Analysis of diversity and function of epiphytic bacterial communities associated with macrophytes using a metagenomic approach.

Epiphytic bacteria constitute a vital component of aquatic ecosystems, pivotal in regulating elemental cycling. Despite their significance, the diversity and functions of epiphytic bacterial communities adhering to various submerged macrophytes remain largely unexplored. In this study, we employed a metagenomic approach to investigate the diversity and function of epiphytic bacterial communities associated with six submerged macrophytes: Ceratophyllum demersum, Hydrilla verticillata, Myriophyllum verticillatum, Potamogeton lucens, Stuckenia pectinata, and Najas marina. The results revealed that the predominant epiphytic bacterial species for each plant type included Pseudomonas spp., Microbacterium spp., and Stenotrophomonas rhizophila. Multiple comparisons and linear discriminant analysis effect size indicated a significant divergence in the community composition of epiphytic bacteria among the six submerged macrophytes, with 0.3-1% of species uniquely identified. Epiphytic bacterial richness associated with S. pectinata significantly differed from that of both C. demersum and H. verticillata, although no significant differences were observed in diversity and evenness. Functionally, notable variations were observed in the relative abundances of genes associated with carbon, nitrogen, and phosphorus cycling within epiphytic bacterial communities on the submerged macrophyte hosts. Among these communities, H. verticillata exhibited enrichment in genes related to the 3-hydroxypropionate bicycle and nitrogen assimilation, translocation, and denitrification. Conversely, M. verticillatum showcased enrichment in genes linked to the reductive citric acid cycle (Arnon-Buchanan cycle), reductive pentose phosphate cycle (Calvin cycle), polyphosphate degradation, and organic nitrogen metabolism. In summary, our findings offer valuable insights into the diversity and function of epiphytic bacteria on submerged macrophyte leaves, shedding light on their roles in lake ecosystems.

Epiphytic microorganisms of submerged macrophytes effectively contribute to nitrogen removal.

Submerged macrophytes play important roles in nutrient cycling and are widely used in ecological restoration to alleviate eutrophication and improve water quality in lakes. Epiphytic microbial communities on leaves of submerged macrophytes might promote nitrogen cycling, but the mechanisms and quantification of their contributions remain unclear. Here, four types of field zones with different nutrient levels and submerged macrophytes, eutrophic + Vallisneria natans (EV), eutrophic + V. natans + Hydrilla verticillata, mesotrophic + V. natans + H. verticillata, and eutrophic without macrophytes were selected to investigate the microbial communities that involved in nitrification and denitrification. The alpha diversity of bacterial community was higher in the phyllosphere than in the water, and that of H. verticillata was higher compared to V. natans. Bacterial community structures differed significantly between the four zones. The highest relative abundance of dominant bacterioplankton genera involved in nitrification and denitrification was observed in the EV zone. Similarly, the alpha diversity of the epiphytic ammonia-oxidizing archaea and nosZI-type denitrifiers were highest in the EV zone. Consist with the diversity patterns, the potential denitrification rates were higher in the phyllosphere than those in the water. Higher potential denitrification rates in the phyllosphere were also found in H. verticillata than those in V. natans. Anammox was not detected in all samples. Nutrient loads, especially nitrogen concentrations were important factors influencing potential nitrification, denitrification rates, and bacterial communities, especially for the epiphytic nosZI-type taxa. Overall, we observed that the phyllosphere harbors more microbes and promotes higher denitrification rates compared to water, and epiphytic bacterial communities are shaped by nitrogen nutrients and macrophyte species, indicating that epiphytic microorganisms of submerged macrophytes can effectively contribute to the N removal in shallow lakes.

Evolution and distribution of antibiotic resistance genes in submerged macrophytes and biofilm systems: From seasonal monitoring to mesocosm experiments.

