Changes in Metabolism and Content of Chlorophyll in Common Duckweed (Lemna minor L.) Caused by Environmental Contamination with Fluorides.

The impact of fluorine on plants remains poorly understood. We examined duckweed growth in extracts of soil contaminated with fluorine leached from chicken manure. Additionally, fluorine levels were analyzed in fresh manure, outdoor-stored manure, and soil samples at varying distances from the manure pile. Fresh manure contained 37-48 mg F- × kg-1, while soil extracts contained 2.1 to 4.9 mg F- × kg-1. We evaluated the physiological effects of fluorine on duckweed cultured on soil extracts or in 50% Murashige-Skoog (MS) medium supplemented with fluorine concentrations matching those in soil samples (2.1 to 4.9 mg F- × L-1), as well as at 0, 4, and 210 mg × L-1. Duckweed exposed to fluorine displayed similar toxicity symptoms whether in soil extracts or supplemented medium. Fluoride at concentrations of 2.1 to 4.9 mg F- × L-1 reduced the intact chlorophyll content, binding the porphyrin ring at position 32 without affecting Mg2+. This reaction resulted in chlorophyll a absorption peak shifted towards shorter wavelengths and formation of a new band of the F--chlorophyll a complex at λ = 421 nm. Moreover, plants exposed to low concentrations of fluorine exhibited increased activities of aminolevulinic acid dehydratase and chlorophyllase, whereas the activities of both enzymes sharply declined when the fluoride concentration exceeded 4.9 mg × L-1. Consequently, fluorine damages chlorophyll a, disrupts the activity of chlorophyll-metabolizing enzymes, and diminishes the plant growth rate, even when the effects of these disruptions are too subtle to be discerned by the naked human eye.

Comparison of mercury (Hg) bioaccumulation with mono- and mixed Lemna minor and Spirodela polyrhiza cultures.

Mercury (Hg) is a prevalent and harmful contaminant that persists in the environment. For phytoremediation, it is important to discover which plants can bioaccumulate meaningful amounts of Hg while also tolerating its toxicity. Additionally, increasing biodiversity could create a more resilient and self-sustaining system for remediation. This study explores whether mixed populations of Lemna minor and Spirodela polyrhiza can better bioaccumulate and tolerate Hg than monocultures. Mono- and mixed cultures of L. minor and S. polyrhiza were grown in mesocosms of 0.5 µg/L or 100 µg/L Hg (HgCl2) spiked water for 96 h. Change in weight of duckweed was used to assess Hg tolerance. Diffusive gradients in thin-films (DGTs) were used as surrogate monitoring devices for bioavailable levels of Hg. For biomass growth, the mixed culture of the L. minor was greater than the monoculture at the high dose. The L. minor accumulated more Hg in the mixed culture at the low dose while the S. polyrhiza was higher in the mixed at the high dose. Hg speciation in water was modeled using Windermere Humic Aqueous Model 7 (WHAM7) to compare the bioavailable species indicated by the DGTs.  Potentially due to the controlled conditions, the WHAM7 output of bioavailable Hg was almost 1:1 to that estimated by the DGTs, indicating good predictive capability of geochemical modeling and passive sampler DGT on metal bioavailability. Overall, the mixed cultures statistically performed as well as or better than the monocultures when tolerating and bioaccumulating Hg. However, there needs to be further work to see if the significant differences translate into practical differences worth the extra resources to maintain multiple species.

miR397 regulates cadmium stress response by coordinating lignin polymerization in the root exodermis in Kandelia obovata.

Secondary lignification of the root exodermis of Kandelia obovata is crucial for its response to adversity such as high salinity and anaerobic environment, and this lignification is also effective in blocking cadmium transport to the roots. However, how the differences in lignification of root exodermis at different developmental stages respond to Cd stress and its regulatory mechanisms have not been revealed. In this study, after analyzing the root structure and cell wall thickness using a Phenom scanning electron microscope as well as measuring cadmium content in the root cell wall, we found that the exodermis of young and mature roots of K. obovata responded to Cd stress through the polymerization of different lignin monomers, forming two different mechanisms: chelation and blocking. Through small RNA sequencing, RLM-5'-RACE and dual luciferase transient expression system, we found that miR397 targets and regulates KoLAC4/17/7 expression. The expression of KoLAC4/17 promoted the accumulation of guaiacyl lignin during lignification and enhanced the binding of cadmium to the cell wall. Meanwhile, KoLAC7 expression promotes the accumulation of syringyl lignin during lignification, which enhances the obstruction of cadmium and improves the tolerance to cadmium. These findings enhance our understanding of the molecular mechanisms underlying the differential lignification of the root exodermis of K. obovata in response to cadmium stress, and provide scientific guidance for the conservation of mangrove forests under heavy metal pollution.

