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The aim of the study was to evaluate the effect of two types of negatively charged quasi-spherical silver nanoparticles (AgNPs) at concentrations of 10, 20 and 30 mg L−1 and silver ions at a concentration of 30 mg L−1 on the growth, selected physiological aspects and yielding of wheat (Triticum aestivum L.) cv. Tybalt, and on plant resistance to seedling blight. Seed germination, α-amylase activity in seeds, morphology and infestation of seedlings by pathogens were assessed in a hydroponic treatment. Growth rate, PSII efficiency, heading and yield of the same plants were then analysed in pot culture. Results showed that the AgNPs and silver ions had a negative effect on roots, but reduced seedling blight and improved leaf area compared to the control. In addition, the AgNPs reduced with sodium borohydride in the presence of trisodium citrate at concentrations of 10 and 20 mg L−1 stimulated germination, α-amylase activity and shoot length, which was not observed in the case of silver ions and the AgNPs reduced with sodium hypophosphite in the presence of sodium hexametaphosphate. In a pot experiment, the AgNPs improved plant growth, PSII efficiency, accelerated heading and increased yield-related parameters compared with the control. Results revealed the interaction strength in the following order: TCSB-AgNPs > SHSH-AgNPs > silver ions. TCSB-AgNPs in the lowest concentration had the most favourable effect, indicating their great potential for use in improving wheat cultivation.

The aim of the research was to evaluate the effect of three types of silver nanoparticles (AgNPs) with different physicochemical properties and silver ions delivered in the form of silver nitrate (AgNO3) at the concentration of 50 mg L−1 on germination and initial growth of monocots (common wheat, sorghum) and dicots (garden cress, white mustard). The AgNPs were prepared using trisodium citrate (TCSB-AgNPs), tannic acid (TA-AgNPs), and cysteamine hydrochloride (CHSB-AgNPs). They exhibited comparable shape, size distribution, and an average size equal to 15 ± 3 nm which was confirmed with the use of transmission electron microscopy. The electrokinetic characteristics revealed that CHSB-AgNPs have positive, whereas TCSB-AgNPs and TA-AgNPs negative surface charge. First, toxicity of the silver compounds was assessed using the Phytotestkit test. Next, after transferring seedlings to pots, shoot length, leaf surface, shoot dry mass, electrolyte leakage measurement, and photosystem II (PSII) efficiency were determined. AgNPs and silver ions delivered in the form of AgNO3 reduced root and shoots length of common wheat, sorghum, and garden cress; leaves surface of garden cress and white mustard; and shoots dry mass of white mustard. The positively charged CHSB-AgNPs and silver ions delivered in the form of AgNO3 showed the greatest inhibition effect. Moreover, silver ions and positively charged CHSB-AgNPs were more toxic to PSII of model plants than negatively charged TCSB-AgNPs and TA-AgNPs. AgNPs impact differed in the case of monocots and dicots, but the size of the changes was not significant, so it concerned individual parameters. The results revealed the interaction strength, which was generally similar in all tested plants, i.e., increasing negative effect in sequence TCSB-AgNPs 3

This study aimed to examine the hypothesis that silver nanoparticles (AgNPs) and silver ions might induce specific changes and thereby affect plant development and final yield. The experiment was performed on spring wheat, cultured hydroponically with two types of negatively charged AgNPs of an average size of 13–15 nm and silver ions for 14 days and then transplanted to pots with soil. Our results indicated that treatment with the AgNPs stabilized by specific compounds resulted in growth promotion and a reduced number of days to flowering, while that with the ionic form of Ag only caused greater growth in height without influencing the time to heading. Accelerated flowering was caused by changes in phytohormone balance, with GA6 found to be especially favorable. Nanoparticles and silver ions affected the function of photosystem II and the transport and partitioning of assimilates. Increases in the transport form of sugars such as sucrose, raffinose and sorbitol were associated with a considerable improvement in wheat yield, especially in the case of plants treated with the nanoparticle forms, which were more stable and resistant to oxidative dissolution.

Plant overwintering may be affected in the future by climate change. Low-temperature waterlogging, associated with a predicted increase in rainfall during autumn and winter, can affect freezing tolerance, which is the main component of winter hardiness. The aim of this study was to elucidate the mechanism of change in freezing tolerance caused by low-temperature waterlogging in Lolium perenne, a cool-season grass that is well adapted to a cold climate. The work included: (i) a freezing tolerance test (plant regrowth after freezing), (ii) analysis of plant phytohormones production (abscisic acid [ABA] content and ethylene emission), (iii) measurement of leaf water content and stomatal conductance, (iv) carbohydrate analysis, and (v) analysis of Aco1, ABF2, and FT1 transcript accumulation. Freezing tolerance may be improved as a result of cold waterlogging. The mechanism of this change is reliant on multifaceted actions of phytohormones and carbohydrates, whereas ethylene may counteract ABA signaling. The regulation of senescence processes triggered by concerted action of phytohormones and glucose signaling may be an essential component of this mechanism.