The aquatic ecosystem has been extensively investigated as a hotspot for the spread of antibiotic resistance genes (ARGs); yet, the evolution and distribution of ARGs profiles in submerged macrophytes biofilms and surrounding water remained unclear. In this study, the dynamic distribution and seasonal variations of microbial communities and ARGs profiles were investigated, alongside their assembly processes and mutual interactions. Bacitracin and multidrug resistance genes were predominant, constituting more than 60% of the total ARGs abundance. The deterministic processes (<65%), influenced by the physicochemical properties of the river environment, governed the assembly and composition of ARGs profiles, exhibiting significant seasonal variation. The peak diversity (21 types) and abundance (0.316 copy ratios) of ARGs were detected during the summer. Proteobacteria and Actinobacteria were the dominant bacterial phyla, accounting for 38.41-85.50% and 4.03-27.09% of the microbial community, respectively. Furthermore, Proteobacteria, especially genera such as Acinetobacter, Burkholderia, and Pseudomonas, with various resistance sequences, were the primary carriers of multiple ARGs. Notably, the genetic exchanges between biofilms and surrounding water facilitated the further propagation of high-risk ARGs, posing greater ecological risks. Redundancy analysis indicated that the total nitrogen and temperature in water determined the fate of pathogenic-resistant species. These findings provided theoretical support for the mitigation of ARGs contamination in aquatic environments.

Eutrophication decreases ecological resilience by reducing species diversity and altering functional traits of submerged macrophytes.

Positive feedback is key to producing alternative stable states and largely determines ecological resilience in response to external perturbations. Understanding the positive feedback mechanisms in macrophyte-dominated lakes is crucial for resilience-based management and restoration. Based on the field investigation of submerged macrophyte communities in 35 lakes in China, we found that morphological complexity (MC) and morphological plasticity (MP) are correlated with the stoichiometric homeostasis of phosphorus (HP ) and are related to ecosystem structure, functioning, and stability. We also found that the positive feedback strength of lakes dominated by macrophytes is biomass- and diversity-dependent. Eutrophication can decrease the community biomass by decreasing community MC, MP, and HP and the species diversity through low-light availability, ultimately decreasing the positive feedback strength and resilience of clear water states. We argue that functional traits and species diversity should be considered to build more resilient ecosystems in future changing environment scenarios.

Temporal Succession of Bacterial Community Structure, Co-occurrence Patterns, and Community Assembly Process in Epiphytic Biofilms of Submerged Plants in a Plateau Lake.

In shallow macrophytic lakes, epiphytic biofilms are formed on the surface of submerged plant stems and leaves because of algae and bacterial accumulation. Epiphytic biofilms significantly impact the health of the host vegetation and the biogeochemical cycling of lake elements. However, community diversity, species interactions, and community assembly mechanisms in epiphytic bacterial communities (EBCs) of plants during different growth periods are not well understood. We investigated the successional dynamics, co-occurrence patterns, and community assembly processes of epiphytic biofilm bacterial communities of submerged plants, Najas marina and Potamogeton lucens, from July to November 2020. The results showed a significant seasonal variation in EBC diversity and richness. Community diversity and richness increased from July to November, and the temperature was the most important driving factor for predicting seasonal changes in EBC community structure. Co-occurrence network analysis revealed that the average degree and graph density of the network increased from July to November, indicating that the complexity of the EBC network increased. The bacterial community co-occurrence network was limited by temperature, pH, and transparency. The phylogeny-based null model analysis showed that deterministic processes dominated the microbial community assembly in different periods, increasing their contribution. In addition, we found that as the dominance of deterministic processes increased, the microbial co-occurrence links increased, and the potential interrelationships between species became stronger. Thus, the findings provide insights into the seasonal variability of EBC assemblage and co-occurrence patterns in lacustrine ecosystems.

Responses of sediment nitrogen forms and bacterial communities to different aquatic nitrogen conditions in three submerged macrophyte-type ecological treatment systems.

