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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.)

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.)

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.)

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.)

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.)

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.

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.

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.)

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.)

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.)

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.)

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.)

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.)

Aquatic macrophytes can absorb heavy metals either from sediments via the root system, from the water phase by leaves, or from both sources. In this study, cadmium accumulation and distribution in the aquatic plant Hydrilla verticillata were investigated, with a focus on the role of roots. Results showed that leaves of H. verticillata had a higher Cd concentration than roots when intact plants were grown in sediments and solutions containing Cd. Cadmium can significantly decrease the leaf chlorophyll content, and the leaves of intact plants with roots had lower chlorophyll contents than the leaves of detached ones without roots due to the transfer effect of roots. The majority of the Cd accumulated in leaves of H. verticillata was bound to the cell walls. When roots were submerged in a solution containing Cd, with shoots in a control solution without Cd, the Cd concentrations in leaves were considerably lower than in roots. In contrast, Cd was almost undetectable in roots when the shoots were submerged in a solution containing Cd, with roots in the control solution. Compared to the leaves and stems of detached shoots without roots, the concentrations of Cd were much higher in the leaves and stems of intact plants with roots. It is suggested that the roots of intact plants absorb Cd and transfer it to leaves and that more Cd is removed from the solution by intact plants.

Submersed freshwater macrophytes are considered as relevant indicators for use in bulk bottom sediment contact tests. The purpose of this study was to estimate the validity of endpoints of aquatic plant Elodea canadensis for laboratory genotoxicity testing of natural bottom sediments. The inherent level of chromosome abnormalities (on artificial sediments) in roots of E. canadensis under laboratory conditions was lower than the percentage of abnormal cells in bulk sediments from the Yenisei River. The percentage of abnormal cells in roots of E. canadensis was more sensitive to the presence of genotoxic agents in laboratory contact tests than in the natural population of the plant. The spectra of chromosomal abnormalities that occur in roots of E. canadensis under natural conditions in the Yenisei River and in laboratory contact tests on the bulk bottom sediments from the Yenisei River were similar. Hence, chromosome abnormalities in roots of E. canadensis can be used as a relevant and sensitive genotoxicity endpoint in bottom sediment-contact tests., (Copyright © 2015 Elsevier Inc. All rights reserved.)

In this study, the uptake and sub-toxic effects of CuO nanoparticles (CuO-NPs), dissolved Cu(II) alone or in combination with UV radiation on the aquatic macrophyte Elodea nuttallii were studied. Emphasis was on Cu accumulation, growth, photosynthesis and the oxidative stress related enzymes peroxidase (POD) and superoxide dismutase (SOD). The results showed stronger Cu accumulation in plants exposed to 10 mg L(-1) CuO-NPs, corresponding to 1.4-2 mg L(-1) dissolved Cu(II), than to 256 μg L(-1) Cu(II). However, the ratio between the accumulated Cu and dissolved Cu in CuO treatments was lower than in Cu(II) treatments. Additional UV exposure increased accumulation in both treatments, with the effect being stronger for Cu accumulation from CuO-NPs than for dissolved Cu(II). Photosynthetic capacity was strongly reduced by UV treatment, whereas remained unaffected by Cu(II) or CuO-NP treatments. Similarly, the increase of SOD activity was more pronounced in the UV treatments. On the other hand, POD activity enhancement was strongest in the plants exposed to CuO-NPs for 24 h. Expression of the copper transporter COPT1 as revealed by RT-qPCR was inhibited by Cu(II) and CuO-NP treatment, limiting the uptake of excess Cu into the cells. Overall, the combined exposure of E. nuttallii to UV radiation with CuO-NPs or Cu(II) has a higher impact than exposure to CuO-NPs or Cu(II) alone. The results imply that heavy pollution of natural water with CuO-NPs or dissolved Cu might have stronger effects in combination with natural UV irradiation on organisms in situ., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

