Metabolomics is an analytical profiling technique that measures and compares large numbers of metabolites in biological samples, providing insight into metabolic mechanisms. There are few studies concerning the effects of xenobiotics and their transformation products on aquatic plant metabolites, which can uptake and detoxify them using untargeted metabolomics. This study investigates how pharmaceuticals, including diclofenac (DCF) and carbamazepine (CBZ), as well as sulfamethoxazole (SMX) and trimethoprim (TRIM), present in aquatic environments, can influence the biosynthetic pathways of Lemna minor.Based on previous research on the effects of DCF, SMX, and TRIM on Lemna pathways, specifically phenylalanine, tyrosine, and tryptophan biosynthesis, folate biosynthesis, and the phenylpropanoid pathway, including flavonoid and anthocyanin metabolism. Lemna was incubated with DCF, CBZ, SMX, and TRIM alone and in a mixture (MIX) at 5 ppb (5 µg/L) for 5 days, at concentrations near environmental levels. The methanolic extract was analysed using a Q Exactive Focus Orbitrap to investigate changes in the aforementioned biosynthetic pathways, as reported in previous studies. Lemna can modulate its pathways to produce more phenolic compounds as a defence mechanism against various drugs. This modulation can be considered an indicator for each drug. The presence of pharmaceuticals in the aquatic environment can affect the biosynthetic pathways of Lemna. Therefore,Lemna minor can be used as a model to study the stress-response of different pharmaceuticals on plant metabolites and their pathways.

Enhanced weathering (EW) is proposed as a key strategy for climate change mitigation and carbon dioxide removal technology. Dissolution of silicate minerals enhances the alkalinity of the pore water, resulting at a shift of the carbonate system towards carbonate and bicarbonate, leading to higher dissolved inorganic carbon when the water is equilibrated with the atmosphere. Here, we evaluated the effects of EW on a crop ecosystem under future climate change conditions within a macro-scale ecotron – an enclosed facility enabling complete quantification of carbon fluxes among the atmosphere, vegetation, soil, and leachates. We monitored all greenhouse gases in deep mesocosms representative of marginal soil conditions and, after liming and fertilization, applied 10 t ha−1 of basalt at the start of the experiment. EW treatment resulted in an almost three-fold enhancement of measured carbon flux into the soil, achieving rates up to 1.5 t ha−1 during the growing season. Moreover, the observed carbon sequestration surpassed the levels expected from weathering processes alone. This is notable because the near-neutral soil pH environment was not favourable to EW kinetics. Therefore, we conclude that EW facilitated significant carbon accrual in our simulated ecosystems via not only carbonate precipitation but also enhanced biogeochemical activities promoting additional carbon storage. Based on these findings, we speculate on the underlying pathways responsible for such outcomes.

Water and N availability are key factors limiting crop yield, particularly in marginal soils. This study evaluated the effects of water and N stress on barley grown in marginal soils using field trials and the AgroC model. Experiments from 2020 to 2022 in Lithuania with spring barley cv. KWS Fantex under two N fertilization treatments on sandy soil provided data for model parameterization. The AgroC model simulated barley growth to assess yield potential and yield gaps due to water and N stress. Potential grain yields (assuming no water or N stress) ranged from 4.8 to 6.02 t DW ha−1, with yield losses up to 54.4% assuming only N stress and 59.2% assuming only water stress, even with the N100 treatment (100 kg N ha−1 yr−1). A synthetic case study varying N fertilization from 0 to 200 kg N ha−1 yr−1 showed that increasing N still enhanced yield, but the optimal rate of 100–120 kg N ha−1 yr−1 depended on climatic conditions, leading to uncertainty in fertilization recommendations. This study underscores the importance of integrating advanced modeling techniques with sustainable agricultural practices to boost yield potential and resilience in marginal soils. Incorporating remote sensing data to capture soil and crop variability is recommended for improving simulation accuracy, contributing to sustainable agriculture strategies in the Baltic–Nordic region.

