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.

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.

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.

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.

Defense Mechanisms of Two Pioneer Submerged Plants during Their Optimal Performance Period in the Bioaccumulation of Lead: A Comparative Study.

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.

Phosphate alleviation of glyphosate-induced toxicity in Hydrocharis dubia (Bl.) Backer.

Glyphosate, as a broad-spectrum herbicide, is frequently detected in water, and phosphorus widely enters the water due to the extensive use of phosphorus-containing substances in agriculture, industries and daily life. Thus, aquatic ecosystems are exposed to both glyphosate and phosphorus, which may affect aquatic organisms. In the present research, we studied the physiological responses of the floating aquatic plant species H. dubia to different concentrations of glyphosate (0, 1, 5, 15 mg/L) with different levels of phosphate (0, 50, 100 mg/L) after 14 days (d) of treatment. We explored glyphosate toxicity in H. dubia and investigated whether phosphate addition mitigates glyphosate toxicity in this species, which will provide a theoretical basis for the ecotoxicological study of aquatic plants. The results show that glyphosate significantly reduced the chlorophyll content, leaf number and root length of H. dubia, while it significantly increased the malondialdehyde (MDA), hydrogen peroxide (H2O2), shikimate, proline, and soluble protein content and enzyme activities (superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and polyphenol oxidase (PPO)) in H. dubia. After phosphate supplement, the MDA, H2O2, proline, and soluble protein contents and enzyme activities in the plants treated with glyphosate decreased. These results indicate that the concentration of glyphosate investigated in our study can cause oxidative stress and affect the growth of H. dubia. Phosphate can alleviate glyphosate-induced oxidative stress in H. dubia.

High-potential accumulation and tolerance in the submerged hydrophyte Hydrilla verticillata (L.f.) Royle for nickel-contaminated water.

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.

Effect of the exogenous anthocyanin extract on key metabolic pathways and antioxidant status of Brazilian elodea (Egeria densa (Planch.) Casp.) exposed to cadmium and manganese.

Present study deals with the effect of 24 h pre-incubation with exogenous anthocyanins (ANTH), extracted from red cabbage leaves, on key metabolic processes (photosynthesis and respiration) and pro-/antioxidant balance in the aquatic macrophyte Egeria densa (Planch.) Casp., Hydrocharitaceae family, treated with Cd and Mn (in sulfate form) at a concentration of 100 µmol. After five days of metal treatments, Cd was accumulated and the damage caused to metabolic processes was stronger than Mn. In Cd-treated leaves, the protein level, chlorophyll concentration and maximal photochemical efficiency of PS II decreased twofold, and net-photosynthesis was significantly inhibited, whereas lipid peroxidation and H2O2 production increased. In turn, protective responses developed, including an increase in the total soluble thiols, alternative respiratory pathway capacity and the activity of superoxide dismutase and peroxidases. Pre-incubation in the ANTH-enriched extract caused an increase in foliar ANTH content, enhanced Cd and reduced Mn uptake into the tissue. A decrease in the level of oxidative reactions, an increase in the protein and chlorophyll concentration compared to the control values and a partial improvement of the photosynthetic parameters confirmed the ability of ANTH to reduce Cd-induced damage effects and to mitigate ROS-driven stress reactions. Stimulation of catalase and ascorbate peroxidase activity, an alternative respiration capacity and non-enzymatic antioxidant (carotenoids, ascorbate and proline) synthesis by ANTH were also revealed. These data suggest that ANTH-enriched extract from red cabbage leaves has a protective action against metal toxicity in Egeria plants.

Effects of two-hour exposure to environmental and high concentrations of methylmercury on the transcriptome of the macrophyte Elodea nuttallii.

The effects of two methylmercury (CH3Hg+, 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.

Influence of the residue from an iron mining dam in the growth of two macrophyte species.

On November 5th, 2015 the worst environmental disaster in Brazil spilled 60 million m3 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.

Impacts of residual aluminum from aluminate flocculant on the morphological and physiological characteristics of Vallisneria natans and Hydrilla verticillata.

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.

The effects of cadmium pulse dosing on physiological traits and growth of the submerged macrophyte Vallisneria spinulosa and phytoplankton biomass: a mesocosm study.

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

Zinc supplementation imparts tolerance to arsenite stress in Hydrilla verticillata (L.f.) Royle.

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.

Elodea nuttallii exposure to mercury exposure under enhanced ultraviolet radiation: Effects on bioaccumulation, transcriptome, pigment content and oxidative stress.

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.

The role of roots in the accumulation and removal of cadmium by the aquatic plant Hydrilla verticillata.

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.

Oxidative effects, nutrients and metabolic changes in aquatic macrophyte, Elodea nuttallii, following exposure to lanthanum.

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.

Effects of copper-oxide nanoparticles, dissolved copper and ultraviolet radiation on copper bioaccumulation, photosynthesis and oxidative stress in the aquatic macrophyte Elodea nuttallii.

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.

Cellular proton dynamics in Elodea canadensis leaves induced by cadmium.

Our earlier investigations showed that Elodea canadensis shoots, grown in the presence of cadmium (Cd), caused basification of the surrounding medium. The present study was aimed to examine the proton dynamics of the apoplastic, cytosolic and vacuolar regions of E. canadensis leaves upon Cd exposure and to establish possible linkage between cellular pH changes and the medium basification. The changes in cytosolic calcium [Ca(2+)]cyt was also investigated as the [Ca(2+)]cyt and [pH]cyt homeostasis are closely linked. The cellular H(+) and Ca(2+) concentrations were monitored by fluorescence microscopy and ion-specific fluorescent dyes. Cadmium concentration of leaf-cell walls was measured after plant cultivation at different fixed levels of starting pH. The protoplasts from E. canadensis leaves were isolated by use of a newly developed enzymatic method. Upon Cd addition, both cytosolic and vacuolar pH of leaf protoplasts increased with a concomitant rise in the cytosolic Ca(2+) concentration. Time course studies revealed that changes in [Ca(2+)]cyt and [pH]cyt followed similar dynamics. Cadmium (0.5 µM) exposure decreased the apoplastic pH by 0.85 units. The maximum cell wall bound Cd-contents were obtained in plants grown at low starting pH. It is concluded that Cd treatment causes apoplastic acidosis in E. canadensis leaves associated with enhanced Cd binding to the cell walls and, consequently, reduced Cd influx into the cytosol.