Warming alters the network of physiological traits and their contribution to plant abundance.

The impact of global warming on plant abundance has been widely discussed, but it remains unclear how warming affects plant physiological traits, and how these traits contribute to the abundance of aquatic plants. We explored the adjustments in physiological traits of two common aquatic plant species (Potamogeton crispus L. and Elodea canadensis Michx.) and their links to plant abundance in three temperature treatments by determining twelve physiological traits and plant abundance over an 11-month period in outdoor mesocosms. This mesocosms facility has been running uninteruptedly for 16 years, rendering the plants a unique opportunity to adapt to the warming differences. We found that 1) warming reduced the starch storage in winter for P. crispus and in summer for E. canadensis while increased the nitrogenous substances (e.g., TN, FAA, and proline) in winter for P. crispus. 2) For E. canadensis, TC, starch, SC, and sucrose contents were higher in summer than in winter regardless of warming, while TC, SC, and sucrose contents were lower in summer for P. crispus. 3) Warming decreased the association strength between physiological traits and plant abundance for P. crispus but enhanced it for E. canadensis. 4) E. canadensis showed increased interaction strength among physiological traits under warming, indicating increased metabolic exertion in the response to warming, which contributed to the reduction in abundance. Trait interaction strength of P. crispus was reduced under warming, but with less impact on plant abundance compared with E. canadensis. Our study emphasizes that warming alters the network of plant physiological traits and their contribution to abundance and that different strengths of susceptibility to warming of the various plant species may alter the composition of plant communities in freshwater ecosystems.

Accumulation of elements by submerged (Stuckenia pectinata (L.) Börner) and emergent (Phragmites australis (Cav.) Trin. ex Steud.) macrophytes under different salinity levels.

Accumulation of essential/beneficial and non-essential chemical elements by submerged (Stuckenia pectinata (L.) Börner) and emergent (Phragmites australis (Cav.) Trin. ex Steud.) rooted macrophytes in three water bodies was studied as dependent on water salinity (specific conductivity) and main environmental factors (temperature, pH of water, growing period, element contents in water and sediments). Inductively coupled plasma optical emission spectrometry (ICP - OES) was used for the element detection in the plants, sediments, and water. Multivariate statistics redundancy analysis (RDA) showed that investigated factors together explained 37.8 - 44.7% and 29.9 - 42.6% of the essential/beneficial and non-essential elements variability, respectively, whereas salinity was one of the main factors determining the accumulation of a number of elements (Mg, S, Na, Li, Sr, Sb) by the macrophytes. A decrease in the essential element contents (K, P, N, Cu, Zn) was observed in both species of the macrophytes during the growing season. Environmental factors (salinity, growing period, and pH) were more significant than the element contents in water and the sediments for the essential/beneficial element accumulation by two species of the macrophytes. The element contents in the sediments to a greater extent determined non-essential element accumulation by P. australis than by S. pectinata. The study showed that the efficiency of using P. australis and S. pectinata for remediation of aquatic ecosystems contaminated by heavy metals should not decrease with a salinity increase.

Allelopathy appraisal of worm metabolites in the synergistic effect between Limnodrilus hoffmeisteri and Potamogeton malaianus on algal suppression.

In Chinese Lake Taihu, the algal quantity was significantly larger in summer than late spring (p < 0.01). In summer, compared with the dredged area including neither zoobenthos nor submerged macrophytes, the algal biomass and density were significantly lower in the area filled with the submerged macrophytes. Interestingly, the minimum algal bloom was observed in the combined area containing submerged macrophytes and zoobenthos, which was due to the synergistic interaction between the zoobenthos and the macrophytes. The metabolite extracts from the numerically dominant zoobenthos Limnodrilus hoffmeisteri had significant algal inhibitory effects of Microcystis aeruginosa, and displayed stimulatory effects on seed germination, seedling growth, and peroxidase activity of the prevalent submerged macrophyte Potamogeton malaianus. 27 active compounds in the worm metabolites were identified by gas chromatography-mass spectrometry (GC-MS). Among these compounds three chemicals arachidonic acid, eicosapentaenoic acid, and linoleic acid with concentrations of 13.92 ±â€¯1.11, 10.57 ±â€¯2.52, 2.75 ±â€¯0.73 mg/kg dry weight, respectively, were confirmed as the typical allelochemicals with algal inhibition potential. In short, the metabolites allelopathy of L. hoffmeisteri can form and assist the synergistic effect between L. hoffmeisteri and P. malaianus on algal suppression. Thus, it is feasible to simultaneously restore submerged macrophytes and zoobenthos community in the disturbed eutrophic lakes for removing harmful algae.

