Novel Gene Clusters for Natural Product Synthesis Are Abundant in the Mangrove Swamp Microbiome.

Natural microbial communities produce a diverse array of secondary metabolites with ecologically and biotechnologically relevant activities. Some of them have been used clinically as drugs, and their production pathways have been identified in a few culturable microorganisms. However, since the vast majority of microorganisms in nature have not been cultured, identifying the synthetic pathways of these metabolites and tracking their hosts remain a challenge. The microbial biosynthetic potential of mangrove swamps remains largely unknown. Here, we examined the diversity and novelty of biosynthetic gene clusters in dominant microbial populations in mangrove wetlands by mining 809 newly reconstructed draft genomes and probing the activities and products of these clusters by using metatranscriptomic and metabolomic techniques. A total of 3,740 biosynthetic gene clusters were identified from these genomes, including 1,065 polyketide and nonribosomal peptide gene clusters, 86% of which showed no similarity to known clusters in the Minimum Information about a Biosynthetic Gene Cluster (MIBiG) repository. Of these gene clusters, 59% were harbored by new species or lineages of Desulfobacterota-related phyla and Chloroflexota, whose members are highly abundant in mangrove wetlands and for which few synthetic natural products have been reported. Metatranscriptomics revealed that most of the identified gene clusters were active in field and microcosm samples. Untargeted metabolomics was also used to identify metabolites from the sediment enrichments, and 98% of the mass spectra generated were unrecognizable, further supporting the novelty of these biosynthetic gene clusters. Our study taps into a corner of the microbial metabolite reservoir in mangrove swamps, providing clues for the discovery of new compounds with valuable activities. IMPORTANCE At present, the majority of known clinical drugs originated from cultivated species of a few bacterial lineages. It is vital for the development of new pharmaceuticals to explore the biosynthetic potential of naturally uncultivable microorganisms using new techniques. Based on the large numbers of genomes reconstructed from mangrove wetlands, we identified abundant and diverse biosynthetic gene clusters in previously unsuspected phylogenetic groups. These gene clusters exhibited a variety of organizational architectures, especially for nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS), implying the presence of new compounds with valuable activities in the mangrove swamp microbiome.

Spatiotemporal Dynamics of Bacterial Taxonomic and Functional Profiles in Estuarine Intertidal Soils of China Coastal Zone.

Bacteria play an important role in regulating carbon (C), nitrogen (N), and sulfur (S) in estuarine intertidal wetlands. To gain insights into the ecological and metabolic modes possessed by bacteria in estuarine intertidal wetlands, a total of 78 surface soil samples were collected from China's coastal intertidal wetlands to examine the spatial and seasonal variations of bacterial taxonomic composition, assembly processes, and ecological system functions through shotgun metagenomic and 16S rRNA gene sequencing. Obvious spatiotemporal dynamic patterns in the bacterial community structure were identified, with more pronounced seasonal rather than spatial variations. Dispersion limitation was observed to act as a critical factor affecting community assembly, explaining approximately half of the total variation in the bacterial community. Functional bacterial community structure exhibited a more significant latitudinal change than seasonal variability, highlighting that functional stability of the bacterial communities differed with their taxonomic variability. Identification of biogeochemically related links between C, N, and S cycles in the soils showed the adaptive routed metabolism of the bacterial communities and the strong interactions between coupled metabolic pathways. Our study broadens the insights into the taxonomic and functional profiles of bacteria in China's estuarine intertidal soils and helps us understand the effects exerted by environmental factors on the ecological health and microbial diversity of estuarine intertidal flats.

Increased Nitrogen Loading Facilitates Nitrous Oxide Production through Fungal and Chemodenitrification in Estuarine and Coastal Sediments.

