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The microplastics in aquatic ecosystems pose a serious threat to ecological security and environmental health, which have received widespread attention. To reveal the response of a water- Vallisneria natans -sediment system to microplastics exposure, the V. natans was exposed to polyethylene microplastics （PE-MPs） with different mass fractions （1%-5%, sediment wet mass fraction）, and the effects of PE-MPs on the physiochemical indicators of water quality, morphological characteristics of submerged plants, physiological characters, antioxidant system, and microbial community structure in sediments were studied respectively. The results showed that the physiochemical properties of the water body were not significantly changed in the PE-MPs treatment group, whereas the plant height, oxidative stress index, and antioxidant system were significantly inhibited. For the plant height, the 1% PE-MPs treatment group height was only 47.44% of that in the control group. Chlorophyll a content was 81.04% of that in the control group, and the activities of catalase （CAT）, malondialdehyde （MDA）, and peroxidase （POD） increased by 233.70%, 117.82%, and 61.62%, respectively. Different mass fractions of PE-MPs had a certain impact on microbial community structure in sediments. The above results are helpful to improve the evaluation system of PE-MPs ecological risk in the water-submerged plant-sediment system.

Microplastic pollution and biological invasion, as two by-products of human civilization, interfere the ecological function of aquatic ecosystem. The restoration of aquatic vegetation has been considered a practical approach to offset the deterioration of aquatic ecosystem. However, a lack of knowledge still lies in the species selection in the revegetation when confronting the interference from microplastic pollution and exotic counterpart. The present study subjected the native submerged species, Hydrilla verticillata and its exotic confamilial, Elodea nuttallii to the current and future scenarios of polyamide microplastic pollution. The plant performance proxies including biomass and ramet number were measured. We found that the native H. verticillata maintained its performance while the exotic E. nuttallii showed decreases in biomass and ramet number under severest pollution conditions. The restoration of native submerged plant such as H. verticillata appeared to be more effective in stabilizing aquatic vegetation in the scenario of accelerating microplastic pollution. In order to explore the underlying driving mechanism of performance differentiation, stress tolerance indicators for plants, sediment enzymatic activity and sediment fungal microbiome were investigated. We found that polyamide microplastic had weak effects on stress tolerance indicators for plants, sediment enzymatic activity and sediment fungal diversity, reflecting the decoupling between these indicators and plant performance. However, the relative abundance of sediment arbuscular mycorrhizal fungi for H. verticillata significantly increased while E. nuttallii gathered "useless" ectomycorrhizal fungi at the presence of severest polyamide microplastic pollution. We speculate that the arbuscular mycorrhizal fungi assisted the stabilization of plant performance for H. verticillata with exposure to the severest polyamide microplastic pollution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Since the Industrial Revolution, increasing atmospheric CO 2 concentrations have had a substantial negative impact influence on coastal ecosystems because of direct effects including ocean acidification and indirect effects such as extreme rainfall events. Using a two-factor crossover indoor simulation experiment, this study examined the combined effects of acidification and hyposaline stress on Thalassia hemprichii. Seawater acidification increased the photosynthetic pigment content of T. hemprichii leaves and promoted seagrass growth rate. Hyposaline stress slowed down seagrass growth and had an impact on the osmotic potential and osmoregulatory substance content of seagrass leaves. Acidification and salinity reduction had significant interaction effects on the photosynthesis rate, photosynthetic pigment content, chlorophyll fluorescence parameters, and osmotic potential of T. hemprichii, but not on the growth rate. Overall, these findings have shown that the hyposaline stress inhibitory effect on the T. hemprichii physiological performance and growth may be reduced by acidification., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Per- and poly-fluoroalkyl substances (PFAS) are an emerging class of persistent organic pollutants that are widespread in aquatic ecosystems and pose a serious threat to aquatic organisms. It is thus crucial to explore the toxicity mechanisms of PFAS to submerged macrophytes and biofilms. In this study, Vallisneria natans (V. natans) was exposed to environmentally relevant concentrations of perfluorooctanoic acid (PFOA) and perfluorooctane sulphonate (PFOS). Results showed that PFAS induced the excessive production of reactive oxygen species, triggering antioxidant responses. V. natans exhibited an improved stress tolerance by altering the biosynthesis of several plant secondary metabolites and the histidine, arginine, proline pathways in response to PFAS exposure. Moreover, PIP1-1, PIP2-2, SLAH1 and SLAH2 genes were upregulated, indicating the activation of aquaporins and slow-type anion channels. The uptake of PFOA and PFOS by V. natans was 41.74 % and 52.31 %, respectively. Notably, PFAS bound to functional proteins (GSTF10), promoting the detoxification of plants. Exposure to PFAS also altered the structure of biofilms by inducing the synthesis of large amounts of polysaccharides and proteins. The diversity and richness of the microbial community within periphytic biofilms changed significantly. These results provide a comprehensive description of the responses of aquatic plants and periphytic biofilms to PFAS and the removal mechanism of PFAS, contributing to the environmental risk assessments and removal of PFAS in aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

