Mitigating sediment cadmium contamination through combining PGPR Enterobacter ludwigii with the submerged macrophyte Vallisneria natans.

Sediment cadmium contamination poses risks to aquatic ecosystems. Phytoremediation is an environmentally sustainable method to mitigate cadmium contamination. Submerged macrophytes are affected by cadmium stress, but plant growth-promoting rhizobacteria (PGPR) can restore the health status of submerged macrophytes. Herein, we aimed to reduce sediment cadmium concentration and reveal the mechanism by which the combined application of the PGPR Enterobacter ludwigii and the submerged macrophyte Vallisneria natans mitigates cadmium contamination. Sediment cadmium concentration decreased by 21.59% after submerged macrophytes were planted with PGPR, probably because the PGPR colonized the rhizosphere and roots of the macrophytes. The PGPR induced a 5.09-fold increase in submerged macrophyte biomass and enhanced plant antioxidant response to cadmium stress, as demonstrated by decreases in oxidative product levels (reactive oxygen species and malondialdehyde), which corresponded to shift in rhizosphere metabolism, notably in antioxidant defence systems (i.e., the peroxidation of linoleic acid into 9-hydroperoxy-10E,12Z-octadecadienoic acid) and in some amino acid metabolism pathways (i.e., arginine and proline). Additionally, PGPR mineralized carbon in the sediment to promote submerged macrophyte growth. Overall, PGPR mitigated sediment cadmium accumulation via a synergistic plantmicrobe mechanism. This work revealed the mechanism by which PGPR and submerged macrophytes control cadmium concentration in contaminated sediment.

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.

Characterization and seasonal variation in biofilms attached to leaves of submerged plant.

The microorganisms and functional predictions of leaf biofilms on submerged plants (Vallisneria natans (Val)) and in water samples (surface water (S) and bottom water (B)) in different seasons were evaluated in this study. S and B groups had 3249 identical operational taxonomic units (OTUs) (50.03%), while the Val group only had 1201 (18.49%) unique OTUs. There was significant overlap between microbial communities of S and B groups in the same season, while Val group showed the greater diversity. The dominant microbial clades were Proteobacteria (18.2-47.3%), Cyanobacteria (3.74-39.3%), Actinobacteria (1.64-29.3%), Bacteroidetes (1.31-21.7%), and Firmicutes (1.10-15.72%). Furthermore, there was a significant relationship between total organic carbon and the distribution of microbial taxa (p = 0.047), and TN may have altered the status of Cyanobacteria by affecting its biological nitrogen fixation capacity and reproductive capacity. The correlation network analysis results showed that the whole system consisted of 249 positive correlations and 111 negative correlations, indicating strong interactions between microbial communities. Functional predictions indicated that microbial functions were related to seasonal variation. These findings would guide the use of submerged plants to improve the diversity and stability of wetland microbial communities.

Coupling of submerged macrophytes and epiphytic biofilms reduced methane emissions from wetlands: Evidenced by an antibiotic inhibition experiment.

Wetlands are the largest natural methane source, but how submerged macrophytes affect methane emission remains controversial. In this study, the impacts of submerged macrophytes on methane fluxes, water purification, and epiphytic microbial community dynamics were investigated in simulated wetlands (with and without Hydrilla verticillata) treated with norfloxacin (NOR) for 24 days. Mean methane fluxes were significantly lower in treatments with Hydrilla verticillata (56.84-90.94 mg/m2/h) than bulks (65.96-113.21 mg/m2/h) (p < 0.05) during the experiment regardless of NOR. The relative conductivity (REC) values, H2O2, and malondialdehyde (MDA) contents increased in plant leaves, while water nutrients removal rates decreased with increasing NOR concentration at the same sampling time. The partial least squares path model analysis revealed that plant physiological indices and water nutrients positively affected methane fluxes (0.72 and 0.49, p < 0.001). According to illumina sequencing results of 16S rRNA and pmoA genes, α-proteobacteria (type II) and γ-proteobacteria (type I) were the dominant methanotroph classes in all epiphytic biofilms. The ratio of type I/type II methanotrophs and pmoA gene abundance in epiphytic biofilm was considerably lower in treatment with 16 mg/L NOR than without it (p < 0.05). pmoA gene abundance was negatively correlated with methane fluxes (p < 0.05). Additionally, the assembly of epiphytic bacterial community was mainly governed by deterministic processes, while stochastic dispersal limitation was the primary assembly process in the epiphytic methanotrophic community under NOR stress. The deterministic process gained more importance with time both in bacterial and methanotrophic community assembly. Network analysis revealed that relationships among bacteria in epiphytic biofilms weakened with time but associations among methanotrophic members were enhanced under NOR stress over time. It could be concluded that submerged macrophytes-epiphytic biofilms symbiotic system exhibited potential prospects to reduce methane emissions from wetlands under reasonable management.