Estuarine and coastal environments are assumed to contribute to nitrous oxide (N2O) emissions under increasing nitrogen loading. However, isotopic and molecular mechanisms underlying N2O production pathways under elevated nitrogen concentration remain poorly understood. Here we used microbial inhibition, isotope mass balance, and molecular approaches to investigate N2O production mechanisms in estuarine and coastal sediments through a series of anoxic incubations. Site preference of the N2O molecule increased due to increasing nitrate concentration, suggesting the changes in N2O production pathways. Enhanced N2O production under high nitrate concentration was not mediated by bacterial denitrification, but instead was mainly regulated by fungal denitrification. Elevated nitrate concentration increased the contribution of fungal denitrification to N2O production by 11-25%, whereas it decreased bacterial N2O production by 16-33%. Chemodenitrification was also enhanced by high nitrate concentration, contributing to 13-28% of N2O production. Elevated nitrate concentration significantly mediated nirK-type denitrifiers structure and abundance, which are the keystone taxa driving N2O production. Collectively, these results suggest that increasing nitrate concentration can shift N2O production pathways from bacterial to fungal and chemodenitrification, which are mainly responsible for the enhanced N2O production and have widespread implications for N2O projections under ongoing nitrogen pollution in estuarine and coastal ecosystems.

Nitrification Regulates the Spatiotemporal Variability of N2O Emissions in a Eutrophic Lake.

Nitrous oxide (N2O) emissions from lakes exhibit significant spatiotemporal heterogeneity, and quantitative identification of the different N2O production processes is greatly limited, causing the role of nitrification to be undervalued or ignored in models of a lake's N2O emissions. Here, the contributions of nitrification and denitrification to N2O production were quantitatively assessed in the eutrophic Lake Taihu using molecular biology and isotope mapping techniques. The N2O fluxes ranged from -41.48 to 28.84 µmol m-2 d-1 in the lake, with lower N2O concentrations being observed in spring and summer and significantly higher N2O emissions being observed in autumn and winter. The 15N site preference and relevant isotopic evidence demonstrated that denitrification contributed approximately 90% of the lake's gross N2O production during summer and autumn, 27-83% of which was simultaneously eliminated via N2O reduction. Surprisingly, nitrification seemed to act as a key process promoting N2O production and contributing to the lake as a source of N2O emissions. A combination of N2O isotopocule-based approaches and molecular techniques can be used to determine the precise characteristics of microbial N2O production and consumption in eutrophic lakes. The results of this study provide a basis for accurately assessing N2O emissions from lakes at the regional and global scales.

N2O emission dynamics along an intertidal elevation gradient in a subtropical estuary: Importance of N2O consumption.

Studying nitrous oxide (N2O) production and consumption processes along an intertidal elevation gradient can improve the understanding of N2O dynamics among coastal wetlands. A natural-abundance isotope technique was applied to characterize the processes responsible for N2O emission in high, middle and low intertidal zones in the Yangtze Estuary. The results showed that N2O emission rates in high tidal zones (0.84 ± 0.35 nmol g-1 h-1) were significantly higher than those in middle (0.21 ± 0.04 nmol g-1 h-1) and low tidal zones (0.26 ± 0.05 nmol g-1 h-1). Gross N2O production and consumption rates were greater in high and low tidal zones than in middle tidal zones, whereas N2O consumption proportions generally increased from high to low tidal zones. N2O consumption was quite pronounced, implying that N2O emission in estuarine wetlands accounts for only a small fraction of the total production. Higher degrees of N2O consumption were the pivotal driver of less N2O emission in low tidal zones. Bacterial denitrification (>84%) was the dominant pathway, although hydroxylamine (NH2OH) oxidation/fungal denitrification contributed substantially to N2O production in high tidal flats. The contribution to N2O production exhibited a decrease in NH2OH oxidation/fungal denitrification and an increase in bacterial denitrification with decreasing elevation. Changes in N2O dynamics along the elevation gradient were affected by carbon and nitrogen substrate availabilities as well as the redox environments. Overall, our findings highlight the importance of N2O consumption in controlling N2O emission in intertidal wetlands, especially with higher inundation frequencies and durations.

Crab bioturbation affects competition between microbial nitrogen removal and retention in estuarine and coastal wetlands.

