Antibiotic and heavy metal pollution of aquatic environments are issues of serious concern, and the macrophyte Myriophyllum aquaticum may provide a viable solution for the removal of these contaminants. However, the toxic effects of coexisting tetracyclines (TCs) and Cu(II) on this plant species are currently unclear. In the present study, we constructed wetland microcosms planted with M. aquaticum and spiked these with three TCs (tetracycline, oxytetracycline, and chlortetracycline) and Cu(II) at concentrations ranging from 100 to 10,000 μg/L to investigate how Cu(II) influences the growth and tolerance responses of plants to TCs. After 12 weeks, we found that TCs had accumulated in the plants, and that plant growth and characteristics were significantly affected by the levels of both TCs and Cu(II). While low Cu(II) levels had a synergistic effect on the accumulation of TCs, high levels were observed to reduce accumulation. However, low levels of TCs and Cu(II) had a hormesis effect on plant growth, with plant biomass and leaf chlorophyll content decreasing and the malondialdehyde content and activities of antioxidant enzymes gradually increasing with an increase in TC dosage. The coexistence of low levels of Cu(II) was, however, found to alleviate these adverse effects. Principal component analysis revealed a close relationship among plant biomass, chlorophyll content, malondialdehyde content, and antioxidant enzyme activities. Considering that the Cu/TC ratio was shown to markedly affect M. aquaticum growth, the respective proportions of these pollutants should be taken into consideration in the future design of constructed wetlands.

In this study, the tips of Myriophyllum aquaticum (M. aquaticum) plants were planted in open-top plastic bins and treated by simulated wastewater with various ammonium-N concentrations for three weeks. The contents of related carbohydrates and key enzyme activities of carbon metabolism were measured, and the mechanisms of carbon metabolism regulation of the ammonia tolerant plant M. aquaticum under different ammonium-N levels were investigated. The decrease in total nonstructural carbohydrates, soluble sugars, sucrose, fructose, reducing sugar and starch content of M. aquaticum were induced after treatment with ammonium-N during the entire stress process. This finding showed that M. aquaticum consumed a lot of carbohydrates to provide energy during the detoxification process of ammonia nitrogen. Moreover, ammonia-N treatment led to the increase in the activitives of invertase (INV) and sucrose synthase (SS), which contributed to breaking down more sucrose to provide substance and energy for plant cells. Meanwhile, the sucrose phosphate synthase (SPS) activity was also enhanced under stress of high concentrations of ammonium-N, especially on day 21. The result indicated that under high-concentration ammonium-N stress, SPS activity can be significantly stimulated by regulating carbon metabolism of M. aquaticum, thereby accumulating sucrose in the plant body. Taken together, M. aquaticum can regulate the transformation of related carbohydrates in vivo by highly efficient expression of INV, SPS and SS, and effectively regulate the osmotic potential, thereby delaying the toxicity of ammonia nitrogen and improving the resistance to stress. It is very important to study carbon metabolism under ammonia stress to understand the ammonia nitrogen tolerance mechanism of M. aquaticum.

Due to a close contact with water column, submerged macrophytes are easily disturbed by environment change in freshwater ecosystems, especially at the seedling stage. In recent decades, freshwater ecosystems have been subject to severe cadmium (Cd) pollution, which can cause toxic effects on the growth of submerged macrophytes. Moreover, the temperature rise resulting from climate warming and water level decline may further aggravate such effect, especially in shallow lakes. Here, we investigated the independent and interaction effects of Cd exposure levels (0, 0.5, 1, and 2.5 mg L⁻¹) and temperature (15, 25, and 30 °C) on morphological and physiological traits of Myriophyllum aquaticum (Vell.) Verd. Seedlings generated from propagules and seeds. The temperature rise and Cd exposure generally resulted in a significant increase of Cd concentrations and antioxidant enzyme activities in leaves, as well as a decrease of chlorophyll a and b concentrations. The number and length of leaves generated from propagules always show a downward trend with the increase of Cd exposure, regardless of the temperature. Moreover, the lowest leaf number and length always occurred at high temperature (i.e. 30 °C) when the Cd exposure level increased to 1 and 2.5 mg L⁻¹. For the seedlings generated from seeds, the temperature rise caused an increase of leaf emergence rate under low Cd exposure levels, but resulted in a significant decrease with the Cd exposure level. This study indicates the negative effects of Cd exposure and temperature rise on submerged macrophytes at the seedling stage, and highlights that temperature rise would enhance Cd toxicity.