Ecological treatment system (ETS) has been recognized as a promising technology for mitigating agricultural non-point pollution, whereas it remains to be seen how nitrogen (N) forms and bacterial communities in ETS sediments respond to different aquatic N conditions. Therefore, a four-month microcosm experiment was conducted to investigate the effects of three aquatic N conditions (2 mg/L NH4+-N, 2 mg/L NO3--N and 1 mg/L NH4+-N + 1 mg/L NO3--N) on sediment N forms and bacterial communities in three ETSs vegetated by Potamogeton malaianus, Vallisneria natans and artificial aquatic plant, respectively. Through analysis of four transferable N fractions, the valence states of N in ion-exchange and weak acid extractable fractions were found to be mainly determined by aquatic N conditions, while significant N accumulation was observed only in strong oxidant extractable and strong alkali extractable fractions. Sediment N profiles were primarily influenced by time and plant types, with N condition having secondary effect, while sediment bacterial community structures experienced a significant shift over time and were slightly influenced by plant types. Sediment functional genes related to N fixation, nitrification, assimilable nitrate reduction, dissimilatory nitrite reduction (DNRA) and denitrification were substantially enriched in month 4, and the bacterial co-occurrence network exhibited less complexity but more stability under NO3- condition compared to others. Furthermore, certain sediment N fractions were found to have strong relationships with specific sediment bacteria, such as nitrifiers, denitrifiers and DNRA bacteria. Our findings highlight the significant influence of aquatic N condition in submerged macrophyte-type ETSs on sediment N forms and bacterial communities.

Response of dissolved organic matter (DOM) and microbial community to submerged macrophytes restoration in lakes: A review.

Microorganisms play a crucial role in the biogeochemical processes of Dissolved Organic Matter (DOM), and the properties of DOM also significantly influence changes in microbial community characteristics. This interdependent relationship is vital for the flow of matter and energy within aquatic ecosystems. The presence, growth state, and community characteristics of submerged macrophytes determine the susceptibility of lakes to eutrophication, and restoring a healthy submerged macrophyte community is an effective way to address this issue. However, the transition from eutrophic lakes dominated by planktic algae to medium or low trophic lakes dominated by submerged macrophytes involves significant changes. Changes in aquatic vegetation have greatly affected the source, composition, and bioavailability of DOM. The adsorption and fixation functions of submerged macrophytes determine the migration and storage of DOM and other substances from water to sediment. Submerged macrophytes regulate the characteristics and distribution of microbial communities by controlling the distribution of carbon sources and nutrients in the lake. They further affect the characteristics of the microbial community in the lake environment through their unique epiphytic microorganisms. The unique process of submerged macrophyte recession or restoration can alter the DOM-microbial interaction pattern in lakes through its dual effects on DOM and microbial commu-----nities, ultimately changing the stability of carbon and mineralization pathways in lakes, such as the release of methane and other greenhouse gases. This review provides a fresh perspective on the dynamic changes of DOM and the role of the microbiome in the future of lake ecosystems.

Diatom community composition on submerged macrophyte species from an Ontario (Canada) lake.

The introduction of invasive macrophyte species can affect submerged macrophyte community composition and abundance, which in turn can alter the functions of lake ecosystems. Knowing when and how invasive macrophytes arrive and spread can help disentangle the effects of invasive species from other stressors on lake ecosystems. This requires a long-term (decades) perspective of macrophyte community composition, which is rarely available. An alternative is paleolimnological inferences of macrophyte community composition from fossil diatom assemblages, which requires knowledge of epiphytic diatom communities. Here, we investigated the epiphytic diatom community composition of three common submerged macrophyte species (Chara sp., Potamogeton robbinsii, and the invasive Myriophyllum spicatum) in a typical temperate, mixed forest lake, Chandos Lake, Ontario, Canada, to provide a basis for future paleolimnological research. Non-parametric, multivariate analysis of variance indicated a statistically significant difference in the epiphytic diatom communities of different macrophyte species, despite principal components analysis showing some overlap among the diatom communities. Diatom community composition of all macrophytes had abundant Achnanthidium minutissimum and Cocconeis placentula. Generalized linear models and univariate analysis of variance identified six diatoms (Encyonopsis microcephala, Epithemia turgida, Gomphonema parvulius, Navicula gerloffi, Rhopalodia gibba, and Rossithidium anastasiae) that were significantly different among macrophyte species. Although it remains uncertain whether these differences are sufficient to infer historical macrophyte community composition from epiphytic diatom fossil assemblages, our results indicate the potential of such an approach and offer suggestions for future research.

Unraveling the ecological mechanisms of bacterial succession in epiphytic biofilms on Vallisneria natans and Hydrilla verticillata during bioremediation of phenanthrene and pyrene polluted wetland.