Metabolic and biochemical responses of Potamogeton anguillanus Koidz. (Potamogetonaceae) to low oxygen conditions.

Oxygen availability in water is considered one of the most important factors for growth and productivity in aquatic submerged macrophytes. In the present study, the growth, stress responses, and metabolic changes in Potamogeton anguillanus Koidz. (Potamogetonaceae) were assessed after a 21-day exposure to low (hypoxia; dissolved oxygen, DO < 1 mg/L) or null (anoxia) oxygen concentrations in water. High growth rates and an increased indole acetic acid (IAA) content in P. anguillanus were observed under the hypoxic conditions (4-fold to control) compared to the anoxic conditions. In addition, the activation of glycolysis and fermentation processes was further recorded, given the increase in alcohol dehydrogenase activity and pyruvate concentration on the studied plants that were exposed to low oxygen concentrations. Moreover, the positive correlations of antioxidative enzyme activities, catalase (CAT) and guaiacol peroxidase (POD) with hydrogen peroxide (H2O2) confirmed the species ability to scavenge excess H2O2 under low oxygen stress. The capillary electrophoresis-mass spectrometry (CE-MS) analysis of the metabolome identified metabolite accumulations (e.g., glutamate, glutamine, aspartate, asparagine, valine, malate, lactate, citrate, isocitrate, proline and γ-amino butyric acid) in response to the anoxia.

Impact of herbivory and competition on lake ecosystem structure: underwater experimental manipulation.

Two basic ecological relationships, herbivory and competition, distinctively influence terrestrial ecosystem characteristics, such as plant cover, species richness and species composition. We conducted a cage experiment under natural conditions in an aquatic ecosystem to test the impacts of two treatments combined in a factorial manner: (i) a pulse treatment - removal of dominant competitors among primary producers (macroalgae Chara sp. and Vaucheria sp.), and (ii) a press treatment - preventing herbivore (fish, crayfish) access to caged plots. The plots were sampled once before the treatments were established and four more times within two years. Both treatments had a significantly positive impact on macrophyte cover and species richness and changed the macrophyte species composition. The effect of the macroalgae removal was immediate with the highest species richness occurrence during the first post-treatment monitoring, but the positive effect vanished with time. In contrast, preventing herbivore access had a gradual but long-lasting effect and reached a more steady-state over time. Two of the most common species showed contrasting responses, the palatable Potamogeton pectinatus was most supported by caging, while the distasteful Myriophyllum spicatum preferred open plots. Our findings may be applicable during the revitalisation of aquatic ecosystems that aims to increase macrophyte biodiversity.

Responses of aquatic macrophytes to anthropogenic pressures: comparison between macrophyte metrics and indices.

Macrophyte responses to anthropogenic pressures in two rivers of Central Spain were assessed to check if simple metrics can exhibit a greater discriminatory and explanatory power than complex indices at small spatial scales. Field surveys were undertaken during the summer of 2014 (Duraton River) and the spring of 2015 (Tajuña River). Aquatic macrophytes were sampled using a sampling square (45 × 45 cm). In the middle Duraton River, macrophytes responded positively to the presence of a hydropower dam and a small weir, with Myriophyllum spicatum and Potamogeton pectinatus being relatively favored. Index of Macrophytes (IM) was better than Macroscopic Aquatic Vegetation Index (MAVI) and Fluvial Macrophyte Index (FMI) in detecting these responses, showing positive and significant correlations with total coverage, species richness, and species diversity. In the upper Tajuña River, macrophytes responded both negatively and positively to the occurrence of a trout farm effluent and a small weir, with Leptodictyum riparium and Veronica anagallis-aquatica being relatively favored. Although IM, MAVI, and FMI detected both negative and positive responses, correlations of IM with total coverage, species richness, and species diversity were higher. Species evenness was not sensitive enough to detect either positive or negative responses of aquatic macrophytes along the study areas. Overall, traditional and simple metrics (species composition, total coverage, species richness, species diversity) exhibited a greater discriminatory and explanatory power than more recent and complex indices (IM, MAVI, FMI) when assessing responses of aquatic macrophytes to anthropogenic pressures at impacted specific sites.

