Co-composting of sewage sludge as an effective technology for the production of substrates with reduced content of pharmaceutical residues.

Sewage sludge is a valuable source of elements such as phosphorus and nitrogen. At the same time, heavy metals, emerging organic compounds, micropollutants (pharmaceuticals, pesticides, PCPs, microplastics), or some potentially dangerous bacteria can be present. In this study, the sewage sludge was aerobically treated by composting with other materials (co-composted), and the resulting substrate was tested for suitability of its use in agriculture. Closer attention was focused on the pharmaceuticals (non-steroidal antiphlogistics, sartanes, antiepileptics, caffeine, and nicotine metabolites) content and ecotoxicity of the resulting substrates in the individual phases of sludge co-composting. It has been verified that during co-composting there is a potential for reduction of the content of pharmaceutical in the substrates up to 90 %. The course of the temperature in the thermophilic phase is decisive. Growth and ecotoxicity experiments demonstrated that with a suitable co-composting procedure, the resulting stabilized matter is suitable as a substrate for use in plant production, and the risk of using sewage sludge on agricultural land is substantially reduced.

Light Drives and Temperature Modulates: Variation of Phenolic Compounds Profile in Relation to Photosynthesis in Spring Barley.

Accumulation and metabolic profile of phenolic compounds (PheCs; serving as UV-screening pigments and antioxidants) as well as carbon fixation rate (An) and plant growth are sensitive to irradiance and temperature. Since these factors are naturally co-acting in the environment, it is worthy to study the combined effects of these environmental factors to assess their possible physiological consequences. We investigated how low and high irradiance in combination with different temperatures modify the metabolic profile of PheCs and expression of genes involved in the antioxidative enzyme and PheCs biosynthesis, in relation to photosynthetic activity and availability of non-structural carbohydrates (NSC) in spring barley seedlings. High irradiance positively affected An, NSC, PheCs content, and antioxidant activity (AOX). High temperature led to decreased An, NSC, and increased dark respiration, whilst low temperature was accompanied by reduction of UV-A shielding but increase of PheCs content and AOX. Besides that, irradiance and temperature caused changes in the metabolic profile of PheCs, particularly alteration in homoorientin/isovitexin derivatives ratio, possibly related to demands on AOX-based protection. Moreover, we also observed changes in the ratio of sinapoyl-/feruloyl- acylated flavonoids, the function of which is not yet known. The data also strongly suggested that the NSC content may support the PheCs production.

A novel approach for measuring membrane permeability for organic compounds via surface plasmon resonance detection.

Well-known methods for measuring permeability of membranes include static or flow diffusion chambers. When studying the effects of organic compounds on plants, the use of such model systems allows to investigate xenobiotic behavior at the cuticular barrier level and obtain an understanding of the initial penetration processes of these substances into plant leaves. However, the use of diffusion chambers has disadvantages, including being time-consuming, requiring sampling, or a sufficiently large membrane area, which cannot be obtained from all types of plants. Therefore, we propose a new method based on surface plasmon resonance imaging (SPRi) to enable rapid membrane permeability evaluation. This study presents the methodology for measuring permeability of isolated cuticles for organic compounds via surface plasmon resonance detection, where the selected model analyte was the widely used pesticide metazachlor. Experiments were performed on the cuticles of Ficus elastica, Citrus pyriformis, and an artificial PES membrane, which is used in passive samplers for the detection of xenobiotics in water and soils. The average permeability for metazachlor was 5.23 × 10-14 m2 s-1 for C. pyriformis, 1.34 × 10-13 m2 s-1 for F. elastica, and 7.74 × 10-12 m2 s-1 for the PES membrane. We confirmed that the combination of a flow-through diffusion cell and real-time optical detection of transposed molecules represents a promising method for determining the permeability of membranes to xenobiotics occurring in the environment. This is necessary for determining a pesticide dosage in agriculture, selecting suitable membranes for passive samplers in analytics, testing membranes for water treatment, or studying material use of impregnated membranes.

Surface plasmon resonance: An innovative method for studying water permeability of plant cuticles.