We investigated the phytoremediation potential of Elodea nuttallii to remove rare earth metals from contaminated water. The laboratory experiments were designed to assess the responses induced by lanthanum (5-20mgL(-1)) in E. nuttallii over a period of 7 days. The results showed that most La (approximately 85%) was associated with the cell wall. The addition of La to the culture medium reduced the concentration of K, Ca, Cu, Mg, and Mn. However, O2(·-) levels increased with a concomitant increase in the malondialdehyde (MDA) concentration as the La concentration increased, which indicated that the cells were under oxidative stress. Significant reductions in the levels of chlorophyll (Chl) a, b, and carotenoids (Car) were observed in a concentration-dependent manner. However, the levels of reduced glutathione (GSH), total non-protein thiols (TNP-SH) and phytochelatins (PCs) increased for all La concentrations. The results suggested that La was toxic to E. nuttallii because it induced oxidative stress and disturbed mineral uptake. However, E. nuttallii was able to combat La induced damage via an immobilization mechanism, which involved the cell wall and the activation of non-enzymatic antioxidant., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Arsenic pollution of groundwater is a serious problem in many regions of Latin America that causes severe risks to human health. As a consequence, non-destructive monitoring methodologies, sensitive to arsenic presence in the environment and able to perform a rapid screening of large polluted areas, are highly sought-after. Both chlorophyll - a fluorescence and reflectance of aquatic plants may be potential indicators to sense toxicity in water media. In this work, the effects of arsenic on the optical and photophysical properties of leaves of different aquatic plants (Vallisneria gigantea, Azolla filiculoides and Lemna minor) were evaluated. Reflectance spectra were recorded for the plant leaves from 300 to 2400 nm. The spectral distribution of the fluorescence was also studied and corrected for light re-absorption processes. Photosynthetic parameters (Fv/Fm and ΦPSII) were additionally calculated from the variable chlorophyll fluorescence recorded with a pulse amplitude modulated fluorometer. Fluorescence and reflectance properties for V. gigantea and A. filiculoides were sensitive to arsenic presence in contrast to the behaviour of L. minor. Observed changes in fluorescence spectra could be interpreted in terms of preferential damage in photosystem II. The quantum efficiency of photosystem II for the first two species was also affected, decreasing upon arsenic treatment. As a result of this research, V. gigantea and A. filiculoides were proposed as bioindicators of arsenic occurrence in aquatic media., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Seagrass is particularly susceptible to environmental degradation. The objective of the study is to develop an effective bioindicator to assess human-induced effects using morphological variation and fluctuating asymmetry (FA) of seagrass. Samples were collected from eight islands situated at different distance from mainland with different human population density and therefore expected to experience different level of anthropogenic pressure. Cd, Pb, Cu, Zn, nitrate, and phosphate were measured. Metals were also measured in tissues of seagrass. Metal concentrations in sediment, water, and seagrass did not exceed the quality standards required for marine life. Heterogeneity of FA was found among sites suggesting that there are some factors changing developmental instability of seagrass which is not associated to particular toxicants. This baseline study indicates that the water condition is still natural and shows no signs of metal contamination, therefore it does not cause a detectable stress on morphological variation and FA of seagrass., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A growing body of evidence suggests that aquatic plants can mitigate the toxicity of insecticides to sensitive aquatic animals. The current paradigm is that this ability is driven primarily by insecticide sorption to plant tissues, especially for hydrophobic compounds. However, recent work shows that submerged plants can strongly mitigate the toxicity of the relatively hydrophilic insecticide malathion, despite the fact that this compound exhibits a slow sorption rate to plants. To examine this disparity, we tested the hypothesis that the mitigating effect of submerged plants on malathion's toxicity is driven primarily by the increased water pH from plant photosynthesis causing the hydrolysis of malathion, rather than by sorption. To do this, we compared zooplankton (Daphnia magna) survival across five environmentally relevant malathion concentrations (0, 1, 4, 6, or 36 μg L(-1)) in test containers where we chemically manipulated water pH in the absence of plants or added the submerged plant (Elodea canadensis) but manipulated plant photosynthetic activity via shading or no shading. We discovered that malathion was equally lethal to Daphnia at all concentrations tested when photosynthetically inactive (i.e. shaded) plants were present (pH at time of dosing=7.8) or when pH was chemically decreased (pH=7.7). In contrast, when photosynthetically active (i.e. unshaded) plants were present (pH=9.8) or when pH was chemically increased (pH=9.5), the effects of 4 and 6 μg L(-1) of malathion on Daphnia were mitigated strongly and to an equal degree. These results demonstrate that the mitigating effect of submerged plants on malathion's toxicity can be explained entirely by a mechanism of photosynthesizing plants causing an increase in water pH, resulting in rapid malathion hydrolysis. Our findings suggest that current ecotoxicological models and phytoremediation strategies may be overlooking a critical mechanism for mitigating pesticides., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Laboratory experiments were conducted to investigate copper (Cu) subcellular distribution and toxicity in Hydrilla verticillata. Fronds were subjected to different concentrations (15, 75, and 150 μM) of Cu for 7 days. Cu grains were found in cell walls, plasmodesmata, and within the nuclei and chloroplasts using the autometallographic technique. Subcellular fractionation of Cu-containing tissues indicated that in leaves subjected to high Cu concentrations, 59-65 % of the element was located in the cell wall fraction, followed by cell organelles (21-30 %) and the soluble fraction (10-14 %). The levels of K, P, Zn, and Mg declined under all Cu concentrations, but Ca, Mn, and Fe contents reached their peak at 15 μM Cu and decreased thereafter. F v/F m, F 0, and F m fell significantly in line with the decrease in pigment content. Cu exposure also caused significant damage to the chloroplasts, mitochondria, and nuclei, including disintegration of the chloroplasts and vacuolization of the mitochondria and nuclei, all of which suggested that Cu hastened plant senescence. The Cu maximum permissible concentration for H. verticillata was 10 μM, which was less than the existing general water quality standard. This suggested that H. verticillata could be used to assess Cu phytotoxicity.