Cultivation of the macrophyte Lemna minor and the microalgae Chlorella sorokiniana in thermal mineral waters: Biomass characteristics, radioisotopes and heavy metals content.

Microalgae and macrophytes are commonly used as human and animal food supplements. We examined the cultivation of the microalgae Chlorella sorokiniana and the duckweed Lemna minor in thermal waters under batch and sequencing batch conditions and we characterized the produced biomass for the presence of essential nutrients as well as for heavy metals and radioisotope content. The highest specific growth rate for the microalgae was observed when 5 or 15 mg/L N were supplemented while the optimal conditions for Lemna minor were observed in the co-presence of 5 mg/L N and 1.7 mg/L P. Lemna minor presented higher concentrations of proteins and lipids comparing to the studied microalgae. Both organisms contained high amounts of lutein (up to 1378 mg/kg for Lemna minor) and chlorophyll (up to 1518 mg/kg for Lemna minor) while ß-carotene and tocopherols were found at lower concentrations, not exceeding a few tens of mg/kg. The heavy metal content varied between the two species. Lemna minor accumulated more Cd, Cu, K, Mn, Na, Ni, and Zn whereas Al, Ca and Mg were higher in Chlorella sorokiniana. Both organisms could be a significant source of essential metals but the occasional exceedance of the statutory levels of toxic metals in food products raises concern for potential risk to either humans or animals. Application of gamma-spectroscopy to quantify the effective dose to humans from 228Ra, 226Ra and 40K showed that Chlorella sorokiniana was well under the radiological limits while the collected mass of Lemna minor was too small for radiological measurements with confidence.

Responses of Vallisneria natans and Pistia stratiotes to Cu2+ and Mn2+ stress: Occurrence of caffeic acid and its degradation kinetics during chlorination.

Macrophytes are crucial in maintaining the equilibrium of aquatic ecosystems. However, the pattern of macrophyte-derived caffeic acid (CA) release under heavy metal stress is yet to be fully understood. More importantly, due to its functional groups, CA may be a precursor to the formation of disinfection by-products, posing threats to water ecology and even safety of human drinking water. This study analyzed the responses of CA released by Vallisneria natans (V. natans) and Pistia stratiotes (P. Stratiotes) when exposed to Cu2+ and Mn2+ stress. Additionally, the CA levels in two constructed wetland ponds were detected and the degradation kinetics of CA during chlorination were investigated. Results indicated that CA occurred in two constructed wetland ponds with the concentrations of 44.727 µg/L (planted with V. natans) and 61.607 µg/L (planted with P. Stratiotes). Notably, heavy metal stress could significantly affect CA release from V. natans and P. Stratiotes. In general, under Cu2+ stress, V. natans secreted far more CA than under Mn2+ stress, the level could reach up to 435.303 µg/L. However, compared to V. natans, P. Stratiotes was less affected by Cu2+ and Mn2+ stress, releasing a maximum CA content of 55.582 µg/L under 5 mg/L Mn2+ stress. Aquatic macrophytes secreted more CA in response to heavy metal stresses and protected macrophytes from harmful heavy metals. CA degradation followed the pseudo first-order kinetics model, and the chlorination of CA conformed to a second-order reaction. The reaction rate significantly accelerated as NaClO, pH, temperature and Br- concentration increased. A new pathway for CA degradation and a new DBP 2, 2, 3, 3-tetrachloropropanal were observed. These findings pointed at a new direction into the adverse effect of CA, potentially paving the way for new strategies to solve drinking water safety problems.

An experimental and prediction modeling study on water lettuce (Pistia stratiotes L.) assisted heavy metals removal from glass industry effluent.