Brassinosteroids (BR) are steroid phytohormones that are involved in the growth and stress response in plants, but the precise mechanisms of their action are still being discovered. In our study we have used BR-deficient barley mutants 522DK and BW084 (which carry missense mutations in the HvDWARF and HvCPD genes, respectively). We have also used a BR-signalling mutant that harbors missense substitutions in the HvBRI1 gene. Our aim was (1) to find out if the content of phytohormones in the mutants grown at 20 °C is different than in the wild types and whether/how the content of phytohormones changes after plant acclimation at temperatures of 5 °C and 27 °C?, (2) to characterise the effectiveness of the light reactions of photosynthesis of the barley mutants in comparison to wild types at various temperatures, and (3) to verify the impact of mutations on the tolerance of barley to high and low temperatures. Hormonal characteristics of the BR mutants of barley show the complexity of the interactions between BR and other plant hormones that are additionally modified by temperature and possibly by other factors. The results suggest the participation of BR in auxin catabolism. Further, BR appears to play a role in maintaining the ABA–ABAGlc balance. As for the gibberellin content in plants at a temperature of 20 °C, more in-depth studies will be required to explain the contradictory effects regarding the accumulation of GA3, GA4 and GA5, which appears to be dependent on the type of mutation and connected to the BR level. A fast-kinetic chlorophyll a fluorescence analysis has revealed that the mutants had lower values of energy absorption than the wild types, but the values of the energy transferred via the electron-transport chain was maintained at the wild-type level. We presumed that BR are involved in regulating plant acclimation to extreme (low/high) temperatures, thus the BR-deficient and BR-signalling mutants should be less tolerant to low/high temperatures when compared to the wild types. Unexpectedly, all of the mutants showed a higher tolerance to high temperatures than the wild types. The BW084 and BW312 mutants were less tolerant to frost than the wild type, but 522DK had a similar frost tolerance as the reference wild-type cultivar.

The applications and benefits of nanotechnology in the agricultural and food sector are now established as a matter of fact, but scientists still have a problem to properly acknowledge the influence of nanocompounds on the environment. We are starting to use nanopesticides and nanofertilizers, but the effects they have on organisms in the environment can be diverse in nature – ranging from the beneficial to the harmful. Environmental microbiome can be strongly modified by nanoparticles. Soil microbiomes are the basis for healthy plants and crops. At the same time, the unique properties of nanomaterials make them particularly suitable for environmental remediation due to their high surface area-to-volume ratio, which often results in higher reactivity. Nanomaterials are starting to be used for the remediation of problematic contaminants (organic and organophosphorus compounds, volatile organic compounds, some heavy metals, or herbicides), and such use offers some very promising prospects. This chapter provides an overview of nanotechnology in general, including a description of the main categories of nanomaterials, the utilization of nanotechnology in the agricultural and food sector, as well as the use of nanomaterials in environmental remediation. Lastly, some toxicological aspects of nanomaterials present in the environment are presented on the basis of the scientific literature in the field.

Climate changes, which result in the occurrence of periods with relatively high temperatures during the winter, can lead to the deacclimation of cold-hardened plants and cause problems with their winter survival. The aim of these studies was to investigate the physiological changes (photosynthesis and water relations including aquaporin expression) that accompany deacclimation process in the economically important winter oilseed rape plants. The effect of deacclimation on frost tolerance was also estimated for two tested cultivars: semi-dwarf and plants of a normal height. After cold acclimation at 4 &deg, C (compared to the unacclimated control) the typical changes that accompany cold acclimation such as an increase in the content of water-soluble sugars or a lower water content in the leaves, which lead to an increased frost tolerance, were observed. Deacclimation partially or completely reversed these changes, which resulted in a decreased frost tolerance that was accompanied by a decrease in the content of sugars and an increase of the osmotic potential. The chemical composition of the leaves, which was measured using FT-Raman spectroscopy also clearly confirmed the metabolic differences between the cold-acclimated and deacclimated plants. The plants were significantly different in regard to the content of the various pigments as well as fatty acids and polysaccharides. The phenomenon of a deacclimation-induced decrease in aquaporin PIP1 accumulation, which was accompanied by unchanged PIP1 transcript accumulation, will be discussed in the aspects of the water relations and decreased frost tolerance in deacclimated plants.

Tolerance to deacclimation is an important physiological feature in plants in the face of global warming, which is resulting in incidents of increases in winter temperatures. The aim of the work was to explore how disturbances in the signalling and synthesis of brassinosteroids (BR) influence the deacclimation tolerance of barley. One group of mutants and their reference cultivars (Bowman and Delisa) was cold-acclimated, deacclimated and then tested for frost tolerance at − 12 °C. After cold acclimation, the second group of plants was additionally exposed to frost (− 6 °C) and then, deacclimated and tested for frost tolerance at − 12 °C. The deacclimated brassinosteroid mutants were characterised by an increased tolerance to frost, and consequently, had a higher tolerance to deacclimation than their wild-type cultivars. The mechanism of this phenomenon may be partly explained by analysing the hormonal homeostasis in the crowns. For all of the tested plants, a characteristic feature of the response to the deacclimation phase was an increase in the growth-promoting hormones and abscisic acid compared to the cold acclimation phase. The increase was greater in the BR-deficient (BW084) and BR-insensitive (BW312) mutants compared to the Bowman reference cultivar. Mutant 522DK was characterised by a lower accumulation of total cytokinins and gibberellins as well as an enhanced auxin deactivation compared to the Delisa. In the second group, when the plants were exposed to a temperature of − 6 °C before deacclimation, the hormonal homeostasis was further altered in both the mutants and reference cultivars, but all of the mutants had a higher frost tolerance than the wild types.

