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Contamination of agricultural soils with heavy metals (HMs) poses a significant environmental threat, especially because industrial discharges often irrigate agricultural lands. A prominent source of HM(s) pollution occurs from tannery effluents containing high concentrations of chromium (Cr) in both Cr3+ and Cr6+ forms along with other toxic materials. Cr is known for its carcinogenic and mutagenic properties in biological systems. Microbe-assisted phytoremediation has emerged as a promising and environmentally friendly approach for detoxifying Cr-contaminated environments. This study aimed to evaluate the performance of citric acid (CA) and a Cr-reducing bacterial strain (Staphylococcus aureus) on the phytoextraction potential of Lemna minor within a Constructed Wetland System treated with tannery wastewater. Various combinations of tannery wastewater (0, 50, and 100 %), CA (0, 5 and 10 mM), and microbial inoculants were applied to the test plants. The mitigative effects of Staphylococcus aureus strain K1 were examined in combination with different concentrations of CA (0, 5, 10 mM). Data on growth and yield attributes highlighted the beneficial effects of bacterial inoculation and CA in ameliorating Cr toxicity in L. minor, as evidenced by increased foliar chlorophyll and carotenoid contents, enhanced antioxidant enzyme activities (SOD, POD, APX, CAT), and improved nutrient uptake. Specifically, CA application resulted in an enhancement of Cr ranging from 12% to 15% and 23%-31% in concentration, and 134%-141% and 322%-337% in Cr accumulation, respectively. When combined with the S. aureus inoculation treatment, CA application (5 and 10 mM) further increased the concentration and accumulation of Cr in L. minor. The enhancement in Cr ranged from 12% to 23% and 27%-41% in concentration, 68%-75%, and 179%-185% in accumulation, respectively. These results demonstrated that L. minor is an effective choice for environmentally friendly Cr remediation due to its continued ability to grow in polluted wastewater. This study suggested that microbial-assisted phytoextraction combined with chelating agents such as CA could be a practical and effective approach for remediating tannery effluents.

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Triclosan (TCS), an emerging pollutant, is a notable contributor to adverse impacts on aquatic organisms due to its widespread use during COVID-19 and hydrophobic properties. There is extensive documented literature on TCS toxicity in commercially important fish species; however, studies on aquatic plants remain limited. In this prelude, the present study aims to evaluate the effect of TCS on Lemna minor, a commercially important aquatic plant species for 7 days. The results showed dose-dependent significant alterations in growth, pigments and stress enzymes of L. minor at varied concentrations of TCS (1 to 8 mg L-1). Median inhibitory concentration (IC50) was found to be 4.813 mg L-1. Total chlorophyll and carotenoid levels decreased 73.11 and 81.83%, respectively after 7 days of TCS exposure. A significant increase in catalase and superoxide dismutase activity was observed in TCS exposed groups as compared to the control. Bioconcentration factor was found to be in the range of 5.855 to 37.129 signifying TCS ability to accumulate and transfer through the food chain. Scanning electron microscopy (SEM) analysis showed deformation in the cell surface and alteration of stroma morphology of TCS exposed groups. Furthermore, the Fourier transform infrared spectroscopy (FTIR) study also revealed that higher concentrations of TCS could cause alteration in the functional groups in the plant. This study demonstrates that TCS negatively impacts the growth and metabolism of primary producers, offering crucial insights into its interactions with aquatic plants and establishing baseline information essential for crafting effective mitigation strategies for TCS contamination in aquatic environments.

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The method of plant micropropagation is widely used to obtain genetically homogeneous and infection-free plants for the needs of various industries and agriculture. Optimization of plant growth and development conditions plays a key role in economically successful micropropagation. Computer technologies have provided researchers with new approaches for modeling and a better understanding of the role of the factors involved in plant growth in vitro. To develop new models for optimizing growth conditions, we used plants with a high speed of vegetative in vitro reproduction, such as duckweed (Wolffia arrhiza and Lemna minor). Using the development of the optimal modeling of the biological processes, we have obtained the prescriptions for an individually balanced culture medium that enabled us to obtain 1.5-2.0 times more duckweed biomass with a 1.5 times higher protein concentration in the dry mass. Thus, we have demonstrated that the method of optimization modeling of the biological processes based on solving multinomial tasks from the series of quadratic equations can be used for the optimization of trophic needs of plants, specifically for micropropagation of duckweeds in vitro.