Morphological and biomechanical response to eutrophication and hydrodynamic stresses.

Eutrophication and hydrodynamics determine the final distribution patterns of aquatic macrophytes; however, there is limited available knowledge regarding their interactive effects. Morphological and biomechanical responses to eutrophication and hydrodynamic stresses were assessed by sampling five abundant and dominant species, Potamogeton maackianus, P. pectinatus, P. lucens, Ceratophyllum demersum and Myriophyllum spicatum, in three macrophyte beds in Lake Erhai, Yunnan Province, China: one exposed to eutrophication and moderate southeast (SE) wind; one with mesotrophication, but sheltered by the lakeshore, with weak wind disturbance; and one with meso-eutrophication and strong SE wind. The results showed significant interactive effects of eutrophication and hydrodynamics on most biomechanical traits and some morphological traits, suggesting that aquatic macrophytes preferentially undergo biomechanical adjustments to resist the coexisting eutrophication and hydrodynamic stresses. In particular, hydrodynamics increased both the tensile force and tensile strain of P. maackianus under meso-eutrophication and dramatically decreased them in eutrophic areas, suggesting that eutrophication triggers mechanical failure in this species. Additionally, P. pectinatus, C. demersum and M. spicatum showed the lowest and highest values for the biomechanical variables (greater values for M. spicatum) in the most eutrophic and hydrodynamic areas, respectively, implying that increases in hydrodynamics primarily induce mechanical damage in eutrophic species. The plants generally exhibited greater tensile strain in both shallow and deep waters and the greatest tensile force at moderate depths. The stem cross-sectional area, plant height, stem length, internode length, and branch traits were all responsible for determining the biomechanical variables. This study reveals that hydrodynamic changes primarily induce mechanical damage in eutrophic species, whereas eutrophication triggers mechanical damage in sensitive species.

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.

Enhancement of surface flow constructed wetlands performance at low temperature through seasonal plant collocation.

In the present study, a novel seasonal plant collocation system (SPCS), specifically the Potamogeton crispus and Phragmites australis series system, was investigated to enhance the performance of surface flow constructed wetlands (SFCWs) at low temperature. Results of a year-round experiment showed that SPCS conquered the adverse effect of low temperature and achieved sustainable nutrients removal. In addition, during winter, removal efficiencies of NH4-N, TP, COD, and TN in SPCS were 18.1%, 17.6%, 10.1% and 5.2% higher than that in the control, respectively. P. crispus and P. australis complemented each other in terms of plant growth and plant uptake during the experiment period. Furthermore, it emerged that P. crispus could increase the quantity of ammonia oxidizing bacteria by 10.2%, due to its high oxygen enrichment ability. It is suggested that seasonal plant collocation has a promising future in SFCWs of areas being affected by climate change, e.g. northern China.

Reproduction capacity of Potamogeton crispus fragments and its role in water purification and algae inhibition in eutrophic lakes.