Effect of erythromycin on epiphytic bacterial communities and water quality in wetlands.

The occurrence of antibiotics such as erythromycin (ERY) under macrolide group, has long been acknowledged for negatively affecting ecosystems in freshwater environments. However, the effects of ERY on water quality and microbial communities in epiphytic biofilms are poorly understood. Here, Scanning Electron Microscopy (SEM), High-throughput sequencing, and physicochemical analytical methods were employed to unravel the impact of ERY on the water quality and bacterial morphology, biodiversity, composition, interaction, and ecological function in epiphytic biofilms attached to Vallisneria natans and artificial plants in mesocosmic wetlands. The study showed that ERY exposure significantly impaired the nutrient removal capacity (TN, TP, and COD) and altered the epiphytic bacterial morphology of V. natans and artificial plants. ERY did not affect the bacterial α-diversity. Notwithstanding ERY decreased the bacterial composition, but the relative abundance of Proteobacteria and Patescibacteria spiked by 62.2 % and 54 %, respectively, in V. natans, while Desulfobacteria and Chloroflexi increased by 8.9 % and 11.2 %, respectively, in artificial plants. Notably, ERY disturbed the food web structure and metabolic pathways such as carbohydrate metabolism, amino acid metabolism, energy metabolism, cofactor and vitamin metabolism, membrane transport, and signal transduction. This study revealed that ERY exposure disrupted the bacterial morphology, composition, interaction or food web structure, and metabolic functions in epiphytic biofilm. These data underlined that ERY negatively impacts epiphytic bacterial communities and nutrient removal in wetlands.

Effect of PAHs on nitrogen-fixing and sulfate-reducing microbial communities in seagrass Enhalus acoroides sediment.

Seagrass meadows are vital ecosystems with high productivity and biodiversity and often in the oligotrophic area. Nitrogen usually limits productivity in this ecosystem as the main nutrient factor. Biological nitrogen fixation by diazotrophs in the rhizosphere sediment can introduce "new" nitrogen into the ecosystem. Previous studies revealed that most sulfate-reducing bacteria (SRB) can also fix nitrogen like the nitrogen-fixing bacteria (NFB). Moreover, both sulfate reduction and nitrogen fixation were affected by the organic pollutant. However, rare information is available regarding the NFB and SRB community composition and their temporal response to the pollutant. The quantitative real-time polymerase chain reaction and polymerase chain reaction denaturing gradient gel electrophoresis have been used to analyze NFB and SRB communities' shifts under different PAHs concentrations. They both experienced a dramatic shift under PAHs stress but exhibited different patterns. SRB could use the low and high concentration PAHs at the early stage of the incubation, while only the low concentration of PAHs could stimulate the growth of NFB through the whole incubation period. The predominant species of NFB communities were Alphaproteobacteria, Gammaproteobacteria, and Deltaproteobacteria; while for SRB communities were class Epsilonproteobacteria. Redundancy analysis indicated the significant environmental factors for the two communities were both ammonium and pH (P < 0.05). There existed nifH sequences related to known nitrogen fixing SRB Desulfatibacillum alkenivorans, which confirmed that microbial N2 fixation and sulfate reduction were coupled in the seagrass ecosystem by molecular technique. Our investigation provides new insight into the NFB and SRB community in the seagrass meadow.

