensive in parts of grains richer in easily oxidized iron species Fe(II), located in inorganic structures. On the contrary, Fe(II) ions situated in embryos were stabilized by organic matrix and did not undergo oxidation by ozone. [ABSTRACT FROM AUTHOR]

Winter crops acquire frost tolerance during the process of cold acclimation when plants are exposed to low but non-freezing temperatures that is connected to specific metabolic adjustments. Warm breaks during/after cold acclimation disturb the natural process of acclimation, thereby decreasing frost tolerance and can even result in a resumption of growth. This phenomenon is called deacclimation. In the last few years, studies that are devoted to deacclimation have become more important (due to climate changes) and necessary to be able to understand the mechanisms that occur during this phenomenon. In the acclimation of plants to low temperatures, the importance of plant membranes is indisputable; that is why the main aim of our studies was to answer the question of whether (and to what extent) deacclimation alters the physicochemical properties of the plant membranes. The studies were focused on chloroplast membranes from non-acclimated, cold-acclimated and deacclimated cultivars of winter oilseed rape. The analysis of the membranes (formed from chloroplast lipid fractions) using the Langmuir technique revealed that cold acclimation increased membrane fluidity (expressed as the A lim values), while deacclimation generally decreased the values that were induced by cold. Moreover, because the chloroplast membranes were penetrated by lipophilic molecules such as carotenoids or tocopherols, the relationships between the structure of the lipids and the content of these antioxidants in the chloroplast membranes during the process of the cold acclimation and deacclimation of oilseed rape are discussed. [Display omitted] • Cold-induced changes in unsaturation of FA in chloroplast were associated with changes in the fluidity of the monolayers. • Cold increased the fluidity of the membranes - deacclimation partly reverse these effect. • Deacclimation causes a decrease in the amount of tocopherols, it may contribute to reduced frost tolerance of plants. • Accumulation of carotenoids in membranes may results in better tolerance of plants to frost, especially after deacclimation. [ABSTRACT FROM AUTHOR]

Highlights: [•] Formation of stable radicals of various character in flours. [•] Influence of the content of transition metal ions on thermal radical generation. [•] Mechanism of thermal radical formation engaging the redox processes. [•] Proteins and carbohydrates as sites of radical stabilization. [•] Participation of starch in thermal radical generation in flours. [ABSTRACT FROM AUTHOR]

Abstract: The behaviour of equimolar mixtures of α-tocopherol with monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) and phospholipids (PL) isolated from wheat calli cultured on media with and without cadmium was investigated at the air–water interface by surface pressure–area (π–A) measurements established using an automated Langmuir-type film balance. It was found that monolayers of all studied compounds were expanded. The additivity rule was not fulfilled and the collapse pressure of mixtures was different from these recorded for pure components. This can be related with the existence of interactions between molecules in mixed monolayers. Tocopherol diminished the differences between parameters of monolayers formed by lipids extracted from objects cultivated on various media (with and without cadmium). [Copyright &y& Elsevier]

The purpose of experiments was to describe the alterations of content of steroid regulators (brassinosteroids, progesterone) during cold hardening of winter wheat. Further we studied physiological and biochemical changes induced by these steroids in cold hardened winter wheat together with estimation of plant frost tolerance. The endogenous brassinosteroid content was elevated in winter wheat during cold hardening while level of progesterone was lowered. A higher content of brassinosteroids (but not progesterone) was connected to better frost tolerance of winter wheat cultivars. Plant supplementation with brassinosteroid (24-epibrassinolide) and progesterone before cold hardening reduced frost damage. Tests with the inhibitors of the biosynthesis of brassinosteroids and progesterone suggested that these steroids are one of players in regulating the antioxidant system in winter wheat during cold hardening. Their role in regulating the expression of Rubisco or the Rubisco activase gene was less clear. Steroid regulators did not affect the content of the stress hormone ABA. Model studies of the membranes, made on a Langmuir bath, showed an increase in the value of the parameter describing differences in membrane compressibility (resulting from stronger interactions among the molecules in the monolayers). This suggests that 24-epibrassinolide and progesterone enter into the lipid layer and - in a similar way to sterols – stabilise the interaction among lipids. It may be significant step for better frost tolerance. The use of steroid regulators (especially brassinosteroids) as agrochemicals improving frost tolerance of winter cereals will be discussed. • Content of brassinosteroids (BR) increased in winter wheat during cold hardening. • Progesterone (P) content decreased in winter wheat during cold hardening. • Higher BR content characterised cultivars with higher frost tolerance. • BR and P were incorporated into the lipid layer and stabilized it. • BR and P regulated the activity of the SOD enzyme. [ABSTRACT FROM AUTHOR]