Heterogeneity of contaminant-stress can be an important environmental factor for clonal plants. We focused on Cu transport among the clones, the foraging or fugitive behavior and biomass allocation of submerged plant, Vallisneria natans (Lour.) Hara, exposed to heterogeneous sediments. This study was carried out in aquatic mesocosms between March and September 2010. Cu accumulated in contaminated ramets was exported horizontally via stolons to other ramets in uncontaminated patches, and then transported both acropetally to leaves and basipetally to belowground structures. There was no indication that V. natans adopted morphological plasticity in response to heterogeneous contaminated habitat. In contrast to predictions, more biomass was allocated to belowground tissues in contaminated patches. We concluded that risk of Cu stress spread among submerged clones, and V. natans did not actively select habitat in contaminated patchy environment. Furthermore, V. natans adopted compensatory investments instead of division of labor to acquire nutrient and survive., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Increasing amounts of lanthanum (La) is released into aquatic environments. However, little information is available on the influence of La on aquatic plants. In this study, physiological and ultrastructural responses of Hydrocharis dubia (Bl.) Backer leaves to elevated concentrations of La (up to 160 μM) were investigated. The accumulation of La was found to be increased in a concentration-dependent manner. La disturbed the intrinsic balance of nutrient elements (P, Mg, Ca, Fe, K, and Zn). Pigment content decreased with the rise of the La concentrations and the EC(50) value for chlorophyll was 20 μM on day 7. The antioxidants (superoxide dismutase, peroxidase, catalase, reduced ascorbate, and reduced glutathione) exhibited varied response to the La treatments. Malondialdehyde content enhanced gradually at all La concentrations. The enhancement in proline content was found in a concentration-dependent manner. The amounts of three polypeptides with apparent molecular weights of 61.9, 51.5, and 16.7 kDa, respectively, were gradually diminished, as well as one existing polypeptides with apparent molecular weight of 22.3 kDa, elevating in response to increasing La concentrations. Significant damage to the chloroplast, mitochondrion, and nucleus was imposed by La indicated a general disarray in the cellular functions. The negative effects of La on H. dubia unequivocally indicate that La could exert an adverse influence on aquatic ecosystem and should lead to a more careful discharge of such elements into water environment.

We studied the antioxidant responses of macrophyte Vallisnerria natans seedlings to combined ammonia (0, 0.21, and 0.85 mg L(-1)) and microcystin-LR (MC-LR) (0, 10, and 50 μg L(-1)) for 7 days. Results showed that superoxide dismutase, catalase (CAT), peroxidase, and glutathione were significantly induced by the mixtures of ammonia and MC-LR, and there were significant interactions between ammonia and MC-LR. Specially, CAT increased about fivefold at ammonia 0.85 mg L(-1) and MC-LR 50 μg L(-1) on day 3. Malondialdehyde fluctuated with both ammonia and MC-LR, and significant interactions were detected between the two stressors. Changes in all the measured variables were time-dependent.