High concentrations of copper can pollute coastal waters, primarily from agricultural runoff and mining activities, which can harm marine organisms, including seagrasses. The molecular mechanism of copper toxicity to seagrass currently remains unclear. To determine the response to copper, physiological and multi-omic analyses were conducted to explore the molecular mechanism by which copper affects the global threatened seagrass Halophila beccarii Asch. Excessive copper stress causes oxidative damage and stimulates the activity of the antioxidant enzyme system to remove excess reactive oxygen species (ROS), thereby reducing the damage caused by copper stress. Cu increases the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7), ascorbate peroxidase (EC 1.11.1.11), glutathione peroxidase (EC 1.11.1.9), ascorbate oxidase (EC 1.10.3.3), glutathione reductase (EC 1.6.4.2), and dehydroascorbate reductase (EC 1.8.5.1) and the content of malondialdehyde and reduces the activity of monodehydroascorbate reductase (EC 1.6.5.4). Under copper stress, H. beccarii upregulates the metabolic pathways of steroid biosynthesis and cutin, suberin, and wax biosynthesis, downregulates the metabolic pathways of arginine and proline metabolism and fructose and mannose metabolism; the levels of expression of the ribosome-related genes; upregulates the levels of expression of circadian rhythm-related proteins and downregulates the levels of glutathione metabolism and the proteins related to carbon fixation. This study provides new insights into the response of seagrass to copper stress and reports potential candidate metabolites, genes, and proteins that can be considered as biomarkers to improve the protection and management of seagrass meadows., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Controlling light qualities have been acknowledged as an effective method to enhance the efficiency of phytoremediation, as light has a significant impact on plant growth. This study examined the effects of light qualities on cadmium (Cd) tolerance in aquatic plant Egeria densa using a combination of biochemical and transcriptomic approaches. The study revealed that E. densa exhibits higher resistance to Cd toxicity under red light (R) compared to blue light (B), as evidenced by a significant decrease in photosynthetic inhibition and damage to organelle ultrastructure. After Cd exposure, there was a significantly reduced Cd accumulation and enhanced levels of both glutathione reductase (GR) activity and glutathione (GSH), along with an increase in jasmonic acid (JA) in R-grown E. densa compared to B. Transcriptional analysis revealed that R caused an up-regulation of Cd transporter genes such as ABCG (G-type ATP-binding cassette transporter), ABCC (C-type ATP-binding cassette transporter), and CAX2 (Cation/H + exchanger 2), while down-regulated the expression of HIPP26 (Heavy metal-associated isoprenylated plant protein 26), resulting in reduced Cd uptake and enhanced Cd exportation and sequestration into vacuoles. Moreover, the expression of genes involved in phytochromes and JA synthesis was up-regulated in Cd treated E. densa under R. In summary, the results suggest that R could limit Cd accumulation and improve antioxidant defense to mitigate Cd toxicity in E. densa, which might be attributed to the enhanced JA and phytochromes. This study provides a foundation for using light control methods with aquatic macrophytes to remediate heavy metal contamination in aquatic systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Perfluorinated compounds (PFCs) are persistent organic contaminants that are highly toxic to the environment and bioaccumulate, but their ecotoxic effects on aquatic plants remain unclear. In this study, the submerged plant Vallisneria natans was treated with short-term (7 days) and long-term (21 days) exposures to perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) at concentrations of 0, 0.01, 0.1, 1.0, 5.0, and 10 mg/L, respectively. The results showed that both high concentrations of PFOA and PFOS inhibited the growth of V. natans and triggered the increase in photosynthetic pigment content in leaves. The oxidative damage occurred mainly in leaves, but both leaves and roots gradually built up tolerance during the stress process without serious membrane damage. Both leaves and roots replied to short-term stress by activating superoxide dismutase (SOD), catalase (CAT) and polyphenol oxidase (PPO), while peroxidase (POD) was involved under high concentration stress with increasing exposure time. Leaves showed a dose-effect relationship in integrated biomarker response (IBR) values under short-term exposure, and the sensitivity of roots and leaves to PFOS was higher than that of PFOA. Our findings help to increase knowledge of the toxic effects of PFCs and have important reference value for risk assessment and environmental remediation of PFCs in the aquatic ecosystem., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The effects of cadmium (Cd) have been investigated in an aquatic plant Ottelia alismoides grown under low CO 2 . Under low CO 2 , no Cd treated O. alismoides operated three carbon dioxide-concentrating mechanisms (CCMs) efficiently, including HCO 3 - acquisition, C4 and CAM photosynthesis. After 4 days of treatment with 200 μM and 2000 μM Cd, O. alismoides exhibited an elevated Cd accumulation along with the increasing Cd concentration. Both Cd treatments induced appreciable phytotoxicities in O. alismoides. The leaves showed chlorosis symptoms and the anatomy as well as chloroplast ultrastructure were obviously damaged. Significant decreases in the content of pigments, chlorophyll fluorescence (Fv/Fm and Yield of PS II) and carbon isotope ratio (δ 13 C) were measured in leaf extracts of O. alismoides grown with both concentrations of Cd. In addition, the pH-drift technique showed that both Cd-treated O. alismoides plants could not uptake HCO 3 - . The maximum and minimum acidity in Cd-exposed O. alismoides were greatly decreased and the diurnal change of acidity was absent in both Cd treated plants. Furthermore, significant decreases in ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), pyruvate phosphate dikinase (PPDK) and phosphoenolpyruvate carboxylase (PEPC) activities were also found at Cd treated O. alismoides plants, indicating the disturbance within C4 cycle. The alterations in the functionality of CCMs in O. alismoides induced by Cd might be related with the inhibition of the enzymes such as carbonic anhydrase (CA) and PEPC involved in inorganic carbon fixation, and the destruction of chloroplasts, as well as the re-allocation of energy and nutrients involved in CCMs and Cd detoxification., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The presence of tetracycline is ubiquitous and has adverse effects on aquatic systems. A hydroponic experiment was conducted to investigate the ecological sensitivity of Hydrocharis dubia (Bl.) Backer and Trapa bispinosa Roxb. Exposed to different concentrations of tetracycline (0, 0.1, 1, 10, 30 and 50 mg/L) for one day (1D) and 14 days (14D). The results showed that after 1D of tetracycline exposure, the physiological indices of H. dubia had no remarkable change except for proline which was significantly stimulated under 0.1 mg/L tetracycline. For T. bispinosa, guaiacol peroxidase (POD), polyphenol oxidase (PPO) and ascorbate peroxidase (APX) activity and protein and proline content were notably promoted under different concentrations of tetracycline, but PPO activity was significantly decreased in 50 mg/L. After 14D, tetracycline caused no harm to the growth and protein content of H. dubia, but negatively influenced lipid peroxidation product and chlorophyll content in H. dubia under high tetracycline concentrations. Superoxide dismutase (SOD) and POD activity of H. dubia significantly increased at high tetracycline concentrations, while catalase (CAT) and PPO activity significantly decreased. APX activity in H. dubia increased with tetracycline concentrations at low tetracycline concentrations. For T. bispinosa, high concentrations of tetracycline application significantly inhibited its growth and the content of protein and chlorophyll. SOD, POD, CAT, and PPO activity of T. bispinosa were induced under different concentrations of tetracycline and no lipid peroxidation was observed. APX activity in T. bispinosa was significantly inhibited at high tetracycline concentrations. The results suggest that tetracycline can cause oxidative damage in H. dubia but harm the metabolism process of T. bispinosa without inducing oxidative damage. Overall, the sensitivity of T. bispinosa exposed to tetracycline exposure is higher than that of H. dubia., (Copyright © 2020 Elsevier Inc. All rights reserved.)