Phytolith-occluded carbon (PhytOC) is highly stable, and constitutes an important source of long-term C storage in agrosystems. This stored carbon is resistant to the processes of oxidation of carbon compounds. In our research phytolith content in barley (Estonia) and oat (Poland) grain and straw was assessed at field trials, with Si as a liquid immune stimulant OPTYSIL and compost fertilisation. We showed that cereals can produce relatively high amounts of phytoliths. PhytOC plays a key role in carbon sequestration, particularly for poor, sandy Polish and Estonian soils. The phytolith content was always higher in straw than in grain regardless of the type of cereals. The phytolith content in oat grains varied from 18.46 to 21.28 mg∙g−1 DM, and in straw 27.89–38.97 mg∙g−1 DM. The phytolith content in barley grain ranged from 17.24 to 19.86 mg∙g−1 DM, and in straw from 22.06 to 49.08 mg∙g−1 DM. Our results suggest that oat ecosystems can absorb from 14.94 to 41.73 kg e-CO2∙ha−1 and barley absorb from 0.32 to 1.60 kg e-CO2∙ha−1. The accumulation rate of PhytOC can be increased 3-fold in Polish conditions through foliar application of silicon, and 5-fold in Estonian conditions. In parallel, the compost fertilisation increased the phytolith content in cereals.

To facilitate nutrient management and the use of manure as a feedstock for biogas production, manure is often separated into a solid and a liquid fraction. The former fraction is usually high in P and low in N, so when incorporated in the soil as fertilizer, it needs to be supplemented by N from, e.g., mineral fertilizers or nitrogen-fixing species. To explore strategies to manage N with solid-separated manure, we examined how the amount of digestate and the N:P ratio of pig digestate, i.e., manure that had partially undergone anaerobic digestion, affected the productivity of Westerwolds ryegrass and red clover in a pot experiment with one soil which was rich and another which was poor in plant nutrients. The soil and plant species treatments were combined with four doses of digestate, which gave plant available phosphorus (P) concentrations of 2, 4, 8, or 16 mg P100 g−1 soil. Ammonium nitrate was dosed to obtain factorial combinations of digestate amount and N:P ratios of 1.8, 4, 8, and 16. Clover was harvested once at the beginning of flowering (15 weeks after seeding), while Westerwolds ryegrass was allowed to regrow three times after being cut at the shooting stage (in total, 4 cuts, 6, 9, 12, and 15 weeks after seeding). Ryegrass yield increased by up to 2.9 times with digestate dosage. Interactions with the N:P ratio and soil type were weak. Hence, the effect of increasing the N:P ratio was additive across digestate dosages. Red clover biomass also increased by up to 39% with digestate dosage. Residual nutrients in the soil after red clover cultivation were affected by the initial differences in soil characteristics but not by digestate treatment or biomass of harvested red clover. A targeted N management is required to benefit from the P-rich digestate in grass cultivation, while the long-term effects of red clover culture on N input need further investigation.

Sewage sludge (biosolids) management represents a worldwide issue. Due to its valuable properties, approximately one half of the EU production is recovered in agriculture. Nevertheless, growing attention is given to potential negative effects deriving from the presence of harmful pollutants. It is recognized that a (even very detailed) chemical characterization is not able to predict ecotoxicity of a mixture. However, this can be directly measured by bioassays. Actually, the choice of the most suitable tests is still under debate. This paper presents a multilevel characterization protocol of sewage sludge and other organic residues, based on bioassays and chemical-physical-microbiological analyses. The detailed description of the experimental procedure includes all the involved steps: the criteria for selecting the organic matrices to be tested and compared; the sample pre-treatment required before the analyses execution; the chemical, physical and microbiological characterisation; the bioassays, grouped in three classes (baseline toxicity; specific mode of action; reactive mode of action); data processing. The novelty of this paper lies in the integrated use of advanced tools, and is based on three pillars: • the direct ecosafety assessment of the matrices to be reused. • the adoption of innovative bioassays and analytical procedures. • the original criteria for data normalization and processing.

