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.

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.

Assessing interactions between environmental factors and aquatic toxicity: Influences of dissolved CO2 and light on Cd toxicity in the aquatic macrophyte Potamogeton crispus.

The objective of this study was to investigate the combined effects of varying dissolved CO2 concentration (ambient CO2, 3˜17 µmol L-1, elevated CO2, 48˜81 µmol L-1) and light intensity (high light, c. 150 µmol photon m-2 s-1, low light, c. 25 µmol photon m-2 s-1) on the bioaccumulation and phytotoxicity of cadmium (Cd) in a macrophyte Potamogeton crispus, under constant Cd exposure. The data confirmed that 100 µM Cd led to adverse changes in morphology, ultrastructure and biochemistry in P. crispus. The toxic effects depended strongly on CO2 concentration and light intensity: elevated CO2 and high light both increased Cd concentrations in P. crispus, and there was a significant interaction between the two factors. Compared to high light grown plants, the photochemical efficiency and chlorophyll content of low light grown P. crispus were much less affected and the MDA content was lower, when exposed to 100 µM Cd. In addition, an antioxidative response was observed with a significant increase in SOD, POD and GST activities, indicating that low light grown P. crispus are more protected against Cd toxicity. When compared with ambient CO2 concentrations, chlorophyll content, chlorophyll fluorescence, photosynthetic rate and starch content, as well as the activity of SOD and GST, were significantly enhanced in Cd treated P. crispus under elevated CO2. This suggests that elevated CO2 reduced Cd toxicity in P. crispus by increasing photosynthesis and enhancing the antioxidant system. Moreover, the statistical results showed that dissolved CO2 and light had additive effects on Cd toxicity in P. crispus, reflected by the physiological parameters of total chlorophyll content, SOD activity and MDA content, indicating that the combination of high CO2 and low light produced more protection against Cd toxicity than did the factors alone. Based on the results of this study, it appears clear that referring to a specific site in aquatic ecosystem, dissolved CO2 concentration and light availability should be considered when assessing and managing Cd impacts on aquatic plants.

Rare-earth element yttrium enhances the tolerance of curly-leaf pondweed (Potamogeton crispus) to acute nickel toxicity.

Nickel is a ubiquitous heavy-metal pollutant in lakes and severely affects aquatic organisms. Aquatic plants are often initially linked to having heavy metal contents and further are proposed as phytoremediation agent to remove heavy metal from water. Although the toxic effects of nickel on aquatic plants are thoroughly explored, the effective investigation to increase Ni tolerance is still in its infancy. The role of rare-earth elements (REEs) in plant resisting heavy-metal pollution has recently received considerable interest. To explore the physiological effects of REEs on Potamogeton crispus under Ni stress, we explored whether or not the additive exposure to low-dose yttrium (Y; 2.5 µM) promotes the polyamine metabolism, antioxidation, and photosynthesis performance of P. crispus under Ni stress values of 0, 50, 100, 150, and 200 µM. Results showed that Y exposure did not influence Ni bioaccumulation in P. crispus. Furthermore, Y exposure alleviated the adverse effects of Ni stress to convergent degrees because Y positively converts putrescine into spermidine and spermine, inhibits oxidative stress, increases the total chlorophyll content, and maximum/potential quantum efficiency of photosystem II. We concluded that low-dose Y can positively regulate polyamine transformation, inhibit oxidative stress, stimulate photosynthesis, and finally promote the resist ability of P. crispus to nickel stress. Thus, REEs have potential to be applied in regulating submerged plant tolerance to aquatic heavy-metal pollution.

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.

Toxicity of linear alkylbenzene sulfonate to aquatic plant Potamogeton perfoliatus L.