The cuticle forms an effective barrier protecting plants from water loss. Its permeability to water and other compounds significantly differs between species, types of cuticle (stomatous, astomatous), and can be affected by a wide variety of ambient conditions. Enzymatic isolation of the leaf cuticle allows obtaining intact cuticles for permeability measurements. However, the most available gravimetric method, which is used for the assessment of water permeability of isolated cuticles, requires a relatively large area of the cuticle and does not allow the determination of membrane heterogeneity. We propose a new method for the determination of water permeance based on an on-line detection of water flux from a liquid phase to the atmosphere through isolated leaf cuticles in semi-flow chambers. This approach is new in using the phenomenon of surface plasmon resonance for the detection of the liquid phase refractive index affected by water vapor. Isolated cuticles of the leaves of Ficus elastica and an artificial polyethersulfone membrane were used for method evaluation. The composition of cuticular wax and its influence on cuticular permeability was also studied. It has been confirmed that the application of the surface plasmon resonance principle can be used for the assessment of leaf cuticle water permeability and heterogeneity.

A modified method for enzymatic isolation of and subsequent wax extraction from Arabidopsis thaliana leaf cuticle.

BACKGROUND: The plant cuticle represents one of the major adaptations of vascular plants to terrestrial life. Cuticular permeability and chemical composition differ among species. Arabidopsis thaliana is a widely used model for biochemical and molecular genetic studies in plants. However, attempts to isolate the intact cuticle from fresh leaves of Arabidopsis have failed so far. The goal of this study was to optimise an enzymatic method for cuticle isolation of species with a thin cuticle and to test it on several A. thaliana wild types and mutants. RESULTS: We developed a method for isolation of thin cuticles that allows reducing the isolation time, the separation of abaxial and adaxial cuticles, and avoids formation of wrinkles. Optical microscopy was used for studying cuticle intactness and scanning electron microscopy for visualisation of external and internal cuticle structures after isolation. Wax extracts were analysed by GC-MS. Isolation of intact cuticle was successful for all tested plants. The wax compositions (very-long-chained fatty acids, alcohols and alkanes) of intact leaves and isolated cuticles of wild type Col-0 were compared. CONCLUSIONS: We conclude that the optimised enzymatic method is suitable for the isolation of A. thaliana adaxial and abaxial cuticles. The isolated cuticles are suitable for microscopic observation. Analysis of wax composition revealed some discrepancies between isolated cuticles and intact leaves with a higher yield of wax in isolated cuticles.

Monochromatic green light induces an aberrant accumulation of geranylgeranyled chlorophylls in plants.

Light quality is an important environmental factor affecting the biosynthesis of photosynthetic pigments whose production seems to be affected not only quantitatively but also qualitatively. In this work, we set out to identify unusual pigment detected in leaves of barley (Hordeum vulgare L.) and explain its presence in plants grown under monochromatic green light (GL; 500-590 nm). The chromatographic analysis (HPLC-DAD) revealed that a peak belonging to this unknown pigment is eluted between chlorophyll (Chl) a and b. This pigment exhibited the same absorption spectrum and fluorescence excitation and emission spectra as Chl a. It was negligible in control plants cultivated under white light of the same irradiance (photosynthetic photon flux density of 240 µmol m-2 s-1). Mass spectrometry analysis of this pigment (ions m/z = 889 [M-H]-; m/z = 949 [M+acetic acid-H]-) indicates that it is Chl a with a tetrahydrogengeranylgeraniol side chain (containing two double bonds in a phytyl side chain; Chl aTHGG), which is an intermediate in Chl a synthesis. In plants grown under GL, the proportion of Chl aTHGG to total Chl content rose to approximately 8% and 16% after 7 and 14 days of cultivation, respectively. Surprisingly, plants cultivated under GL exhibited drastically increased concentration of the enzyme geranylgeranyl reductase, which is responsible for the reduction of phytyl chain double bonds in the Chl synthesis pathway. This indicates impaired activity of this enzyme in GL-grown plants. A similar effect of GL on Chl synthesis was observed for distinct higher plant species.

Protective effect of UV-A radiation during acclimation of the photosynthetic apparatus to UV-B treatment.

We examined the acclimation response of the photosynthetic apparatus of barley (Hordeum vulgare L.) to a combination of UV-A and UV-B radiation (UVAB) and to UV-B radiation alone. Our aim was to evaluate whether UV-A radiation prevents UV-B-induced damage to the photosynthetic apparatus and whether UV-A pre-acclimation is required to mitigate the negative influence of UV-B radiation. Barley plants were grown from seeds under low photosynthetically active radiation (50 µmol m(-2) s(-1)) either in the absence or presence of UV-A radiation (UVA- and UVA+ plants, respectively). After 8 days of development, plants were exposed simultaneously to UV-A and UV-B radiation for the next 6 days. Additionally, UVA- plants were exposed to UV-B radiation alone. The UVA+ plants had a higher CO2 assimilation rate near the light-saturation region (A(N)) and a higher content of both total chlorophylls (Chls) and total carotenoids than the UVA- plants. Chls content, A(N), the potential quantum yield of photosystem II (PSII) photochemistry (F(V)/F(M)), the capacity of light-induced thermal energy dissipation and the efficiency of excitation energy transfer within PSII remained the same or even increased in both UVA+ and UVA- plants after UVAB treatment. On the contrary, exposure of UVA- plants to UV-B radiation itself led to a reduction in all these characteristics. We revealed that the presence of UV-A radiation during UVAB treatment not only mitigated but completely eliminated the negative effect of UV-B radiation on the functioning of the photosynthetic apparatus and that UV-A pre-acclimation was not crucial for development of this UV-A-induced resistance against UV-B irradiation.