Hunting the eagle killer: A cyanobacterial neurotoxin causes vacuolar myelinopathy.

Vacuolar myelinopathy is a fatal neurological disease that was initially discovered during a mysterious mass mortality of bald eagles in Arkansas in the United States. The cause of this wildlife disease has eluded scientists for decades while its occurrence has continued to spread throughout freshwater reservoirs in the southeastern United States. Recent studies have demonstrated that vacuolar myelinopathy is induced by consumption of the epiphytic cyanobacterial species Aetokthonos hydrillicola growing on aquatic vegetation, primarily the invasive Hydrilla verticillata Here, we describe the identification, biosynthetic gene cluster, and biological activity of aetokthonotoxin, a pentabrominated biindole alkaloid that is produced by the cyanobacterium A. hydrillicola We identify this cyanobacterial neurotoxin as the causal agent of vacuolar myelinopathy and discuss environmental factors-especially bromide availability-that promote toxin production.

Negative effects on the leaves of submerged macrophyte and associated biofilms growth at high nitrate induced-stress.

High nitrate (NO3--N) concentration is a growing aquatic risk concern worldwide. However, adverse effects of high NO3--N concentration on submerged macrophytes-epiphytic biofilms are unclear. In this study, the alterations in physiological changes, biofilms formation and chemical compositions were investigated on leaves of Vallisneria asiatica exposed to different NO3--N concentrations. The findings showed that 10 mg L-1NO3--N resulted in low photosynthetic efficiency by inhibiting chlorophyll content 26.2 % and decreased intrinsic efficiency of photosystem II significantly at 14th day post treatment. Malondialdehyde, several antioxidant enzyme activities (i.e., superoxide dismutase, peroxidase and catalase), and secondary metabolites (i.e., phenolic compounds and anthocyanin) were all significantly up-regulated with 10 mg L-1NO3--N, implied oxidative stress were stimulated. However, no significant alterations in these indicators were observed with 5 mg L-1NO3--N. Compared to control, 10 mg L-1NO3--N concentration significantly stimulated microbes growth in biofilm and reduced the roughness of leaf-biofilms surface, but it had little effect on the biofilms distribution (from single clone to blocks) as revealed by scanning electron microscope and multifractal analysis. Results from X-ray photoelectron spectroscopy analysis showed that the percentage of P, Cl, K and the ratio of O1 (-O-) /O2 (C = O) were higher in leaves of control than treatments with 10 mg L-1NO3--N, indicating that 10 mg L-1NO3--N concentration exhibited significant inhibition of chemical activity and nutrient uptake of the leaf surfaces. Overall, these results demonstrated that high NO3--N does stimulate the biofilm growth and can cause negative impacts on submerged macrophytes growth.

Host and environmental determinants of microbial community structure in the marine phyllosphere.

Although seagrasses are economically and ecologically critical species, little is known about their blade surface microbial communities and how these communities relate to the plant host. To determine microbial community composition and diversity on seagrass blade surfaces and in the surrounding seawater,16S rRNA gene sequencing (iTag) was used for samples collected at five sites along a gradient of freshwater input in the northern Gulf of Mexico on three separate sampling dates. Additionally, seagrass surveys were performed and environmental parameters were measured to characterize host characteristics and the abiotic conditions at each site. Results showed that Thalassia testudinum (turtle grass) blades hosted unique microbial communities that were distinct in composition and diversity from the water column. Environmental conditions, including water depth, salinity, and temperature, influenced community structure as blade surface microbial communities varied among sites and sampling dates in correlation with changes in environmental parameters. Microbial community composition also correlated with seagrass host characteristics, including growth rates and blade nutrient composition. There is some evidence for a core community for T. testudinum as 21 microorganisms from five phyla (Cyanobacteria, Proteobacteria, Planctomycetes, Chloroflexi, and Bacteroidetes) were present in all blade surface samples. This study provides new insights and understanding of the processes that influence the structure of marine phyllosphere communities, how these microbial communities relate to their host, and their role as a part of the seagrass holobiont, which is an important contribution given the current decline of seagrass coverage worldwide.