Aquatic plants play an important role in maintaining the health of water environment in nature. Studies have shown that linear alkylbenzene sulfonate (LAS), a type of omnipresent pollutant, can cause toxic damage to aquatic plants. In the present research, we studied the physiological and growth response of submerged plant Potamogeton perfoliatus L. to different concentrations of LAS (0.1, 1.0, 10.0, 20.0, and 50.0 mg l-1). The results showed that LAS is toxic to P. perfoliatus, and the toxicity is dose-dependent. Only slightly reversible oxidative damages were observed in the physiological parameters of P. perfoliatus when P. perfoliatus was exposed to lower LAS doses (< 10 mg l-1): soluble sugar, soluble protein, H2O2, and malondialdehyde (MDA) content in P. perfoliatus increased significantly at 0.1 mg l-1 and then returned to normal levels at 1.0 mg l-1. Antioxidant enzymes were activated before the LAS concentration reached 10 mg l-1, and the activities of superoxide dismutase (SOD), catalase (CAT), and photosynthesis pigment content declined significantly when the concentration of LAS exceeded 10 mg l-1. In addition, at higher concentrations (20-50 mg l-1) of LAS, dry weight and fresh weight of P. perfoliatus showed significant declines. The results indicate that LAS above 10 mg l-1 can cause serious physiological and growth damage to P. perfoliatus.

Accumulation and effects of copper on aquatic macrophytes Potamogeton pectinatus L.: Potential application to environmental monitoring and phytoremediation.

This study investigated the ability of Potamogeton pectinatus L. to accumulate copper and its effects on plants. In accumulation tests, macrophytes were exposed (96 h) to different copper concentrations (0-1000 µM) and the metal was measured in media and plant tissues (roots, stems and leaves) to determine the bioconcentration factor (BCF). Plants accumulated high concentrations of copper in a dose-dependent manner and roots was the main organ for copper accumulation. However, the more copper increased in water, the more BCF values decreased. It may be due to either saturation of copper uptake or down-regulation of metal uptake by plants. In the physiological and morphological analyses, plants were kept (96 h) in Hoagland nutrient solution without copper, in full Hoagland solution (0.5 µM Cu) and in Hoagland medium with copper from 1 to 100 µM. The absence and the presence of copper above to 1 µM inhibited photosynthesis. Chlorophylls and carotenoid levels also decreased with the excess of copper, a fact that may have affected the photosystem II-dependent of chlorophyll and caused photosynthesis suppression. Only macrophytes at 10 µM Cu showed decrease in length and number of leaves on the 10th day of the test, when they died. Chlorosis and necrosis were observed in control groups and groups with extra copper, but not in Hoalgand group. Overall, the macrophyte P. pectinatus can be considered a suitable plant for monitoring environments contaminated by copper, based on results of copper accumulation in the plant, decrease in pigment concentration and presence of chlorosis and necrosis. However, values of BCF based on fresh water tissues was not proper to indicate the use of P. pectinatus for cleaning environments contaminated by copper.

Spatial (bio)accumulation of pharmaceuticals, illicit drugs, plasticisers, perfluorinated compounds and metabolites in river sediment, aquatic plants and benthic organisms.

Organic contaminants such as pharmaceuticals, personal care products (PPCPs) and other emerging contaminants (ECs) are known to persist in the aquatic environment and many are indicated as endocrine, epigenetic, or other toxicants. Typically, the study of PPCPs/ECs in the aquatic environment is limited to their occurrence dissolved in river water. In this study, accumulation and spatial distribution of thirteen PPCPs/ECs were assessed in aquatic sediment (n = 23), periphyton (biofilm, n = 8), plants Callitriche sp. (n = 8) and Potamogeton sp. (n = 7) as well as amphipod crustaceans (Gammarus pulex, n = 10) and aquatic snails (Bithynia tentaculata, n = 9). All samples (n = 65) were collected from the Hogsmill, Blackwater and Bourne Rivers in southern England. Targeted PPCPs/ECs included pharmaceuticals, plasticisers, perfluorinated compounds, illicit drugs and metabolites. Extraction from solid matrices occurred using ultrasonic-assisted extraction followed by an in-house validated method for solid-phase extraction and subsequent liquid-chromatography tandem mass-spectrometry. Field-derived bioconcentration-factors and biota-sediment accumulation-factors were determined for all studied biota. Residues of studied contaminants were found in all sediment and biota. Concentrations of contaminants were generally higher in biota than sediment. Evidence suggests that the studied aquatic plants may effectively degrade bisphenol-A into its main transformation product hydroxyacetophenone, potentially mediated by cytochrome p450 and internalisation of contaminants into the cellular vacuole. A positive association between both hydrophobicity and PFC chain length and contaminant accumulation was observed in this work. Only PFCs, plasticisers and HAP were classified as either 'bioaccumulative' or 'very bioaccumulative' using BCF criteria established by guidelines of four governments. Contaminants appeared to be differentially bioaccumulative in biota, indicating there may be a need for a species-specific BCF/BSAF classification system. These data form a detailed accounting of PPCP/EC fate and distribution in the aquatic environment highlighting accumulation at lower trophic levels, a potential source for higher organisms.