As the important benthic animal in coastal wetlands, crab bioturbation may significantly affect the nitrogen (N) budgets by regulating microbial N transformation processes. However, the response of interaction between different microbial N processes to crab bioturbation remains poorly understood. Here, a 30-day microcosmic experiment was conducted using sediment collected from the Yangtze Estuary wetland, followed by the determination of temporal variations of physicochemical parameters, N removal (denitrification plus anammox, which is defined as N2 production) and retention rates (nitrate dissimilatory reduction to ammonium, DNRA) as well as relevant gene abundances in response to different crabs densities. The results showed that crab bioturbation simultaneously promoted the rates of N2 production and DNRA processes. These two process rates were positively associated with the intensity of crab bioturbation, which was supported by molecular analysis of relevant functional gene abundance. Crab bioturbation was more beneficial to DNRA than N2 production. Due to this disproportionate stimulation, crab bioturbation increased the importance of DNRA, indicating that N retention was becoming more significant under crab bioturbation in estuarine and coastal wetlands. The variations of sediment total organic carbon and oxygen availability driven by crab bioturbation were the critical factors mediating the changes in microbial N removal and retention. Overall, our findings highlighted that crab bioturbation can affect the N budgets in estuarine and coastal wetlands by altering the competition between microbial N removal and retention.

Anaerobic oxidation of methane with denitrification in sediments of a subtropical estuary: Rates, controlling factors and environmental implications.

Anaerobic oxidation of methane with denitrification (DAMO), as an important microbial process regulating methane emission, has been widely reported in freshwater ecosystems. However, the DAMO process and associated biogeochemical controls in estuaries remain poorly understood. Here, we used 13C- and 15N-labelling experiments to quantify the potential rates of DAMO and determined the crucial factors controlling the DAMO rates in the sediment of Yangtze Estuary. Potential rates of DAMO varied greatly across the estuary, ranging from 0.07 to 0.28 nmol CO2 g-1 d-1. Salinity negatively affected the DAMO and also showed an indirectly negative influence on DAMO process by high salinity inhibition on NO3- availability and denitrification. Nitrate concentrations were significantly correlated with the DAMO rates. Denitrification rates showed positive correlation with DAMO rates, implying that nitrate reduction drives the DAMO process. Sediment total organic carbon and NH4+ had important effects on DAMO rates. These results together indicate that DAMO process can occur and the DAMO rates were mainly controlled by sediment NO3- and denitrification in estuary. We further conclude that increasing NO3- load can drive the DAMO process with more important implications on methane sink in estuarine ecosystems.

In situ nitrogen removal processes in intertidal wetlands of the Yangtze Estuary.

Estuarine and intertidal wetlands are important sites for nitrogen transformation and elimination. However, the factors controlling nitrogen removal processes remain largely uncertain in the highly dynamic environments. In this study, continuous-flow experiment combined with 15N isotope pairing technique was used to investigate in situ rates of denitrification and anaerobic ammonium oxidation (anammox) and their coupling with nitrification in intertidal wetlands of the Yangtze Estuary. The measured rates varied from below the detection limit to 152.39 µmol N/(m2·hr) for denitrification and from below the detection limit to 43.06 µmol N/(m2·hr) for anammox. The coupling links of nitrogen removal processes with nitrification were mainly dependent on nitrate, organic carbon, sulfide, dissolved oxygen and ferric iron in the estuarine and intertidal wetlands. Additionally, it was estimated that the actual nitrogen removal processes annually removed approximately 5% of the terrigenous inorganic nitrogen discharged into the Yangtze Estuary. This study gives new insights into nitrogen transformation and fate in the estuarine and intertidal wetlands.

Trophodynamics and parabolic behaviors of polycyclic aromatic hydrocarbons in an urbanized lake food web, Shanghai.

Understanding the trophic movement and trophic magnification factor (TMF) of polycyclic aromatic hydrocarbons (PAHs) is an important criterion to assess their fate and potential effects in an aquatic ecosystem. This study investigated concentrations and trophodynamics of 16 priority PAHs in food webwb (food web comprising whole bodies) and food webm (food web containing only muscles) of total 14 species ((including plankton, shrimp, whitebait, mussels, snails, red-eared terrapin and seven other fish species) from the Dianshan Lake. Values of δ13C and δ15N were determined to assign trophic levels to organisms. In general, there was no evidence of biomagnification because of biological degradation of PAHs compounds, but only 9 PAH compounds in food webm showed a statistically significant negative relationship between trophic level and lipid normalized concentration compared to the 6 PAHs congeners in food webwb. However, TMF values of PAHs in food webm ranged from 0.32 for pyrene to 0.68 for phenanthrene compared to TMF values of food webwb ranged from 0.34 for pyrene to 0.74 for fluorene. Because of two opposing scientific views for biomagnification and biodilution of PAHs in the food web, albeit based on a rather limited number of studies, our study investigated that there is parabolic behavior of most of the PAHs. Concentrations of PAHs in the red-eared terrapin (Trachemys scripta elegans) were biodiluted which showed that this species metabolises such compounds. There was no particular relationship between Kow and TMFs of PAHs in the Dianshan Lake. Hepatobiliary system (such as gall bladder and liver) in the whole body considered to have higher bioaccumulation potential of organic compounds than extrahepatic tissue (muscles).