Damage of heavy metals to Vallisneria natans (V. natans) and characterization of microbial community in biofilm.

Heavy metals can cause a significant damage to submerged macrophytes and affect its periphyton biofilms in aquatic environments. This study investigated the effects of heavy metals such as copper (Cu), lead (Pb), cadmium (Cd) and their mixture on physiological and biochemical responses and ultrastructure characteristics of Vallisneria natans (V. natans). Furthermore, differences in structures of microbial communities were observed in biofilms. The results showed that Cu2+, Pb2+, Cd2+ and their mixture could destroy cell structure and photosynthetic system, and directly caused oxidative damage to submerged macrophyte and induced antioxidant enzyme system. In general, biomass and total chlorophyll content of V. natans noticeably decreased, while the activities of superoxide dismutase, peroxidase and catalase were enhanced by heavy metal stress inducement in restricted range, and the malondialdehyde content increased with the aggravation of the damage. The single heavy metal stress played a negative impact, however, the combined stress was not always synergistic effects on plants. High-throughput sequencing analysis suggested that heavy metals changed the abundance and structure of the microbial biofilm community. Proteobacteria and Bacteroidete were the dominant bacteria under heavy metal stress and other species and abundance of bacteria such as Firmicute, Cyanobacteria, Chloroflexi, Actinobacteria, Verrucomicrobia, Acidobacteria, Deinococcus-Thermus, Chlamydiae were also present. These findings provided useful information for further understanding about submerged macrophytes and periphyton biofilms responsed to heavy metal stress in aquatic environments in the future.

Influence of epiphytic bacteria on arsenic metabolism in Hydrilla verticillata.

Microbial assemblages such as biofilms around aquatic plants play a major role in arsenic (As) cycling, which has often been overlooked in previous studies. In this study, arsenite (As(III))-oxidizing, arsenate (As(V))-reducing and As(III)-methylating bacteria were found to coexist in the phyllosphere of Hydrilla verticillata, and their relative activities were shown to determine As speciation, accumulation and efflux. When exposed to As(III), As(III) oxidation was not observed in treatment H(III)-B, whereas treatment H(III)+B showed a significant As(III) oxidation ability, thereby indicating that epiphytic bacteria displayed a substantial As(III) oxidation ability. When exposed to As(V), the medium only contained 5.89% As(III) after 48 h of treatment H(V)-B, while an As(III) content of 86.72% was observed after treatment H(V)+B, thereby indicating that the elevated As(III) in the medium probably originated from As(V) reduction by epiphytic bacteria. Our data also indicated that oxidizing bacteria decreased the As accumulation (by approximately 64.44% compared with that of treatment H(III)-B) in plants, while reducing bacteria played a critical role in increasing As accumulation (by approximately 3.31-fold compared with that of treatment H(V)-B) in plants. Regardless of whether As(III) or As(V) was supplied, As(III) was dominant in the plant tissue (over 75%). Furthermore, the presence of epiphytic bacteria enhanced As efflux by approximately 9-fold. Metagenomic analysis revealed highly diverse As metabolism genes in epiphytic bacterial community, particularly those related to energetic metabolism (aioAB), and As resistance (arsABCR, acr3, arsM). Phylogenetic analysis of As metabolism genes revealed evidence of both vertical inheritance and horizontal gene transfer, which might have contributed to the evolution of the As metabolism genes. Taken together, our research suggested that the diversity of As metabolism genes in epiphytic bacterial community is associated with aquatic submerged macrophytes which may play an important role in As biogeochemistry in aquatic environments.

Root microbiomes as indicators of seagrass health.