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With the advent of technological advancements post the industrial revolution, thousands of chemicals are introduced into the market annually to enhance different facets of human life. Among these, pharmaceutical and personal care products (PPCPs), including antibiotics and disinfectants, such as benzalkonium chlorides (BACs), are prominent. BACs, often used for surface and hand disinfection in high concentrations or as preservatives in health products such as nasal sprays and eye drops, may present environmental risks if they seep into irrigation water through prolonged exposure or improper application. The primary objective of this study is to elucidate the tolerance mechanisms that may arise in Lemna minor plants, known for their remarkable capability to accumulate substances efficiently, in response to exogenously applied BACs at varying concentrations. The study applied six different concentrations of BACs, ranging from 0.25 to 10 mg L-1. The experimental period spanned seven days, during which the treatments were conducted in triplicate to ensure reliability and reproducibility of the results. It was observed that low concentrations of BACs (0.25, 0.5 and 1 mg L-1) did not elicit any statistically significant changes in growth parameters. However, higher concentrations of BACs (2.5, 5, and 10 mg L-1) resulted in a reduction in RGR by 20%, 28%, and 36%, respectively. Chlorophyll fluorescence declined significantly at BAC doses of 5 and 10 mg L-1, with Fv/Fm ratios decreasing by 9% and 15%, and Fv/Fo ratios by 40% and 39%, respectively. Proline content decreased in all treatment groups, with a 46% reduction at 10 mg L-1 BAC. TBARS and H2O2 contents increased proportionally with BAC dosage, showing the highest increases of 30% and 40% at 10 mg L-1, respectively. The noticeable increase in SOD enzyme activity at BAC concentrations of 0.5, 1, and 2.5 mg L-1, with increases of 2.7-fold, 2.2-fold, and 1.7-fold respectively, along with minimal accumulation of H2O2, suggests that L. minor plants have a strong tolerance to BAC. This is supported by the efficient functioning of the CAT and GST enzymes, especially evident at the same concentrations, where increased activities effectively reduce the buildup of H2O2. In the AsA-GSH cycle, although variations were observed between groups, the contribution of the GR enzyme to the preservation of GSH content by recycling GSSG likely maintained redox homeostasis in the plant, especially at low concentrations of BACs. The study revealed that L. minor effectively accumulates BAC alongside its tolerance mechanisms and high antioxidant activity. These results underscore the potential for environmental cleanup efforts through phytoremediation.

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Microbiomes often benefit plants, conferring resistance to pathogens, improving stress tolerance, or promoting plant growth. As potential plant mutualists, however, microbiomes are not a single organism but a community of species with complex interactions among microbial taxa and between microbes and their shared host. The nature of ecological interactions among microbes in the microbiome can have important consequences for the net effects of microbiomes on hosts. Here, we compared the effects of individual microbial strains and 10-strain synthetic communities on microbial productivity and host growth using the common duckweed Lemna minor and a synthetic, simplified version of its native microbiome. Except for Pseudomonas protegens, which was a mutualist when tested alone, all of the single strains we tested were commensals on hosts, benefiting from plant presence but not increasing host growth relative to uninoculated controls. However, 10-strain synthetic microbial communities increased both microbial productivity and duckweed growth more than the average single-strain inoculation and uninoculated controls, meaning that host-microbiome mutualisms can emerge from community interactions among microbes on hosts. The effects of community inoculation were sub-additive, suggesting at least some competition among microbes in the duckweed microbiome. We also investigated the relationship between L. minor fitness and that of its microbes, providing some of the first empirical estimates of broad fitness alignment between plants and members of their microbiomes; hosts grew faster with more productive microbes or microbiomes. IMPORTANCE: There is currently substantial interest in engineering synthetic microbiomes for health or agricultural applications. One key question is how multi-strain microbial communities differ from single microbial strains in their productivity and effects on hosts. We tested 20 single bacterial strains and 2 distinct 10-strain synthetic communities on plant hosts and found that 10-strain communities led to faster host growth and greater microbial productivity than the average, but not the best, single strain. Furthermore, the microbial strains or communities that achieved the greatest cell densities were also the most beneficial to their hosts, showing that both specific single strains and multi-strain synthetic communities can engage in high-quality mutualisms with their hosts. Our results suggest that ~5% of single strains, as well as multi-strain synthetic communities comprised largely of commensal microbes, can benefit hosts and result in effective host-microbe mutualisms.