The glass manufacturing industry produces hazardous effluent that is difficult to manage and causes numerous environmental problems when disposed of in the open. In this study, an attempt was made to study the phytoremediation feasibility of water lettuce (Pistia stratiotes L.), a free-floating aquatic macrophyte, for the removal of six heavy metals from glass industry effluent (GIE) at varying concentrations (0, 25, 50, 75, and 100%). After a 40-day experiment, the results showed that 25% GIE dilution showed maximum removal of heavy metals i.e., Cu (91.74%), Cr (95.29%), Fe (86.47%), Mn (92.95%), Pb (87.10%), and Zn (91.34%), respectively. The bioaccumulation, translocation, and Pearson correlation studies showed that the amount of heavy metals absorbed by vegetative parts of P. stratiotes was significantly correlated with concentrations. The highest biomass production, chlorophyll content, relative growth rate, and biomass productivity were also noted in the 25% GIE treatment. Moreover, the multiple linear regression models developed for the prediction of heavy metal uptake by P. stratiotes also showed good performance in determining the impact of GIE properties. The models showed a high coefficient of determination (R2 > 0.99), low mean average normalizing error (MANE = 0.01), and high model efficiency (ME > 0.99) supporting the robustness of the developed equations. This study outlined an efficient method for the biological treatment of GIE using P. stratiotes to reduce risks associated with its unsafe disposal.

Mechanisms of Spirodela polyrhiza tolerance to FGD wastewater-induced heavy-metal stress: Lipidomics, transcriptomics, and functional validation.

Unlike terrestrial angiosperm plants, the freshwater aquatic angiosperm duckweed (Spirodela polyrhiza) grows directly in water and has distinct responses to heavy-metal stress. Plantlets accumulate metabolites, including lipids and carbohydrates, under heavy-metal stress, but how they balance metabolite levels is unclear, and the gene networks that mediate heavy-metal stress responses remain unknown. Here, we show that heavy-metal stress induced by flue gas desulfurization (FGD) wastewater reduces chlorophyll contents, inhibits growth, reduces membrane lipid biosynthesis, and stimulates membrane lipid degradation in S. polyrhiza, leading to triacylglycerol and carbohydrate accumulation. In FGD wastewater-treated plantlets, the degraded products of monogalactosyldiacylglycerol, primarily polyunsaturated fatty acids (18:3), were incorporated into triacylglycerols. Genes involved in early fatty acid biosynthesis, ß-oxidation, and lipid degradation were upregulated while genes involved in cuticular wax biosynthesis were downregulated by treatment. The transcription factor gene WRINKLED3 (SpWRI3) was upregulated in FGD wastewater-treated plantlets, and its ectopic expression increased tolerance to FGD wastewater in transgenic Arabidopsis (Arabidopsis thaliana). Transgenic Arabidopsis plants showed enhanced glutathione and lower malondialdehyde contents under stress, suggesting that SpWRI3 functions in S. polyrhiza tolerance of FGD wastewater-induced heavy-metal stress. These results provide a basis for improving heavy metal-stress tolerance in plants for industrial applications.

Duckweed application in nature-based system for water phytoremediation and high-value coproducts at family agrisystem from a circular economy perspective.

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.

The evolution of the duckweed ionome mirrors losses in structural complexity.

BACKGROUND AND AIMS: The duckweeds (Lemnaceae) consist of 36 species exhibiting impressive phenotypic variation, including the progressive evolutionary loss of a fundamental plant organ, the root. Loss of roots and reduction of vascular tissues in recently derived taxa occur in concert with genome expansions of ≤14-fold. Given the paired loss of roots and reduction in structural complexity in derived taxa, we focus on the evolution of the ionome (whole-plant elemental contents) in the context of these fundamental changes in body plan. We expect that progressive vestigiality and eventual loss of roots might have both adaptive and maladaptive consequences that are hitherto unknown. METHODS: We quantified the ionomes of 34 accessions in 21 species across all duckweed genera, spanning 70 Myr in this rapidly cycling plant (doubling times are as rapid as ~24 h). We related both micro- and macroevolutionary ionome contrasts to body plan remodelling and showed nimble microevolutionary shifts in elemental accumulation and exclusion in novel accessions. KEY RESULTS: We observed a robust directional trend in calcium and magnesium levels, decreasing from the ancestral representative Spirodela genus towards the derived rootless Wolffia, with the latter also accumulating cadmium. We also identified abundant within-species variation and hyperaccumulators of specific elements, with this extensive variation at the fine (as opposed to broad) scale. CONCLUSIONS: These data underscore the impact of root loss and reveal the very fine scale of microevolutionary variation in hyperaccumulation and exclusion of a wide range of elements. Broadly, they might point to trade-offs not well recognized in ionomes.

Heavy metals/-metalloids (As) phytoremediation with Landoltia punctata and Lemna sp. (duckweeds): coupling with biorefinery prospects for sustainable phytotechnologies.