The development of early warning indicators that identify ecosystem stress is a priority for improving ecosystem management. As microbial communities respond rapidly to environmental disturbance, monitoring their composition could prove one such early indicator of environmental stress. We combined 16S rRNA gene sequencing of the seagrass root microbiome of Halophila ovalis with seagrass health metrics (biomass, productivity and Fsulphide) to develop microbial indicators for seagrass condition across the Swan-Canning Estuary and the Leschenault Estuary (south-west Western Australia); the former had experienced an unseasonal rainfall event leading to declines in seagrass health. Microbial indicators detected sites of potential stress that other seagrass health metrics failed to detect. Genera that were more abundant in 'healthy' seagrasses included putative methylotrophic bacteria (e.g. Methylotenera and Methylophaga), iron cycling bacteria (e.g. Deferrisoma and Geothermobacter) and N2 fixing bacteria (e.g. Rhizobium). Conversely, genera that were more abundant in 'stressed' seagrasses were dominated by putative sulphur-cycling bacteria, both sulphide-oxidising (e.g. Candidatus Thiodiazotropha and Candidatus Electrothrix) and sulphate-reducing (e.g. SEEP-SRB1, Desulfomonile and Desulfonema). The sensitivity of the microbial indicators developed here highlights their potential to be further developed for use in adaptive seagrass management, and emphasises their capacity to be effective early warning indicators of stress.

The dissipation and risk alleviation mechanism of PAHs and nitrogen in constructed wetlands: The role of submerged macrophytes and their biofilms-leaves.

The role of submerged macrophytes (Vallisneria natans, Hydrilla verticillata and artificial plant) and their biofilms-leaves for the dissipation and risk alleviation mechanism of PAHs (phenanthrene and pyrene) and nitrogen in constructed wetland systems with PAH-polluted sediments were investigated. Biofilms-leaves/surface might contribute to PAHs degradation, which was positively correlated with PAHs degrading bacteria. Nitrogen-fixing bacteria in biofilms on surface might cause total nitrogen in sediment (TNs) increasing by 4% from 14th d to 28th d indirectly when suffering PAHs pollution. The relative abundance of nitrogen-fixing bacteria significantly increased with the increase of PAHs concentrations in early period (p < 0.01), which might lead to risk of nitrogen accumulation further. Heat maps showed that the relative abundance of functional bacteria were influenced in order of attached surface > incubation time > spiking concentration of PAHs. Interestingly, differences of deduced bacterial functions were affected in order of incubation time > attached surface > spiking concentration. Thus, submerged macrophytes and their biofilms on leaves not only played an important role in PAHs degradation, but also regulated the nitrogen cycling in constructed wetland systems, which could reduce these pollutants risk for natural environment, organisms and human health.

Ammonium loading disturbed the microbial food webs in biofilms attached to submersed macrophyte Vallisneria natans.

The knowledge about the impacts of ammonium loading on microbial food webs in biofilms attached to submersed macrophytes is limited. In the present study, Illumina sequencing method was employed to investigate bacterial and eukaryotic communities in biofilms attached to leaves of Vallisneria natans (V. natans) exposed to 1-16 mg L-1 NH4+-N for 10 days, and 8 mg L-1 NH4+-N for 21 days. Ammonium loading stimulated biofilms growth, enhanced the relative abundance of nitrifying genus Nitrospira and several denitrifying genera. Eukaryotic kingdom Metazoa, Viridiplantae, Chromista, Fungi and super group SARNU (Stramenopiles, Alveolata, Rhizaria, Nucleariidae and Fonticula group and unknown eukaryotes) were obtained. Relative abundance of Metazoa decreased with the increased ammonium concentration and exposure time. Redundancy analysis revealed that ammonium, dissolved oxygen (DO) and pH had a key role in determining microbial community structure. Network analyses revealed that there were complex interactions including feeding, parasitism and predatism among organism in biofilms, and the microbial food webs were disturbed by inhibiting metazoan growth but stimulating bacteria and algae growth. These results suggest that ammonium-disturbed microbial food webs in biofilms may contribute to the growth of biofilms and algae, and thus contribute to the decline of submersed macrophyte and provide "algal seeds" for the algae burst in water column. These data will be helpful in understanding the macrophytic region transform into algal region in water column polluted by ammonium.

Oxygen loss from seagrass roots coincides with colonisation of sulphide-oxidising cable bacteria and reduces sulphide stress.