In wetland ecosystem, the microbial succession in epiphytic biofilms of submerged macrophytes remains to be fully elucidated, especially submerged macrophytes used to remediate organic pollutants contaminated sediment. Herein, 16 S rRNA gene sequencing was used to investigate the bacterial dynamics and ecological processes in the biofilms of two typical submerged macrophytes (Vallisneria natans and Hydrilla verticillata) settled in sediment polluted by polycyclic aromatic hydrocarbons (PAHs) at two growth periods. The results presented that the variations of bacterial community in the biofilms were influenced by attached surfaces (explanation ratio: 17.30%), incubation time (32.30%) and environmental factors (39.10%). Bacterial community assembly was mainly driven by dispersal limitation which triggered more positive co-occurrence associations in microbial networks, maintaining ecological stability in the process of bioremediation of PAHs. Additionally, the functional redundancy strength of bacterial community was more affected by attached surface than incubation time. The structural equation model illustrated that community assembly drove ß-diversity and explained a part of ecological functions. Environmental factors, community assembly, and ß-diversity jointly affected microbial networks. Overall, our study offers new insights into the microbial ecology in biofilms attached on the submerged macrophytes settled in PAH-polluted sediment, providing important information for deeply understanding submerged macrophyte-biofilm complex and promoting sustainable phytoremediation in shallow lacustrine and marshy ecosystems.

Transcriptomic profiling of atrazine phytotoxicity and comparative study of atrazine uptake, movement, and metabolism in Potamogeton crispus and Myriophyllum spicatum.

The accumulation of atrazine in sediments raises wide concern due to its potential negative effects on aquatic environments. Here we collected sediments and different submerged macrophytes to simulate natural shallow lakes and to measure atrazine levels and submerged macrophyte biomass. We determined gene expressions in submerged macrophytes treated with or without atrazine. We also examined atrazine concentrations and its metabolite structures in submerged macrophytes. When the initial concentration of atrazine in sediments ranged from 0.1 to 2.0 mg kg-1 dry weight (DW), atrazine levels in the pore water of the sediments ranged from 0.003 to 0.05 mg L-1 in 90 days. Atrazine did not show obvious long-term effects on the biomass of Potamogeton crispus and Myriophyllum spicatum (P > 0.05). On day 90, gene expressions related to cell wall in P. crispus were changed by atrazine phytotoxicity. Moreover, the decrease in the number genes controlling light-harvesting chlorophyll a/b-binding proteins verified the toxic effects of atrazine on the photosynthesis of M. spicatum. Compared with unexposed plants on day 90, ribosome pathway was significantly enriched with differentially expressed genes after submerged macrophytes were exposed to 2.0 mg kg-1 DW atrazine (P < 0.05). In addition, shoots and roots of P. crispus and M. spicatum could absorb the equal amount of atrazine (P > 0.05). Once absorbed by submerged macrophytes, atrazine was degraded into 1-hydroxyisopropylatrazine, hydroxyatrazine, deethylatrazine, didealkylatrazine, cyanuric acid, and biuret, and some of its metabolites could conjugate with organic acids, cysteinyl ß-alanine, and glucose. This study establishes a foundation for aquatic ecological risk assessments and the phytoremediation of atrazine in sediments.

Shallow plant-dominated lakes - extreme environmental variability, carbon cycling and ecological species challenges.

BACKGROUND: Submerged plants composed of charophytes (green algae) and angiosperms develop dense vegetation in small, shallow lakes and in littoral zones of large lakes. Many small, oligotrophic plant species have declined due to drainage and fertilization of lakes, while some tall, eutrophic species have increased. Although plant distribution has been thoroughly studied, the physiochemical dynamics and biological challenges in plant-dominated lakes have been grossly understudied, even though they may offer the key to species persistence. SCOPE: Small plant-dominated lakes function as natural field laboratories with eco-physiological processes in dense vegetation dictating extreme environmental variability, intensive photosynthesis and carbon cycling. Those processes can be quantified on a whole lake basis at high temporal resolution by continuously operating sensors for light, temperature, oxygen, etc. We explore this hitherto hidden world. CONCLUSIONS: Dense plant canopies attenuate light and wind-driven turbulence and generate separation between warm surface water and colder bottom waters. Daytime vertical stratification becomes particularly strong in dense charophyte vegetation, but stratification is a common feature in small, shallow lakes also without plants. Surface cooling at night induces mixing of the water column. Daytime stratification in plant stands may induce hypoxia or anoxia in dark bottom waters by respiration, while surface waters develop oxygen supersaturation by photosynthesis. Intensive photosynthesis and calcification in shallow charophyte lakes depletes dissolved inorganic carbon (DIC) in surface waters, whereas DIC is replenished by respiration and carbonate dissolution in bottom waters and returned to surface waters before sunrise. Extreme diel changes in temperature, DIC and oxygen in dense vegetation can induce extensive rhythmicity of photosynthesis and respiration and become a severe challenge to the survival of organisms. Large phosphorus pools are bound in plant tissue and carbonate precipitates. Future studies should test the importance of this phosphorus sink for ecosystem processes and competition between phytoplankton and plants.