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Little is known about the effect of surface coatings on the fate and toxicity of CeO2 nanoparticles (NPs) to aquatic plants. In this study, we modified nCeO2 with chitosan (Cs) and alginate (Al) to obtain positively charged nCeO2@Cs and negatively charged nCeO2@Al, respectively, and exposed them to a representative aquatic plant, duckweed (Lemna minor L.). Uncoated nCeO2 could significantly inhibit the growth of duckweed, induce oxidative damage and lead to cell death, whereas nCeO2@Cs and nCeO2@Al exhibited lower toxicity to duckweed. ICP-MS analysis revealed that the Ce content in duckweed from the nCeO2 group was 1.74 and 2.85 times higher than that in the nCeO2@Cs and nCeO2@Al groups, respectively. Microscopic observations indicated that the positively charged nCeO2@Cs was more readily adsorbed on the root surface of duckweed than the negatively charged nCeO2@Al. The results of XANES and LCF demonstrated that a certain percentage of Ce(Ⅳ) was reduced to Ce(Ⅲ) after the interaction of the three NPs with duckweed, but the degree of biotransformation differed among the treatments. Specifically, the absolute contents of Ce(III) produced of nCeO2@Cs and nCeO2@Al through biotransformation were reduced by 55.5% and 83.5%, respectively, compared with that of the nCeO2 group, which might be the key factor for the diminished phytotoxicity of the coated nCeO2 to the duckweed. These findings were valuable for understanding the toxicity of metal-based NPs to aquatic plants and for the synthesis of environmentally friendly nanomaterials.

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Duckweed (Lemna minor L.) is a small floating aquatic plant that has an important economic impact in several industrial areas. With its high biomass production, reasonable protein content, and resilience to several climates, it has been attracting increasing interest for potential use in animal and human food systems. Historically consumed in southwest Asia, this duckweed is now gaining attention as a potential novel food in Europe. This manuscript explores the contributions of duckweed to various food and feed industries, including aquaculture and livestock, while also pointing out the incipient research carried out for human consumption. Most importantly, it highlights the potential of Lemna minor as a vegetable for future human consumption whether eaten whole or through extraction of its nutrients.

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Isotope labeling coupled with mass spectrometry imaging (MSI) presents a potent strategy for elucidating the dynamics of metabolism at cellular resolution, yet its application to plant systems is scarce. It has the potential to reveal the spatio-temporal dynamics of lipid biosynthesis during plant development. In this study, we explore its application to galactolipid biosynthesis of an aquatic plant, Lemna minor, with D2O labeling. Specifically, matrix-assisted laser desorption/ionization-MSI data of two major galactolipids in L. minor, monogalactosyldiacylglycerol and digalactosyldiacylglycerol, were studied after growing in 50% D2O media over a 15-day time period. When they were partially labeled after 5 d, three distinct binomial isotopologue distributions were observed corresponding to the labeling of partial structural moieties: galactose only, galactose and a fatty acyl chain and the entire molecule. The temporal change in the relative abundance of these distributions follows the expected linear pathway of galactolipid biosynthesis. Notably, their mass spectrometry images revealed the localization of each isotopologue group to the old parent frond, the intermediate tissues and the newly grown daughter fronds. Besides, two additional labeling experiments, (i) 13CO2 labeling and (ii) backward labeling of completely 50% D2O-labeled L. minor in H2O media, confirm the observations in forward labeling. Furthermore, these experiments unveiled hidden isotopologue distributions indicative of membrane lipid restructuring. This study suggests the potential of isotope labeling using MSI to provide spatio-temporal details in lipid biosynthesis in plant development.