The role of fragments in restoring eutrophic lakes remains unclear despite the importance of re-establishing submerged macrophytes via fragments. This study established a manipulative experiment using different biomass fragments of Potamogeton crispus. This approach was adapted to study the reproductive capacity, nutrient removal efficiency, and algae inhibitory effect of fragments. Results showed that fragments could grow throughout a 49-day experiment by maintaining the stable photosynthesis efficiency of leaves and lengthening the stems. These floating fragments could regenerate by producing turions for the maintenance of their species. Moreover, the increasing removal efficiency of TP, TN, NH4+-N, and NO3--N in water with the increase of fragment biomass indicates that the fragments could effectively purify water quality. Floating fragments competed with algae for nutrients, occupied a favorable ecological niche, and reduced algae biomass. They altered the structure of algae community and shifted the dominated green algae to cyanobacteria, the green algae of phytoplankton, and benthic algae. Findings imply that the postponable regulation of fragments is necessary for the ecological restoration of eutrophic lakes.

Reproductive Allocation in Three Macrophyte Species from Different Lakes with Variable Eutrophic Conditions.

Reproductive allocation is a key process in the plant life cycle and aquatic plants exhibit great diversity in their reproductive systems. In the present study, we conduct a field investigation of three aquatic macrophytes: Stuckenia pectinata, Myriophyllum spicatum, and Potamogeton perfoliatus. Our results showed that widespread species, including S. pectinata and M. spicatum had greater plasticity in their allocation patterns in the form of increased sexual and asexual reproduction, and greater potential to set seeds and increase fitness in more eutrophic environments. P. perfoliatus also exhibited a capacity to adopt varied sexual reproductive strategies such as setting more offspring for the future, although only in clear conditions with low nutrient levels. Our results establish strategies and mechanisms of some species for tolerating and surviving in varied eutrophic lake conditions.

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.

Optimal methods for fitting probability distributions to propagule retention time in studies of zoochorous dispersal.

BACKGROUND: Propagule retention time is a key factor in determining propagule dispersal distance and the shape of "seed shadows". Propagules dispersed by animal vectors are either ingested and retained in the gut until defecation or attached externally to the body until detachment. Retention time is a continuous variable, but it is commonly measured at discrete time points, according to pre-established sampling time-intervals. Although parametric continuous distributions have been widely fitted to these interval-censored data, the performance of different fitting methods has not been evaluated. To investigate the performance of five different fitting methods, we fitted parametric probability distributions to typical discretized retention-time data with known distribution using as data-points either the lower, mid or upper bounds of sampling intervals, as well as the cumulative distribution of observed values (using either maximum likelihood or non-linear least squares for parameter estimation); then compared the estimated and original distributions to assess the accuracy of each method. We also assessed the robustness of these methods to variations in the sampling procedure (sample size and length of sampling time-intervals). RESULTS: Fittings to the cumulative distribution performed better for all types of parametric distributions (lognormal, gamma and Weibull distributions) and were more robust to variations in sample size and sampling time-intervals. These estimated distributions had negligible deviations of up to 0.045 in cumulative probability of retention times (according to the Kolmogorov-Smirnov statistic) in relation to original distributions from which propagule retention time was simulated, supporting the overall accuracy of this fitting method. In contrast, fitting the sampling-interval bounds resulted in greater deviations that ranged from 0.058 to 0.273 in cumulative probability of retention times, which may introduce considerable biases in parameter estimates. CONCLUSIONS: We recommend the use of cumulative probability to fit parametric probability distributions to propagule retention time, specifically using maximum likelihood for parameter estimation. Furthermore, the experimental design for an optimal characterization of unimodal propagule retention time should contemplate at least 500 recovered propagules and sampling time-intervals not larger than the time peak of propagule retrieval, except in the tail of the distribution where broader sampling time-intervals may also produce accurate fits.

Mitochondrial electron transport protects floating leaves of long leaf pondweed (Potamogeton nodosus Poir) against photoinhibition: comparison with submerged leaves.