High nitrate (NO 3 - -N) concentration is a growing aquatic risk concern worldwide. However, adverse effects of high NO 3 - -N concentration on submerged macrophytes-epiphytic biofilms are unclear. In this study, the alterations in physiological changes, biofilms formation and chemical compositions were investigated on leaves of Vallisneria asiatica exposed to different NO 3 - -N concentrations. The findings showed that 10 mg L -1 NO 3 - -N resulted in low photosynthetic efficiency by inhibiting chlorophyll content 26.2 % and decreased intrinsic efficiency of photosystem II significantly at 14th day post treatment. Malondialdehyde, several antioxidant enzyme activities (i.e., superoxide dismutase, peroxidase and catalase), and secondary metabolites (i.e., phenolic compounds and anthocyanin) were all significantly up-regulated with 10 mg L -1 NO 3 - -N, implied oxidative stress were stimulated. However, no significant alterations in these indicators were observed with 5 mg L -1 NO 3 - -N. Compared to control, 10 mg L -1 NO 3 - -N concentration significantly stimulated microbes growth in biofilm and reduced the roughness of leaf-biofilms surface, but it had little effect on the biofilms distribution (from single clone to blocks) as revealed by scanning electron microscope and multifractal analysis. Results from X-ray photoelectron spectroscopy analysis showed that the percentage of P, Cl, K and the ratio of O1 (-O-) /O2 (C = O) were higher in leaves of control than treatments with 10 mg L -1 NO 3 - -N, indicating that 10 mg L -1 NO 3 - -N concentration exhibited significant inhibition of chemical activity and nutrient uptake of the leaf surfaces. Overall, these results demonstrated that high NO 3 - -N does stimulate the biofilm growth and can cause negative impacts on submerged macrophytes growth., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Although excess ammoniacal-nitrogen (NH 4 + -N) results in the disturbance of various important biochemical and physiological processes, a detailed study on the effects of NH 4 + -N stress on the photosynthesis and global changes in protein levels in submerged macrophytes is still lacking. Here, the changes of excess NH 4 + -N on physiological parameters in Hydrilla verticillata (L.f.) Royle, a submerged macrophyte were investigated, including the contents of photosynthetic pigments, soluble sugars, net photosynthesis and respiration, glutamine synthetase (GS) and glutamate synthase (GOGAT) activities, chloroplast ultrastructure, chloroplast reactive oxygen species (ROS) accumulation and protein levels. Our results showed that the net photosynthetic rate and pigment content reached maximum values when the plants were treated with 1 and 2 mg L -1 NH 4 + -N, respectively, and decreased at NH 4 + -N concentrations at 5, 10, 15 and 20 mg L -1 . This decrease might be caused by ROS accumulation. Compared that in 0.02 mg L -1 NH 4 + -N as a control, ROS generation in chloroplasts significantly increased in the presence of more than 2 mg L -1 NH 4 + -N. Consistently, the damages caused by over-accumulated ROS were observed in chloroplast ultrastructure, showing a loose thylakoid membranes and swollen grana/stroma lamellae. Furthermore, through proteomic analysis, we identified 91 differentially expressed protein spots. Among them, six proteins involved in photosynthesis decreased in abundance in response to excess NH 4 + -N. Surprisingly, the abundance of all the identified proteins that were involved in nitrogen assimilation and amino acid metabolism tended to increase under excess NH 4 + -N compared with the control, suggestive of the imbalanced carbon and nitrogen (C-N) metabolisms. In support, activated GS and GOGAT cycle was observed, evidenced by higher activities of GS and GOGAT enzymes. To our knowledge, this work is the first description that excess NH 4 + -N results in chloroplast ultrastructural damages and the first proteomic evidence to support that excess NH 4 + -N can lead to a decline in photosynthesis and imbalance of C-N metabolism in submerged macrophytes., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The presence of surfactants may affect the bioavailability of polycyclic aromatic hydrocarbons. A hydroponic experiment was conducted to investigate the response of Hydrocharis dubia (Bl.) Backer to different concentrations of linear alkylbenzene sulfonate (LAS), naphthalene (NAP) and their mixture (0.5, 5, 10, and 20 mg/L) for 14 days and 28 days. The results showed that LAS had a greater toxic effect on H. dubia growth than NAP at treatment concentrations of 0.5-20 mg/L. The combined effect of LAS and NAP was damaging to H. dubia at concentrations of LAS + NAP ≥5 + 5 mg/L. When LAS + NAP ≥10 + 10 mg/L, the underground parts of H. dubia suffered more significant damage than the aboveground parts. Under the treatments with LAS, NAP and their mixture, H. dubia experienced oxidative stress. Soluble proteins and antioxidant enzymes were the main substances protecting H. dubia from LAS stress, and superoxide dismutase (SOD) and peroxidase (POD) were the main protective enzymes. When exposed to NAP, H. dubia growth was stimulated and promoted at the same time. In the short-term treatment (14 d), catalase (CAT) activity was sensitive to NAP stimulation, and soluble proteins and SOD were the main protective substances produced. Soluble sugars, SOD and ascorbate peroxidase (APX) played important protective roles during the longer exposure time (28 d). The physiological response of H. dubia exposed to the combined toxicants was weaker than the response to exposure to individual toxicants. The responses of SOD and CAT activity were positive in the short term (14 d), and these were the main protective enzymes. As the exposure time increased (28 d), the plant antioxidant system responded negatively., Competing Interests: Declaration of competing interest All authors have no any potential sources of conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Heavy metals can cause a significant damage to submerged macrophytes and affect its periphyton biofilms in aquatic environments. This study investigated the effects of heavy metals such as copper (Cu), lead (Pb), cadmium (Cd) and their mixture on physiological and biochemical responses and ultrastructure characteristics of Vallisneria natans (V. natans). Furthermore, differences in structures of microbial communities were observed in biofilms. The results showed that Cu 2+ , Pb 2+ , Cd 2+ and their mixture could destroy cell structure and photosynthetic system, and directly caused oxidative damage to submerged macrophyte and induced antioxidant enzyme system. In general, biomass and total chlorophyll content of V. natans noticeably decreased, while the activities of superoxide dismutase, peroxidase and catalase were enhanced by heavy metal stress inducement in restricted range, and the malondialdehyde content increased with the aggravation of the damage. The single heavy metal stress played a negative impact, however, the combined stress was not always synergistic effects on plants. High-throughput sequencing analysis suggested that heavy metals changed the abundance and structure of the microbial biofilm community. Proteobacteria and Bacteroidete were the dominant bacteria under heavy metal stress and other species and abundance of bacteria such as Firmicute, Cyanobacteria, Chloroflexi, Actinobacteria, Verrucomicrobia, Acidobacteria, Deinococcus-Thermus, Chlamydiae were also present. These findings provided useful information for further understanding about submerged macrophytes and periphyton biofilms responsed to heavy metal stress in aquatic environments in the future., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A piscicide, rotenone (RT), is frequently used for clear and management of aquatic systems such as fish pond, and even for illegal fishing throughout the world. The effects of RT on submerged macrophytes remain elusive although the effects of RT on many kinds of animals are well documented. We wanted to determine the effects of RT on the growth and metabolism of three submerged plants (Vallisneria natans, Myriophyllum spicatum, Potamogeton maackianus) and try to find the reasons of these effects. The results showed that the shoot height, shoot dry weight, root dry weight, root:shoot ratios, contents of soluble protein and soluble carbohydrate of the three tested submerged plants were significantly negatively affected by RT and the effects were different among the studied species. Furthermore, pH rised a little and light transmission was greatly reduced in the water with RT treatment. We think that the negative effects of RT on the growth and metabolism of submerged species is partially attributing to the lower light caused by RT application. Accordingly, we highlight that submerged species may be greatly suppressed by RT, and we should apply RT in water ecosystems with great caution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The restoration of submerged plants is critical for the reconstruction of eutrophic lake ecosystems. The growth of submerged plants is influenced by many factors. For the first time in this study, the effects of silicate-mineral maifanite supplement on the growth, physiological and phytochemical process of Vallisneria spiralis (V. spiralis) were investigated by an outdoor PVC barrel experiment, to provide a technical reference for further applications in aquatic ecological restoration. The results show that the maifanite could significantly promote the growth of V. spiralis. Specifically, the biomass, height, number of leaves, leaf width, root length, and root activity of V. spiralis in the maifanite-supplemented group were better than those of the control (P < 0.05). Moreover, the modified maifanite group performed better than the raw maifanite group (P < 0.05). The photosynthetic pigment, root activity, and the malondialdehyde and peroxidase activity of the maifanite-treated V. spiralis were better than those of the control to some extent. It was found that maifanite contained abundant major and trace elements, which are required for the growth of V. spiralis. It is concluded that maifanite is beneficial to the growth of V. spiralis and can be further applied to the ecological restoration of eutrophic lakes., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Information on silver nanoparticle (AgNP) phytotoxicity on seagrasses is provided for the first time. Toxic effects of environmentally relevant AgNP concentrations on Halophila stipulacea were assessed to identify sensitive biomarkers, to determine threshold effect concentrations and to evaluate potential risks. Potential alterations in the cytoskeleton, endoplasmic reticulum, cell ultrastructure and viability, oxidative stress parameters and elongation in H. stipulacea leaves exposed to AgNP concentrations ranging from 0.0002 to 0.2 mg L -1 for 8 days were examined. The first signs of actin filament (AF) response in differentiating cells, exhibiting disorientation and slight bundling, were observed on the 4th day at 0.0002 mg L -1 , while at the end of the experiment and at the higher concentrations, AFs were extremely bundled. Endoplasmic reticulum was affected in meristematic and differentiating cells; massive aggregations and loss of the "grainy" structure were observed, initially on the 6th day at 0.002 mg L -1 . Effects on microtubules were detected on the last day at 0.2 mg L -1 . An increase in H 2 O 2 levels on the 4th and/or 6th day even at 0.0002 mg L -1 was followed by a decrease on, or up to the last day. On the 6th day at the lowest concentration, elevated malondialdehyde content, and superoxide dismutase and peroxidase activity were detected, indicating oxidative damage and antioxidant defense mechanism activation. Dead epidermal cells mainly occurred at 0.02 and 0.2 mg L -1 , while no dead vein cells were detected. A significant inhibition in leaf elongation was observed only at 0.2 mg L -1 . Therefore, AF disturbance in differentiating leaf cells, being a susceptible response parameter, could be regarded as an early warning indicator of risk posed by AgNPs to H. stipulacea meadows, while most of the remaining parameters examined also constitute useful biomarkers. The lowest observed effect concentration (0.0002 mg L -1 ), being within the range of environmentally relevant AgNPs concentrations, suggests the possibility of negative impacts of AgNPs on seagrass health. A risk quotient of 1.33 was calculated, indicating that AgNPs may pose a significant potential risk to the coastal environment. The data presented highlight the importance of future research to further investigate the seagrass-AgNP interactions, stress the need for a refinement of the environmental risk assessment of AgNPs and could be utilized for the design of biomonitoring programs for rational management of the coastal environment., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The symbiosis between Vallisneria denseserrulata and indigenous Bacillus sp. XZM was investigated for arsenic removal for the first time. It was found that the native bacterium was able to reduce arsenic toxicity to the plant by producing higher amount of extra cellular polymeric substances (EPS), indole-3-acetic acid (IAA) and siderosphore. Interestingly, V. denseserrulata-Bacillus sp. XZM partnership showed significantly higher arsenic uptake and removal efficiency. The shift in FT-IR spectra indicated the involvement of amide, carboxyl, hydroxyl and thiol groups in detoxification of arsenic, and the existence of an arsenic metabolizing process in V. denseserrulata leaves. The scanning electron microscopy (SEM) images further confirmed that the bacterium colonized on plant roots and facilitated arsenic uptake by plant under inoculation condition. In plant, most of the arsenic existed as As(III) (85%) and was massively (>77%) found in vacuole of particularly leaves cells. Thus, these findings are highly suggested for arsenic remediation in the constructed wetlands., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Aquatic plants play a vital role in maintaining the health and stability of ecosystems and in ecological restoration of contaminated water bodies. Herein, a 21-day-long laboratory-scale experiment was designed to explore the growth and physiological responses of Vallisneria natans (Lour.) Hara (V. natans) to the combined effects of manganese (Mn, 5, 20, and 80 mg L -1 ) and pH (pH 4.0, 5.5, and 7.0). Our results showed the combined toxicity intensity was closely related to Mn concentration and the toxicity exhibited by Mn gradually strengthened with the decrease of pH level. High concentration of Mn stress significantly reduced plants leaf area, final leaf number, photosynthetic pigment content, RGR (relative growth rate) and biomass accumulation, but significantly increased the contents of malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ). At the same time, V. natans plants can resist the adverse stress by activating the antioxidant defense system, including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and polyphenol oxidase (PPO) activities. Besides, V. natans tended to adjust the biomass allocation strategy and transferred more energy to the subsurface and the ramets and stolons parts under the combined stress. This experiment also showed that the increasing pH within a certain range could largely improve the removal rate of Mn (at highest by 84.28%). This may indicate the V. natans plant species can act as a promising tool for the Mn phytoremediation in aquatic environments which needs to be further explored by longer cycle field studies.