Biomonitoring of nutrient and toxic element concentrations in the Sarno River through aquatic plants.

The Sarno River is considered the most polluted river in Europe and one of the ten most polluted rivers in the world. So far, its quality has been usually evaluated by water and sediment analyses of either inorganic or organic pollutants. However, a biomonitoring approach would be of paramount importance in the evaluation of river quality, since it integrates pollutant temporal fluctuations, as in the case of discontinuous inputs from urban, industrial and agricultural activities. To this end, a passive biomonitoring study of the Sarno River was carried out, using two native aquatic plants accumulators of inorganic pollutants. The spring area was monitored analysing the roots of the semi-submerged Apium nodiflorum, whereas the whole river course was monitored analysing the shoots of the submerged Potamogeton pectinatus. The information on the four macronutrient (Ca, K, Mg, P), the six micronutrient (Cu, Fe, Mn, Na, Ni, Zn) and the four toxic element (Cd, Cr, Pb, V) concentrations were separately combined in the Nemerow Pollution Index. Results evidenced a severe pollution degree of the Sarno River, attributable to toxic elements > micronutrients > macronutrients. In particular, the spring area showed high K concentrations, as well as high concentrations of several micronutrients and toxic elements. A generalized Zn contamination and a progressive macronutrient (above all Ca and P), micronutrient (above all Ni, Cu and Fe) and toxic element (above all Cr and Pb) accumulation toward the mouth was related to pollution from agricultural and urban activities. Industrial sources, especially tanneries along the Solofrana tributary, accounted for high Mn concentrations, whereas the volcanic origin of the substrate accounted for a generalized V contamination.

Effects of aluminate flocculant on turion germination and seedling growth of Potamogeton crispus.

Aluminate flocculants are employed widely in water treatment for precipitating suspended solids and emergency treatment of algal blooms in eutrophic lake, but the residual aluminum (Al) may have phytotoxic effects on aquatic organisms after entering aquatic ecosystems. To elucidate the potential impacts of Al on turion germination and early growth in Potamogeton crispus, we conducted a mesocosm experiment using five Al concentrations (0 (control group), 0.3, 0.6, 1.2, and 1.5mg/L) in alum solutions. The results showed that the germination of turions and the early growth of P. crispus were reduced and inhibited by Al. The maximum numbers of germinating turions and newly-formed seedlings occurred in the control group, and their numbers declined in the end of the experiment as the Al concentration increased. Al at a concentration of 1.5mg/L decreased the number of germinating turions 3.0 times and the number of newly-formed seedlings 30.7 times compared with the control. The chlorophyll content and root activity decreased when the Al concentration increased. The maximum soluble protein contents in seedling tissues (1.953mg/g fresh weight) occurred in the 0.6mg/L treatment group, which differed significantly from the other treatment groups. The Al contents in the seedling tissues had a significant positive correlation with the Al treatment concentrations (P<0.05, r=0.763), but negative correlations with the biomass, root number, stem weight, soluble protein, and root activity (r=-0.935, -0.975, -0.907, -0.721, -0.944, respectively). Persistent Al concentration ≥1.2mg/L significantly decreased the germination of turions and seedling growth in P. crispus. These results may facilitate the restoration of aquatic macrophytes and ecological risk assessments in Al-exposed lakes.

Loss of heterophylly in aquatic plants: not ABA-mediated stress but exogenous ABA treatment induces stomatal leaves in Potamogeton perfoliatus.