Heavy metals/-metalloids can result in serious human health hazards. Phytoremediation is green bioresource technology for the remediation of heavy metals and arsenic (As). However, there exists a knowledge gap and systematic information on duckweed-based metal phytoremediation in an eco-sustainable way. Therefore, the present review offers a critical discussion on the effective use of duckweeds (genera Landoltia and Lemna)-based phytoremediation to decontaminate metallic contaminants from wastewater. Phytoextraction and rhizofiltration were the major mechanism in 'duckweed bioreactors' that can be dependent on physico-chemical factors and plant-microbe interactions. The biotechnological advances such as gene manipulations can accelerate the duckweed-based phytoremediation process. High starch and protein contents of the metal-loaded duckweed biomass facilitate their use as feedstock in biorefinery. Biorefinery prospects such as bioenergy production, value-added products, and biofertilizers can augment the circular economy approach. Coupling duckweed-based phytoremediation with biorefinery can help achieve Sustainable Development Goals (SDGs) and human well-being.

Insights of phytoremediation mechanisms for viruses based on in-vitro, in-vivo and in-silico assessments of selected herbal plants.

Waterborne pathogenic viruses present unrelenting challenges to the global health and wastewater treatment industry. Phytoremediation offers promising solutions for wastewater treatment through plant-based technologies. This study investigated antiviral mechanisms in-vivo using bacteriophages MS2 and T4 as surrogates for effective herbs screened in-vitro from three embryophytes (Ocimum basilicum, Mentha sp., Plectranthus amboinicus), two macrophytes (Eichhornia crassipes, Pistia stratiotes) and a perennial grass (Cyperus rotundas). In-silico virtual screening predicted antiviral phytochemicals for further antiviral potency assessment. Results suggested in-vitro antiviral activities of embryophytes and macrophytes were higher (43-62%) than grass (21-26%). O. basilicum (OB, 57-62%) and P. stratiotes (PS, 59-60%) exhibited the highest antiviral activities. In-vivo tests showed notable virus reduction (>60%) in culture solution, attributed to rhizofiltration (66-74%) and phytoinactivation/phytodegradation (63-84%). In-silico analysis identified rutin as a primary antiviral phytochemical for MS2 (-9.7 kcal/mol) and T4 (-10.9 kcal/mol), correlating with dose-response inactivation (∼58-62%). In-vivo tests suggested additional phytocompounds may contribute to viral inactivation, presenting new opportunities for herb-based wastewater treatment solutions. Consequently, this study not only demonstrates the antiviral capabilities of OB and PS but also introduces an innovative approach for addressing viral contaminants in water.

Effects of dissolved organic matter, pH and nutrient on ciprofloxacin bioaccumulation and toxicity in duckweed.

Water pollution induced by antibiotics has garnered considerable concern, necessitating urgent and effective removal methods. This study focused on exploring ciprofloxacin (CIP) removal by duckweed and assessing CIP bioaccumulation and toxic effects within duckweed under varying dissolved organic matter categories, pH levels, and nutrient (nitrogen (N) and phosphorus (P)) levels. The results revealed the proficient and rapid elimination of CIP from water by duckweed, resulting in 86.17 % to 92.82 % removal efficiency at the end of the 7-day experiment. Across all exposure groups, varying degrees of CIP bioaccumulation in duckweed were evident, with uptake established as a primary pathway for CIP elimination within this plant. Additionally, five CIP metabolites were identified in duckweed tissues. Interestingly, the presence of humic acid (HA) and fulvic acid (FA) reduced CIP absorption by duckweed, with FA yielding a more pronounced impact. Optimal CIP removal was recorded at a pH of 7.5, while duckweed displayed heightened physiological stress induced by CIP at pH 8.5. Although the influence of N and P concentrations on CIP removal by duckweed was modest, excessive N and P levels intensified the physiological strain of CIP on duckweed.

The "Duckweed Dip": Aquatic Spirodela polyrhiza Plants Can Efficiently Uptake Dissolved, DNA-Wrapped Carbon Nanotubes from Their Environment for Transient Gene Expression.