A mesocosm study with three photosystem-II inhibitors and an equipotent mixture was performed to address the value of functional and structural endpoints in evaluating the impact of herbicides on aquatic systems. The herbicides atrazine, diuron, and isoproturon were dosed in the ratio of their relative potencies as HC30 for the single substance treatments and as 1/3 HC30 for the mixture treatment to obtain comparable effect concentrations. To investigate the effects of the three herbicides and their mixture on photosynthesis of the whole system, the physical-chemical parameters pH, dissolved oxygen, and conductivity were monitored. To address effects on photosynthesis more specifically, the photosynthetic efficiency of phytoplankton and three submersed macrophytes (Elodea canadensis, Myriophyllum spicatum, and Potamogeton lucens) were investigated applying in vivo chlorophyll fluorescence as an indicator for their activity. As a structural endpoint, the species abundance and community structure of the phytoplankton community was determined. Effects were continuously monitored over a five week period of constant exposure, and during a 3 month post-exposure period. The sensitivity, expressed as maximum effect during constant exposure, was higher for the structural parameters (total and single species abundances and PRC) than for the functional parameters. The mean coefficient of variation (CV) for the physical-chemical parameters was below 10%, for the photosynthesis measurement of the phytoplankton and macrophytes below 10 and 30%, respectively. Structural parameters, however, yielded higher variability with mean CVs for phytoplankton abundance data and single sensitive species reaching up to 96%. Effects on the phytoplankton photosynthesis measured via in vivo chlorophyll fluorescence were constant during the exposure period; whereas macrophytes recovered quickly from photosynthesis inhibition despite constant exposure. Effects on total system photosynthesis, determined via physical-chemical parameters, lasted for a shorter period than for the phytoplankton photosynthesis demonstrating the importance of the macrophytes for total primary production. Thus, the evaluation of effects on communities in model ecosystems such as micro- and mesocosms should not be based on structural endpoints only due to their comparably high inherent variability. Instead, we recommend complementing the risk assessment with data obtained from sensitive functional endpoints addressing the specific mode of action of the respective compound for the most sensitive group of organisms to avoid over-estimation of the recovery potential of the aquatic system., (Copyright © 2011 Elsevier Inc. All rights reserved.)

A comprehensive understanding of the uptake, tolerance, and transport of heavy metals by plants will be essential for the development of phytoremediation technologies. In the present paper, we investigated accumulation, tissue and intracellular localization, and toxic effects of cadmium (Cd), lead (Pb), zinc (Zn), and copper (Cu) in three aquatic macrophytes (the angiosperms Lemna minor and Elodea canadensis, and the moss Leptodictyum riparium). We also tested and compared their capacity to absorb heavy metal from water under laboratory conditions. Our data showed that all the three species examined could be considered good bioaccumulators for the heavy metals tested. L. riparium was the most resistant species and the most effective in accumulating Cu, Zn, and Pb, whereas L. minor was the most effective in accumulating Cd. Cd was the most toxic metal, followed by Pb, Cu, and Zn. At the ultrastructural level, sublethal concentrations of the heavy metals tested caused induced cell plasmolysis and alterations of the chloroplast arrangement. Heavy metal removal experiments revealed that the three macrophytes showed excellent performance in removing the selected metals from the solutions in which they are maintained, thus suggesting that they could be considered good candidates for wastewaters remediation purpose.

A study was conducted to determine the response of the rooted submersed macrophyte, Vallisneria spiralis to phenanthrene in freshwater sediments with initial phenanthrene concentrations from 0 to 80 mg kg(-1) dry sediment. The sensitivity of various morphological endpoints was evaluated after 90 days of exposure. The most sensitive toxicity test endpoints were those that reflected root growth. Toxicological sensitivity of the endpoints changed with the effect level selected. The toxicity threshold from a plot of the EC(10) values was 1-2 orders of magnitude lower than those calculated for the threshold from plots of the EC(25) or EC(50) values. In addition, stimulatory responses (hormesis) on root growth were observed at subtoxic concentrations of phenanthrene, and a hormetic model should thus be incorporated for ecological risk assessment.

For better understanding the metabolic adaptations to Pb stress in submerged plants, the alterations in mineral elements uptake and in secondary metabolism were studied in leaves of Vallisneria natans (Lour.) Hara exposed to 0-100μM Pb for 0-7d. Pb content increased in leaves in a dose-dependent way. The increase of calcium, magnesium and iron content and the decrease of phosphorus, potassium and manganese content were detected in leaves of V. natans under Pb stress, while no significant changes were detected in copper and zinc concentration. Meanwhile, there was an increase in the concentrations of total phenolic and flavonoids. Pb treatment caused an increase in the catalytic activities of shikimate dehydrogenase, phenylalanine ammonialyase and polyphenol oxidase. The results suggest that nutrient uptake and secondary metabolism were actively regulated by V. natans plants in response to Pb stress., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The following study was carried out to investigate the cadmium (Cd) accumulating potential of Vallisneria. After subjecting plants to different concentrations of Cd, it was observed that plants are able to accumulate ample amount of metal in their roots (5,542 microg g(-1) dw) and leaves (4,368 microg g(-1) dw) in a concentration- and duration-dependent manner. Thus, it is evident that the accumulation in roots was 1.3 times higher than the shoots. It was also noted that with increasing Cd accumulation, roots of the plant appeared darker in color and harder in texture. In response to metal exposure, amount of low molecular weight antioxidants such as cysteine and nonprotein thiols (NP-SH) and activity of enzymes such as APX and GPX were significantly enhanced at lower concentrations of Cd, followed by decline at higher doses. It was also observed that in exposed plants, activity of APX enzyme was higher in roots (ca. 3 times) as compared to leaves. However, chlorophyll and protein content was found to decline significantly in a dose-dependent manner. Results suggested that due to its high accumulation potential, Vallisneria may be effectively grown in water bodies moderately contaminated with Cd.

