Andrzej Skoczowski

Salt tolerance of barley induced by the root endophyte Piriformospora indica is associated with a strong increase in antioxidants.

The root endophytic basidiomycete Piriformospora indica has been shown to increase resistance against biotic stress and tolerance to abiotic stress in many plants. Biochemical mechanisms underlying P. indica-mediated salt tolerance were studied in barley (Hordeum vulgare) with special focus on antioxidants. Physiological markers for salt stress, such as metabolic activity, fatty acid composition, lipid peroxidation, ascorbate concentration and activities of catalase, ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase and glutathione reductase enzymes were assessed. Root colonization by P. indica increased plant growth and attenuated the NaCl-induced lipid peroxidation, metabolic heat efflux and fatty acid desaturation in leaves of the salt-sensitive barley cultivar Ingrid. The endophyte significantly elevated the amount of ascorbic acid and increased the activities of antioxidant enzymes in barley roots under salt stress conditions. Likewise, a sustained up-regulation of the antioxidative system was demonstrated in NaCl-treated roots of the salt-tolerant barley cultivar California Mariout, irrespective of plant colonization by P. indica. These findings suggest that antioxidants might play a role in both inherited and endophyte-mediated plant tolerance to salinity.

Protection of winter rape photosystem 2 by 24-epibrassinolide under cadmium stress

Seedlings of winter rape were cultured in vitro on media containing 24-epibrassinolide, EBR (100 nM) and cadmium (300 µM). After 14 d of growth, fast fluorescence kinetics of chlorophyll (Chl) a and contents of photosynthetic pigments and Cd in cotyledons were measured. Cd was strongly accumulated but its content in cotyledons was 14.7 % smaller in the presence of EBR. Neither Cd nor EBR influenced the contents of Chl a and b and carotenoids. Cd lowered the specific energy fluxes per excited cross section (CS) of cotyledon. The number of active reaction centres (RC) of photosystem 2 (RC/CS) decreased by about 21.0 % and the transport of photosynthetic electrons (ET0/CS) by about 17.1 %. Simultaneously, under the influence of Cd, the activity of O2 evolving centres (OEC) diminished by about 19.5 % and energy dissipation (DI0/CS) increased by about 14.6 %. In the cotyledons of seedlings grown on media without Cd, EBR induced only a small increase in the activity of most photochemical reactions per CS. However, EBR strongly affected seedlings cultured with cadmium. Specific energy fluxes TR0/CS and ET0/CS of the cotyledons of plants Cd+EBR media were about 10.9 and 20.9 % higher, respectively, than values obtained for plants grown with Cd only. EBR also limited the increase of DI0/CS induced by Cd and simultaneously protected the complex of OEC against a decrease of activity. Hence EBR reduces the toxic effect of Cd on photochemical processes by diminishing the damage of photochemical RCs and OECs as well as maintaining efficient photosynthetic electron transport.

Effects of brassinosteroid infiltration prior to cold treatment on ion leakage and pigment contents in rape leaves

The effect of 24-epibrassinolide (BR27) on cold resistance of rape seedlings was studied by ion leakage and photosynthetic pigment degradation measurements. Aqueous solutions of BR27 were injected into cotyledons or primary leaves of rape plants and these plants were incubated at 2 °C or 20 °C. Cold treatment (2 °C) without BR27 injection elevated the membrane permeability in both primary leaves and cotyledons significantly. Surprisingly, injection of leaves with water or 0.467 % aqueous ethanol solution led to a massive increase in membrane permeability after cold stress at 2 °C. The synergistic effect of leaf infiltration and cold on permeability was abolished by 0.05 and 1.00 µM of BR27 in primary leaves and by 1.00 µM of BR27 in cotyledons. On the other hand, BR27 solutions strongly elevated the membrane permeability at 20 °C, while water and ethanol solutions brought about only negligible increases. Water or ethanol infiltrations strongly reduced the leaf contents of chlorophyll (Chl) a, Chl b and carotenoids at 2 °C but less markedly at 20 °C. However, in seedlings exposed to 2 °C pigments content was significantly higher in BR27-treated leaves as compared to water/ethanol control. There were no differences between pigment contents of leaves injected with BR27 solutions or only water/ethanol at 20 °C. The above data strongly support the stress protecting effect of BR27.