Natural chalcopyrite mitigates nitrous oxide emissions in sediment from coastal wetlands.

Coastal wetlands are one of the most important natural sources of nitrous oxide (N2O). Previous studies have shown that copper-containing chemicals are able to reduce N2O emissions from these ecosystems. However, these chemicals may harm organisms present in coastal waters and sediment, and disturb the ecological balance of these areas. Here, we first investigated the physiological characteristics and genetic potential of denitrifying bacteria isolated from coastal wetlands. Based on an isolated denitrifier carrying a complete denitrification pathway, we tested the effect of the natural mineral chalcopyrite on N2O production by the bacteria. The results demonstrated that chalcopyrite addition lowers N2O emissions from the bacteria while increasing its N2 production rate. Among the four denitrification genes of the isolate, only nosZ gene expression was significantly upregulated following the addition of 2 mg L-1 chalcopyrite. Furthermore, chalcopyrite was applied to coastal wetland sediments. The N2O flux was significantly reduced in 50-100 mg L-1 chalcopyrite-amended sets relative to the controls. Notably, the dissolved Cu concentration in chalcopyrite-amended sediment remained within the limit set by the National Sewage Treatment Discharge Standard. qPCR and metagenomic analysis revealed that the abundance of N2O-reducing bacteria with the nosZ or nirK + nosZ genotype increased significantly in the chalcopyrite-amended groups relative to the controls, suggesting their active involvement in the reduction of N2O emissions. Our findings offer valuable insights for the use of natural chalcopyrite in large-scale field applications to reduce N2O emissions.

Changes in sediment greenhouse gases production dynamics in an estuarine wetland following invasion by Spartina alterniflora.

Invasive Spartina alterniflora (S. alterniflora) has significant impacts on sediment biogeochemical cycling in the tidal wetlands of estuaries and coasts. However, the impact of exotic Spartina alterniflora invasion on greenhouse gases (GHGs) production dynamics in sediments remain limited. Here, we investigated the dynamics of sediment physicochemical properties, GHGs production rates, and microbial gene abundances in a native Cyperus malacensis habitat and three invasive S. alterniflora habitats (6-, 10-, and 14-year) in the Minjiang River Estuary, China. The methane (CH4), carbon dioxide (CO2), and nitrous oxide (N2O) production rates varied both spatially and seasonally, while microbial gene abundances (bacterial and fungal gene abundances) and organic matter (TOC and TN) only varied spatially. GHGs production rates were also characterized by higher values in surface sediment (0-10 cm) compared to subsurface sediment (10-20 cm) and by seasonal variations with higher values in summer than in winter. S. alterniflora invasion can significantly increase CH4 and CO2 production rates, organic matter, and microbial gene abundances (p < 0.05). Temperature, organic matter and microbial gene abundances were the most dominating factor controlling the spatio-temporal variations of CH4 and CO2 production rates. Overall, our findings highlighted the significant role of S. alterniflora invasion in regulating GHGs production rates in coastal wetland sediments and provided fundamental data for estimating GHGs emissions and carbon sequestration in the complex tidal wetlands.

Impact of soil inorganic nitrogen on bacterial phylogeny in estuarine intertidal zones: a study of nitrogen metabolism.