Ammonium pulse attributed to runoff of urban surface and agriculture following heavy rain is common in inland aquatic systems and can cause profoundly effects on the growth of macrophytes, especially when combined with low light. In this study, three patterns of NH4-N pulse (differing in magnitude and frequency) were applied to examine their effects on the growth of three submersed macrophytes, namely, Myriophyllum spicatum, Potamogeton maackianus, and Vallisneria natans, in terms of biomass, height, branch/ramet number, root length, leaf number, and total branch length under high and low light. Results showed that NH4-N pulse caused negative effects on the biomass of the submerged macrphytes even on the 13th day after releasing NH4-N pulse. The negative effects on M. spicatum were significantly greater than that on V. natans and P. maackianus. The effects of NH4-N pulse on specific species depended on the ammonium loading patterns. The negative effects of NH4-N pulse on P. maackianus were the strongest at high loading with low frequency, and on V. natans at moderate loading with moderate frequency. For M. spicatum, no significant differences were found among the three NH4-N pulse patterns. Low light availability did not significantly aggregate the negative effects of NH4-N pulse on the growth of the submersed macrophytes. Our study contributes to revealing the roles of NH4-N pulse on the growth of aquatic plants and its species specific effects on the dynamics of submerged macrophytes in lakes., Competing Interests: The authors have declared that no competing interests exist.

Metal-contaminated sediments in lakes is a global concern that poses toxicological risk to aquatic organisms. This study performed bioassays using the submerged macrophyte, Vallisneria natans (Lour.) Hara, exposed to contaminated sediments collected from five locations in Dianchi Lake, Yunnan, China. Among the sediments collected, I geo showed enrichment of As and Cd in Dianchi Lake sediments. In spite of enriched toxic metals at some locations, laboratory bioassays found no significant difference in leaf biomass or leaf photosynthesis rate between the sites. Root biomass and root activity showed significant differences between locations and were negatively correlated with the concentration of As, Cd, Hg, and Pb in sediment but not related to Cr. The above correlations were strongest for Hg and As, respectively. Accumulation of Cd and Pb to leaves of bioassay plants was observed, but this was not evident for As and Cr. Overall, the results indicate that V. natans can be used as a bioassay organism and measures of root toxicity are sensitive to metal concentrations present in Dianchi Lake sediments. Furthermore, the study species holds promise for use as a biomonitor of Cd and Pb sediment metal content.

The co-occurrence of aquatic contaminants, such as antibiotics and herbicides, has motivated investigations into their interactive effects on aquatic organisms. We examined the combined effects of environmental concentrations of the antibiotic Enrofloxacin (Enro; 0-2.25 μg l -1 ) and Roundup Original DI (Roundup ® ; 0-0.75 μg active ingredient l -1 ), a glyphosate based-herbicide, on Elodea canadensis. Enro alone was not toxic, but the plants were highly sensitive to Roundup ® whose toxicity is related to the induction of oxidative stress. The metabolism of Enro by plants into Ciprofloxacin (Cipro) was observed, and although former is not phytotoxic, oxidative events associated with Cipro generation were observed. The activity of cytochrome P450 was shown to be involved in Enro degradation in E. canadensis. As a cytochrome P450 inhibitor, Roundup ® decreases Enro metabolism in plants. Enro, in turn, increases glyphosate uptake and toxicity, so that Enro and Roundup ® have synergistic effects, disrupting the physiological processes of E. canadensis. Our results suggest E. canadensis as a potential candidate for the reclamation of Enro in contaminated waters, but not for Roundup ® due to its high sensitivity to that herbicide., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This study investigated the allelopathic effects of Microcystis aeruginosa (M. aeruginosa) extracts and exudates on the physiological responses, photosynthetic activity, and microbial structure of biofilms on leaves of Vallisneria natans (V. natans). By measuring physiological and photosynthetic indices, the results showed that M. aeruginosa allelochemicals inhibited photosynthesis, oxidative stress and antioxidant system stress response in the biofilms of V. natans leaves. Multifractal analysis found that the surface topography of V. natans leaves was altered due to the allelochemicals found in M. aeruginosa. Microbial diversity on the leaves was analyzed using high-throughput sequencing, and the results showed that M. aeruginosa exudates had a stronger effects on the microbial community structure of biofilms compared to extracts. These findings highlight how cyanobacterial allelochemicals induce negative effects on submerged macrophytes., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Plastic debris of all sizes has been detected in marine, terrestrial and freshwater habitats. Effects of plastic debris on macrophytes have hardly been studied, despite their importance in aquatic ecosystems. We provide the first experimental study exploring nano- and microplastic effects on the growth of sediment-rooted macrophytes. Myriophyllum spicatum and Elodea sp. were exposed to sediments amended with six doses of polystyrene (PS) nanoplastic (50-190 nm, up to 3% sediment dry weight) and PS microplastic (20-500 μm, up to 10% dry weight) under laboratory conditions. Both macrophyte species were tested for changes in root and shoot dry weight (DW), relative growth rate (RGR), shoot to root ratio (S:R), main shoot length and side shoot length. Microplastics did not produce consistent dose-effect relationships on the endpoints tested, except that main shoot length was reduced for M. spicatum with increasing microplastic concentration. Nanoplastic significantly reduced S:R for both macrophytes as a result of increased root biomass compared to shoot biomass. Nanoplastic also caused a decrease in M. spicatum main shoot length; however, shoot biomass was not affected. Elodea sp. side shoot length, root and shoot biomass and RGR were positively correlated to the nanoplastic concentration. All effects occurred at higher than environmentally realistic concentrations, suggesting no immediate implications for ecological risks. Our study did not aim for the elucidation of the exact mechanistic processes that cause the effects, however, particle size seems to play an important factor. CAPSULE: Nano- and microplastics affect growth of sediment-rooted macrophytes., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Submerged macrophytes are considered the main primary producers in shallow lakes. Recently, they have experienced a decline due to increasing environmental impacts, e.g., excessive heavy metal loads. Compared to extensive studies on vegetative growth, reports on effects of heavy metals on propagule germination and reproduction remain scarce. In this study, three experiments were conducted to investigate the effects of cadmium (Cd) on the propagule germination and sexual reproduction of submerged macrophytes. In Experiment I, six Cd concentrations were used (0, 0.05, 0.5, 1, 2.5, and 5 mg L -1 ), with seed germination found to be marginally affected by Cd treatment. In Experiment II, Cd exposure (5 d) at the six Cd concentrations was performed 15, 30, 60, 90, and 120 d prior to the designated germination date for turions/tubers. The Vallisneria spinulosa tubers did not germinate at ≥ 2.5 mg Cd L -1 when exposed to Cd 90 and 120 d prior to germination, whereas the Potamogeton crispus turions remained viable but with a low germination rate at ≥ 2.5 mg Cd L -1 . In Experiment III, with an increase from 0 to 0.5 Cd mg L -1 , the fruit weight of Ottelia alismoides and V. spinulosa decreased, whereas the fruit number increased for O. alismoides but not for V. spinulosa. Furthermore, the phenology of sexual reproduction for both species advanced under Cd exposure. In summary, Cd exposure affected the germination of asexual propagules and sexual reproduction of submerged macrophytes, with seeds found to be tolerant of Cd treatment up to 5 mg L -1 ., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In this study, the effects of excess NH 4 + -N on the subcellular accumulation of O 2 - and H 2 O 2 in submerged plant Hydrilla verticillata (L.f.) Royle were investigated using both histochemical and cytochemical methods. Treatments with ≥ 2.00 and ≥ 5.00 mg L -1 NH 4 + -N for 5 d significantly increased production of O 2 - and H 2 O 2 , respectively. The activities of plasma membrane-bound NADPH (nicotinamide adenine dinucleotide phosphate) oxidases and antioxidant enzymes (superoxide dismutase, peroxidase, ascorbate peroxidase, catalase, dehydroascorbate reductase and glutathione reductase) were also increased correspondingly. This study also provides the first cytochemical evidence of subcellular accumulation of O 2 - and H 2 O 2 in the submerged plants. In the leaves of H. verticillata treated with 20.0 mg L -1 NH 4 + -N, O 2 - dependent DAB precipitates were found primarily on the inner side of the plasma membrane, extracellular space and chloroplasts. H 2 O 2 -CeCl 3 precipitates were mainly localized on the inner side of the plasma membrane and extracellular space of the mesophyll cells. Treatments with the inhibitors of NADPH oxidase (diphenylene iodonium and imidazole) indicate that NH 4 + -N-induced production of O 2 - and H 2 O 2 in H. verticillata leaves may involve plasma membrane-bound NADPH oxidase. Moreover, low-light treatment decreased NH 4 + -induced O 2 - production, suggesting that alterations in the photosynthetic electron transfer chain due to NH 4 + toxicity could lead to O 2 - production., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Arsenic contamination of water is a global concern due to its heavy threat to human health. In this study, the submerged macrophyte Vallisneria natans (Lour.) Hara was used to remove environmentally relevant concentrations of arsenic in the binary As(III)/As(V) system. The concentrations of total arsenic (tAs) and As(III) in water dropped rapidly within 3 days, while As(V) first increased slightly within 3 days and then gradually decreased. About 1.2% dimethylarsinate (DMA) was detected at the 14th day of treatment. These findings indicated that As(III) could be oxidized to As(V) and methylated to DMA in water with V. natans. In relation to V. natans, both tAs and As(V) were much higher in roots compared to leaves. Arsenate was the predominant species (≥ 95.65 ± 0.10%) in roots, and As(III) was only found at the 14th day (3.45-6.96 mg kg -1 ). In leaves, As(III) significantly increased (P < 0.05) as the treatment duration increased. The proportions of As(V) (27.99-40.03%) were lower than those of As(III) and arsenobetaine (AsB) was detected (0.52-1.87 mg kg -1 ) after 7 d. The results of arsenic speciation demonstrated that the transformation of arsenic species in V. natans included As(V) reduction and As(III) methylation to AsB. There were a decrease in chlorophyll content, and an increase in MDA level and antioxidant enzymes (SOD, CAT, and POD) activities. The MDA level was much higher in leaves than roots, whereas the activities of SOD, CAT, and POD were the opposite, suggesting their possible role in arsenic resistance and detoxification. Our results indicate the potential of V. natans in phytoremediation of arsenic-contaminated water., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The herbicide diquat dibromide is used in North America to manage nuisance macrophytes. However, its effect on native macrophytes is less clear and it could cause indirect effects on other aquatic biota. This study determined the sensitivity of both native and non-native macrophytes grown in test systems with varying complexity to diquat dibromide applied directly to water following label directions. In an outdoor mesocosm experiment and single species greenhouse concentration-response tests, Elodea canadensis Michx., Myriophyllum spicatum L., Ceratophyllum demersum L. and Hydrocharis morsus-ranae L. were exposed to a range of diquat dibromide concentrations (4.7 - 1153 µg/L), corresponding to 0.4 - 100% of the recommended label rate of the formulated product. The mesocosm experiment contained all four plant taxa in the same system along with caged amphipods (Hyalella azteca Saus.), tadpoles (Lithobates pipiens Schreber), phytoplankton and periphyton; however, this study focuses on the macrophytes only. In both test systems, severe direct effects of diquat dibromide on macrophytes were detected, with almost 100% mortality of all macrophytes in both test systems at 74 µg/L. The most sensitive species in the single species tests, E. canadensis, showed almost 100% mortality at concentrations below the HPLC-based method detection limit of 5 µg/L. Effects occurred very rapidly and showed no difference in severity between native and non-native macrophytes or complexity of test systems. These results suggest that diquat dibromide could be applied at a considerably lower label rate, depending on the characteristics of the waterbody, while still achieving effective control of nuisance macrophytes., (Crown Copyright © 2018. Published by Elsevier Inc. All rights reserved.)