Duckweeds (Lemnaceae) are aquatic nongrass monocots that are the smallest and fastest-growing flowering plants in the world. While having simplified morphologies, relatively small genomes, and many other ideal traits for emerging applications in plant biotechnology, duckweeds have been largely overlooked in this era of synthetic biology. Here, we report that Greater Duckweed (Spirodela polyrhiza), when simply incubated in a solution containing plasmid-wrapped carbon nanotubes (DNA-CNTs), can directly uptake the DNA-CNTs from their growth media with high efficiency and that transgenes encoded within the plasmids are expressed by the plants─without the usual need for large doses of nanomaterials or agrobacterium to be directly infiltrated into plant tissue. This process, called the "duckweed dip", represents a streamlined, "hands-off" tool for transgene delivery to a higher plant that we expect will enhance the throughput of duckweed engineering and help to realize duckweed's potential as a powerhouse for plant synthetic biology.

Characterization of the IAA-Producing and -Degrading Pseudomonas Strains Regulating Growth of the Common Duckweed (Lemna minor L.).

The rhizosphere represents a center of complex and dynamic interactions between plants and microbes, resulting in various positive effects on plant growth and development. However, less is known about the effects of indole-3-acetic acid (IAA) on aquatic plants. In this study, we report the characterization of four Pseudomonas strains isolated from the rhizosphere of the common duckweed (Lemna minor) with IAA-degradation and -utilization ability. Our results confirm previous reports on the negative effect of IAA on aquatic plants, contrary to the effect on terrestrial plants. P. putida A3-104/5 demonstrated particularly beneficial traits, as it exhibited not only IAA-degrading and -producing activity but also a positive effect on the doubling time of duckweeds in the presence of IAA, positive chemotaxis in the presence of IAA, increased tolerance to oxidative stress in the presence of IAA and increased biofilm formation related to IAA. Similarly, P. gessardii C31-106/3 significantly shortened the doubling time of duckweeds in the presence of IAA, while having a neutral effect in the absence of IAA. These traits are important in the context of plant-bacteria interactions and highlight the role of IAA as a common metabolite in these interactions, especially in aquatic environments where plants are facing unique challenges compared to their terrestrial counterparts. We conclude that IAA-degrading and -producing strains presented in this study might regulate IAA effects on aquatic plants and confer evolutionary benefits under adverse conditions (e.g., under oxidative stress, excess of IAA or nutrient scarcity).

Impact of pharmaceutical industry wastewater on stress physiological responses of Spirodela polyrhiza (L.) Schleiden.

Impact of pharmaceutical wastewater collected from aeration tank on aquatic macrophyte Spirodela polyrhiza (L.) Schleiden was studied in the present study. Various plant parameters such as chlorophyll, protein, and proline content of wastewater-exposed plants were examined to determine the toxic impacts of pharmaceutical wastewater. In addition to these parameters, electrolyte leakage (EL) and catalase (CAT) activities in S. polyrhiza were assessed for each day of exposure to the wastewater. S. polyrhiza was exposed to four different wastewater concentrations (25%, 50%, 75%, and 100%) under laboratory conditions for 7 days. The plants experienced severe toxicity as revealed by the reduction in photosynthetic pigments, increase in electrolyte leakage, and enhancement of antioxidant enzyme (CAT) activity in S. polyrhiza with an increase in concentration and time of exposure to pharmaceutical effluent.

Pharmaceutical-contaminated irrigation water: implications for ornamental plant production and phytoremediation using enrofloxacin-accumulating species.

Enrofloxacin (Enro) has been widely encountered in natural water sources, and that water is often used for irrigation in crop production systems. Due to its phytotoxicity and accumulation in plant tissues, the presence of Enro in water used for crop irrigation may represent economical and toxicological concerns. Here, we irrigated two ornamental plant species (Zantedeschia rehmannii Engl. and Spathiphyllum wallisii Regel.) with water artificially contaminated with the antimicrobial enrofloxacin (Enro; 0, 5, 10, 100, and 1000 µg L-1) to evaluate its effects on ornamental plant production, as well as its accumulation and distribution among different plant organs (roots, leaves, bulbs, and flower stems), and examined the economic and environmental safety of commercializing plants produced under conditions of pharmaceutical contamination. The presence of Enro in irrigation water was not found to disrupt plant growth (biomass) or flower production. Both species accumulated Enro, with its internal concentrations distributed as the following: roots > leaves > bulbs > flower stems. In addition to plant tolerance, the content of Enro in plant organs indicated that both Z. rehmannii and S. wallisii could be safety produced under Enro-contaminated conditions and would not significantly contribute to contaminant transfer. The high capacity of those plants to accumulate Enro in their tissues, associated with their tolerance to it, indicates them for use in Enro-phytoremediation programs.