Here we investigated the potential impacts of soil inorganic nitrogen (SIN) content on the phylogenetic characteristics and ecological functions of soil bacterial communities in estuarine intertidal zones in China, aiming to comprehend the response mechanism of soil microorganisms to variations in SIN content within estuarine wetlands. Our results show that SIN in estuarine areas has a significant spatiotemporal variation on spatial and seasonal scales, in this study and is significantly associated with the phylogenetic diversity and phylogenetic turnover of soil bacterial communities. In addition, the results of the metagenomic analysis showed that the relative abundance of nitrogen-cycling functional genes in bacterial communities did not differ significantly in sampling sites and seasons, and weakly correlated with SIN content. Further, the results based on structural equation modeling (SEM) analysis showed that SIN directly and significantly regulated the phylogenetic characteristics of bacterial communities, thereby indirectly affecting the potential of bacterial nitrogen metabolism. This study emphasizes the key influence of SIN variations on the phylogenetic dissimilarity in soil bacterial communities. Moreover, although there was a weak direct relationship between the functional characteristics of the bacterial nitrogen metabolism and SIN content, the spatiotemporal variation of bacterial nitrogen metabolic potential may be indirectly regulated by SIN content by influencing the phylogenetic diversity in bacterial communities. Our study unravels the pivotal mechanisms through which SIN content influences bacterial communities, thereby offering novel insights into the microbial intricacies governing nitrogen metabolism within estuaries.

Active pathways of anaerobic methane oxidization in deep-sea cold seeps of the South China Sea.

IMPORTANCE: Cold seeps occur in continental margins worldwide and are deep-sea oases. Anaerobic oxidation of methane is an important microbial process in the cold seeps and plays an important role in regulating methane content. This study elucidates the diversity and potential activities of major microbial groups in dependent anaerobic methane oxidation and sulfate-dependent anaerobic methane oxidation processes and provides direct evidence for the occurrence of nitrate-/nitrite-dependent anaerobic methane oxidation (Nr-/N-DAMO) as a previously overlooked microbial methane sink in the hydrate-bearing sediments of the South China Sea. This study provides direct evidence for occurrence of Nr-/N-DAMO as an important methane sink in the deep-sea cold seeps.

Effects of periodic drying-wetting on microbial dynamics and activity of nitrite/nitrate-dependent anaerobic methane oxidizers in intertidal wetland sediments.

Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) plays an important role in methane (CH4) consumption in intertidal wetlands. However, little is known about the responses of n-DAMO in intertidal wetlands to periodic drying-wetting caused by tidal cycling. Here, comparative experiments (waterlogged, desiccated, reflooded) with the Yangtze estuarine intertidal sediments were performed to examine the effects of periodic tidal changes on n-DAMO microbial communities, abundances, and potential activities. Functional gene sequencing indicated the coexistence of n-DAMO bacteria and archaea in the tide-fluctuating environments and generally higher biodiversity under reflooded conditions than consecutive inundation or emersion. The n-DAMO microbial abundance and associated activity varied significantly during alternative exposure and inundation, with higher abundance and activity under the waterlogged than desiccated conditions. Reflooding of intertidal wetlands might intensify n-DAMO activities, indicating the resilience of n-DAMO microbial metabolisms to the wetting-drying events. Structural equation modeling and correlation analysis showed that n-DAMO activity was highly related to n-DAMO microbial abundance and substrate availability under inundation, whereas salt accumulation in sediment was the primary factor restraining n-DAMO activity under the desiccation. Overall, this study reveals tidal-induced shifts of n-DAMO activity and associated contribution to mitigating CH4, which may help accurately project CH4 emission from intertidal wetlands under different tidal scenarios.

Tidal dynamics regulates potential coupling of carbon­nitrogen­sulfur cycling microbes in intertidal flats.

Tide-driven hydrodynamic process causes significant geochemical gradients that influence biogeochemical cycling and ecological functioning of estuarine and coastal ecosystems. However, the effects of tidal dynamics on microbial communities, particularly at the functional gene level, remain unclear even though microorganisms play critical roles in biogeochemical carbon (C), nitrogen (N) and sulfur (S) cycling. Here, we used 16S rRNA gene amplicon sequencing and microarray-based approach to reveal the stratification of microorganisms related to C, N and S cycles along vertical redox gradients in intertidal wetlands. Alpha-diversity of bacteria and archaea was generally higher at the deep groundwater-sediment interface. Microbial compositions were markedly altered along the sediment profile, and these shifts were largely due to changes in nutrient availability and redox potential. Furthermore, functional genes exhibited redox partitioning between interfaces and transition layer, with abundant genes involved in C decomposition, methanogenesis, heterotrophic denitrification, sulfite reduction and sulfide oxidation existed in the middle anoxic zone. The influence of tidal dynamics on sediment function was highly associated with redox state, sediment texture, and substrates availability, leading to distinct distribution pattern of metabolic coupling of microbes involved in energy flux and elemental cycling in intertidal wetlands. These results indicate that tidal cycles are critical in determining microbial community and functional structure, and they provide new insights into sediment microbe-mediated biogeochemical cycling in intertidal habitats.