Ceratophyllum demersum L. and Hydrilla verticillata (L.f.) Royle, two pioneer, submerged plants, effectively remove heavy metals from contaminated water. The present work evaluates the bioaccumulation and defense mechanisms of these plants in the accumulation of lead from contaminated water during their optimal performance period. C. demersum and H. verticillata were investigated after 14 days of exposure to various lead concentrations (5⁻80 μM). The lead accumulation in both C. demersum and H. verticillata increased with an increasing lead concentration, reaching maximum values of 2462.7 and 1792 mg kg -1 dw, respectively, at 80 μM. The biomass and protein content decreased significantly in C. demersum when exposed to lead. The biomass of H. verticillata exposed to lead had no significant difference from that of the controls, and the protein content increased for the 5⁻10 μM exposure groups. The malondialdehyde (MDA) content and superoxide dismutase (SOD), peroxidase (POD), and polyphenol oxidase (PPO) activities were much higher in C. demersum , suggesting considerable damage from lipid peroxidation and sensitivity to lead stress. Enzyme inhibition and inactivation were also observed in C. demersum at high lead concentrations (40⁻80 μM). The excellent growth status, low damage from lipid peroxidation, and high activity of catalase (CAT) and phenylalanine ammonia-lyase (PAL) observed in H. verticillata illustrate its better tolerance under the same lead stress.

Aquatic plants in agricultural landscapes play a vital role in maintaining the ecological integrity within the aquatic systems while facing an array of disturbances. Among them, information on herbicide exposure on non-target aquatic plants is scarce. The present study was designed to fill this information gap by detecting the impacts of 2-methyl-4-chlorophenoxyacetic acid (MCPA) on Hydrilla verticillata using morpho-anatomical and physiological biomarkers and assessing the environmental risk of MCPA to the non-target environment. H. verticillata was exposed to different MCPA concentrations (10, 100, 500, 1000 μg/L) and control (0 μg/L) for 7 days. At the end of the experiment, plant growth, pigments, H 2 O 2 content, peroxidase activity (POD) and plant anatomy were compared. The environmental risk was assessed using predicted environmental concentration/predicted no effect concentration (PEC:PNEC) ratio, hazard quotient (HQ) and hazard index (HI). Control plants exhibited the highest growth, and a growth decline was noted in parallel to MCPA exposure, where a similar trend was detected for the plant pigment contents. MCPA induced chlorosis and oxidative stress in H. verticillata. Risk analysis detected high values for PEC:PNEC ratios (3-9), HQ (1.92-5.79) and HI (28.15). MCPA-exposed H. verticillata could recover once those plants received natural conditions. Overall, present findings showed the negative impacts of MCPA on non-target aquatic plant H. verticillata. These findings will be useful to clarify the interaction between agrochemicals and non-target aquatic plants. Such information would benefit to decide the criteria in aquatic ecosystem management.