One active ingredient can be a component of different types of formulations of pesticides, while the toxicity of its formulations may vary depending on various constituents used in the mixture. The present study focuses on evaluating the effects of the active ingredient clomazone and its formulations (Rampa® EC and GAT Cenit 36 CS, both containing 360 g a.i./l of clomazone) on non-target aquatic macrophytes. The two formulation types differ in their active ingredient release and presumed environmental impact. In order to cover different ecological traits, two species of aquatic macrophytes – the floating monocot Lemna minor and the rooted dicot Myriophyllum aquaticum, were used as test models. The results of this study revealed differences in the sensitivity of tested plants to clomazone. Based on the most sensitive parameters, M. aquaticum proved to be more sensitive than L. minor to the technical ingredient and both formulations. The species sensitivity distribution (SSD) approach that was tried out in an attempt to create a higher tier step of risk assessment of clomazone for primary producers indicates that tests on rooted macrophytes can add value in risk assessment of plant protection products. The capsule formulation of clomazone was less toxic than the emulsion for L. minor, but more toxic for M. aquaticum. The most toxic for L. minor was the emulsifiable concentrate formulation Rampa® EC, followed by technical clomazone (EC₅₀ 33.3 and 54.0 mg a.i./l, respectively), while the aqueous capsule suspension formulation GAT Cenit 36 CS did not cause adverse effects. On the other hand, the most toxic for M. aquaticum was the formulation GAT Cenit 36 CS, followed by technical clomazone and the formulation Rampa® EC, demonstrating a greater effect of the capsule formulation.

Aquatic macrophytes play a significant role in nutrients removal in constructed wetlands, yet nutrients could be re-released due to plant debris decomposition. In this study, Myriophyllum aquaticum was used as a model plant debris and three debris biomass levels of 3 g, 9 g dry biomass, and 20 g fresh biomass (D3, D9, and F20, respectively) were used to simulate 120-d plant debris decomposition in a sediment-water system. The biomass first-order decomposition rate constants of D3, D9, and F20 treatments were 0.0058, 0.0117, and 0.0201 d⁻¹, respectively with no significant difference of decomposition rate among three mass groups (p > 0.05). Plant debris decomposition decreased nitrate and total nitrogen concentrations but increased ammonium, organic nitrogen, and dissolved organic carbon (DOC) concentrations in overlying water. The parallel factor analysis confirms that three components of DOC in overlying water changed over decomposition time. Emission fluxes of methane and nitrous oxide in the plant debris treatments were several to thousands of times higher than the control group within the initial 0–45 d, which was mainly attributed to DOC released from the plant debris. Plant debris decomposition can affect the gas emission fluxes for relatively shorter time (30–60 d) than water quality (>120 d). The 16S rRNA, nirK, nirS and hazA gene abundance increased in the early stage for plant debris treatments, and then decreased to the end of 120-d incubation time while ammonia monooxygenase α-subunit A gene abundance of ammonia-oxidizing archaea and bacteria had no large variations during the entire decay time compared with no plant debris treatment. The results demonstrate that decomposition of M. aquaticum debris could affect greenhouse gas emission fluxes and microbial gene abundance in the sediment-water system besides overlying water quality.