Seagrasses thrive in anoxic sediments where sulphide can accumulate to phytotoxic levels. So how do seagrasses persist in this environment? Here, we propose that radial oxygen loss (ROL) from actively growing root tips protects seagrasses from sulphide intrusion not only by abiotically oxidising sulphides in the rhizosphere of young roots, but also by influencing the abundance and spatial distribution of sulphate-reducing and sulphide-oxidising bacteria. We used a novel multifaceted approach combining imaging techniques (confocal fluorescence in situ hybridisation, oxygen planar optodes, and sulphide diffusive gradients in thin films) with microbial community profiling to build a complete picture of the microenvironment of growing roots of the seagrasses Halophila ovalis and Zostera muelleri. ROL was restricted to young root tips, indicating that seagrasses will have limited ability to influence sulphide oxidation in bulk sediments. On the microscale, however, ROL corresponded with decreased abundance of potential sulphate-reducing bacteria and decreased sulphide concentrations in the rhizosphere surrounding young roots. Furthermore, roots leaking oxygen had a higher abundance of sulphide-oxidising cable bacteria; which is the first direct observation of these bacteria on seagrass roots. Thus, ROL may enhance both abiotic and bacterial sulphide oxidation and restrict bacterial sulphide production around vulnerable roots, thereby helping seagrasses to colonise sulphide-rich anoxic sediments.

Allelopathic effects of harmful algal extracts and exudates on biofilms on leaves of Vallisneria natans.

This study investigated the allelopathic effects of Microcystis aeruginosa (M. aeruginosa) extracts and exudates on the physiological responses, photosynthetic activity, and microbial structure of biofilms on leaves of Vallisneria natans (V. natans). By measuring physiological and photosynthetic indices, the results showed that M. aeruginosa allelochemicals inhibited photosynthesis, oxidative stress and antioxidant system stress response in the biofilms of V. natans leaves. Multifractal analysis found that the surface topography of V. natans leaves was altered due to the allelochemicals found in M. aeruginosa. Microbial diversity on the leaves was analyzed using high-throughput sequencing, and the results showed that M. aeruginosa exudates had a stronger effects on the microbial community structure of biofilms compared to extracts. These findings highlight how cyanobacterial allelochemicals induce negative effects on submerged macrophytes.

Effects of nutrient loading on sediment bacterial and pathogen communities within seagrass meadows.

Eutrophication can play a significant role in seagrass decline and habitat loss. Microorganisms in seagrass sediments are essential to many important ecosystem processes, including nutrient cycling and seagrass ecosystem health. However, current knowledge of the bacterial communities, both beneficial and detrimental, within seagrass meadows in response to nutrient loading is limited. We studied the response of sediment bacterial and pathogen communities to nutrient enrichment on a tropical seagrass meadow in Xincun Bay, South China Sea. The bacterial taxonomic groups across all sites were dominated by the Gammaproteobacteria and Firmicutes. Sites nearest to the nutrient source and with the highest NH4+ and PO43- content had approximately double the relative abundance of putative denitrifiers Vibrionales, Alteromonadales, and Pseudomonadales. Additionally, the relative abundance of potential pathogen groups, especially Vibrio spp. and Pseudoalteromonas spp., was approximately 2-fold greater at the sites with the highest nutrient loads compared to sites further from the source. These results suggest that proximity to sources of nutrient pollution increases the occurrence of potential bacterial pathogens that could affect fishes, invertebrates and humans. This study shows that nutrient enrichment does elicit shifts in bacterial community diversity and likely their function in local biogeochemical cycling and as a potential source of infectious diseases within seagrass meadows.

Planctopirus hydrillae sp. nov., an antibiotic producing Planctomycete isolated from the aquatic plant Hydrilla and its whole genome shotgun sequence analysis.