Growing crops on marginal lands is a promising solution to alleviate the increasing pressure on agricultural land in Europe. Such crops will however be at the same time exposed to increased drought and pathogen prevalence, on already challenging soil conditions. Some sustainable practices, such as Silicon (Si) foliar fertilization, have been proposed to alleviate these two stress factors, but have not been tested under controlled, future climate conditions. We hypothesized that Si foliar fertilization would be beneficial for crops under future climate, and would have cascading beneficial effects on ecosystem processes, as many of them are directly dependent on plant health. We tested this hypothesis by exposing spring barley growing on marginal soil macrocosms (three with, three without Si treatment) to 2070 climate projections in an ecotron facility. Using the high-capacity monitoring of the ecotron, we estimated C, water, and N budgets of every macrocosm. Additionally, we measured crop yield, the biomass of each plant organ, and characterized bacterial communities using metabarcoding. Despite being exposed to water stress conditions, plants did not produce more biomass with the foliar Si fertilization, whatever the organ considered. Evapotranspiration (ET) was unaffected, as well as water quality and bacterial communities. However, in the 10-day period following two of the three Si applications, we measured a significant increase in C sequestration, when climate conditions where significantly drier, while ET remained the same. We interpreted these results as a less significant effect of Si treatment than expected as compared with literature, which could be explained by the high CO2 levels under future climate, that reduces need for stomata opening, and therefore sensitivity to drought. We conclude that making marginal soils climate proof using foliar Si treatments may not be a sufficient strategy, at least in this type of nutrient-poor, dry, sandy soil.

GATAs are evolutionarily conserved zinc-finger transcription factors from eukaryotes. In plants, GATAs can be subdivided into four classes, A-D, based on their DNA-binding domain, and into further subclasses based on additional protein motifs. B-GATAs with a so-called leucine-leucine-methionine (LLM)-domain can already be found in algae. In angiosperms, the B-GATA family is expanded and can be subdivided in to LLM- or HAN-domain B-GATAs. Both, the LLM- and the HAN-domain are conserved domains of unknown biochemical function. Interestingly, the B-GATA family in the liverwort Marchantia polymorpha and the moss Physcomitrium patens is restricted to one and four family members, respectively. And, in contrast to vascular plants, the bryophyte B-GATAs contain a HAN- as well as an LLM-domain. Here, we characterise mutants of the single B-GATA from Marchantia polymorpha. We reveal that this mutant has defects in thallus growth and in gemma formation. Transcriptomic studies uncover that the B-GATA mutant displays a constitutive high-light (HL) stress response, a phenotype that we then also confirm in mutants of Arabidopsis thaliana LLM-domain B-GATAs, suggesting that the B-GATAs have a protective role towards HL stress.

Insufficiently treated reclaimed water can act as a source of contamination by introducing recalcitrant contaminants (e.g., pharmaceutical compounds) to various water bodies and/or agricultural soils after irrigation. Tramadol (TRD) is one of these pharmaceuticals that can be detected in influents and effluents of wastewater treatment plants, at dis-charge points as well as in surface waters in Europe. While the uptake of TRD by plants through irrigation water has been shown, plant responses towards this compound are still unclear. Therefore, this study aims to evaluate the effects of TRD on selected plant enzymes as well as on the root bacterial community structure. A hydroponic experiment was conducted to test the effects of TRD (100 & mu;g L-1 TRD) on barley plants, at two harvesting time points after treatment. Accumulation of TRD in root tissues over time was observed reaching concentrations of 111.74 and 138.39 & mu;g g-1 in total root FW after 12 and 24 days of exposure, respectively. Furthermore, noticeable inductions in guaiacol peroxidase (5.47-fold), catalase (1.83-fold) and glutathione S-transferase (3.23-and 2.09-fold) were recorded in roots of TRD-treated plants compared to controls after 24 days. A significant alteration in the beta diversity of root-associated bac-teria due to TRD treatment was observed. Three amplicon sequence variants assigned to Hydrogenophaga, U. Xanthobacteraceae and Pseudacidovorax were differentially abundant in TRD-treated compared to control plants at both harvesting time points. This study reveals the resilience of plants through the induction of the antioxidative system and changes in the root-associated bacterial community to cope with the TRD metabolization/detoxification process.