The roles of endogenous brassinosteroids (BRs) in the modulation of reaction to drought and genetic regulation of this process are still obscure. In this study, a multidirectional analysis was performed on semi-dwarf barley (Hordeum vulgare) Near-Isogenic Lines (NILs) and the reference cultivar &ldquo, Bowman&rdquo, to get insights into various aspects of metabolic reaction to drought. The NILs are defective in BR biosynthesis or signaling and displayed an enhanced tolerance to drought. The BR metabolism perturbations affected the glucose and fructose accumulation under the control and stress conditions. The BR metabolism abnormalities negatively affected the sucrose accumulation as well. However, during drought, the BR-deficient NILs accumulated higher contents of sucrose than the &ldquo, cultivar. Under the control conditions, accumulation of transcripts encoding antioxidant enzymes ascorbate peroxidase (HvAPX) and superoxide dismutase (HvSOD) was BR-dependent. However, during drought, the accumulation of HvAPX transcript was BR-dependent, whereas accumulations of transcripts encoding catalase (HvCAT) and HvSOD were not affected by the BR metabolism perturbations. The obtained results reveal a significant role of BRs in regulation of the HvAPX and HvCAT enzymatic activities under control conditions and the HvAPX and HvSOD activities during physiological reactions to drought.

The potential of resistance to Microdochium nivale is still not recognized for numerous plant species. The forage grasses of Lolium-Festuca complex are important for grass-biomass production in the temperate regions. Lolium multiflorum is a grass with a high forage quality and productivity but also a relatively low resistance to M. nivale. On the contrary, F. arundinacea has a higher potential of resistance but simultaneously a significantly lower forage quality. These two species cross with each other and the intergeneric hybrids possess complementary characters of both genera. Herein, for the first time, we perform the research on L. multiflorum/F. arundinacea introgression forms to decipher mechanisms of resistance to M. nivale in that group of plants. Two forms with distinct levels of resistance were used as models in cytogenetic and biochemical studies. The resistant plant was shown to be a tetraploid with 28 L. multiflorum chromosomes, including one with three F. arundinacea introgressions. The susceptible introgression form revealed the unbalanced genomic structure and only 25 chromosomes. Twenty four chromosomes were shown to be L. multiflorum chromosomes, including one chromosome with F. arundinacea segment. One Festuca chromosome with additional two interstitial F. arundinacea segments, was also revealed in the susceptible form. The selected introgression forms differed in the accumulation profiles of total soluble carbohydrates, phytohormones, and phenolics in the leaf and crown tissue under the control and infection conditions. The higher amount of carbohydrates and salicylic acid in the leaves and crowns as well as a lower amount of abscisic acid in both studied organs and jasmonic acid in the crowns, were shown to be crucial for the expression of resistance to M. nivale in the analyzed hybrids.

Drought is one of the most adverse stresses that affect plant growth and yield. Disturbances in metabolic activity resulting from drought cause overproduction of reactive oxygen species. It is postulated that brassinosteroids (BRs) regulate plant tolerance to the stress conditions, but the underlying mechanisms remain largely unknown. An involvement of endogenous BRs in regulation of the antioxidant homeostasis is not fully clarified either. Therefore, the aim of this study was to elucidate the role of endogenous BRs in regulation of non-enzymatic antioxidants in barley (Hordeum vulgare) under control and drought conditions. The plant material included the 'Bowman' cultivar and a group of semi-dwarf near-isogenic lines (NILs), representing mutants deficient in BR biosynthesis or signaling. In general, accumulations of 11 compounds representing various types of non-enzymatic antioxidants were analyzed under both conditions. The analyses of accumulations of reduced and oxidized forms of ascorbate indicated that the BR mutants contain significantly higher contents of dehydroascorbic acid under drought conditions when compared with the 'Bowman' cultivar. The analysis of glutathione accumulation indicated that under the control conditions the BR-insensitive NILs contained significantly lower concentrations of this antioxidant when compared with the rest of genotypes. Therefore, we postulate that BR sensitivity is required for normal accumulation of glutathione. A complete accumulation profile of various tocopherols indicated that functional BR biosynthesis and signaling are required for their normal accumulation under both conditions. Results of this study provided an insight into the role of endogenous BRs in regulation of the non-enzymatic antioxidant homeostasis.

Photosynthetic acclimation to cold conditions is an important factor influencing freezing tolerance of plants. Photosynthetic enzyme activities increase as part of a photochemical mechanism underlying photosynthetic acclimation to low temperatures. Additionally, a non-photochemical mechanism may be activated to minimize photooxidative damage. The aim of this study was to test the hypothesis that differences in stomatal conductance in Hordeum vulgare plants with contrasting freezing tolerances induce various strategies for photosynthetic acclimation to cold stress. Different stomatal behaviors during the prehardening step resulted in diverse plant reactions to low-temperature stress. Plants with a relatively low freezing tolerance exhibited decreased stomatal conductance, resulting in decreased photochemical activity, faster induction of the non-photochemical mechanism, and downregulated expression of two Rubisco activase (RcaA) splicing variants. In contrast, plants with a relatively high freezing tolerance that underwent a prehardening step maintained the stomatal conductance at control level and exhibited delayed photochemical activity and RcaA expression decrease, and increased Rubisco activity, which increased net photosynthetic rate. Thus, in barley, the induction of photoinhibition avoidance (i.e., non-photochemical photoacclimation mechanism) is insufficient for an effective cold acclimation. An increase in cold-induced net photosynthetic rate due to open stomata is also necessary.

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.