Brassinosteroids (BR) are plant steroid hormones that were discovered more than thirty years ago, but their physiological function has yet to be fully explained. The aim of the study was to answer the question of whether/how disturbances in the production of BR in barley affects the plant's metabolism and development under conditions of optimal watering and drought. Mutants with an impaired production of BR are one of the best tools in research aimed at understanding the mechanisms of action of these hormones. The study used barley cultivars with a normal BR synthesis (wild type) and semi-dwarf allelic mutants with an impaired activity of C6-oxidase (mutation in HvDWARF ), which resulted in a decreased BR synthesis. Half of the plants were subjected to drought stress in the seedling stage and the other half were watered optimally. Plants with impaired BR production were characterised by a lower height and developmental retardation. Under both optimal watering and drought, BR synthesis disorders caused the reduced production of ABA and cytokinins, but not auxins. The BR mutants also produced less osmoprotectant (proline). The optimally watered and drought-stressed mutants accumulated less sucrose, which was accompanied by changes in the production of other soluble sugars. The increased content of fructooligosaccharide (kestose) in optimally watered mutants would suggest that BR is a negative regulator of kestose production. The decreased level of nystose in the drought-stressed mutants also suggests BR involvement in the regulation of the production of this fructooligosaccharide. The accumulation of the transcripts of genes associated with stress response ( hsp90 ) was lower in the watered and drought-stressed BR-deficient mutants. In turn, the lower efficiency of photosystem II and the net photosynthetic rate in mutants was revealed only under drought conditions. The presented research allows for the physiological and biochemical traits of two BR-barley mutants to be characterised, which helps BR function to be understood. The knowledge can also be a good starting point for some breeding companies that are interested in introducing new semi-dwarf barley cultivars. [ABSTRACT FROM AUTHOR]

The aim of this study was to uncover the specific species in grains that might differentiate the wheat genotypes according to their tolerance to oxidative stress. Measurements by EPR and Raman spectroscopy techniques were used to examine whole grains and their parts (embryo, endosperm, seed coat) originating from four wheat genotypes with differing tolerance to drought stress. Raman spectra showed that, in spite of the similar amounts of proteins in whole grains from tolerant and sensitive genotypes, in tolerant ones they were accumulated mainly in embryos. Moreover, in embryos from these grains, a higher content of unsaturated fatty acids was observed. Endosperm of grains from the tolerant genotype, richer with starch than that of sensitive one, exhibited higher content of amylopectin. Detailed analysis of EPR signals and simulation procedures of the spectra allowed the estimation of the nature of interactions of Fe(III) and Mn(II) with organic and inorganic structures of grains and the character of organic stable radicals. Three types of these radicals: carbohydrate, semiquinone and phenoxyl, were identified. The amounts of these radicals were higher in grains of sensitive genotypes, mostly because of differences in carbohydrate radical content in endosperm. Taking into account the level of radical concentration and greater capacity for radical formation in grains from plants of lower tolerance to stress, the content of radicals, especially of a carbohydrate nature, was considered as a marker of the plant resistance to stress conditions. [ABSTRACT FROM AUTHOR]