Invasive Spartina alterniflora accelerates the increase in microbial nitrogen fixation over nitrogen removal in coastal wetlands of China.

Salt marsh plants play a vital role in mediating nitrogen (N) biogeochemical cycle in estuarine and coastal ecosystems. However, the effects of invasive Spartina alterniflora on N fixation and removal, as well as how these two processes balance to determine the N budget, remain unclear. Here, simultaneous quantifications of N fixation and removal via 15N tracing experiment with native Phragmites australis, invasive S. alterniflora, and bare flats as well as corresponding functional gene abundance by qPCR were carried out to explore the response of N dynamics to S. alterniflora invasion. Our results showed that N fixation and removal rates ranged from 0.77 ± 0.08 to 16.12 ± 1.13 nmol/(g·h) and from 1.42 ± 0.14 to 16.35 ± 1.10 nmol/(g·h), respectively, and invasive S. alterniflora generally facilitated the two processes rates. Based on the difference between N removal and fixation rates, net N2 fluxes were estimated in the range of -0.39 ± 0.14 to 8.24 ± 2.23 nmol/(g·h). Estimated net N2 fluxes in S. alterniflora stands were lower than those in bare flats and P. australis stands, indicating that the increase in N removal caused by S. alterniflora invasion may be more than offset by N fixation process. Random forest analysis revealed that functional microorganisms were the most important factor associated with the corresponding N transformation process. Overall, our results highlight the importance of N fixation in evaluating N budget of estuarine and coastal wetlands, providing valuable insights into the ecological effect of S. alterniflora invasion.

Effects of acidification on nitrification and associated nitrous oxide emission in estuarine and coastal waters.

In the context of an increasing atmospheric carbon dioxide (CO2) level, acidification of estuarine and coastal waters is greatly exacerbated by land-derived nutrient inputs, coastal upwelling, and complex biogeochemical processes. A deeper understanding of how nitrifiers respond to intensifying acidification is thus crucial to predict the response of estuarine and coastal ecosystems and their contribution to global climate change. Here, we show that acidification can significantly decrease nitrification rate but stimulate generation of byproduct nitrous oxide (N2O) in estuarine and coastal waters. By varying CO2 concentration and pH independently, an expected beneficial effect of elevated CO2 on activity of nitrifiers ("CO2-fertilization" effect) is excluded under acidification. Metatranscriptome data further demonstrate that nitrifiers could significantly up-regulate gene expressions associated with intracellular pH homeostasis to cope with acidification stress. This study highlights the molecular underpinnings of acidification effects on nitrification and associated greenhouse gas N2O emission, and helps predict the response and evolution of estuarine and coastal ecosystems under climate change and human activities.

Nitrous oxide emissions from subtropical estuaries: Insights for environmental controls and implications.

Estuaries are expected to contribute large nitrous oxide (N2O) emissions, however the environmental controls and implications of N2O emissions have not been well understood. Here we investigated water N2O concentrations, fluxes and sources in wet and dry seasons for 2019-2020 in five subtropical estuaries spanning hydrologic characteristics and nitrogen concentrations gradient. Water dissolved N2O concentrations and fluxes were in a range of 15.8-84.9 nmol L-1 and 0.66-22.2 µg m-2 h-1, respectively. These studied estuaries were oversaturated in N2O, with the saturations of 118-615%. Water dissolved N2O concentrations, saturations and fluxes increased significantly as nitrogen concentrations increase, whereas they did not differ significantly between the wet and dry seasons. Water N2O emissions, however, were also lower in the estuaries characterized by large discharge and water flow. N2O saturations and fluxes were determined directly by water nitrogen and oxygen concentrations and more indirectly by water temperature and velocity. The δ15N-N2O and site preference-N2O varied respectively from 2.86 to 11.31‰ and from 1.58 to 11.72‰, which overlapped the values between nitrification and denitrification. Nitrification and denitrification were responsible for 18.7-38.1% and 61.9-81.3% of N2O emissions, respectively. Indirect N2O emission factors were 0.08-0.14% and decreased with increasing total nitrogen concentrations. It is estimated that water N2O emissions in CO2 equiv could offset approximately 4.9% of average CO2 sink of China estuaries. Therefore, these results suggest that nitrogen concentrations and hydrologic characteristics together modify N2O emissions and that estuaries may be the important contributors to N2O emissions.