Stomatal and pavement cell density linked to leaf internal CO2 concentration.

BACKGROUND AND AIMS: Stomatal density (SD) generally decreases with rising atmospheric CO2 concentration, Ca. However, SD is also affected by light, air humidity and drought, all under systemic signalling from older leaves. This makes our understanding of how Ca controls SD incomplete. This study tested the hypotheses that SD is affected by the internal CO2 concentration of the leaf, Ci, rather than Ca, and that cotyledons, as the first plant assimilation organs, lack the systemic signal. METHODS: Sunflower (Helianthus annuus), beech (Fagus sylvatica), arabidopsis (Arabidopsis thaliana) and garden cress (Lepidium sativum) were grown under contrasting environmental conditions that affected Ci while Ca was kept constant. The SD, pavement cell density (PCD) and stomatal index (SI) responses to Ci in cotyledons and the first leaves of garden cress were compared. (13)C abundance (δ(13)C) in leaf dry matter was used to estimate the effective Ci during leaf development. The SD was estimated from leaf imprints. KEY RESULTS: SD correlated negatively with Ci in leaves of all four species and under three different treatments (irradiance, abscisic acid and osmotic stress). PCD in arabidopsis and garden cress responded similarly, so that SI was largely unaffected. However, SD and PCD of cotyledons were insensitive to Ci, indicating an essential role for systemic signalling. CONCLUSIONS: It is proposed that Ci or a Ci-linked factor plays an important role in modulating SD and PCD during epidermis development and leaf expansion. The absence of a Ci-SD relationship in the cotyledons of garden cress indicates the key role of lower-insertion CO2 assimilation organs in signal perception and its long-distance transport.

Acclimation of Norway spruce photosynthetic apparatus to the combined effect of high irradiance and temperature.

Diurnal courses of photosynthetic gas exchange parameters, chlorophyll a fluorescence characteristics and the de-epoxidation state of the xanthophyll cycle pigments (DEPS) were measured during the gradual acclimation of 4-year-old Norway spruce seedlings to different photosynthetic photon flux density (PPFD) and air temperature (T(air)) regimes, simulating cloudy days with moderate T(air) (LI, maximum PPFD 300 micromol m(-2)s(-1), T(air) range 15-25 degrees C), sunny days with moderate T(air) (HI, maximum PPFD 1000 micromol m(-2)s(-1), T(air) range 15-25 degrees C) and hot sunny days (HI-HT, maximum PPFD 1000 micromol m(-2)s(-1), T(air) range 20-35 degrees C). The plants were acclimated inside a growth chamber and each acclimation regime lasted for 13d. Acclimation to HI conditions led to a strong depression of the net CO(2) assimilation rates (A(N)), particularly during noon and afternoon periods. Exposure to the HI-HT regime led to a further decrease of A(N) even during the morning period. Insufficient stomatal conductance was found to be the main reason for depressed A(N) under HI and HI-HT conditions. Only slight changes of the maximum photosystem II (PSII) photochemical efficiency (F(V)/F(M)), in the range of 0.78-0.82, supported the resistance of the Norway spruce photosynthetic apparatus against PSII photoinhibition during acclimation to both HI and HI-HT conditions. The HI plants showed increased content of xanthophyll cycle pigments (VAZ) and enhanced efficiency of thermal energy dissipation within PSII (D) that closely correlated with the increased DEPS. In contrast, acclimation to the HI-HT regime resulted in a slight reduction of VAZ content and significantly diminished D and DEPS values during the entire day in comparison with HI plants. These results indicate a minor role of the xanthophyll cycle-mediated thermal dissipation in PSII photoprotection under elevated temperatures. The different contributions of the thermal dissipation and non-assimilatory electron transport pathways in PSII photoprotection during acclimation of the Norway spruce photosynthetic apparatus to excess irradiance and heat stresses are discussed.