Alternative splicing of the Rubisco activase gene was shown to be a point for optimization of photosynthetic carbon assimilation. It can be expected to be a stress-regulated event that depends on plant freezing tolerance. The aim of the study was to examine the relationships among Rubisco activity, the expression of two Rubisco activase splicing variants and photoacclimation to low temperature. The experiment was performed on two Lolium perenne genotypes with contrasting levels of freezing tolerance. The study investigated the effect of pre-hardening (15°C) and cold acclimation (4°C) on net photosynthesis, photosystem II photochemical activity, Rubisco activity and the expression of two splicing variants of the Rubisco activase gene. The results showed an induction of Rubisco activity at both 15°C and 4°C only in a highly freezing-tolerant genotype. The enhanced Rubisco activity after pre-hardening corresponded to increased expression of the splicing variant representing the large isoform, while the increase in Rubisco activity during cold acclimation was due to the activation of both transcript variants. These boosts in Rubisco activity also corresponded to an activation of non-photochemical mechanism of photoacclimation induced at low temperature exclusively in the highly freezing-tolerant genotype. In conclusion, enhanced expression of Rubisco activase splicing variants caused an increase in Rubisco activity during pre-hardening and cold acclimation in the more freezing-tolerant Lolium perenne genotype. The induction of the transcript variant representing the large isoform may be an important element of increasing the carbon assimilation rate supporting the photochemical mechanism of photosynthetic acclimation to cold.

Increased precipitation expected during autumn and winter at higher latitudes in the northern hemisphere may lead to low temperature short-term waterlogging. The temperature of waterlogging was shown to be an important factor controlling plant reaction to this stress. Photosynthetic apparatus response to water excess in the soil at low temperature was examined in Festuca pratensis genotypes with contrasting freezing tolerance. The study was aimed to test the hypothesis whether changes in leaf water-soluble carbohydrate concentration brought about alterations in Rubisco activity may affect the photoacclimation to cold under water excess in the soil. The study investigated the effects of waterlogging during cold acclimation process on a set of chlorophyll a fluorescence parameters, water-soluble carbohydrates, expression of Rubisco activase gene, and Rubisco activity. It was shown that carbohydrate status affected by Rubisco activity might be crucial for the activation of non-photochemical mechanism of photoacclimation to cold under waterlogging. Altered Rubisco activity was only partially attributed to the expression of Rubisco activase gene. Additionally, low carbohydrate concentration in the leaves of waterlogged plants was the condition preventing sugar repression of photosynthesis, including RcaA expression. This indicates that sugar de-repression of photosynthetic genes may be considered a component of photosynthetic acclimation to cold under waterlogging.

Photosynthetic acclimation to cold conditions is an important factor influencing freezing tolerance of plants. Photosynthetic enzyme activities increase as part of a photochemical mechanism underlying photosynthetic acclimation to low temperatures. Additionally, a non-photochemical mechanism may be activated to minimize photooxidative damage. The aim of this study was to test the hypothesis that differences in stomatal conductance in Hordeum vulgare plants with contrasting freezing tolerances induce various strategies for photosynthetic acclimation to cold stress. Different stomatal behaviors during the prehardening step resulted in diverse plant reactions to low-temperature stress. Plants with a relatively low freezing tolerance exhibited decreased stomatal conductance, resulting in decreased photochemical activity, faster induction of the non-photochemical mechanism, and downregulated expression of two Rubisco activase (RcaA) splicing variants. In contrast, plants with a relatively high freezing tolerance that underwent a prehardening step maintained the stomatal conductance at control level and exhibited delayed photochemical activity and RcaA expression decrease, and increased Rubisco activity, which increased net photosynthetic rate. Thus, in barley, the induction of photoinhibition avoidance (i.e., non-photochemical photoacclimation mechanism) is insufficient for an effective cold acclimation. An increase in cold-induced net photosynthetic rate due to open stomata is also necessary.

The purpose of this study was to examine the effect of silver nanoparticles (AgNPs) produced using the high-voltage arc discharge method on the growth and metabolism of common wheat seedlings. Additionally, a simultaneous assessment of the AgNP-induced reduction in seedling infection by Fusarium culmorum (Fc) was performed. AgNP- and Fc-treated seedlings indicated that both factors considerably inhibited their growth. A significant Fc-induced reduction in seedling blight was observed following treatment with AgNPs; however, treatment with nanoparticles was also accompanied by a serious disintegration of the cell membranes of roots. Moreover, treatment with AgNPs increased the quantum efficiency of energy trapping in the PSII reaction centre (Fv/Fm) with a simultaneous decrease in energy dissipation in the form of heat. Induction of photosynthesis in the presence of AgNPs did not affect height but was reflected in higher total dry weight. Moreover, analysis of antioxidant enzyme activity typical for the stress response indicated the toxicity of AgNPs treatment compared to Fc treatment. Seedlings exposed to AgNP activity demonstrated accumulation of Ag in roots and its translocation to aerial parts, while the pathogen reduced both accumulation and translocation of this element.

We studied the effect of Trichoderma viridae and Trichoderma harcianum applied both before and after cold acclimation treatment on carbohydrate accumulation as well as on the course of pathogenesis caused by Microdochium nivale . Trichoderma applied prior to cold acclimation treatment decreased the resistance of winter rye to M. nivale . The process of cold acclimation was disrupted as levels of soluble carbohydrates in leaves and crowns decreased despite the stimulation of photosynthesis. In contrast to the shoots, the level of carbohydrates in the roots, especially sucrose and nystose, increased considerably. It can be assumed that the sink activity of roots colonized with Trichoderma affected photosynthesis and carbon partitioning. Sucrose, a substrate for the synthesis of fructans and a transport sugar, was probably consumed by Trichoderma fungi, contributing to decreased levels of soluble carbohydrates and, as a result, reduced resistance to snow mould. Comparing the effect of T.viridae applied before and after cold acclimation treatment, the survival rate was significantly higher in the latter. It suggests interactions between both the saprophytic T. viridae and the pathogenic fungus M. nivale when applied together. However, changes induced in fructan metabolism may also contribute to increased resistance to pink snow mould.