In the greenhouse and container nursery production industry there is potential for runoff of nitrogen (N) and phosphorus (P), which may contaminate surface and groundwater. Since the 1950s constructed wetlands (CWs), as a simple, low-technology method, have been shown to effectively treat agricultural, industrial, and municipal wastewater. We investigated the N and P attenuating potential of three floating hydrophytes planted in a laboratory-scale subsurface flow (SSF) CW system. Over an 8-week period plants were supplied with N and P (0.39 to 36.81 mg·L⁻¹ N and 0.07 to 6.77 mg·L⁻¹ P) that spanned the rates detected in nursery runoff between the discharge and inflow locations of a commercial nursery currently employing CWs. Whole plant dry weight was positively correlated with N and P supplied. Highest N recovery rates were exhibited by water hyacinth (Eichhornia crassipes [Mart.] Solms.) and water lettuce (Pistia stratiotes L.). P recovery rates were similar for water hyacinth, water lettuce, and dwarf redstemmed parrotfeather (Myriophyllum aquaticum [Vell.] Verdc.). These floating hydrophytes can be cultivated in a SSF CW to remediate runoff losses of N and P. The possibility exists for integrating them into a polycultural remediation system that includes emergent aquatic macrophytes for processing and polishing nursery/greenhouse wastewater.

In this work, we evaluated whether the species Myriophyllum aquaticum (Vell.) Verdc. can be a promising material for devising reliable eco-toxicological tests for Cd-contaminated waters. Plants of M. aquaticum were exposed to Cd, using different concentrations (1 mg L⁻¹, 2.5 mg L⁻¹, 5 mg L⁻¹, and 10 mg L⁻¹; experiment 1) and exposure times (2.5 mg L⁻¹ for 3 days, 7 days, 14 days, and 21 days; experiment 2). Plant growth and Cd accumulation were monitored during the treatment period, and Cd genotoxicity was assessed by analyzing Cd-induced changes in the AFLP fingerprinting profiles using famEcoRI₍TAC₎/MseI₍ATG₎ and hexEcoRI₍ACG₎/MseI₍ATG₎ pairs of primers. Root and shoot growth was reduced already at the lowest Cd concentration used (about 20% reduction for roots and 60% for shoots at 1 mg L⁻¹; experiment 1) and after 7 days (about 50% reduction for roots and 70% for shoots; experiment 2). The primer combinations produced 154 and 191 polymorphic loci for experiments 1 and 2, respectively. Mean genetic diversity (He) reduction among the treatment groups was observed starting from 2.5 mg L⁻¹ (He 0.211 treated vs 0.236 control; experiment 1) and after 3 days (He 0.169 treated vs 0.261 control; experiment 2), indicating that results obtained from AFLP profiles did not match with plant growth measurements. Therefore, our results showed that M. aquaticum proved to be a suitable model system for the investigation of Cd genotoxicity through AFLP fingerprinting profile, whereas the more classic eco-toxicological tests based only on biometric parameters could not correctly estimate the risk associated with undetected Cd genotoxicity.

Feeding a growing population requires striking a balance between increasing production and decreasing environmental impacts in agricultural settings. We established 12 experimental mesocosms with silt loam atop a base of sand and examined the ability of three emergent aquatic plants common to the USA to remediate pesticides and nutrients in agricultural runoff. Mesocosms were planted in monocultures of Myriophyllum aquaticum, Polygonum amphibium, and Typha latifolia, or left unvegetated to serve as controls. All mesocosms were amended with target concentrations of 10 mg L⁻¹ (each) nitrate, ammonium, and orthophosphate; 20 μg L⁻¹ (each) of the pesticides propanil and clomazone; and 10 μg L⁻¹ of the pesticide cyfluthrin. After a 6-h-simulated agricultural runoff with amended water, mesocosms sat idle for 48 h before flushing with unamended water for another 6 h. Outflow water samples were collected and analyzed for contaminant concentrations. Most significant differences between vegetated mesocosms and controls occurred when comparing mean contaminant transfer/transformation rates post-amendment. Differences among plant species occurred regarding retention of dissolved nutrients orthophosphate, ammonium, and nitrate. Similarly, all three plant species retained more propanil than controls during post-amendment (8–48 h), but individual plant differences occurred with regard to clomazone and cyfluthrin retention. While variation in mitigation of specific dissolved components of nutrients suggests different mechanisms involved in nutrient cycling within our mesocosms, consistent overall total nutrient and pesticide reduction during the post-amendment period indicate that holding runoff in vegetated ditches may reduce transport of agricultural contaminants to downstream aquatic ecosystems.