Accelerated eutrophication is common to many freshwater and marine environments and often co-occurs with the presence of anthropogenic chemicals. However, the toxic effects of common chemical stressors such as herbicides in the presence of elevated nutrients are not well understood for most aquatic flora, particularly vascular species. To provide insight, field-collected Vallisneria americana Michx. (wild celery) were sequentially exposed to three nutrient concentrations for 3 months and then to nominal 11 and 110 microg L(-1) atrazine for 96 h. Nutrient concentrations (combined NH(4)(+), NO(2)(-), NO(3)(-), PO(4)(-)) were based on ambient concentrations in the St. Johns River (FL) and ranged from 0.013 to 0.668 mg L(-1). Nutrient pretreatment potentiated the toxicity of atrazine as determined by chlorophyll fluorescence activity. Electron transport rates (ETR) were significantly less (48-59%) for plants pretreated with low and ambient nutrient levels in the presence of an average of 107.5-128.1 microg L(-1) atrazine. Significant ETR reductions were also observed for plants exposed to an average of 11.4 microg L(-1) atrazine after exposure to nutrients three times the ambient concentration in the St. Johns River. The results indicate the importance of considering the presence of nutrients in chemical hazard assessments, particularly for phytotoxicants and nontarget vascular plants.

To investigate ammonium toxicity, the submerged plant Hydrilla verticillata (L.f.) Royle was treated with 0.1-3.0mM ammonium for 12h and 4d. After exposure to ammonium for 4d, content of O2(-) and H2O2 increased in leaves of H. verticillata exposed to 3mM ammonium compared with control (0mM NH4Cl), while the malondialdehyde content decreased. The chlorophyll (a+b) and carotenoid concentrations decreased in H. verticillata plants exposed to 1.5-3mM ammonium for 12h and 4d. Compared with controls, the activity of superoxide dismutase, peroxidase, catalase, glutathione reductase, ascorbate peroxidase, and glutathione S-transferase increased in plants treated with ammonium for 12h, and the activity of most enzymes was further enhanced at 4d. The changes in nonprotein thiols, total glutathione, ascorbic acid, and dehydroascorbate content were also assayed. Our results suggest that ammonium induced the oxidative stress and the heated antioxidant response in H. verticillata.

Vallisneria americana Michx. (tapegrass) is an ecologically important submersed, vascular aquatic plant that provides food and shelter for many aquatic and waterfowl species. This plant often occurs close to land areas where herbicides are used. Nontarget exposure of these plants to herbicides may compromise ecological structure and function. The objective of the present study was to evaluate the suitability of several endpoint measurements for determining no-observable-adverse effect concentrations (NOAECs), lowest-observable-adverse effect concentrations (LOAECs), and median effective concentration values (EC50s) for tapegrass exposed to the herbicides bromacil (0-0.092 mg/L) and simazine (0-0.592 mg/L) following a 13-d single-pulse exposure and 15-d (bromacil) or 14-d (simazine) postexposure periods. The NOAEC/LOAEC/EC50 for fresh weight gains, new leaf production, and total leaf growth after 13-d exposure to bromacil were 0.020/0.036/0.032, 0.036/0.054/0.036, and 0.036/0.054/0.043 mg/L, respectively. The same respective NOAEC/LOAEC/EC50s for simazine were <0.058/0.058/0.067, 0.229/0.344/0.154, and 0.058/0.116/0.081 mg/L. Reductions in quantity and fresh weight of daughter plants produced and stolon fresh weights occurred at bromacil concentrations > or = 77, 0.020, and 0.036 mg/L, respectively; and simazine concentrations > or = 0.344, >0.592, and > or = 0.116 mg/L, respectively. Neither herbicide affected leaf greenness, total chlorophyll concentrations, or carbohydrate allocation. Although toxicity was shown for many endpoints, most EC50 values were greater than aquatic life benchmark values for algae used by the U.S. Environmental Protection Agency (U.S. EPA), but less than for aquatic plants, indicating that V. americana would likely be protected by use of the algal benchmark criteria.