Quantitative PCR studies need proper reference genes with expression stability exclusively validated under certain experimental conditions. The expression stability of several genes commonly used as references was tested under 24-epibrassinolide (EBR) and temperature treatment. Different statistical approaches (qBase PLUS , BestKeeper, NormFinder) were used to prepare rankings of expression stability in two species of an economic importance: common wheat ( Triticum aestivum ) and perennial ryegrass ( Lolium perenne ). Candidate reference genes were shown to be regulated differentially in these two plant species. The maximum stability values indicated that the expression stability was higher in T. aestivum . Taking into account of all ranks it seems that TBP-1 and UBI in ryegrass and ACT , ADP and EF1A in wheat should be used as reference genes in the brassinosteroids and temperature involving studies.

Resistance to snow mould is a feature determined by multiple genes. Therefore, determining the phenotype of resistant plants is difficult as it requires an investigation over a long period of time from cold acclimation through pathogenesis. The aim of the present study was (i) to determine the characteristics of the resistant genotype and (ii) to clarify the connections between photosynthesis during cold acclimation and then pathogenesis caused by Microdochium nivale. Two inbred lines of winter rye (Secale cereale L.) differing in their susceptibility to snow mould were used in the study. After cold acclimation snow mould resistant (SMR) line was characterised by higher values of CO2 assimilation and electron transport efficiency but did not differ from snow mould susceptible (SMS) line in carboxylation rate of RuBisCO (Vcmax). Higher soluble carbohydrate accumulation, due to higher photosynthesis intensity, as well as an ABA increase at 5 days post infection (DPI) in leaves and crowns were found in SMR line during the pathogenesis period. Callose deposition was found around non-infected bundle sheets and in cortex cells at 5 DPI (at the same time point as ABA peak) only in SMR line, which probably prevented the infection of leaf initials. Early leaf initials infection in SMS line may be responsible for inhibiting leaf growth and plant regeneration after stress cessation. The results show different physiological and biochemical characteristics of the investigated lines, which can be applied in the selection of resistant genotypes and identifying genomic regions responsible for metabolic pathways increasing pathogen resistance.

Frost tolerance, resistance to Microdochium nivale and associated biochemical changes were investigated in leaves and crowns of six winter rye inbred lines. Investigated lines differed in their susceptibility to snow mould but not in frost tolerance. It was shown that winter rye resistance is linked to defence response in crowns expressed by higher activity of antioxidant enzymes e CAT, SOD and total peroxidases e on the 1st day after inoculation and b-D-glucosidase activation during the course of pathogenesis. It could be postulated that changes in leaves are attributed to reaction to cold while changes in crowns are attributed to pathogen reaction.

Plants at various stages of growth are differentiated in their susceptibility to environmental stresses and possibility of regeneration during recovery period after cessation of stress. The role of antioxidative enzymes in the stabilisation of the physiological state of field bean plants was studied. Catalase, peroxidises activity, and hydrogen peroxide accumulation were determined after 1 and 7 days of flooding and after 1 and 7 days of recovery from flooding. The influence of flooding on plants growth was analysed on the basis of leaf area index (LAI), relative growth rate (RGR) and net assimilation rate (NAR). Changes in antioxidative enzymes caused by flooding applied at vegetative stage were reversible. In generative plants, antioxidative enzymes were activated to a greater extent during recovery period and in contrast to vegetative plants did not attain the control level after that period. The reduction in growth was greater when flooding stress was applied at vegetative stage. Vegetative plants regardless of the stabilisation of antioxidant system were not able to restore the physiological function to attain the optimal growth level after flooding. Probably other injuries caused by flooding had a dominant effect on enzymatic antioxidant changes during stress period.

According to predicted changes in climate, waterlogging events may occur more frequently in the future during autumn and winter at high latitudes of the Northern Hemisphere. If excess soil water coincides with the process of cold acclimation for plants, winter survival may potentially be affected. The effects of waterlogging during cold acclimation on stomatal aperture, relative water content, photochemical activity of photosystem II, freezing tolerance and plant regrowth after freezing were compared for two prehardened overwintering forage grasses, Lolium perenne and Festuca pratensis. The experiment was performed to test the hypothesis that changes in photochemical activity initiated by waterlogging-triggered modifications in the stomatal aperture contribute to changes in freezing tolerance. Principal component analysis showed that waterlogging activated different adaptive strategies in the two species studied. The increased freezing tolerance of F. pratensis was associated with increased photochemical activity connected with stomatal opening, whereas freezing tolerance of L. perenne was associated with a decrease in stomatal aperture. In conclusion, waterlogging-triggered stomatal behavior contributed to the efficiency of the cold acclimation process in L. perenne and F. pratensis.