Changes of leaf water potential and gas exchange during and after drought in triticale and maize genotypes differing in drought tolerance

Influence of drought (D) on changes of leaf water potential (Ψ) and parameters of gas exchange in D-resistant and D-sensitive genotypes of triticale and maize was compared. Soil D (from −0.01 to −2.45 MPa) was simulated by mannitol solutions. At −0.013 MPa significant differences in Ψ, net photosynthetic rate (PN), transpiration rate (E), stomatal conductance (gs), and internal CO2 concentration (Ci) of D-resistant and D-sensitive triticale and maize genotypes were not found. Together with the increase in concentration of the mannitol solution the impact of D on E and gs for D-sensitive genotypes (CHD-12, Ankora) became lower than for the D-resistant ones (CHD-247, Tina). Inversely, impact of D on Ψ was higher in D-sensitive than D-resistant genotypes. From 1 to 3 d of D, a higher decrease in PN was observed in D-resistant genotypes than in the D-sensitive ones. Under prolonged D (5–14 d) and simultaneous more severe D the decrease in PN was lower in D-resistant than in D-sensitive genotypes. Changes in Ψ, PN, E, and gs caused by D in genotypes differing in the drought susceptibility were similar for triticale and maize. Compared to control plants, increase of Ci was different for triticale and maize genotypes. Hence one of the physiological reasons of different susceptibility to D between sensitive and resistant genotypes is more efficient protection of tissue water status in resistant genotypes reflected in higher decrease in gs and limiting E compared to the sensitive ones. Other reason, observed in D-resistant genotypes during the recovery from D-stress, was more efficient removal of detrimental effects of D.

Impact of Sunflower (Helianthus annuus L.) Extracts Upon Reserve Mobilization and Energy Metabolism in Germinating Mustard (Sinapis alba L.) Seeds

One commonly observed effect of phytotoxic compounds is the inhibition or delay of germination of sensitive seeds. Mustard (Sinapis alba L.) seeds were incubated with aqueous extracts of sunflower (Helianthus annuus L.) leaves. Although sunflower phytotoxins did not influence seed viability, extracts completely inhibited seed germination. Inhibition of germination was associated with alterations in reserve mobilization and generation of energy in the catabolic phase of germination. Degradation of lipids was suppressed by sunflower foliar extracts resulting in insufficient carbohydrate supply. The lack of respiratory substrates and decrease in energy (ATP) generation resulted in suppression of the anabolic phase of seed germination and ultimately growth inhibition.

Changes in response to drought stress of triticale and maize genotypes differing in drought tolerance

Direct effects and after-effects of soil drought for 7 and 14 d were examined on seedling dry matter, leaf water potential (ψ), leaf injury index (LI), and chlorophyll (Chl) content of drought (D) resistant and sensitive triticale and maize genotypes. D caused higher decrease in number of developed leaves and dry matter of shoots and roots in the sensitive genotypes than in the resistant ones. Soil D caused lower decrease of ψ in the triticale than maize leaves. Influence of D on the Chl b content was considerably lower than on the Chl a content. In triticale the most harmful D impact was observed for physiologically younger leaves, in maize for the older ones. A period of 7-d-long recovery was too short for a complete removal of an adverse influence of D.

Effects of Ozone Fumigation on Photosynthesis and Membrane Permeability in Leaves of Spring Barley, Meadow Fescue, and Winter Rape

Seedlings of spring barley, meadow fescue, and winter rape were fumigated with 180 μg kg−1 of ozone for 12 d, and effect of O3 on photosynthesis and cell membrane permeability of fumigated plants was determined. Electrolyte leakage and chlorophyll fluorescence were measured after 6, 9, and 12 d of fumigation, while net photosynthetic rate (PN) and stomatal conductance (gs) were measured 9 d after the start of ozone exposure. O3 treatment did not change membrane permeability in fescue and barley leaves, while in rape a significant decrease in ion leakage was noted within the whole experiment. O3 did not change the photochemical efficiency of photosystem 2 (PS2), i.e., Fv/Fm, and the initial fluorescence (F0). The values of half-rise time (t1/2) from F0 to maximal fluorescence (Fm) decreased in fescue and barley after 6 and 9 d of fumigation. PN decreased significantly in ozonated plants, in the three species. The greatest decrease in PN was observed in ozonated barley plants (17 % of the control). The ozone-induced decrease in PN was due to the closure of stomata. Rape was more resistant to ozone than fescue or barley. Apparently, the rape plants show a large adaptation to ozone and prevent loss of membrane integrity leading to ion leakage.