Constructed wetlands (CWs) cultivated with Myriophyllum aquaticum showed great potential for total nitrogen (TN) removal from aquatic ecosystems in previous studies. To evaluate the growth characteristics, photosynthetic pigment content, and antioxidative responses of M. aquaticum, as well as its TN removal efficiency in CWs, M. aquaticum was treated with different levels of ammonium (NH₄⁺) and nitrate (NO₃⁻) for 28 days. The results indicated that M. aquaticum had strong nitrogen stress tolerance and was more likely to be suppressed by high levels of NH₄⁺ than NO₃⁻. High levels of NH₄⁺ also led to inhibition of synthesis of photosynthetic pigments and increased peroxidase activity in plant leaves, which was not found in the NO₃⁻ treatments. High levels of both NH₄⁺ and NO₃⁻ generated obvious oxidative stress through elevation of malondialdehyde content while decreasing superoxide dismutase activity in the early stage. A sustainable increase of TN removal efficiency in most of the CWs indicated that M. aquaticum was a candidate species for treating wastewater with high levels of nitrogen because of its higher tolerance for NH₄⁺ and NO₃⁻ stress. However, the increase of TN removal efficiency was hindered in the late stage when treated with high levels of NH₄⁺ of 26 and 36 mmol/L, indicating that its tolerance to NH₄⁺ stress might have a threshold. The results of this study will enrich the studies on detoxification of high ammonium ion content in NH₄⁺-tolerant submerged plants and supply valuable reference data for proper vegetation of M. aquaticum in CWs.

Submerged macrophytes have been found to be promising in removing cadmium (Cd) from aquatic ecosystems; however, the mechanism of Cd detoxification in these plants is still poorly understood. In the present study, Cd chemical forms and subcellular distributing behaviors in Myriophyllum aquaticum and the physiological mechanism underlying M. aquaticum in response to Cd stress were explored. During the study, M. aquaticum was grown in a hydroponic system and was treated under different concentrations of Cd (0, 0.01, 0.05, 0.25, and 1.25 mg/L) for 14 days. The differential centrifugation suggested that most Cd was split in the soluble fraction (57.40–66.25%) and bound to the cell wall (24.92–38.57%). Furthermore, Cd in M. aquaticum was primarily present in NaCl-extractable Cd (51.76–91.15% in leaves and 58.71–84.76% in stems), followed by acetic acid–extractable Cd (5.17–22.42% in leaves and 9.54–16.56% in stems) and HCl-extractable Cd (0.80–12.23% in leaves and 3.56–18.87% in stems). The malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations in M. aquaticum were noticeably increased under each Cd concentration. The activities of catalase (CAT), guaiacol peroxidase (POD), and superoxide dismutase (SOD) in leaves were initially increased under relatively low concentrations of Cd but were decreased further with the increasing concentrations of Cd. The ascorbate (AsA), glutathione (GSH), and nitric oxide (NO) concentrations in stems increased with increasing Cd concentrations. Taken together, our results indicate that M. aquaticum can be used successfully for phytoremediation of Cd-contaminated water, and the detoxification mechanisms in M. aquaticum include enzymatic and non-enzymatic antioxidants, subcellular partitioning, and the formation of different chemical forms of Cd.