Brassinosteroids (BRs) are a class of steroid phytohormones, which regulate various processes of morphogenesis and physiology – from seed development to regulation of flowering and senescence. An accumulating body of evidence indicates that BRs take part in regulation of physiological reactions to various stress conditions, including drought. Many of the physiological functions of BRs are regulated by a complicated, and not fully elucidated network of interactions with metabolic pathways of other phytohormones. Therefore, the aim of this study was to characterize phytohormonal homeostasis in barley (Hordeum vulgare) in reaction to drought and validate role of BRs in regulation of this process. Material of this study included the barley cultivar ‘Bowman’ and five Near-Isogenic Lines (NILs) representing characterized semi-dwarf mutants of several genes encoding enzymes participating in BR biosynthesis and signaling. Analysis of endogenous BRs concentrations in these NILs confirmed that their phenotypes result from abnormalities in BR metabolism. In general, concentrations of eighteen compounds, representing various classes of phytohormones, including brassinosteroids, auxins, cytokinins, gibberellins, abscisic acid, salicylic acid and jasmonic acid were analyzed under control and drought conditions in the ‘Bowman’ cultivar and the BR-deficient NILs. Drought induced a significant increase in accumulation of the biologically active form of BRs – castasterone in all analyzed genotypes. Another biologically active form of BRs – 24-epi-brassinolide - was identified in one, BR-insensitive NIL under normal condition, but its accumulation was drought-induced in all analyzed genotypes. Analysis of concentration profiles of several compounds representing gibberellins allowed an insight into the BR-dependent regulation of gibberellin biosynthesis. The concentration of the gibberellic acid GA7 was significantly lower in all NILs when compared with the ‘Bowman’ cultivar, indicating that GA7 biosynthesis represents an enzymatic step at which the stimulating effect of BRs on gibberellin biosynthesis occurs. Moreover, the accumulation of GA7 is significantly induced by drought in all the genotypes. Biosynthesis of jasmonic acid is also a BR-dependent process, as all the NILs accumulated much lower concentrations of this hormone when compared with the ‘Bowman’ cultivar under normal condition, however the accumulation of jasmonic acid, abscisic acid and salicylic acid were significantly stimulated by drought.

Pink snow mould caused by Microdochium nivale can lead to serious yield loss in many winter cereals and turf grasses. The study examined the infection mechanism in four inbred lines and one cultivar of rye (Secale cereale L.) with different sensitivities to this pathogenic fungus. Fungus development was analyzed at the early stages of the infection, up to the 13th day after inoculation, in hardened and non-hardened seedlings, using light/fluorescence microscopy. The cytological analysis was supplemented with a spectrophotometrical assay of the phenolics. Significantly higher phenolic content was revealed in the leaves of the lines more resistant to M. nivale, as compared to those which are more sensitive.

Winter rye plants of three Polish inbred lines and cv. Stach differing in Microdochium nivale resistance were studied relating to their frost and pink snow mould tolerance. The plants were prehardened at 12°C for 2 weeks and hardened at 2°C for 3 weeks. Control plants were grown in the greenhouse at 20°C. Frost resistance expressed as LT50 was determined for leaves and crowns of the hardened and control plants. Cold-hardened were inoculated with mycelium of M. nivale and incubated for 35 days at 2°C in the dark. After this time, their pink snow mould resistance was evaluated and expressed as an average regrowth index (ARI). During 13 days of pathogenesis, changes in the total soluble carbohydrate (TSC) and ketose content were analysed. Moreover, changes in abscisic acid (ABA) and water content (WC) during 9 days of pathogenesis were determined. All analyses were carried out in leaves and crowns of inoculated and non-inoculated (control) plants. Cold acclimation increased frost resistance of the leaves and crowns; however, the crowns were less frost tolerant than the leaves. In the studied lines, there was a negative correlation between frost tolerance of leaves and pink snow mould resistance of plants. Plants of lines more resistant to M. nivale exhibited higher TSC and ketose concentrations in the leaves and crowns as well as lower ABA levels in comparison with the less resistant plants. The role of ABA in the defence response of rye to pink snow mould is still unclear. It seems that ABA concentration does not determine rye resistance to M. nivale, although a higher level of this hormone could decrease it.

This paper presents a study of the metabolic response (dark respiration intensity, photosystem II efficiency, metabolic activity) and the yield of barley treated with 24-epibrassinolide and subjected to high-temperature stress. Transport of exogenously applied 24-epibrassinolide in barley and changes in the profile of brassinosteroids that may occur in tissues after 24-epibrassinolide application were also studied. The water solution of 24-epibrassinolide (0.005 and 0.25 mg dm−3) was applied via infiltration of the first and second leaves of 12-day-old seedlings. Control plants were treated with water solution of hormone solvent (ethanol). Fifteen-day-old plants were subjected to high-temperature stress (42°C for 3 h). The influence of hormone treatment and stress conditions was investigated in the first and second leaves based on measurements of PSII efficiency. The aftereffect of plant treatment was investigated in the seventh leaf (measurements of PS II efficiency, dark respiration intensity, metabolic activity). The transport efficiency of 24-epibrassinolide exogenously applied to the first and second leaves, as well as the profile of other brassinosteroids, was also measured on the seventh leaf. Finally, yield formation was estimated. 24-epibrassinolide showed protective action, which manifested itself in the improved functioning of PSII, but this was observed in case of higher hormone concentration and only for the first, older leaf. The PSII efficiency of the seventh leaf was similar in plants treated with brassinosteroid and in the control plants, whereas the respiration intensity and metabolic activity decreased in plants previously treated with higher concentration of 24-epibrassinolide. The use of a higher hormone concentration at the seedling phase ultimately resulted also in lower crop yield. Brassinosteroids—brassinolide and castasterone—were detected in barley leaves. 24-epibrassinolide was found only in trace amounts in control plants. Its exogenous application directly to the apoplast of the first and second leaves resulted in an increase in the 24-epibrassinolide content in the seventh leaf, but did not depend on whether a high or low concentration had been applied to the plants.