Water contamination by nickel (Ni) has become an increasing concern in recent decades. Hydrilla verticillata (L.f.) Royle has been recognized as a promising accumulator of several potentially toxic elements (PTEs) in phytoremediation, but its Ni-accumulation characteristics and its mechanisms of tolerance to Ni remain largely unknown. This research investigated the biochemical responses of leaves and stems of H. verticillata to various concentrations of Ni (5, 10, 15, 20, and 40 μM) over periods of 7, 14, or 21 days. Plants accumulated considerable Ni to a maximum amount of 1080 mg kg -1 dry weight (DW) with a maximum bioconcentration factor of 1100; thus, high Ni accumulation was detected in H. verticillata. Low concentrations (5-15 μM) or short durations (less than 14 days) of Ni exposure might promote plant growth without adversely affecting normal metabolism. After peaking at day 14, a decline in bioaccumulation was unexpectedly observed as a long-term effect of Ni toxicity. Malondialdehyde content and the activities of defense-related enzymes changed in a similar pattern after treatment with Ni, increasing with both Ni concentration and exposure time to a peak (often at 5-15 μM on day 14), followed by a decline. Through a comprehensive analysis of all the test parameters, the tolerance thresholds were determined to be > 40.0 μM, 24.0 μM, and 15.8 μM at days 7, 14, and 21, respectively. Hydrilla verticillata could be a "high-potential accumulator" capable of decontaminating aquatic bodies polluted by Ni within the threshold range., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Present study deals with the effect of 24 h pre-incubation with exogenous anthocyanins (ANTH), extracted from red cabbage leaves, on key metabolic processes (photosynthesis and respiration) and pro-/antioxidant balance in the aquatic macrophyte Egeria densa (Planch.) Casp., Hydrocharitaceae family, treated with Cd and Mn (in sulfate form) at a concentration of 100 μmol. After five days of metal treatments, Cd was accumulated and the damage caused to metabolic processes was stronger than Mn. In Cd-treated leaves, the protein level, chlorophyll concentration and maximal photochemical efficiency of PS II decreased twofold, and net-photosynthesis was significantly inhibited, whereas lipid peroxidation and H 2 O 2 production increased. In turn, protective responses developed, including an increase in the total soluble thiols, alternative respiratory pathway capacity and the activity of superoxide dismutase and peroxidases. Pre-incubation in the ANTH-enriched extract caused an increase in foliar ANTH content, enhanced Cd and reduced Mn uptake into the tissue. A decrease in the level of oxidative reactions, an increase in the protein and chlorophyll concentration compared to the control values and a partial improvement of the photosynthetic parameters confirmed the ability of ANTH to reduce Cd-induced damage effects and to mitigate ROS-driven stress reactions. Stimulation of catalase and ascorbate peroxidase activity, an alternative respiration capacity and non-enzymatic antioxidant (carotenoids, ascorbate and proline) synthesis by ANTH were also revealed. These data suggest that ANTH-enriched extract from red cabbage leaves has a protective action against metal toxicity in Egeria plants., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Glyphosate, as a broad-spectrum herbicide, is frequently detected in water, and phosphorus widely enters the water due to the extensive use of phosphorus-containing substances in agriculture, industries and daily life. Thus, aquatic ecosystems are exposed to both glyphosate and phosphorus, which may affect aquatic organisms. In the present research, we studied the physiological responses of the floating aquatic plant species H. dubia to different concentrations of glyphosate (0, 1, 5, 15 mg/L) with different levels of phosphate (0, 50, 100 mg/L) after 14 days (d) of treatment. We explored glyphosate toxicity in H. dubia and investigated whether phosphate addition mitigates glyphosate toxicity in this species, which will provide a theoretical basis for the ecotoxicological study of aquatic plants. The results show that glyphosate significantly reduced the chlorophyll content, leaf number and root length of H. dubia, while it significantly increased the malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ), shikimate, proline, and soluble protein content and enzyme activities (superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and polyphenol oxidase (PPO)) in H. dubia. After phosphate supplement, the MDA, H 2 O 2 , proline, and soluble protein contents and enzyme activities in the plants treated with glyphosate decreased. These results indicate that the concentration of glyphosate investigated in our study can cause oxidative stress and affect the growth of H. dubia. Phosphate can alleviate glyphosate-induced oxidative stress in H. dubia., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Seagrasses play an important role in coastal marine ecosystems, but they have been increasingly threatened by human activities. In recent years, seagrass communities have rapidly degenerated in the coastal marine ecosystems of China. To identify the reasons for the decline in seagrasses, the phytotoxic effects of trace metals (Cu, Cd and Zn) on the seagrass Thalassia hemprichii were investigated, and the environmental contents of the metals were analyzed where the seagrass grows. The results showed that leaf necrosis in T. hemprichii exposed to 0.01-0.1 mg L -1 of Cu 2+ for 5 days was more serious than that in plants exposed to the same concentrations of Cd 2+ and Zn 2+ . The chlorophyll content in T. hemprichii declined in a concentration-dependent manner after 5 days of exposure to Cu 2+ , Cd 2+ and Zn 2+ . The evident reduction in ΔF/F m ' in T. hemprichii leaves was observed at day 1 of exposure to 0.01-1.0 mg L -1 of Cu 2+ and at day 3 of exposure to 0.1-1.0 mg L -1 of Cd 2+ . The antioxidant enzyme activities (SOD, POD and CAT) in T. hemprichii leaves exposed to the three metal ions also showed significant changes. In seawater from Xincun Bay (Hainan, China), where T. hemprichii grows, Cu had reached a concentration (i.e., 0.01 mg L -1 ) that could significantly reduce chlorophyll content and ΔF/F m ' in T. hemprichii leaves. Our results indicate that Cu influences the deterioration of seagrasses in Xincun Bay.

Accumulation and bioindication of trace elements were compared in three seagrasses growing in the Mediterranean Sea: Posidonia oceanica, Cymodocea nodosa and Halophila stipulacea. The levels of the elements As, Cd, Cr, Cu, Hg, Mn, Ni, Pb and Zn were investigated in water, sediments, and roots, rhizomes and leaves of seagrasses. Results showed that seagrasses can accumulate comparable levels of trace elements, but P. oceanica and C. nodosa showed higher mean values of element accumulation. Moreover, P. oceanica and C. nodosa may accumulate high element concentrations in their leaves, whereas in H. stipulacea restricted with the bulk of trace elements in roots and rhizomes. Seagrasses reflected to a different degree the levels of several trace elements in sediments, especially P. oceanica and C. nodosa, whose use as bioindicators is recommended. The future step for an effective use of seagrasses as bioindicators of marine pollution is to set up biomonitoring networks on a large scale., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Research into sediment-seagrass tissue metal relationships has been undertaken in Sydney estuary due to the recognized role contamination plays in threats to seagrass health. Seagrass (Halophila ovalis) leaf tissue concentrations are elevated in Cu, Pb and Zn and contain the highest reported root Cr concentrations. Seagrass metal concentrations were significantly different between species H. ovalis and Zostera capricorni; between root and leaf tissue; and between sampling locations. Greatest tissue enrichment was for Pb, however metals were not enriched in seagrass relative to surficial sediment. Fine and total sediment metal concentrations were temporally consistent between collection years 2013/15, whereas root tissue metals changed between years and sites and leaf metal contents were temporally inconsistent. Extractable metal concentrations in fine sediment (<62.5 μm) showed moderate significant correlation with root tissue and a weak significant relationship with leaf tissue, whereas total sediment metal showed no such relationships. Management implications are provided., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A hydroponic experiment was conducted to investigate the effects of oxalic acid (OA) on arsenic (As) uptake and the physiological responses of Hydrilla verticillata exposed to 3 mg L -1 of As in different forms. Plant As(III) uptake was significantly increased by 200-2000 µg L -1 OA. However, an increase of As(V) uptake was only shown with 1000 µg L -1 OA, and no significant difference was observed with dimethylarsinate treatment. Peroxidase and catalase activities, and the contents of photosynthetic pigments, soluble sugar and proline, were significantly increased by 1000 µg L -1 OA during As(III) treatment. Superoxide dismutase and proline were also increased significantly by 1000 µg L -1 OA when plants were exposed to As(V). In DMA treatment, proline was significantly increased by 500 µg L -1 OA. Therefore, As-induced oxidative stress is relieved by OA, but it depends on OA concentration and the form of As. Our results may be useful for the phytoremediation of waste water containing As and OA.

Glyphosate is a broad-spectrum herbicide that is frequently detected in water bodies and is harmful to aquatic systems. We conducted an experiment to explore the ecological sensitivity of Hydrilla verticillata (L.f.) Royle and Vallisneria natans (Lour.) Hara to glyphosate. Our research focused on the physiological responses of H. verticillata and V. natans after exposure to various concentrations of glyphosate (0, 1, 10, 20, 30, 40, 50 and 80 mg/L) in hydroponic culture after one day (1D) and seven days (7D). The results show that after 1D, the soluble protein content of H. verticillata was significantly stimulated under low herbicide concentrations. Other indices for H. verticillata and V. natans had no remarkable changes at 1D. After 7D of treatment, the soluble protein content of H. verticillata showed no significant differences, while the malondialdehyde (MDA), pigment contents and catalase (CAT) activity significantly increased at low glyphosate concentrations. Guaiacol peroxidase (POD) activity in H. verticillata significantly increased with increasing herbicide concentrations. The chlorophyll a/b ratio of H. verticillata sharply decreased above 10 mg/L. For V. natans, soluble protein, chlorophyll a, and carotenoid content; and CAT activity declined significantly after glyphosate application, while other indicators showed no significant changes. Our results indicate that glyphosate concentrations from 0 to 80 mg/L can induce oxidative stress in H. verticillate and may impede metabolism processes for protein and pigments without causing oxidative stress in V. natans. Taken together, our results suggest that the sensitivity of H. verticillata to glyphosate exposure is higher than that of V. natans., (Copyright © 2017. Published by Elsevier Ltd.)