Investigations were carried to unravel mechanism(s) for higher tolerance of floating over submerged leaves of long leaf pondweed (Potamogeton nodosus Poir) against photoinhibition. Chloroplasts from floating leaves showed ~5- and ~6.4-fold higher Photosystem (PS) I (reduced dichlorophenol-indophenol â†’ methyl viologen â†’ O2) and PS II (H2O â†’ parabenzoquine) activities over those from submerged leaves. The saturating rate (V max) of PS II activity of chloroplasts from floating and submerged leaves reached at ~600 and ~230 µmol photons m(-2) s(-1), respectively. Photosynthetic electron transport rate in floating leaves was over 5-fold higher than in submerged leaves. Further, floating leaves, as compared to submerged leaves, showed higher F v/F m (variable to maximum chlorophyll fluorescence, a reflection of PS II efficiency), as well as a higher potential to withstand photoinhibitory damage by high light (1,200 µmol photons m(-2) s(-1)). Cells of floating leaves had not only higher mitochondria to chloroplast ratio, but also showed many mitochondria in close vicinity of chloroplasts. Electron transport (NADH â†’ O2; succinate â†’ O2) in isolated mitochondria of floating leaves was sensitive to both cyanide (CN(-)) and salicylhydroxamic acid (SHAM), whereas those in submerged leaves were sensitive to CN(-), but virtually insensitive to SHAM, revealing the presence of alternative oxidase in mitochondria of floating, but not of submerged, leaves. Further, the potential of floating leaves to withstand photoinhibitory damage was significantly reduced in the presence of CN(-) and SHAM, individually and in combination. Our experimental results establish that floating leaves possess better photosynthetic efficiency and capacity to withstand photoinhibition compared to submerged leaves; and mitochondria play a pivotal role in protecting photosynthetic machinery of floating leaves against photoinhibition, most likely by oxidation of NAD(P)H and reduction of O2.

Ceratophyllum demersum L. and Potamogeton alpinus Balb. from Iset' river, Ural region, Russia differ in adaptive strategies to heavy metals exposure--a comparative study.

We examined the uptake of five heavy metals (Cu, Fe, Ni, Zn, and Mn) in Ceratophyllum demersum L. (hornwort) and Potamogeton alpinus Balb. (pondweed) from Iset' river, Ural region, Russia. This study was conducted in a territory that is highly urbanized where the surface waters are contaminated by a wide spectrum of pollutants. The environmental situation in this territory drastically deteriorated due to anthropogenic activity. The water quality in most of the water bodies in the Ural region is rather poor. In a comparative study of C. demersum and P. alpinus, differential accumulation pattern was noted for heavy metals (HMs). Higher amounts of HMs accumulated in C. demersum compared to P. alpinus. Also it was shown that in leaves of C. demersum there were high amount of total phosphorus, nitrogen, organics acids and ash; high activity of guaiacol peroxidase; high content of nonenzymatic antioxidants viz., flavonoids, ascorbate, glutathione and proline; high amount of thiols (soluble and membrane bound) compared to P. alpinus.

Testing the stress gradient hypothesis in herbivore communities facilitation peaks at intermediate nutrient levels.

The role of positive interactions in structuring plant and animal communities is increasingly recognized, but the generality of current theoretical models has remained practically unexplored in animal communities. The stress gradient hypothesis predicts a linear increase in the intensity of facilitation as environmental conditions become increasingly stressful, whereas other theoretical models predict a maximum at intermediate environmental stress. We tested how competition and facilitation between herbivores change over a manipulated gradient of nutrient availability. We studied the effect of grazing by pond snails (Lymnaea stagnalis L.) as bulk grazers on aquatic caterpillars (Acentria ephemerella Denis and Schiffermüller) as small specialist grazers along an experimental gradient of environmental nutrient concentration. Higher nutrient levels increased overall total plant biomass but induced a shift toward dominance of filamentous algae at the expense of macrophytes. Facilitation of caterpillars by snail presence peaked at intermediate nutrient levels. Both caterpillar biomass and caterpillar grazing on macrophytes were highest at intermediate nutrient levels. Snails facilitated caterpillars possibly by removing filamentous algae and increasing access to the macrophyte resource, whereas they did not affect macrophyte biomass or C: nutrient ratios, a measure of food quality. We conclude that competition and facilitation in herbivore communities change along nutrient availability gradients that affect plant biomass and community composition. Understanding how interspecific interactions may change in strength and direction along environmental gradients is important to predict how the diversity and structure of communities may respond to the introduction or removal of herbivore species in ecosystems.