The intensive use of antibiotics (for human, veterinary, and agricultural purposes) has steadily increased over the last 30 years. Large amounts of antibiotic residues are released into aquatic systems, mostly due to inefficient wastewater treatment. Conventional wastewater treatments are not designed to remove emerging contaminants (such as antibiotics) from wastewater. Therefore, algae treatment (phycoremediation) has emerged as a promising choice for cost-effective, eco-friendly, and sustainable wastewater treatment. For this reason, we investigated the removal performance of a well-established algal consortia (Chlorella protothecoides and Chlorella vulgaris) used in passive wastewater treatment ponds (Mosselbay, South Africa). Five antibiotics (sulfamethoxazole, amoxicillin, trimethoprim, ofloxacin, and clarithromycin) were selected for their ubiquity and/or low removal efficiency in conventional wastewater treatment plants (WWTPs). For each antibiotic, two concentrations were used: one environmentally relevant (10 ppb) and another 10 times higher (100 ppb), tested in triplicate and collected at two-time points (7 and 10 days). The algae remained viable over the exposure period (which is similar to the retention time within maturation ponds) and exhibited the capacity to remove sulfamethoxazole (77.3% ± 3.0 and 46.5% ± 5.3) and ofloxacin (43.5% ± 18.9 and 55.1% ± 12.0) from samples spiked with 10 and 100 ppb, respectively. This study demonstrates the potential and innovation of algal remediation for contaminants in a developing country context, where minimal infrastructure is available.

Tramadol (TRD) is widely detected in aquatic ecosystems as a result of massive abuse and insufficient removal from wastewater facilities. As a result, TRD can contaminate groundwater sources and/or agricultural soils. While TRD toxicity has been reported from aquatic biota, data about TRD detection in plants are scarce. Moreover, information regarding plant capability for TRD removal is lacking. To understand the fate of this opioid, we have investigated the uptake, translocation and removal capacity of TRD by plants, addressing short-term and longterm uptake. The uptake rates of TRD, in excised barley and cattail roots, were 5.18 and 5.79 mu g g(-1) root fresh weight day(-1), respectively. However, TRD uptake was strongly inhibited after co-exposing these roots either with the drug venlafaxine (similar molecular structure as TRD) or with quinidine (an inhibitor of cellular organic cation transporters). When barley seedlings were exposed to TRD in a hydroponic experiment a removal efficiency up to 90% (within 15 days) was obtained, with bioconcentration and translocation factors close to 9 and 1, respectively. The combination of results from both plants and the inhibition observed after treatment with quinidine revealed that organic cation transporters may be involved in the uptake of TRD by plants. (C) 2021 Elsevier B.V. All rights reserved.

The reuse of water for agricultural practices becomes progressively more important due to increasing demands for a transition to a circular economy. Treated wastewater can be an alternative option of blue water used for the irrigation of crops but its risks need to be evaluated. This study assesses the uptake and metabolization of pharmaceuticals and personal care products (PPCPs) derived from treated wastewater into lettuce as well as the impact on root-associated bacteria under a realistic and worst-case scenario. Lettuce was grown in a controlled greenhouse and irrigated with water or treated wastewater spiked with and without a mixture of fourteen different PPCPs at 10 μg/L or 100 μg/L. After harvesting the plants, the same soil was reused for a consecutive cultivation campaign to test for the accumulation of PPCPs. Twelve out of fourteen spiked PPCPs were detected in lettuce roots, and thirteen in leaves. In roots, highest concentrations were measured for sucralose, sulfamethoxazole and citalopram, while sucralose, acesulfame and carbamazepine were the highest in leaves. Higher PPCP concentrations were found in lettuce roots irrigated with spiked treated wastewater than in those irrigated with spiked water. The absolute bacterial abundance remained stable over both cultivation campaigns and was not affected by any of the treatments (type of irrigation water (water vs. wastewater) nor concentration of PPCPs). However, the irrigation of lettuce with treated wastewater had a significant effect on the microbial α-diversity indices at the end of the second cultivation campaign, and modified the structure and community composition of root-associated bacteria at the end of both campaigns. Five and fourteen bacterial families were shown to be responsible for the observed changes at the end of the first and second cultivation campaign, respectively. Relative abundance of Haliangium and the clade Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium was significantly affected in response to PCPPs exposure. Caulobacter, Cellvibrio, Hydrogenophaga and Rhizobacter were significantly affected in microcosms irrigated with wastewater.