The aim of the experiment was to verify whether changes in photosynthetic activity measured by means of chlorophyll fluorescence techniques and soluble carbohydrate level induced during pre-hardening and cold acclimation in androgenic forms of Festulolium are related to their winter hardiness and pathogen resistance. The study was performed on six androgenic genotypes of Festulolium, which differ in their degrees of winter hardiness. The chlorophyll fluorescence parameters and sugar content were measured in the control plants, pre-hardened at 12 °C and cold-acclimated at 2 °C. Cold acclimation at 2 °C, preceded by pre-hardening at 12 °C, induced a decline in maximum PSII quantum efficiency (F v /F m ) and in non-photochemical quenching (NPQ). Exposure to 2 °C also resulted in reduced efficiency of energy conversion in photochemical processes (qp). The soluble carbohydrate amount increased in the susceptible genotypes and was associated with a decline in NPQ and current quantum yield of PSII (φ PSII ). In resistant plants, a decrease in the sugar content was observed, which could be explained by their consumption in metabolic processes initiated during cold acclimation. The changes in sugar content were associated with a decline in NPQ and q P .

The aim of our work is to investigate the changes in phenolic level, PAL activity and heat production rate induced during pre-hardening at 12°C and cold acclimation at 2°C of the forage grasses Festulolium, meadow fescue, tall fescue and Italian ryegrass in relation to their resistance to snow mould caused by Microdochium nivale. Meadow fescue and tall fescue were most resistant to M. nivale infection, while Italian ryegrass demonstrated the least resistance to this fungus inoculation. Festulolium, meadow fescue and tall fescue responded similarly to low temperature, while Italian ryegrass demonstrated considerable disturbance of energy balance and lower phenolic concentration, which could explain a higher susceptibility of the latter species to infection by M. nivale. The enhanced level of phenolic compounds, probably utilised for cell wall lignification as well as equilibrium of the metabolic activity observed in meadow fescue and tall fescue, is very important for both cold and pathogen-resistance mechanisms. The studied Festulolium cultivar ‘Felopa’, a hybrid of the Loliummultiflorum and Festucapratensis genomes, was characterised by changes in biochemical parameters similar to the resistant meadow fescue and tall fescue.

We investigated changes in concentrations of abscisic (ABA) and salicylic acid (SA), phenolic compounds and phenylalanine ammonia-lyase (PAL) activity in relation to cold-induced tolerance of four androgenic genotypes of Festulolium (Festuca × Lolium hybrids) to frost and to the snow mould fungus Microdochium nivale. Cold acclimation increased frost tolerance and resistance to snow mould. Resistant genotypes were characterized by higher ABA concentrations during the first 54 h of cold acclimation and lower concentrations of SA than susceptible genotypes. After cold acclimation, the content of phenolics was significantly lower in genotypes tolerant to frost and M. nivale infection than in susceptible genotypes, while PAL activity was significantly higher. Signalling networks controlling cold acclimation to frost (abiotic) and mould infection (biotic) appears to involve increases in foliar concentrations of ABA and decreases in the SA level during successful cold acclimation. Higher PAL activity and lower concentrations of phenolic compounds also appear to be associated with enhanced tolerance to frost and fungal attack.

The metabolic changes in foliar abscisic acid, proline and phenolic concentration, and PAL activity induced during cold acclimation in androgenic forms of Festulolium contrasting in terms of their resistance to frost and Microdochium nivale were studied. The frost-resistant and snow mould-resistant (cross-tolerant) genotype (561) was characterized by a higher ABA amount and higher PAL activity compared to those of the frost and snow mould-susceptible genotype (621) during cold acclimation. The obtained results suggest that both these parameters are involved in a cross-tolerance mechanism relating to frost and snow mould resistance.

The aim of this study was to investigate the physiological basis for differences in resistance to pink snow mould (Microdochium nivale) in two androgenic genotypes of Festulolium (Festuca pratensis x Lolium multiflorum) which differed in terms of their resistance to M. nivale. Genotype 716 was more resistant than genotype 729. The study consisted of two experiments. The aim of the first experiment was to estimate the ability of the plants to survive winter conditions. The aim of the second experiment was to find physiological markers of resistance to snow mould. Festulolium plants were infected with M. nivale mycelium after pre-hardening and hardening. After 2 weeks in the dark at 2°C, there was a sharp increase in the phenolic content in both genotypes. The increase was greater in the more resistant genotype 716 than in genotype 729. Phenolics therefore may play a very important role in overwintering in grasses, similar to carbohydrates. Based on the differences between the two genotypes, potential indicators of resistance to M. nivale in Festulolium include increased soluble carbohydrate content, increased phenolic content, increased hydrogen peroxide accumulation, decreased catalase activity, increased abscisic acid content and reduced heat emission.