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Fungicides play an important role in crop protection, but they have also been shown to adversely affect non-target organisms, including those living in the aquatic environment. The aim of the present study is to combine experimental and computational approaches to evaluate the effects of flutriafol, metconazole, myclobutanil, tebuconazole, tetraconazole and triticonazole on aquatic model organisms and to obtain information on the effects of these fungicides on Lemna minor, a freshwater plant, at the molecular level. The EC50 (the half-maximum effective concentration) values for the growth inhibition of Lemna minor in the presence of the investigated fungicides show that metconazole (EC50 = 0.132 mg/L) and tetraconazole (EC50 = 0.539 mg/L) are highly toxic, tebuconazole (EC50 = 1.552 mg/L), flutriafol (EC50 = 3.428 mg/L) and myclobutanil (EC50 = 9.134 mg/L) are moderately toxic, and triticonazole (EC50 = 11.631 mg/L) is slightly toxic to this plant. The results obtained with the computational tools TEST, ADMETLab2.0 and admetSAR2.0 also show that metconazole and tetraconazole are toxic to other aquatic organisms: Pimephales promelas, Daphnia magna and Tetrahymena pyriformis. A molecular docking study shows that triazole fungicides can affect photosynthesis in Lemna minor because they strongly bind to C43 (binding energies between -7.44 kcal/mol and -7.99 kcal/mol) and C47 proteins (binding energies between -7.44 kcal/mol and -8.28 kcal/mol) in the reaction center of photosystem II, inhibiting the binding of chlorophyll a to these enzymes. In addition, they can also inhibit glutathione S-transferase, an enzyme involved in the cellular detoxification of Lemna minor.

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Introduction: The exploitation of anthracite A in the Pejão mining complex (Douro Coalfield, North Portugal) resulted in the formation of several coal waste piles without proper environmental control. In 2017, a new pedological zonation emerged in the Fojo area, after the ignition and self-burning of some of the coal waste piles, namely: unburned coal waste (UW); burned coal waste, and a cover layer (BW and CL, respectively); uphill soil (US); mixed burned coal waste (MBW); downhill soil (DS). This study aimed to evaluate the toxic effects of 25 soil elutriates from different pedological materials. Methods: Allivibrio fischeri bioluminescence inhibition assay, Lemna minor growth inhibition assay, and Daphnia magna acute assay were used to assess the toxicity effects. Additionally, total chlorophyll and malondialdehyde (MDA) content and catalase (CAT) activity were also evaluated in L. minor. Results and Discussion: The results obtained from each endpoint demonstrated the extremely heterogeneous nature of soil properties, and the species showed different sensibilities to soil elutriates, however, in general, the species showed the same sensitivity trend (A. fischeri > L. minor > D. magna). The potentially toxic elements (PTE) present in the soil elutriates (e.g., Al, Pb, Cd, Ni, Zn) affected significantly the species understudy. All elutriates revealed toxicity for A. fischeri, while US1 and UW5 were the most toxic for L. minor (growth inhibition and significant alterations in CAT activity) and D. magna (100% mortality). This study highlights the importance of studying soil aqueous phase toxicity since the mobilization and percolation of bioavailable PTE can cause environmental impacts on aquatic ecosystems and biota.