The response of epiphytic microbes to habitat and growth status of Potamogeton malaianus Miq. in Lake Taihu.

To investigate the effects of different habitats and plant growth status on abundance, biomass and community structure of epiphytic microbes, Potamogeton malaianus Miq. at two different habitats (Gonghu Bay and East Taihu) in Lake Taihu were collected in June, August and November (corresponding to the period of development of submerged macrophytes). The relative abundance of major epiphytic algae groups was determined with high performance liquid chromatography (HPLC), and the structures and dynamics of epiphytic bacteria were assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis. Results showed that the biomass of epiphytic microbes was not significant difference between the two sites, and the analysis of similarity found no significant intra-lake heterogeneity in community structure, but the temporal heterogeneity of epiphytic microbes was significant, which linked to the growth state of submerged macrophytes and water temperature. The difference in community structure between June and August was larger than that between August and November at each site, indicating that the growth status of submerged macrophytes has a greater impact on the community structure of epiphytic microbes than the seasonal variation of environmental conditions.

Effects of altered estuarine submerged macrophyte bed cover on the omnivorous Cape stumpnose Rhabdosargus holubi.

The ecological importance of submerged macrophyte beds to fishes within estuaries was investigated through the example of the ubiquitous Cape stumpnose Rhabdosargus holubi, an omnivorous, vegetation and estuary-dependent species, using stable-isotope techniques and long-term abundance (catch-per-unit-effort) data from the East Kleinemonde Estuary, South Africa. Outputs from a Bayesian mixing model using δ(13) C and δ(15) N signatures indicated that the submerged macrophytes Ruppia cirrhosa and Potamogeton pectinatus were not a primary source of nutrition for R. holubi, confirming previous work that revealed that macrophytes are consumed but not digested. Long-term seine netting data showed reduced abundance of R. holubi during a prolonged period of macrophyte senescence, suggesting that submerged macrophyte habitats provide shelter that reduces mortality (predation risk) and a food-rich foraging area.

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.

Comparative studies of thermotolerance: different modes of heat acclimation between tolerant and intolerant aquatic plants of the genus Potamogeton.

BACKGROUND AND AIMS: Molecular-based studies of thermotolerance have rarely been performed on wild plants, although this trait is critical for summer survival. Here, we focused on thermotolerance and expression of heat shock transcription factor A2 (HSFA2) and its putative target gene (chloroplast-localized small heat shock protein, CP-sHSP) in two allied aquatic species of the genus Potamogeton (pondweeds) that differ in survival on land. METHODS: The degree of thermotolerance was examined using a chlorophyll bioassay to assess heat injury in plants cultivated under non- and heat-acclimation conditions. Potamogeton HSFA2 and CP-sHSP genes were identified and their heat-induction was quantified by real-time PCR. KEY RESULTS: The inhibition of chlorophyll accumulation after heat stress showed that Potamogeton malaianus had a higher basal thermotolerance and developed acquired thermotolerance, whereas Potamogeton perfoliatus was heat sensitive and unable to acquire thermotolerance. We found two duplicated HSFA2 and CP-sHSP genes in each species. These genes were induced by heat shock in P. malaianus, while one HSFA2a gene was not induced in P. perfoliatus. In non-heat-acclimated plants, transcript levels of HSFA2 and CP-sHSP were transiently elevated after heat shock. In heat-acclimated plants, transcripts were continuously induced during sublethal heat shock in P. malaianus, but not in P. perfoliatus. Instead, the minimum threshold temperature for heat induction of the CP-sHSP genes was elevated in P. perfoliatus. CONCLUSIONS: Our comparative study of thermotolerance showed that heat acclimation leads to species-specific changes in heat response. The development of acquired thermotolerance is beneficial for survival at extreme temperatures. However, the loss of acquired thermotolerance and plasticity in the minimum threshold temperature of heat response may be favourable for plants growing in moderate habitats with limited daily and seasonal temperature fluctuations.