Acetylcholine (ACh) enhances Cd tolerance through transporting ACh in vesicles and modifying Cd absorption in duckweed (Lemna turionifera 5511).

Acetylcholine (ACh), an important neurotransmitter, plays a role in resistance to abiotic stress. However, the role of ACh during cadmium (Cd) resistance in duckweed (Lemna turionifera 5511) remains uncharacterized. In this study, the changes of endogenous ACh in duckweed under Cd stress has been investigated. Also, how exogenous ACh affects duckweed's ability to withstand Cd stress was studied. The ACh sensor transgenic duckweed (ACh 3.0) showed the ACh signal response under Cd stress. And ACh was wrapped and released in vesicles. Cd stress promoted ACh content in duckweed. The gene expression analysis showed an improved fatty acid metabolism and choline transport. Moreover, exogenous ACh addition enhanced Cd tolerance and decreased Cd accumulation in duckweed. ACh supplement reduced the root abscission rate, alleviated leaf etiolation, and improved chlorophyll fluorescence parameters under Cd stress. A modified calcium (Ca2+) flux and improved Cd2+ absorption were present in conjunction with it. Thus, we speculate that ACh could improve Cd resistance by promoting the uptake and accumulation of Cd, as well as the response of the Ca2+ signaling pathway. Also, plant-derived extracellular vesicles (PDEVs) were extracted during Cd stress. Therefore, these results provide new insights into the response of ACh during Cd stress.

Transcriptomic and Functional Analyses of Two Cadmium Hyper-Enriched Duckweed Strains Reveal Putative Cadmium Tolerance Mechanisms.

Cadmium (Cd) is one of the most toxic metals in the environment and exerts deleterious effects on plant growth and production. Duckweed has been reported as a promising candidate for Cd phytoremediation. In this study, the growth, Cd enrichment, and antioxidant enzyme activity of duckweed were investigated. We found that both high-Cd-tolerance duckweed (HCD) and low-Cd-tolerance duckweed (LCD) strains exposed to Cd were hyper-enriched with Cd. To further explore the underlying molecular mechanisms, a genome-wide transcriptome analysis was performed. The results showed that the growth rate, chlorophyll content, and antioxidant enzyme activities of duckweed were significantly affected by Cd stress and differed between the two strains. In the genome-wide transcriptome analysis, the RNA-seq library generated 544,347,670 clean reads, and 1608 and 2045 differentially expressed genes were identified between HCD and LCD, respectively. The antioxidant system was significantly expressed during ribosomal biosynthesis in HCD but not in LCD. Fatty acid metabolism and ethanol production were significantly increased in LCD. Alpha-linolenic acid metabolism likely plays an important role in Cd detoxification in duckweed. These findings contribute to the understanding of Cd tolerance mechanisms in hyperaccumulator plants and lay the foundation for future phytoremediation studies.

Constructed wetlands and duckweed ponds as a treatment step in liquid manure handling - A life cycle assessment.

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.

Integrated transcriptomics and metabolomics reveal key metabolic pathway responses in Pistia stratiotes under Cd stress.

Cadmium (Cd) can interfere with plant gene expression, change the content of metabolites and affect plant growth. In this study, untargeted metabolomics (LC-MS) and RNA-Seq sequencing were performed on root tissues of Pistia stratiotes exposed to Cd stress. The results showed that cadmium stress affected the accumulation and transport of cadmium in plants and increased the content of soluble sugar, the activities of ascorbate peroxidase (APX), and peroxidase (POD) by 34.89%, 41.45%, and 6.71% on average, and decreased the activity of superoxide dismutase (SOD) by 51.51% on average. At the same time, the contents of carotenoid, chlorophyll a, and chlorophyll b decreased by 29.52%, 20.11%, and 13.14%, respectively, Thus affecting the growth and development of plants. Metabolomic analysis showed that Cd stress affected eight metabolic pathways, involving 27 differentially expressed metabolites, mainly including unsaturated fatty acids, amino acids (phenylalanine), nucleotides, sulfur compounds, and flavonoids. By transcriptome analysis, a total of 3107 differentially expressed genes (DEGs, 2666 up-regulated genes, and 441 down-regulated genes) were identified, which were mainly involved in four pathways, among which glutathione metabolism and lignin biosynthesis were the key metabolic pathways. In conclusion, this study reveals the metabolic and transcriptional response mechanisms of P. stratiotes to Cd stress through multi-omics, providing the theoretical basis for the phytoremediation of water contaminated by Cd.