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Lemnaceae taxonomy is challenged by the particular morphology of these tiny free-floating angiosperms. Although molecular taxonomy has helped clarify the phylogenetic history of this family, some inconsistency with morphological data leads to frequent misclassifications in the genus Lemna. Recently, the finding that Lemna japonica is an interspecific hybrid between Lemna minor and Lemna turionifera provided a clear explanation for one such taxonomic question. Here we demonstrated that L. minor is also capable of hybridizing with Lemna gibba, generating a cryptic but widespread taxon in the Mediterranean area. The nothotaxon Lemna ×mediterranea is described and compared with clones of the putative parental species L. minor and L. gibba. Genetic analysis by nuclear and plastid markers, as well as genome size measurement, revealed that two different cytotypes, diploid and triploid, originated by at least two independent hybridization events. Despite high overall similarity, morphometrical, physiological, and biochemical analyses showed an intermediate position of L. ×mediterranea between its parental species in most qualitative and quantitative characters, and also separation of the two hybrid cytotypes by some criteria. These data provide evidence that hybridization and polyploidization, driving forces of terrestrial plant evolution, contribute to duckweed genetic diversity and may have shaped the phylogenetic history of these mainly asexual, aquatic plants.

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Duckweeds are widely recognized for their efficiency in the phytoremediation of agricultural and industrial effluents. This study had two main objectives: 1) Implement a Nature-based Solutions (NBS) utilizing the environmental services of duckweeds to improve water quality through phytoremediation in small fish farms; 2) Analysis of duckweeds biomass produced in these fish farms to develop coproducts from a circular economy perspective in family agrisystem in Brazilian Atlantic Forest. The effectiveness of the phytoremediation system was assessed by the reduction of the Trophic State Index (TSI). Phytoremediation in small fish farming NBS was implemented using Clarias gariepinus, employing two different managements approaches: (i) System I - L. minor cultured every 15 days, with biomass harvest and effluent analysis conducted in each cycle over 60 days; (ii) System II - L. minor cultured every 30 days, following a similar cycle and analysis. Additionally, effluent from fish production underwent testing for phytoremediation in a batch system within a climate-controlled laboratory. L. minor demonstrated efficiency in System II, leading to a reduction of the TSI. The dry biomass of the plants emerged as a viable source of amino acid for application in functional foods and feed or nutraceuticals. The findings underscore the potential integration of L. minor into the NBS system and the generation of new co-products from circular production. In addition to its effective phyto- remediation properties, L. minor's dry biomass exhibited appealing characteristics, with elevated levels of crude protein, minerals, fatty acids, and carotenoids. This positions L. minor as a promising candidate for developing bioproducts tailored for functional foods and nutraceuticals. This underscores the potential of duckweeds to produce valuable nutritional compounds beyond their remediation capabilities.

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The uptake of sodium selenite (Se(IV)) and sodium selenate (Se(VI)) from aqueous medium by Lemna minor L. and the influence of different Se concentrations on its growth, morphological and ultrastructural characteristics were studied. L. minor was grown at different concentrations (1, 3, 5 and 10 mg L-1) of Se(IV) and Se(IV). The Se(IV) concentration in the plant tissue ranged between 77.7 (± 4.3) to 453 (± 0) mg kg-1 DW. The Se(VI) concentration in plant tissues ranged between 117 (± 11) to 417 (± 2) mg kg-1 DW. The highest bioconcentration factor for Se(VI) was 127 (± 7) at 3 mg/L, with a Se removal efficiency of 44%. For Se(IV), the highest bioconcentration factor was 77.7 (± 4.3) at 1 mg L-1, which had a Se removal efficiency of 23%. Growth of L. minor was suppressed at 10 mg L-1 Se in both forms. The addition of Se promoted the formation of starch granules in L. minor which occupied a chloroplast area of 74% for Se(IV) and 77% for Se(VI). The efficient uptake of both Se forms by L. minor indicates the potential application of this species for phytoremediation of Se laden wastewaters and its use as an alternative feedstock in biofuel production.