Bacterial Communities and Their Predicted Functions Explain the Sediment Nitrogen Changes Along with Submerged Macrophyte Restoration.

Submerged vegetation biomass fluctuation usually occurs during the preliminary stage of vegetation restoration in shallow lakes, which impacts the final status and duration for achieving a macrophyte-dominant state. This study uncovered the sediment N characteristics and the sediment bacterial community and their predicted functions during the preliminary stage of vegetation recovery in the West Lake, a typical subtropical degenerated shallow lake in China. Results showed increased amounts of sediment TN and NH4-N, reaching 3425.76 and 345.5 mg kg-1, respectively, when the vegetation biomass decreased from its maximum to its minimum. The maximum concentration of sediment NH4-N reached 508.60 mg kg-1 with the decline in vegetation, which might restrict further growth of the submerged macrophytes. The bacterial community structure during the high macrophyte biomass (HMB) period was distinct from that observed during the low macrophyte biomass (LMB) period. Specific taxa such as the phyla Chloroflexi and Acidobacteria and the genus Anaerolineaceae that are related to organic carbon degradation were significantly higher during the LMB period. Potential denitrifiers, such as Lactococcus and Bacillus genera decreased during the LMB period. Accumulation of sediment ammonia could be attributed to the enhanced production by assimilatory nitrate reduction, organic N degradation, and/or the decreased consumption by nitrification. Our findings highlight that the unstable preliminary stage of vegetation restoration brings drastic fluctuation of sediment N loading, of which NH4-N accumulation caused by bacterial communities prevents further growth of the submerged macrophytes. Therefore, extra management measures for the vegetation recovery areas should be taken to avoid excess NH4-N accumulation in sediments.

Heavy metals contamination and accumulation in submerged macrophytes in an urban river in China.

Deteriorating urban water quality has attracted considerable attention in China. We investigated the contamination levels and distribution of heavy metals (As, Cd, Cu, Ni, Pb, and Zn) in Yuxi River water and sediments, and assessed the heavy metal accumulation capability of five species of submerged macrophytes: Vallisneria natans (Lour.) Hara, Potamogeton pectinatus L., Hydrilla verticillata (L. f.) Royle, Myriophyllum spicatum L., and Potamogeton crispus L. Samples were collected from upstream and downstream locations in different season. The results showed that the levels of heavy metals in the downstream areas were higher than in the upstream areas. Heavy metal concentrations in the river water during the dry seasons were higher than those during the rainy seasons, and the opposite results appeared in sediments and submerged macrophytes. In general, the river was slightly contaminated by heavy metals, and the concentrations of Pb and Ni in this river should serve as a warning, while Cd and Zn pollution in the sediments desperately needs to be removed. Furthermore, Potamogeton pectinatus L. showed a higher accumulation capacity for these metals among the five native submerged macrophytes and could be defined as a hyperaccumulator for Cd. Therefore, the potential use of native aquatic plants in contaminated rivers is worth further exploration.

Climate warming and heat waves affect reproductive strategies and interactions between submerged macrophytes.