Microbial dynamics and activity of denitrifying anaerobic methane oxidizers in China's estuarine and coastal wetlands.

Estuarine and coastal wetlands, which act as large sources of methane (CH4) and undergo substantial loading of anthropogenic nitrogen (N), provide ideal conditions for denitrifying anaerobic methane oxidation (DAMO) to occur. Yet the microbial mechanisms governing DAMO and the main driving factors in estuarine and coastal ecosystems remain unclear. This study investigated the spatiotemporal distribution and associated activity of DAMO microorganisms along a wide swath of China's coastline (latitudinal range: 22-41°N) using molecular assays and isotope tracing techniques. We uncovered significant spatial and seasonal variation in DAMO bacterial community structure, whereas DAMO archaeal community structure exhibited no seasonal differences. The abundance of DAMO bacterial pmoA gene (2.2 × 105-1.0 × 107 copies g-1) was almost one order of magnitude higher than that of DAMO archaeal mcrA gene (8.7 × 104 -1.8 × 106 copies g-1). A significant positive correlation between pmoA and mcrA gene abundances (p < 0.01) was observed, indicating that DAMO bacteria and archaea may cooperate closely and thus complete nitrate elimination. Potential DAMO rates, in the range of 0.09-23.4 nmol 13CO2 g-1 day-1 for nitrite-DAMO and 0.03-43.7 nmol 13CO2 g-1 day-1 for nitrate-DAMO, tended to be greater in the relatively warmer low-latitudes. Potential DAMO rates were weakly positively correlated with gene abundances, suggesting that DAMO microbial activity could not be predicted directly by gene abundance alone. The heterogeneous variability of DAMO was shaped by interactions among key environmental characteristics (sediment texture, N availability, TOC, Fe3+, salinity of water, and temperature). On a broader continental scale, potential N removal rates of 0.1-11.2 g N m-2 yr-1 were estimated via nitrite-DAMO activity in China's coastal wetlands. Overall, our results highlight the widespread distribution of DAMO microbes and their potential role in eliminating excess N inputs and reducing CH4 emissions in estuarine and coastal ecosystems, which could help mitigate global warming.

High importance of coupled nitrification-denitrification for nitrogen removal in a large periodically low-oxygen estuary.

The coupling between nitrification and denitrification/anammox (nitrate/nitrite used in denitrification/anammox derives from nitrification) is a significant process of reactive nitrogen (N) removal that has attracted much attention. However, the dynamics of coupled nitrification-denitrification/anammox in the periodically low-oxygen estuaries and coasts remain unclear. Here, continuous-flow experiments combined with isotope tracing techniques were conducted in periodically low-oxygen areas of the Yangtze Estuary to reveal the changes in benthic sediment denitrification and anammox as well as their coupling with nitrification. Our results showed that denitrification increased but anammox decreased during low-oxygen summer. The occurrence of low oxygen also promoted coupled nitrification-denitrification but decreased coupled nitrification-anammox. These results implied that decreased dissolved oxygen in summer did not largely restrict nitrification activity, and anaerobic denitrification/anammox regulated the magnitude of coupled nitrification-denitrification/anammox rates. Denitrification (74.95-100 %) was the dominant process in total N removal, while coupled nitrification-denitrification accounted for a higher proportion (45.68-97.05 %) of denitrification, indicating that coupling between nitrification and denitrification played a dominant role in N removal. In addition to dissolved oxygen levels, carbon and N substrate availabilities were also important variables to regulate N transformations. Overall, this study advanced our knowledge of the distribution patterns and controlling factors of N removal processes and highlighted that coupled nitrification-denitrification might have a significant but neglected role in N removal from periodically low-oxygen estuaries.