The effects of two methylmercury (CH 3 Hg + , MeHg) concentrations, representative of environmental level and extreme contamination, were investigated on the macrophyte Elodea nuttallii during a 2h-exposure combining transcriptomic (RNA-Seq), physiological endpoints (pigment contents, activity of anti-oxidative stress enzymes) and bioaccumulation. Exposure to MeHg induced the up- and down-regulation of numerous genes (4389 and 16853 for 10ngL -1 and 10μgL -1 MeHg exposure, respectively) involved in sugar, amino acid and secondary metabolism (e.g. cinnamic acid, flavonoids) at both concentrations. Genes coding for photosynthesis, membrane integrity, metal homeostasis, water transport and anti-oxidative enzymes were additionally up- and down-regulated at the higher concentration. At the physiological level, exposure to both MeHg concentrations resulted in a strong increase of anthocyanin content in shoots. Chlorophyll content and antioxidant enzyme activities were unchanged. The data suggest that the macrophyte was able to efficiently cope with the stress resulting from MeHg exposure, possibly by using anthocyanin as anti-oxidant and S-rich amino acids (such as cysteine and methionine) as chelators. Transcriptomics analysis enabled gaining novel insights on molecular effects of MeHg in primary producers, which are one of the main entry pathway of hazardous MeHg in aquatic food webs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Because of the accumulation of heavy metals, Hydrilla verticillata (L.f.) Royle, a rooted submerged perennial aquatic herb, is being developed as a potential tool to clean the aquatic ecosystem polluted by heavy metals. However, its physiological responses for heavy metal remain to be elucidated. Here, through employing proteomics approach, we found that excess Cu significantly induced the expressions of four DNA methylation related proteins in H. verticillata, which were the homologues of two domains rearranged methyltransferases (DRM), a methyltransferases chromomethylase (CMT) and a histone H3 lysine-9 specific SUVH6-like (SUVH6). Consistently, a dramatic change in DNA methylation patterns was detected in excess Cu-exposed H. verticillata. Surprisingly, administration of the NADPH oxidase inhibitors, diphenylene iodonium (DPI) and imidazole (IMZ) that block production of reactive oxygen species (ROS) could trigger the remethylation of genomic sites that were demethylated by excess Cu, indicating that Cu-induced ROS might be another way to affect DNA methylation. Further analysis suggested this changed DNA methylation may be owing to the ROS-induced DNA damage. Taken together, our findings demonstrate that two different ways to influence DNA methylation in excess Cu-treated H. verticillata., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Aluminate is generally used as a flocculant in water and wastewater treatment processes, but the residual aluminum (Al) may have toxic effects on aquatic organisms when the concentration accumulates beyond a threshold level. The in situ and laboratory tests were conducted to evaluate the impact of residual Al on submerged macrophytes in West Lake, Hangzhou, China, which receives Al flocculant-purified water diverted from the Qiantang River. The responses of Vallisneria natans and Hydrilla verticillata were investigated based on their morphological and physiological parameters in pot culture and aquarium simulation experiments. In the pot culture experiments, the biomass, seedling number, plant height, stolon number, stolon length, and root weight were significantly higher at a site located 150m from the inlet compared with those at a site located 15m from the inlet (P < 0.05), thereby indicating that the residual Al significantly inhibited the morphological development of V. natans and H. verticillata. The variations in the chlorophyll-a, protein, and malondialdehyde contents of the two species in both the pot culture and aquarium simulation experiments also demonstrated that the two submerged macrophytes were stressed by residual Al. V. natans and H. verticillata accumulated 0.052-0.227mg of Al per gram of plant biomass (fresh weight, mg/g FW) and 0.045-0.205mg Al/g FW in the in situ experiments, respectively, where the amounts of Al were significantly higher in the plants in the treatment aquaria during the laboratory experiments than those in the controls. These results may have important implications for the restoration of submerged macrophytes and ecological risk assessments in Al-exposed lakes. It is recommended that the Al salt concentration used for the control of lake eutrophication should be reduced to an appropriate level., (Copyright © 2017 Elsevier Inc. All rights reserved.)

On November 5th, 2015 the worst environmental disaster in Brazil spilled 60 million m 3 of iron mining residue into Gualaxo do Norte River (Minas Gerais State), an affluent of the highest River Basin of the Brazilian Southeast (Doce River Basin), reaching the Atlantic Ocean. To assess the impact of the iron residue on the aquatic plant metabolism, we performed macrophyte growth experiments under controlled light and temperature conditions using two species (Egeria densa and Chara sp.). The plants' growth data were fitted in a kinetic model to obtain the biomass yields (K) and growth rates (μ). Turbidity and electrical conductivity of the water were measured over time. Both plants showed the highest growth rates in the contaminated condition (0.056 d -1 for E. densa and 0.45 d -1 for Chara sp.) and the biomass increased in the short-term (≈20 days). The control condition (i.e. no impacted water) supported the biomass increasing over time and the development of vegetative buddings with high daily rates (1.75 cm d -1 for E. densa and 0.13 cm d -1 for Chara sp). Turbidity showed a sharp decrease in 48 h and had no effects in the plants growth in the contaminated condition. The contamination affected the plants' yields in the long-term affecting the biomass development. This study provides preliminary information about the ecological consequences of a mining dam rupture aiming to collaborate with monitoring and risk assessments., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The spatio-temporal distribution of perchlorate in water sources around an ammonium perchlorate production unit and its toxicity response on a dominant aquatic plant, Hydrilla verticillata are reported in this study. Samples (n=453) from ground water (open well) and surface water sources within 5km from the production unit over a period of 12 months (2014, June - 2015, May) were screened for ClO 4 - . During the period, ClO 4 - concentration in ground water samples close to the production unit increased to >40,000μg/L, and ClO 4 - was detected at 1740μg/L in well water 1.6km away from the production unit. A community pond in the area also showed an increase in ClO 4 - level up to 29,000μg/L. In all water sources, ClO 4 - level was maximum during the rainy season (July, monsoon). A natural degradation of ClO 4 - was not observed in the area as evident from its persistent level and spreading to more areas. H. verticillata, the dominant vegetation in the pond exhibited severe toxic response like massive decay and loss of photosynthetic pigments such as chlorophyll and carotene due to ClO 4 - exposure. The plant accumulated ClO 4 - up to 60 ± 0.8mg/kg wet weight with a Bio-Concentration Factor 2.06±0.005. This is the first report on spatio-temporal distribution of ClO 4 - at higher levels in a natural environment and its toxicity response to plants under natural condition., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The present study focused on oxidative stress effects in the aquatic macrophyte Hydrilla verticillata after exposure to titanium dioxide nanoparticles (TiO 2 -NPs). Experiments were conducted with different TiO 2 -NPs and concentrations (0.1 mg/L and 10 mg/L) in a time-dependent manner (0 h, 24 h, 48 h, 96 h, 168 h). To assess various levels of the oxidative stress response in H. verticillata, the level of hydrogen peroxide (H 2 O 2 ), the ratio of reduced to oxidized glutathione (GSH/GSSG), and activities of the antioxidative enzymes catalase (CAT) and glutathione reductase (GR) were evaluated. Study results imply oxidative stress effects after TiO 2 -NP exposure as adaptations in plant metabolism became apparent to counteract increased ROS formation. All TiO 2 -NPs caused elevated activities of the enzymes CAT and GR. Moreover, decreased ratios of GSH/GSSG indicated an activation of GSH-dependent pathways counteracting ROS formation. Plants exposed to a bulk-sized control revealed a size-dependent influence on the antioxidative stress response. As H 2 O 2 level increases were solely detected after exposure to 10 mg/L TiO 2 -NPs and nano-exposed plants showed normalization in its antioxidative stress response after 168h of exposure, it can be suggested that macrophytes are able to cope with currently predicted low-level exposures to TiO 2 -NPs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Pulse inputs of heavy metals are expected to increase with a higher frequency of extreme climate events (heavy rain), leading to stronger erosion of contaminated and fertilized farmland soils to freshwaters, with potentially adverse effects on lake ecosystems. We conducted a 5-month mesocosm study to elucidate the responses of the submerged macrophyte Vallisneria spinulosa and phytoplankton to four different doses of cadmium (Cd): 0 (control), 0.05, 0.5, and 5 g m -2 (CK, I, II, and III, respectively) under mesotrophic conditions. We found that total phosphorus concentrations were larger in the three Cd pulse treatments, whereas total nitrogen concentrations did not differ among the four treatments. The contents of chlorophyll a and soluble sugar in macrophyte leaves decreased in III, and total biomass, ramet number, plant height, and total stolon length of macrophytes were lower in both II and III. In contrast, abundances of the three main phytoplankton taxa-Cyanophyta, Chlorophyta, and Bacillariophyta-did not differ among treatments. Total phytoplankton biomass was, however, marginally lower in CK than in the Cd treatments. We conclude that exposure to strong Cd pulses led to significantly reduced growth of macrophytes, while no obvious effect appeared for phytoplankton.