In terms of climate changes and global warming, winter hardiness could be determined by unfavorable environmental conditions other than frost. These could include flooding from melting snow and/or rain, coincident with fungal diseases. Therefore, we designed an experiment to identify potential common mechanisms of flooding tolerance and snow mold resistance, involving the regulation of photosynthetic efficiency and carbohydrate metabolism at low temperatures. Snow mold-resistant and susceptible winter rye (Secale cereale) plants were characterized by considerably different patterns of response to flooding. These differences were clearer at low temperature, thus confirming a possible role of the observed changes in snow mold tolerance. The resistant plants were characterized by lower PSII quantum yields at low temperature, combined with much higher energy flux for energy dissipation from the PSII reaction center. During flooding, the level of soluble carbohydrates increased in the resistant plants and decreased in the susceptible ones. Thus increase in resistant line was connected with a decrease in the energy dissipation rate in PSII/increased photosynthetic activity (energy flux for electron transport), a lower rate of starch degradation and higher rates of sucrose metabolism in leaves. The resistant lines accumulated larger amounts of total soluble carbohydrates in the crowns than in the leaves. Irrespective of flooding treatment, the resistant lines allocated more sugars for cell wall composition, both in the leaves and crowns. Our results clearly indicated that studies on carbohydrate changes at low temperatures or during anoxia should investigate not only the alterations in water-soluble and storage carbohydrates, but also cell wall carbohydrates. The patterns of changes observed after low and high-temperature flooding were different, indicating separate control mechanisms of these responses. These included changes in the photosynthetic apparatus, starch accumulation and cell wall carbohydrate accumulation.

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.

The beneficial effects of trans-resveratrol on pathogen-response and its physiological mechanisms were studied in two wheat cultivars differing in the level of resistance against powdery mildew (Blumeria graminis (DC.) Speer). Resveratrol induced phenolics metabolism in a manner that lead to increased phenylalanine ammonia-lyase (PAL) and β-D-glucosidase activity in leaf regions showing no symptoms of pathogen infection. In less-resistant cultivar, similar alterations were additionally observed in regions invaded by the pathogen. Resveratrol also stimulated photosynthetic efficiency during pathogenesis and this effect was more prominent in the resistant cultivar. Regulatory function of resveratrol on wheat growth and development was also observed (stimulation of generative induction). High performance liquid chromatography (HPLC) analysis detected a low amount of compound with retention time corresponding to trans-resveratrol in healthy leaves and increased the content of this compound in infected foliage...

Due to climate changes and global warming the knowledge about the role of hormones promoting cold acclimation will become increasingly important. Brassinosteroids (BR) are plant steroid hormones which control photosynthesis, affect carbohydrate metabolism and protect against environmental factors (Janeczko et al., 2011). In order to demonstrate if BR are essential for the cold acclimation of perennial ryegrass, we studied the effect 24- epibrassinolide (EBR) and brassinazole (BZ) a specific inhibitor of BR biosynthesis, on parameters associated with cold acclimation (CA). We also analyzed the influence of the length of CA on carbohydrate quality and quantity under EBR treatment. The endogenous BR level during cold acclimation was also studied. We had demonstrated that EBR treatment improved frost tolerance of perennial ryegrass. EBR upregulated while BZ downregulated the expression of Rubisco activase ( Rca A ) in leaves and crowns of Lolium perenne . EBR treatment decreased contents of total soluble carbohydrates (TSC) in leaves which coincided with increased level of TSC in crowns. At the same time the expression of sucrose-1-fructosyltransferase (1- SST ) and sucrose synthase ( SUS ) genes in leaves and crowns were reduced by BZ. Gene expression level corresponded to changes in Rubisco and sucrose synthase activities. Endogenous level of castasterone (CS), the only detected BR in plants, increased significantly after 3 weeks of CA and decreased to the control level after next 3 weeks at 4°C. Changes of different soluble sugars concentration including fructooligosacharides was dependent on lenght of CA. Concluding, EBR enhanced frost tolerance of perennial ryegrass through changes during cold acclimation. Particulary EBR upregulated RcaA, SUS and 1-SST genes and activated of Rubisco and SPS enzymes during cold acclimation, leading to increased carbohydrate accumulation in crowns which coincided with decline of TSC level in leaves Founding: National Science Centre (Poland) DEC-2011/03/D/NZ9/05548

Photosynthetic apparatus acclimation to low temperature is a prerequisite for proper hardening and acquired freezing tolerance. It has been shown that Lolium perenne plants have two isoforms of Rubisco activase, products of two splicing variants (RcaA1, RcaA2) of common pre-mRNA (Jurczyk et al., 2015). The aim of the study was to determine the relationship between the activation of mechanisms of photosynthetic acclimation to cold, the Rubisco activity and the expression of alternatively spliced Rubisco activase isoforms in Lolium perenne. Transcript levels of alternatively spliced mRNAs coding for two Rubisco activase isoforms were measured using real-time PCR. Rubisco activity has been determined spectrophotometrically. The measurements of chlorophyll-a fluorescence parameters, including photochemical and non-photochemical quenching, were made with a FMS2 fluorymeter. Gas exchange measurements were performed using an infrared gas analyser. All measurements were taken before cold acclimation, after 2 weeks of prehardening, and after 1 and 3 weeks of cold acclimation in two Lolium perenne genotypes with contrasting levels of freezing tolerance. In more freezing tolerant genotype the Rubisco activity was higher in treated plants relative to control. The increased Rubisco activity during cold acclimation was due to higher expression of both transcripts variants, representing two isoforms of Rubisco activase, but the shorter isoform seemed to be more active. The photochemical, but also non-photochemical mechanism of photosynthetic activation to cold has been activated in freezing tolerant genotype. It may be concluded that the increase in photochemical mechanism of photosynthetic acclimation to cold observed in Lolium perenne may be attributed to enhanced expression of Rubisco activase leading to increased Rubisco activity.The research was supported under a project DEC-2011/03/B/NZ2/00536 “The role of alternate mRNA splicing of Rubisco activase gene in acclimation of photosynthetic apparatus to cold” financed by the Polish National Research Centre.