Red light mitigates Cd toxicity in Egeria densa by restricting Cd accumulation and modulating antioxidant defense system.

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.

Ecotoxicity of perfluorooctanoic acid and perfluorooctane sulfonate on aquatic plant Vallisneria natans.

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.

Responses of CO2-concentrating mechanisms and photosynthetic characteristics in aquatic plant Ottelia alismoides following cadmium stress under low CO2.

The effects of cadmium (Cd) have been investigated in an aquatic plant Ottelia alismoides grown under low CO2. Under low CO2, no Cd treated O. alismoides operated three carbon dioxide-concentrating mechanisms (CCMs) efficiently, including HCO3- 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 (δ13C) 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 HCO3-. 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.

Effects of excess ammoniacal nitrogen (NH4+-N) on pigments, photosynthetic rates, chloroplast ultrastructure, proteomics, formation of reactive oxygen species and enzymatic activity in submerged plant Hydrilla verticillata (L.f.) Royle.

Although excess ammoniacal-nitrogen (NH4+-N) results in the disturbance of various important biochemical and physiological processes, a detailed study on the effects of NH4+-N stress on the photosynthesis and global changes in protein levels in submerged macrophytes is still lacking. Here, the changes of excess NH4+-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 NH4+-N, respectively, and decreased at NH4+-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 NH4+-N as a control, ROS generation in chloroplasts significantly increased in the presence of more than 2 mg L-1 NH4+-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 NH4+-N. Surprisingly, the abundance of all the identified proteins that were involved in nitrogen assimilation and amino acid metabolism tended to increase under excess NH4+-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 NH4+-N results in chloroplast ultrastructural damages and the first proteomic evidence to support that excess NH4+-N can lead to a decline in photosynthesis and imbalance of C-N metabolism in submerged macrophytes.

Negative effects on the leaves of submerged macrophyte and associated biofilms growth at high nitrate induced-stress.

High nitrate (NO3--N) concentration is a growing aquatic risk concern worldwide. However, adverse effects of high NO3--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 NO3--N concentrations. The findings showed that 10 mg L-1NO3--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-1NO3--N, implied oxidative stress were stimulated. However, no significant alterations in these indicators were observed with 5 mg L-1NO3--N. Compared to control, 10 mg L-1NO3--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-1NO3--N, indicating that 10 mg L-1NO3--N concentration exhibited significant inhibition of chemical activity and nutrient uptake of the leaf surfaces. Overall, these results demonstrated that high NO3--N does stimulate the biofilm growth and can cause negative impacts on submerged macrophytes growth.

Damage of heavy metals to Vallisneria natans (V. natans) and characterization of microbial community in biofilm.

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 Cu2+, Pb2+, Cd2+ 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.

Responses of Hydrocharis dubia (Bl.) Backer and Trapa bispinosa roxb. to tetracycline exposure.

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.

Antagonism or synergism? Responses of Hydrocharis dubia (Bl.) Backer to linear alkylbenzene sulfonate, naphthalene and their joint exposure.

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.

Negative effects of a piscicide, rotenone, on the growth and metabolism of three submerged macophytes.

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.

Indigenous strain Bacillus XZM assisted phytoremediation and detoxification of arsenic in Vallisneria denseserrulata.

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.

Silver nanoparticle toxicity effect on the seagrass Halophila stipulacea.

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 H2O2 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.

Effects of maifanite on growth, physiological and phytochemical process of submerged macrophytes Vallisneria spiralis.

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.

Responses of Vallisneria natans (Lour.) Hara to the combined effects of Mn and pH.

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 (H2O2). 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.

Effects of ammonium pulse on the growth of three submerged macrophytes.

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.

Bioaccumulation and growth characteristics of Vallisneria natans (Lour.) Hara after chronic exposure to metal-contaminated sediments.

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, Igeo 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.

Enrofloxacin and Roundup® interactive effects on the aquatic macrophyte Elodea canadensis physiology.

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 OriginalDI (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.

Effects of nanoplastics and microplastics on the growth of sediment-rooted macrophytes.

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.

Responses of propagule germination and sexual reproduction of submerged macrophytes exposed to cadmium.

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.

Allelopathic effects of harmful algal extracts and exudates on biofilms on leaves of Vallisneria natans.

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.

Subcellular accumulation and source of O2- and H2O2 in submerged plant Hydrilla verticillata (L.f.) Royle under NH4+-N stress condition.

In this study, the effects of excess NH4+-N on the subcellular accumulation of O2- and H2O2 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 NH4+-N for 5 d significantly increased production of O2- and H2O2, 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 O2- and H2O2 in the submerged plants. In the leaves of H. verticillata treated with 20.0 mg L-1 NH4+-N, O2- dependent DAB precipitates were found primarily on the inner side of the plasma membrane, extracellular space and chloroplasts. H2O2-CeCl3 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 NH4+-N-induced production of O2- and H2O2 in H. verticillata leaves may involve plasma membrane-bound NADPH oxidase. Moreover, low-light treatment decreased NH4+-induced O2- production, suggesting that alterations in the photosynthetic electron transfer chain due to NH4+ toxicity could lead to O2- production.