Microcystin-LR (MCLR) and copper are commonly found in eutrophic water bodies because of eutrophic run-offs, cyanobacterial blooms, and copper algicide applications. However, the ecotoxicological risk of their combination remains unknown. This study investigated the effect of MCLR, Cu, and their mixture on the growth and physiological responses of Vallisneria natans. Results showed that the combined toxicity of them was concentration dependent. Synergistic effects were elicited at low concentrations of MCLR and Cu exposure (≤0.25+0.64mg/L). Additive or antagonistic effects were induced at higher concentrations. Single and combined exposures could induce oxidative stress, such as increased superoxide anion radical levels. To cope with oxidative stress, V. natans could activate their antioxidant defense systems, such as enhanced superoxide dismutase production and changes in peroxidase activities. Exposure to combined MCLR and Cu (even only with 0.005+0.041mg/L) adversely affected their antioxidant defense systems. As a consequence, malondialdehyde levels significantly increased. The interaction of MCLR and Cu could also significantly increase the bioaccumulations of MCLR and Cu. This increase could be accounted for their synergistic toxic effects on V. natans. Our results suggested that the exacerbated ecological hazard of MCLR and Cu with environmental concentrations may harm aquatic ecosystems., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The present study was aimed to analyze the effects of external Zn supply on arsenic (As) toxicity in Hydrilla verticillata (L.f.) Royle. The plants were exposed to arsenite (AsIII; 10 μM) with or without 50 and 100 μM Zn. The level of As accumulation (μg g -1 dw) after 2 and 4 days was not significantly affected by Zn supply. The plants showed a significant stimulation of the thiol metabolism (nonprotein thiols, cysteine, glutathione-S-transferase activity) upon As(III) exposure in the presence of Zn as compared to As(III) alone treatment. Besides, they did not experience significant toxicity, measured in terms of hydrogen peroxide and malondialdehyde accumulation, which are the indicators of oxidative stress. The minus Zn plants suffered from oxidative stress probably due to insufficient increase in thiols to counteract the stress. Stress amelioration by Zn supply was also evident from antioxidant enzyme activities, which came close to control levels with increasing Zn supply as compared to the increase observed in As(III) alone treatment. Variable Zn supply also modulated the level of photosynthetic pigments and restored them to control levels. In conclusion, an improved supply of Zn to plants was found to augment their ability to withstand As toxicity through enhanced thiol metabolism.

Interest in stream mesocosms has recently revived for higher tier aquatic macrophyte risk assessment of plant protection products mainly because 1) the highest predicted environmental concentrations for the assessment of effects are frequently derived from stream scenarios, and 2) they allow an effect assessment using stream-typical pulse exposures. Therefore, the present stream mesocosm study used an herbicide pulse exposure and evaluated the responses of Elodea canadensis and Myriophyllum spicatum. Macrophytes were exposed for 24 h to 1 μg/L, 3 μg/L, 10 μg/L, and 30 μg/L of the herbicide iofensulfuron-sodium with a subsequent recovery period of 42 d. Biological endpoints were growth rates of the main, side, and total shoot length, the shoot number, the maximum root length, and the dry weight. The total shoot length was identified as the most sensitive endpoint; the growth rate of the total shoot length was inhibited by up to 66% and 45% in M. spicatum and E. canadensis, respectively. The lowest no observed effect concentrations (NOECs) were observed at day 7 and/or day 14 after herbicide treatment and were 1 μg/L for M. spicatum and 3 μg/L for E. canadensis. The no-observed-ecologically-adverse-effect concentrations (NOEAECs) were 10 μg/L and 30 μg/L for M. spicatum and E. canadensis, respectively. Such or similar mesocosm designs are useful to simulate typical stream exposures and estimate herbicide effects on aquatic macrophytes in stream systems. Environ Toxicol Chem 2017;36:1090-1100. © 2016 SETAC., (© 2016 SETAC.)

Changes of diffusive permeability of membranes of Elodea nuttallii cells following a short-term (60 min) treatment with high concentrations of lanthanides were recorded by the 1 H NMR-diffusometry and conductometry methods. The 1-h infiltration of segments of Elodea nuttallii internodes in 10 mM solutions of nitrates of La, Nd and Lu resulted in the increased leakage of electrolytes from cells, but has no effect on a water diffusive permeability of membranes. In samples subjected to a 30 min pretreatment with a water channel inhibitor HgCl 2 the water diffusive permeability of membranes (P d ) drops down under the influence of lanthanides, as well as an outcome of electrolytes. To explain the observed effects the change of spontaneous curvature of membrane lipid layer has been taken into consideration. The interaction of lanthanides with lipids of plasmalemma leads to the negative spontaneous curvature of lipid layer at which membrane channels are unclosed. Blocking of the ionic and water channels by mercury ions compensate the effect of change of spontaneous curvature of lipid layer., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The hypothesis that increased UV radiation result in co-tolerance to Hg toxicity in aquatic plants was studied at the physiological and transcriptomic level in Elodea nuttallii. At the transcriptomic level, combined exposure to UV+Hg enhanced the stress response in comparison with single treatments, affecting the expression level of transcripts involved in energy metabolism, lipid metabolism, nutrition, and redox homeostasis. Single and combined UV and Hg treatments dysregulated different genes but with similar functions, suggesting a fine regulation of the plant to stresses triggered by Hg, UV and their combination but lack of co-tolerance. At the physiological level, UV+Hg treatment reduced chlorophyll content and depleted antioxidative compounds such as anthocyanin and GSH/GSSG in E. nuttallii. Nonetheless, combined exposure to UV+Hg resulted in about 30% reduction of Hg accumulation into shoots vs exposure to Hg alone, which was congruent with the level of expression of several transporter genes, as well as the UV effect on Hg bioavailability in water. The findings of the present work underlined the importance of performing experimentation under environmentally realistic conditions and to consider the interplay between contaminants and environmental variables such as light that might have confounding effects to better understand and anticipate the effects of multiple stressors in aquatic environment., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The present study investigated the effects of titanium dioxide nanoparticles (TiO 2 -NPs) on the oxidative stress response in Hydrilla verticillata. Macrophytes were exposed to different concentrations of TiO 2 -NPs (0 mg/L, 0.01 mg/L, 0.1 mg/L, 1 mg/L, and 10 mg/L) for 24 h, based on currently predicted levels of nano-TiO 2 in surface waters. In addition, TiO 2 -NPs with varying crystalline status were used to assess the potential influence of crystalline phases on oxidative stress responses. The level of hydrogen peroxide (H 2 O 2 ), reduced and oxidized glutathione (GSH and GSSG), and activities of the antioxidative enzymes peroxidase (POD), catalase (CAT), and glutathione reductase (GR) were measured and compared with a bulk counterpart. Although POD was not considered to be active, the results imply an activation of the enzymatic defense system, because increased CAT and GR activities were observed. Exposure to bulk TiO 2 revealed lower enzyme activities at all exposure concentrations, suggesting a nano-specific influence on the antioxidative defense mechanisms in H. verticillata. Moreover, all TiO 2 -NP concentrations resulted in a decreased GSH/GSSG ratio, indicating high GSH-dependent metabolic activity to protect against the destructive effects of reactive oxygen species (ROS) generated during nano-TiO 2 exposure. As the level of H 2 O 2 was solely elevated after exposure to 10 mg/L of P25, it appears plausible that the adaptive metabolic mechanisms of H. verticillata are able to cope with environmentally relevant concentrations of TiO 2 -NPs. Environ Toxicol Chem 2016;35:2859-2866. © 2016 SETAC., (© 2016 SETAC.)