Metabolic activity, the chemical composition of biomass and photosynthetic activity of Chlorella vulgaris under different light spectra in photobioreactors.

Omitting the far‐red in LED lighting for bioreactors is inexplicable because it affects photosynthetic efficiency and photomorphogenetic activity. Therefore, this work compares three light sources (fluorescent—white light (WL), and LED: blue + red—BRL and blue + red + far‐red—BRFRL) for cultures of Chlorella vulgaris. Metabolic activity was determined by isothermal calorimetry. Changes in the chemical composition of biomass were examined by spectrofluorimetry and Raman spectroscopy. Maximum quantum yield of photosystem II was determined on the basis of chlorophyll a fluorescence parameters. The algae grown under BRL produced significantly more thermal energy than those cultured under BRFRL and WL. The Raman spectra of cultures showed characteristic bands for carotenoids, chlorophyll, phenolics, lipids, aliphatic carbohydrates, pectins, and disaccharides. According to the cluster analysis, the chemical composition of cultures grown under BRL and BRFRL was very similar, whereas the WL represented a distinct group. BRL and BRFRL stimulated the biosynthesis of an unidentified compound(s) with fluorescence maximum at 614 nm. At the beginning of the cultivation, photosystem II had very weak photochemical activity. Under BRFRL, ratios of Fv/Fm reached the maximum after 4 days, whereas under BRL and WL, after 6 days of cultivation. The results point to the favorable influence of the far‐red on the metabolism of microalgae.

Exogenous carbohydrate utilisation by explants of Brassica napus cultured in vitro

The influence of carbohydrate type (sucrose, maltose, glucose and fructose) used as carbon source in the medium on particular carbohydrates content in hypocotyls of Brassica napus cv. Topas during in vitro culture was studied. The variability in sugar composition in the explants depending on carbon source in the medium and duration of culture was found. The type of sugar did not influence the growth of cultured tissues during the first 15–20 days of culture. Sugars in the medium were main source of substrates for sucrose synthesis in growing explants. Significant re-synthesis of sucrose in the hypocotyls after the uptake of hexoses was found. Maltose was detected only in traces in tissues cultured on media devoid of this sugar. In the case of glucose and fructose, medium was the main source of these monosaccharides for tissues. In explants cultured on maltose-based medium, glucose content was lower comparing to hypocotyls cultured on sucrose medium. The highest fresh weight (FW) was noticed on the media supplied with sucrose and glucose. Changes in sugar content in culture media were also studied. Autoclaving hydrolysed ∼7% of the initial sucrose into glucose and fructose. The most distinct changes were noticed in the media supplied with sucrose and maltose. Glucose- and fructose-based media remained stable during whole period of culture.

The long-term effect of cold on the metabolism of winter wheat seedlings. II. composition of fatty acids of phospholipids

Abstract 1. 1.|In seedlings of winter and spring (controls) wheat the changes in the composition of phospholipid fatty acids induced by long-term cold treatment and by subsequent adaptation to 20°C are examined. 2. 2.|In both genotypes grown at 2°C, as compared to 20°C, linolenic acid (18:3) was increased and oleic (18:1) and linoleic acids (18:2) decreased in content, and the ratio of 18:3/18:2 also increased. 3. 3.|In the winter variety, during growth at 20°C, changes in 18:3/18:2 ratio were small but at 2°C, after a cold treatment sufficient for vernalization (56 days), rapid decrease in the ratio was observed. This change however, did not influence the double bond index (DBI). 4. 4.|After 4 days adaptation to 20°C winter wheat seedlings, subjected to prior cold treatment, retained a fatty acid composition resembling that formed in the cold. In the spring variety 2 days adaptation to 20°C caused changes in fatty acid composition characteristic for seedlings grown at 20°C.