Heterophyllous aquatic plants produce aerial (i.e., floating and terrestrial) and submerged leaves-the latter lack stomata-while homophyllous plants contain only submerged leaves, and cannot survive on land. To identify whether differences in morphogenetic potential and/or physiological stress responses are responsible for variation in phenotypic plasticity between two plants types, responses to abscisic acid (ABA) and salinity stress were compared between the closely related, but ecologically diverse pondweeds, Potamogeton wrightii (heterophyllous) and P. perfoliatus (homophyllous). The ABA-treated (1 or 10 µM) P. wrightii plants exhibited heterophylly and produced leaves with stomata. The obligate submerged P. perfoliatus plants were able to produce stomata on their leaves, but there were no changes to leaf shape, and stomatal production occurred only at a high ABA concentration (10 µM). Under salinity stress conditions, only P. wrightii leaves formed stomata. Additionally, the expression of stress-responsive NCED genes, which encode a key enzyme in ABA biosynthesis, was consistently up-regulated in P. wrightii, but only temporarily in P. perfoliatus. The observed species-specific gene expression patterns may be responsible for the induction or suppression of stomatal production during exposure to salinity stress. These results suggest that the two Potamogeton species have an innate morphogenetic ability to form stomata, but the actual production of stomata depends on ABA-mediated stress responses specific to each species and habitat.

Lead tolerance mechanism in sterilized seedlings of Potamogeton crispus L.: subcellular distribution, polyamines and proline.

The effects of increasing concentrations of lead (Pb) on malondialdehyde (MDA) content, soluble protein, Pb accumulation, nutrients, polyamines (PAs) and proline metabolism were investigated in sterilized seedlings of Potamogeton crispus L. after 5d exposure. Significant oxidative stress was not caused, indicated by a little induction of MDA content and soluble proteins. Pb accumulation increased in a concentration-dependent manner and most of Pb was stored in the cell wall. Total P, Mg, Na and Zn rose and total Fe fell; total Ca increased at 25 µM Pb but then declined. The nutrients in cell wall fraction changed in the same pattern as total nutrients, whereas those in soluble and organelle fraction declined. Total putrescine (Put) decreased markedly, while total spermidine (Spd), spermine (Spm) and (Spd+Spm)/Put ratio increased progressively but then declined. The trends for free, perchloric acid soluble conjugated (PS-conjugated) and perchloric acid insoluble bound (PIS-bound) PAs were similar to those on total PAs, except that PIS-bound Spm increased significantly. Arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) activities rose gradually, while diamine oxidase (DAO) initially increased but then declined. Proline content increased initially only to decline later, due to the increase of r-glutamyl kinase (GK) activity. Meanwhile, ornithine-d-aminotransferase (OAT) activity gradually reduced, while no significant change was observed in proline dehydrogenase (PDH) activity. Our results indicated that the tolerance of P. crispus to Pb stress was based on cell wall compartmentalization combined with increase of nutrients, alterations of PAs, and induction of proline.

Turion morphological responses to water nutrient concentrations and plant density in the submerged macrophyte Potamogeton crispus.

Asexual propagules are the dominant means of propagation in most submerged macrophytes. To improve the understanding of how water nutrient concentrations and population density influence the turion production of Potamogeton crispus L., the turions were planted in mesocosms with three water nutrient conditions (ambient lake water, high P and high N) and two plant density levels (4 and 15 turions m(-2)). After a 9-month experiment, the +P in the water column significantly increased the total turion number per plant under both of the plant density treatments. However, the +N in the water column did not affect the turion number per plant under low plant density. The +P in the water and high plant density significantly reduced the turion individual biomass. An examination of 3210 turion individuals from all treatments revealed that the increased water nutrient concentrations and plant density impacted the turion size by producing different stem diameters of individual turions. Most of the scale leaf morphological traits of the turions were significantly increased under higher water nutrients, but these traits were similar between the different plant density treatments. These results demonstrate that the water P concentration interacts with plant density, affecting both the production and traits of turions.

The uptake and distribution of selenium in three aquatic plants grown in Se(IV) solution.