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This study investigated the effect of Pseudomonas stutzeri inoculation on Lemna minor treated with Cu(OH)2 nanopesticide (NP). The results showed that P. stutzeri inoculation increased the relative growth rate (RGR) in NP-treated plants. Although chlorophyll and carotenoid contents decreased significantly in NP-treated plants, P. stutzeri inoculation led to an increase in chlorophyll and carotenoid contents in NP-treated plants. Copper (Cu) content increased with increasing NP concentration, but it decreased significantly in the presence of P. stutzeri. NP treatment caused increased H2O2 and TBARS levels, as well as proline levels. However, P. stutzeri inoculation led to decreased H2O2 and TBARS levels and increased SOD, POX, GST, GR, GPX, and DHAR activities. The expression of genes encoding SOD, GST, metacaspase-1, and chlorophyllase was upregulated by NP treatment alone. Additionally, when plants were inoculated with P. stutzeri, the expression of these genes was further enhanced. In conclusion, P. stutzeri inoculation had a positive effect on the growth and antioxidant system of L. minor treated with NP as it enhanced RGR, increased chlorophyll and carotenoid contents, and decreased Cu content and oxidative stress. These findings suggested that P. stutzeri has the potential to promote aquatic plant growth and counteract the negative impacts of NP on these plants.

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The present research was carried out to investigate the performance of oxalic acid (OA) through Lemna minor L. (duckweed) in the phytoextraction of lead (Pb) from metal contaminated water. Zero, 100 µM, 250 µM, and 500 µM Pb concentration and combinations with 2.5 ml of OA were provided to the plants in the form of solution after defining intervals. Continuous aeration was provided to the plants and kept a pH level at 6.5. Results from this research depicted that increasing concentration level of Pb inhibited the overall plant growth, biomass, frond area, chlorophyll, and antioxidant enzyme activities like peroxidase (POD), superoxide-dismutase (SOD), catalases (CAT), and ascorbate-peroxidase (APX). Moreover, Pb stress enhances the concentration, hydrogen peroxide, malondialdehyde, and electrolyte leakage substances in plants. Furthermore, the addition of OA alleviated the Pb-induced toxicity in the plants, increasing the Pb accumulation and its endorsement in the L. minor. The OA addition increased the Pb accumulation in plants at 0, 100, 250 and 500 µM. At higher concentration, Pb showed harmful effect as related to the other low doses. Under the application of OA, Pb higher accumulation and concentration in L. minor were measured, which showed that it could be the most suitable plant for the phytoextraction of lead-contaminated soil and wastewater.

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The toxicity of two pesticides, diazinon (DAZ) and atrazine (ATR), before and after montmorillonite-catalyzed ozonation was comparatively investigated on the duckweed Lemna minor. The results allowed demonstrating the role of clay-containing media in the evolution in time of pesticide negative impact on L. minor plants. Pesticides conversion exceeded 94% after 30 min of ozonation in the presence of both Na+ and Fe2+ exchanged montmorillonites. Toxicity testing using L. minor permitted us to evaluate the change in pesticide ecotoxicity. The plant growth inhibition involved excessive oxidative stress depending on the pesticide concentration, molecular structure, and degradation degree. Pesticide adsorption and/or conversion by ozonation on clay surfaces significantly reduced the toxicity towards L. minor plants, more particularly in the presence of Fe(II)-exchanged montmorillonite. The results showed a strong correlation between the pesticide toxicity towards L. minor and the level of reactive oxygen species, which was found to depend on the catalytic activity of the clay minerals, pesticide exposure time to ozone, and formation of harmful derivatives. These findings open promising prospects for developing a method to monitor pesticide ecotoxicity according to clay-containing host-media and exposure time to ambient factors.

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Glitters are primary microplastics which are directly littered into the environment, yet the ecological effects have seldom been tested. When microplastics enter the environment, their physical presence and chemical leachate may alter the physiology of primary producers. Glitter can be composed of plastic or natural and/or biodegradable materials, often with additives. Three experiments were run for 14 days to separate chemical and physical effects of different types of glitter: polyethylene terephthalate (PET), biodegradable modified regenerated cellulose (MRC), synthetic mica, and a natural particle control (kaolinite) on several physical characteristics of Lemna minor (common duckweed). L. minor was exposed to either fresh (chemical and physical effects), leachate from glitter (chemical) or aged glitter (physical). Overall, there was little effect of PET, synthetic mica, kaolinite or of any aged glitter. High concentrations of fresh MRC glitters, however, decreased root length, biomass and chlorophyll content of L. minor. Some of these effects were also present when exposed to leachate from MRC glitters, but were less pronounced. Elemental analysis revealed the presence of metals in MRC glitters which may explain these responses. Short-term ecotoxicity of biodegradable glitters can arise due to their physical and chemical properties, but may lessen over time as their surface coating degrades.