Extreme climatic events, such as heat waves, are predicted to increase in frequency and intensity during the next hundred years, which may accelerate shifts in hydrological regimes and submerged macrophyte composition in freshwater ecosystems. Since macrophytes are profound components of aquatic systems, predicting their response to extreme climatic events is crucial for implementation of climate change adaptation strategies. We therefore performed an experiment in 24 outdoor enclosures (400 L) separating the impact of a 4 °C increase in mean temperature with the same increase, that is the same total amount of energy input, but resembling a climate scenario with extreme variability, oscillating between 0 °C and 8 °C above present conditions. We show that at the moderate nutrient conditions provided in our study, neither an increase in mean temperature nor heat waves lead to a shift from a plant-dominated to an algal-dominated system. Instead, we show that species-specific responses to climate change among submerged macrophytes may critically influence species composition and thereby ecosystem functioning. Our results also imply that more fluctuating temperatures affect the number of flowers produced per plant leading to less sexual reproduction. Our findings therefore suggest that predicted alterations in climate regimes may influence both plant interactions and reproductive strategies, which have the potential to inflict changes in biodiversity, community structure and ecosystem functioning.

Effects of Planted Versus Naturally Growing Vallisneria natans on the Sediment Microbial Community in West Lake, China.

Submerged macrophytes play an important role in aquatic ecosystems, which has led to an increase in studies on vegetation recovery in polluted lakes from which submerged macrophytes have disappeared. The comparison of microbial communities in sediment cloned with planted and naturally growing submerged macrophytes is an interesting but rarely studied topic. In this investigation, Maojiabu and Xilihu, two adjacent sublakes of West Lake (Hangzhou, China), were selected as aquatic areas with planted and naturally growing macrophytes, respectively. Sediment samples from sites with/without Vallisneria natans were collected from both sublakes. The results showed that sediment total nitrogen and organic matter were significantly lower in the plant-covered sites than that in the non-plant sites in Maojiabu. Additionally, the sediment microbial community characterized by 16S ribosomal RNA (rRNA) sequencing differed more significantly for Maojiabu than for Xilihu. The relative abundances of microbes involved in C, N, and S elemental cycling were significantly higher in the sediments with plants than in those without. Results from both fatty acid methyl ester analysis and 16S rRNA sequencing indicated that vegetation significantly influenced the sulfate-reducing bacteria (SRB). Thus, the gene copies and composition of SRB were explored further. The relative gene abundance of SRB was 66% higher with natural vegetation colonization but was not influenced by artificial colonization. An increase in dominant SRB members from the families Syntrophobacteraceae and Thermodesulfovibrionaceae contributed to the increase of total SRB. Thus, macrophyte planting influences sediment nutrient levels and microbial community more than natural growth does, whereas the latter is more beneficial to sediment SRB.

An ultrasonic method for separation of epiphytic microbes from freshwater submerged macrophytes.

Epiphytic microbes are common inhabitants of freshwater submerged macrophytes, which play an important role in aquatic ecosystems. An important precondition for studying the epiphytic microbes is having an effective method of separating the attached microbes from the host macrophytes. We developed an ultrasound-based method for separating epiphytic microbes from freshwater submerged macrophytes, optimized the conditions of ultrasonic separation with an orthogonal experimental design, and compared the optimized ultrasonic method with manual separation. This method can be particularly useful for freshwater submerged macrophytes having a complex morphology.

The distinctive level of interaction between carbon and nitrogen metabolisms in the leaves of submerged macrophytes plays a key role in ammonium detoxification.

Possible ammonium detoxification mechanisms have been proposed recently, on submerged macrophytes, evidently illustrating that glutamate dehydrogenase (GDH) plays a greater role in ammonium detoxification compared to the primary glutamine synthetase/glutamate oxaloacetate transaminase (GS/GOGAT) pathway. In the current investigation, we cultured three submerged macrophytes to extreme concentrations of [NH4+-N] of up to 50 mg/L with the aim of clarifying the interaction between carbon and nitrogen metabolisms. The activities of carboxylation enzymes pyruvate orthophosphate dikinase (PPDK) and phosphoenolpyruvate carboxylase (PEPC), in lieu of Rubisco, increased almost two-fold for ammonium tolerant species P. maackianus and M. spicatum, compared with the sensitive species P. lucens. While these enzymes are well known for their central role in CO2 fixation, their inference in conferring resistance to ammonium stress has not been well elucidated before. In this study, we demonstrate that the overproduction of PEPC and PPDK led to improved photosynthesis, better ammonium assimilation and overall ammonium detoxification in M. spicatum and P. maackianus. These findings propose likelihood for the existence of a complementary ammonium detoxification pathway that targets carbon metabolism, thus, presenting a relatively efficient linkage between nitrogen and carbon metabolisms and identify candidate species for practical restoration of fresh water resources.