The uptake of Se(IV) by Myriophyllum spicatum, Ceratophyllum demersum and Potamogeton perfoliatus, and the effects of Se(IV) on their physiological and biochemical characteristics were studied. Plants were cultivated outdoors under semi-controlled conditions in water solution containing Na selenite (20 µg Se L(-1) and 10 mg Se L(-1)). The higher concentration of Se lowered the photochemical efficiency of PSII in all species studied, while the lower concentration had no effect on any species. The higher concentration of Se lowered respiratory potential in M. spicatum. The response of M. spicatum and C. demersum to Se(IV) regarding chlorophylls was variable, however in the majority of cases, there was a trend of increasing the amount of chlorophylls, while in P. perfoliatus the amount of chlorophyll a decreased. The concentration of Se in plants cultured in 10 mg Se(IV) L(-1) ranged from 436 to 839 µg Se g(-1) DM in M. spicatum, 319 to 988 µg Se g(-1) DM in C. demersum and 310 to 661 µg Se g(-1) DM in P. perfoliatus. The amount of soluble Se compounds in enzyme extracts of high Se treatment was 27% in M. spicatum, 41% in C. demersum and 35% in P. perfoliatus. Se compounds were determined using HPLC-ICP-MS. It was observed that the applied Se(IV) was mainly transformed to insoluble Se.

Subcellular distribution and toxicity of cadmium in Potamogeton crispus L.

The submerged macrophyte Potamogeton crispus L. was subjected to varying doses of cadmium (0, 20, 40, 60 and 80 µM) for 7 d, and the plants were analyzed for subcellular distribution of Cd, accumulation of mineral nutrients, photosynthesis, oxidative stress, protein content, and ultrastructural distribution of calcium (Ca). Leaf fractionation by differential centrifugation indicated that 48-69% of Cd was accumulated in the cell wall. At all doses of Cd, the levels of Ca and B rose and the level of Mn fell; the levels of Fe, Mg, Zn, Cu, Mo, and P rose initially only to decline later. Exposure to Cd caused oxidative stress as evident by increased content of malondialdehyde and decreased contents of chlorophyll and protein. Photosynthetic efficiency, as indicated by the quenching of chlorophyll a fluorescence (Fv/Fm, Fo and Fm), decreased significantly, the extent of decrease being directly proportional to the concentration of Cd. Increased amounts of precipitates of calcium were noticed in the treated plants, located either outside the cell membrane or in chloroplasts, mitochondria, the nucleus, and the cytoplasm whereas control plants showed small deposits of the precipitates around surface of the vacuole membrane and in the intercellular space but rarely in the cytoplasm. Photosynthetic efficiency and oxidative stress could be used as indicators of physiological end-points in determining the extent of Cd phytotoxicity.

Sensitivity, variability, and recovery of functional and structural endpoints of an aquatic community exposed to herbicides.

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.

Involvement of plasma membrane H+-ATPase in anoxic elongation of stems in pondweed (Potamogeton distinctus) turions.

• Pondweed (Potamogeton distinctus) turions can elongate in the absence of O(2). Alcoholic fermentation serves to produce energy for anoxic elongation via the breakdown of starch stored in cells. However, the mechanism of cell growth during anoxic elongation is not fully understood. • Changes in pH, H(+) equivalent and lactate content of the incubation medium were measured during anoxic elongation. The effects of fusicoccin (FC), indole-3-acetic acid (IAA), vanadate, erythrosine B and K(+) channel blockers on anoxic elongation were examined. Cytoplasmic pH and vacuolar pH were measured by (31)P nuclear magnetic resonance (NMR) spectroscopy. • Acidification of the incubation medium occurred during anoxic elongation. The contribution of CO(2) and lactic acid was not sufficient to explain the acidification. FC and IAA enhanced the elongation of stem segments. Vanadate and erythrosine B inhibited anoxic elongation. Acid growth of notched segments was observed. The activity of plasma membrane H(+)-ATPase extracted from pondweed turions was increased slightly in anoxic conditions, but that from pea epicotyls sensitive to anoxic conditions was decreased by incubation in anoxic conditions. Both the cytoplasmic pH and vacuolar pH of pondweed turion cells chased by (32)P NMR spectroscopy were stabilized during a short period < 3 h after anoxic conditions. • We propose that the enhancement of H(+) extrusion by anoxic conditions induces acidification in the apoplast and may contribute to the stabilization of pH in the cytoplasm.