Nanomaterials can induce plant tolerance to abiotic environmental stresses, whereas the sensing mechanism and the resulting response at the cellular level need further exploration.

The potential environmental toxicity of nano-TiO2to rat fetal development is associated with retention of thyroid hormones in the placenta.

In previous two-tier experiments designed to test agronomical (treated wastewater) and worst-case scenario (wastewater spiked with a mixture of 14 pharmaceuticals and personal care products (PPCPs) at 10 and 100 μg/L), 14 different wastewater-borne PPCPs accumulated in soil, lettuce roots, and leaves leading to a significant ecotoxicological impact on soil and root-associated bacteria. Here, we assessed the effects of wastewater irrigation on the arbuscular mycorrhizal fungal (AMF) colonization, diversity, and composition in lettuce roots. Neither the wastewater nor the concentration of spiked PPCPs had an impact on the colonization, alpha-diversity indices (Chao1, PD whole tree, Simpson reciprocal, and Shannon), and composition of root-associated AMF communities. Taxonomical analysis of the fungi revealed the presence of 14 major phylotypes, 13 of which belonged to the Glomeromycota division. As for the alpha and beta diversity indices, none of the phylotypes was affected by either the wastewater or the PPCPs. This indicates that under both agronomical and worst-case scenario, the irrigation of lettuce with wastewater had no effect on the root-associated AMF community.

The composition of root exudates is modulated by several environmental factors, and it remains unclear how that affects beneficial rhizosphere or inoculated microorganisms under heavy metal (HM) contamination. Therefore, we evaluated the transcriptional response of Pseudomonas putida E36 (a Miscanthus x giganteus isolate with plant growth promotion-related properties) to Cd, Pb and Zn in an in vitro study implementing root exudates from M. x giganteus. To collect root exudates and analyse their composition plants were grown in a pot experiment under HM and control conditions. Our results indicated higher exudation rate for plants challenged with HM. Further, out of 29 organic acids identified and quantified in the root exudates, 8 of them were significantly influenced by HM (e.g., salicylic and terephthalic acid). The transcriptional response of P. putida E36 was significantly affected by the HM addition to the growth medium, increasing the expression of several efflux pumps and stress response-related functional units. The additional supplementation of the growth medium with root exudates from HM-challenged plants resulted in a downregulation of 29% of the functional units upregulated in P. putida E36 as a result of HM addition to the growth medium. Surprisingly, root exudates + HM downregulated the expression of P. putida E36 functional units related to plant colonization (e.g., chemotaxis, motility, biofilm formation) but upregulated its antibiotic and biocide resistance compared to the control treatment without HM. Our findings suggest that HM-induced changes in root exudation pattern may attract beneficial bacteria that are in turn awarded with organic nutrients, helping them cope with HM stress. However, it might affect the ability of these bacteria to colonize plants growing in HM polluted areas. Those findings may offer an insight for future in vivo studies contributing to improvements in phytoremediation measures.