Response strategies of stem/leaves endophyte communities to nano-plastics regulate growth performance of submerged macrophytes.

Research on the toxicity effects of nano-plastics on submerged macrophytes has been increasing over the past several years. However, how the endophytic bacteria of submerged macrophytes respond to nano-plastics remains unknown, although they have been widely shown to help terrestrial plants cope with various environmental stressors. Here, a microcosm experiment was performed to unravel the effects of high concentration of nano-plastics (20 mg/L) on three submerged macrophyte (Vallisneria natans, Potamogeton maackianus, Myriophyllum spicatum) and their endophytic bacterial communities. Results indicated that nano-plastics induced antioxidative stress in plants, but significantly reduction in relative growth rate (RGR) only occurred in V. natans (from 0.0034 to -0.0029 day-1), accompanied by change in the stem/leaves endophyte community composition. Further analysis suggested nano-plastics caused a reduction in environmental nutrient availability and the proportion of positive interactions between endophyte communities (43%), resulting in the lowest RGR of V. natans. In contrast, endophytes may help P. maackianus and M. spicatum cope with nano-plastic stress by increasing the proportion of positive correlations among communities (70% and 75%), leaving their RGR unaffected. Collectively, our study elucidates the species-specific response strategies of submerged macrophyte-endophyte to nano-plastics, which helps to reveal the different phytoremediation potential of submerged macrophytes against nano-plastic pollution.

Growth and physiological responses of submerged macrophytes to linear alkylbenzene sulfonate (LAS).

The potential toxic effects of linear alkylbenzene sulfonate (LAS), widely used in commercial detergents and cleaners, on submerged macrophytes remain unclear. We conducted a two-week exposure experiment to investigate LAS toxicity on five submerged macrophytes (four native and one exotic), focusing on their growth and physiological responses. The results showed that lower concentrations of LAS (< 5 mg/L) slightly stimulated the growth of submerged macrophytes, while higher doses inhibited it. Increasing LAS concentration resulted in decreased chlorophyll content, increased MDA content and POD activity, and initially increased SOD and CAT activities before declining. Moreover, Elodea nuttallii required a higher effective concentration for growth compared to native macrophytes. These findings suggest that different species of submerged macrophytes exhibited specific responses to LAS, with high doses (exceeding 5 ∼ 10 mg/L) inhibited plant growth and physiology. However, LAS may promote the dominance of surfactant-tolerant exotic submerged macrophytes in polluted aquatic environments.

Effect of luminescent materials on the aquatic macrophyte Vallisneria natans and periphytic biofilm.

Luminescent materials can adjust the spectrum of light energy utilization by plants. However, current research on the effects of luminescent materials on aquatic plants and periphytic biofilms is limited. This study investigated the effects of the luminescent materials 4-(di-p-tolylamino) benzaldehyde-A (DTB-A) and 4-(di-p-tolylamino) benzaldehyde-M (DTB-M) on the submerged macrophyte Vallisneria natans (V. natans) and periphytic biofilm. Result demonstrated that low concentrations of DTB (0.1 µM) significantly promoted the growth and photosynthetic rate of V. natans. In terms of enzyme activity, exposure to a higher concentration of DTB (10 µM) increased the activities of peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT). A combination of DTB-A and DTB-M treatment significantly changed the V. natans morphology and physiological characteristics, reducing the thickness of the cell wall and subsequently, promoting protein accumulation in leaves. There was no difference in the removal of ammonia or phosphate by V. natans at the 0.1 µM concentration, and the removal of ammonia and phosphate by V. natans decreased significantly as the concentration of luminescent material increased. A total of 3563 OTUs were identified in the biofilm community. The microbial community was dominated by Pseudomonas and Fusobacteria. Furthermore, results showed that an obvious decrease in diversity in the DTB-A and DTB-M mixed treatment group. In addition, the migratory aggregation of DTB molecules in plants was observed by fluorescence imaging. Overall, these findings extend our understanding of the mechanism of effect of luminescent materials on submerged macrophytes and their periphytic microorganisms.