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Lemnaceae are small freshwater plants with extraordinary high growth rates. We aimed to test whether this correlates with a more efficient photosynthesis, the primary energy source for growth. To this end, we compared photosynthesis properties of the duckweed Lemna minor and the terrestrial model plant Arabidopsis thaliana. Chlorophyll fluorescence analyses revealed high similarity in principle photosynthesis characteristics; however, Lemna exhibited a more effective light energy transfer into photochemistry and more stable photosynthesis parameters especially under high light intensities. Western immunoblot analyses of representative photosynthesis proteins suggested potential post-translational modifications in Lemna proteins that are possibly connected to this. Phospho-threonine phosphorylation patterns of thylakoid membrane proteins displayed a few differences between the two species. However, phosphorylation-dependent processes in Lemna such as photosystem II antenna association and the recovery from high-light-induced photoinhibition were not different from responses known from terrestrial plants. We thus hypothesize that molecular differences in Lemna photosynthesis proteins are associated with yet unidentified mechanisms that improve photosynthesis and growth efficiencies. We also developed a high-magnification video imaging approach for Lemna multiplication which is useful to assess the impact of external factors on Lemna photosynthesis and growth.

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Plants live in close association with microbial organisms that inhabit the environment in which they grow. Much recent work has aimed to characterize these plant-microbiome interactions, identifying those associations that increase growth. Although most work has focused on terrestrial plants, Lemna minor, a floating aquatic angiosperm, is increasingly used as a model in host-microbe interactions and many bacterial associations have been shown to play an important role in supporting plant fitness. However, the ubiquity and stability of these interactions as well as their dependence on specific abiotic environmental conditions remain unclear. Here, we assess the impact of a full L. minor microbiome on plant fitness and phenotype by assaying plants from eight natural sites, with and without their microbiomes, over a range of abiotic environmental conditions. We find that the microbiome systematically suppressed plant fitness, although the magnitude of this effect varied among plant genotypes and depended on the abiotic environment. Presence of the microbiome also resulted in phenotypic changes, with plants forming smaller colonies and producing smaller fronds and shorter roots. Differences in phenotype among plant genotypes were reduced when the microbiome was removed, as were genotype by environment interactions, suggesting that the microbiome plays a role in mediating the plant phenotypic response to the environment.

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Life cycle assessment (LCA) was applied to evaluate duckweed ponds and constructed wetlands as polishing steps in pig manure liquid fraction treatment. Using nitrification-denitrification (NDN) of the liquid fraction as the starting point, the LCA compared direct land application of the NDN effluent with different combinations of duckweed ponds, constructed wetlands and discharge into natural waterbodies. Duckweed ponds and constructed wetlands are viewed as a viable tertiary treatment option and potential remedy for nutrient imbalances in areas of intense livestock farming, such as in Belgium. As the effluent stays in the duckweed pond, settling and microbial degradation reduce the remaining phosphorous and nitrogen concentrations. Combined with duckweed and/or wetland plants that take up nutrients in their plant body, this approach can reduce over-fertilisation and prevent excessive nitrogen losses to aquatic environments. In addition, duckweed could serve as an alternative livestock feed and replace imports of protein destined for animal consumption. The environmental performance of the overall treatment systems studied was found to depend greatly on assumptions about the possible avoidance of potassium fertiliser production through the field application of effluents. If it is assumed that the potassium contained in the effluent replaces mineral fertiliser, direct field application of the NDN effluent performed best. If the application of NDN effluent does not lead to mineral fertiliser savings or if the replaced K fertiliser is of low grade, duckweed ponds seem to be a viable additional step in the manure treatment chain. Consequently, whenever background concentrations of N and/or P in fields allow for effluent application and potassium fertiliser substitution, direct application should be favoured over further treatment. If direct land application of the NDN effluent is not an option, the focus should be on long residence times in duckweed ponds to allow for maximum nutrient uptake and feed production.

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