The reuse of water for agricultural practices becomes progressively more important due to the increasing demand for a transition to a circular economy. Treated wastewater can be an alternative option of blue water used for irrigation of crops but its risks need to be evaluated. This study assesses the uptake and metabolization of pharmaceuticals and personal care products (PPCPs) derived from treated wastewater into lettuce as well as the ecotoxicological impact on root-associated bacteria under a realistic and worst-case scenario. Lettuce was grown in a controlled greenhouse and irrigated with water or treated wastewater spiked with and without a mixture of fourteen different PPCPs at 10 µg/L or 100 µg/L. After harvesting the plants, the same soil was reused for a consecutive cultivation campaign to test for accumulation of PPCPs. Twelve out of fourteen spiked PPCPs were detected in lettuce roots, and thirteen in leaves. In roots, highest concentrations were measured for sucralose, sulfamethoxazole and citalopram, while sucralose, acesulfame and carbamazepine were the highest in the leaves. Higher PPCP concentrations were found in lettuce roots irrigated with spiked treated wastewater than in those irrigated with spiked water. Absolute bacterial abundance remained stable over both cultivation campaigns and was not affected by any of the treatments. However, the irrigation of lettuce with treated wastewater had a significant effect on microbial α-diversity indices at the end of the second cultivation campaign, and modified the structure and community composition of root-associated bacteria at the end of both campaigns. Five and fourteen bacterial families were shown to be responsible for the observed changes at the end of the first and second cultivation campaign, respectively. Relative abundance of Haliangium and the clade Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium was significantly affected in response to PCPPs exposure. Caulobacter, Cellvibrio, Hydrogenophaga and Rhizobacter were significantly affected in microcosms irrigated with wastewater.

Metabolomics is an emerging approach that investigates the changes in the metabolome profile. In the present study, Lemna minor -considered as an experimental aquatic plant model- was incubated with 10 and 100 μM diclofenac (DCF) for 96 h, respectively. Knowing that DCF is internationally often problematic in wastewater effluents and that it might affect particularly the metabolic profiles in aquatic plants, mainly the oxidoreductase, dehydrogenase, peroxidase, and glutathione reductase activities, here it was hypothesized (H) that in the common duckweed, DCF might increase the phenolic and flavonoids pathways, as an antioxidant response to this stress (H1). Also, it was expected DCF to alternate the physiological characteristics, especially the molecular interaction and biochemical properties, of Lemna (H2). Metabolic changes were investigated with target and untargeted screening analysis using RPLC-HILIC-ESI-TOF-MS. Twelve amino acids were identified in all treatments, together with three organic acids (p-coumaric, cinnamic, and sinapic acids). In untargeted screening, the important metabolites to discriminate between different treatments were assigned to Lemna such as organic acids, lignin, sugars, amino acids, dipeptides, flavonoids, biflavonoids, fatty acids, among others. In resume, Lemna responded to both DCF concentrations, showing different stress patterns. A similar metabolic response had already been identified in other studies in exposing Lemna to other anthropogenic stressors (like pesticides).

Metabolomics approaches provide a vast array of analytical datasets, which require a comprehensive analytical, statistical, and biochemical workflow to reveal changes in metabolic profiles. The biological interpretation of mass spectrometric metabolomics results is still obstructed by the reliable identification of the metabolites as well as annotation and/or classification. In this work, the whole Lemna minor (common duckweed) was extracted using various solvents and analyzed utilizing polarity-extended liquid chromatography (reversed-phase liquid chromatography (RPLC)-hydrophilic interaction liquid chromatography (HILIC)) connected to two time-of-flight (TOF) mass spectrometer types, individually. This study (introduces and) discusses three relevant topics for the untargeted workflow: (1) A comparison study of metabolome samples was performed with an untargeted data handling workflow in two different labs with two different mass spectrometers using the same plant material type. (2) A statistical procedure was observed prioritizing significant detected features (dependent and independent of the mass spectrometer using the predictive methodology Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA). (3) Relevant features were transferred to a prioritization tool (the FOR-IDENT platform (FI)) and were compared with the implemented compound database PLANT-IDENT (PI). This compound database is filled with relevant compounds of the Lemnaceae, Poaceae, Brassicaceae, and Nymphaceae families according to analytical criteria such as retention time (polarity and LogD (pH 7)) and accurate mass (empirical formula). Thus, an untargeted analysis was performed using the new tool as a prioritization and identification source for a hidden-target screening strategy. Consequently, forty-two compounds (amino acids, vitamins, flavonoids) could be recognized and subsequently validated in Lemna metabolic profile using reference standards. The class of flavonoids includes free aglycons and their glycosides. Further, according to our knowledge, the validated flavonoids robinetin and norwogonin were for the first time identified in the Lemna minor extracts.