An antibiotic producing novel Planctomycete strain, designated JC280T, was isolated from the surface of the plant Hydrilla verticillata collected from an alkaline lake (Buffalo lake), University of Hyderabad, Hyderabad, India. The morphological and chemotaxonomic properties of strain JC280T were in agreement with the characteristics of the genus Planctopirus. The cell shape was spherical to ovoid and some were tear drop shaped. The cells were Gram-stain-negative divided by budding presenting stalks and rosette formation and were non-sporulating. Crateriform structures with a sub-polar flagellum were observed. Characteristic polyamines were putrescine and spermidine. Diagnostic polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, an unidentified phospholipid (PL1), unidentified glycolipids (GL1-2), an unidentified aminophospholipid (APL), and an unidentified lipid (L3). Major (>10%) fatty acids were C16:0, C17:1ω8c, C18:1ω9c, and summed feature-3. Major (88%) respiratory quinone was MK-6 with minor amount (12%) of MK-7. Strain JC280T showed 99.7% 16S rRNA gene sequence similarity with Planctopirus limnophila DSM 3776T. To resolve their full taxonomic position, the genome sequence was obtained and compared with the available P. limnophila DSM 3776T genome. The genome sequence of strain JC280T was 5,750,243 bp in size with a total of 4490 protein-coding genes, 66 RNA genes, and 2 CRISPR repeats. Based on whole-genome statistics, ANI value, in silico DDH, diversity of secondary metabolite biosynthetic gene clusters, distinct physiological, biochemical and chemotaxonomic differences, strain JC280T represents a new species in the genus Planctopirus, for which the name Planctopirus hydrillae sp. nov. is proposed. The type strain is JC280T (=KCTC 42880T = LMG 29153T).

Nitrogen loading affects microbes, nitrifiers and denitrifiers attached to submerged macrophyte in constructed wetlands.

Submerged macrophytes and biofilms are important components of wetlands. However, little is known about the changes of microbes in biofilms attached to submerged macrophytes upon nitrogen loading. This study investigated the changes of microbes, algae, nitrifiers and denitrifiers in biofilms attached to the leaves of artificial plants (AP), Potamogeton malaianus (PM), Vallisneria natans (VN) and Hydrilla verticillata (HV) under varied initial concentrations of total nitrogen (TN). Nitrogen addition increased biofilm biomass and changed dissolved oxygen concentrations and pH values in overlaying water. Epiphytic algal densities showed the same trend at the same N level:AP>PM>VN>HV. As revealed by cluster analysis at phylum level, algae compositions in biofilm from four plants showed some host-specific at 2 and 12mgL-1 TN, but was clustered in the same group at 22mgL-1 TN regardless of plant species. Submerged macrophytes had better performance in total N removal than AP. In general, N application significantly increased the abundance of amoA, nirK, nirS, napA and cnorB in biofilm. The abundance of the denitrification genes (nirK, nirS, napA, narG and cnorB) was positively correlated with nitrogen application, while amoA was correlated with concentration of dissolved oxygen. These results indicate that N loadings stimulated the growth of biofilms attached to submerged macrophyte and the removal of total N can be partially ascribed to the synergistic interactions of submerged macrophyte and biofilms in wetlands. These results highlight the ecological role of submerged macrophyte-biofilm system in nitrogen removal in wetlands.

Sediment microbes mediate the impact of nutrient loading on blue carbon sequestration by mixed seagrass meadows.

Recent studies have reported significant variability in sediment organic carbon (SOC) storage capacity among seagrass species, but the factors driving this variability are poorly understood, limiting our ability to make informed decisions about which seagrass types are optimal for carbon offsetting and why. Here we show that differences in SOC storage capacity among species within the same geomorphic environment can be explained (in part) by below-ground processes in response to nutrient load; specifically, differences in the activity of microbes harboured by morphologically-different seagrass species. We found that increasing nutrient load enhanced the relative contribution of seagrass and algal sources to SOC pools, boosting sediment microbial biomass and extracellular enzyme activity within mixed seagrass meadows composed of Thalassia hemprichii and Enhalus acoroides, and thus possibly weaken the seagrass blue carbon sequestration capacity. The relative contribution of seagrass plant material to sediment bacterial organic carbon (BOC) and the influencing SOC-decomposing enzymes in E. acoroides meadows were half that of T. hemprichii meadows living side-by-side, even though the mixed seagrass meadows received SOC from the same sources. Overall this research suggests that microbial activity can vary significantly among seagrass species, thereby causing fine-scale (within-meadow) variability in SOC sequestration capacity in response to nutrient load.