Involvement of polyamines in adaptation of Potamogeton crispus L. to cadmium stress.

The effects of increasing concentrations of cadmium (Cd) on the ultrastructure, Cd accumulation, generation of O(2)(-), contents of ascorbate (AsA), reduced glutathione (GSH) and polyamines (PAs), as well as the activities of polyamine oxidase (PAO) (EC 1.5.3.3) and diamine oxidase (DAO) (EC 1.4.3.6) were investigated in the leaves of Potamogeton crispus. Cd exposure resulted in significant damage in chloroplasts and mitochondria, suggesting that Cd hastened the senescence of the tested plants. The accumulation of Cd was found to increase in a concentration-dependent manner, accompanied by increased production of O(2)(-). AsA content increased progressively up to 70µM Cd, followed by a decline at higher concentration. GSH content slightly increased up to 70µM Cd and then declined. In addition, Cd treatment increased the putrescine (Put) content, while decreasing spermidine (Spd) and spermine (Spm) contents, which reduced the ratio of free (Spd+Spm)/Put in the leaves. PS-conjugated PAs changed in the same pattern as free PAs, while PIS-bound PAs was different. PIS-bound Put content enhanced with the increase of Cd concentration up to 50µM and then decreased, and PIS-bound Spd and Spm contents decreased to a lesser extent. Moreover, the activities of PAO and DAO increased significantly with the increase of the Cd concentrations, reaching the peak values at 70µM Cd. Our results suggested that certain PAs and PAs forms could play a significant role in the adaptation mechanism of P. crispus under Cd stress.

Phytotoxicity of atrazine, isoproturon, and diuron to submersed macrophytes in outdoor mesocosms.

The submersed macrophytes Elodea canadensis, Myriophyllum spicatum and Potamogeton lucens were constantly exposed over a five-week period to environmentally relevant concentrations of atrazine, isoproturon, diuron, and their mixture in outdoor mesocosms. Effects were evaluated investigating photosynthetic efficiency (PE) of the three macrophytes and growth of M. spicatum and E. canadensis. Adverse effects on PE were observed on days 2 and 5 after application. M. spicatum was found to be the more sensitive macrophyte. E. canadensis and P. lucens were less sensitive to atrazine, diuron and the mixture and insensitive to isoproturon. PE of M. spicatum was similarly affected by the single herbicides and the mixture demonstrating concentration addition. Growth of E. canadensis and M. spicatum was not reduced indicating that herbicide exposure did not impair plant development. Although PE measurements turned out to be a sensitive method to monitor PSII herbicides, plant growth remains the more relevant ecological endpoint in risk assessment.

Copper-induced response of physiological parameters and antioxidant enzymes in the aquatic macrophyte Potamogeton pusillus.

Bioaccumulation and toxicity of copper was evaluated on Potamogeton pusillus L. The effect of copper (5-100 microg L(-1)) applied for several days was assessed by measuring changes in the chlorophyll's, phaeophytin's, malondialdehyde, electrical conductivity, glutathione peroxidase (GPX), glutathione reductase (GR) and guaiacol peroxidase (POD) activities. Plants accumulated copper with a maximum of 162 microg g(-1) dw after 7-days exposure at 100 microg L(-1), however most of the metal was accumulated after 1-day exposure. The toxic effect caused by Cu was evident by the reduction of photosynthetic pigments, increase of malondialdehyde and electrical conductivity. P. pusillus shows Cu-induced oxidative stress by modulating antioxidant enzymes like GPX, GR and POD. Antioxidant enzymes activity increased significantly after exposure to 40 microg L(-1) during 24 h, followed by a drop at longer times. Thus, P. pusillus is proposed as a good biomonitor for the assessment of metal pollution in aquatic ecosystems.