Assimilatory Nitrate Reduction Research Articles

Bacterial community composition and metabolic characteristics of three representative Marine areas in northern China

Bacteria are essential components of ecosystems, participating in nutrient cycling and biogeochemical processes, and playing a crucial role in maintaining the stability of marine ecosystems. However, the biogeographic distribution patterns of bacterial diversity and metabolic functions in the estuarine and coastal areas of northern China remain unclear. Here, we used metagenomic sequencing to investigate the bacterial community composition and metabolic functions in sediments from the adjacent waters of the Yellow River Estuary, the Yellow Sea Cold Water Mass, and the adjacent waters of the Yangtze River Estuary. Among the 9164 species that were found, the most dominant microbial communities are Pseudomonadota, Actinomycetota, Bacteroidota, and Bacillota, but there are significant differences in the species composition in these three typical habitats. Amino acid metabolism and carbohydrate metabolic pathways were highly enriched. Glycoside hydrolases (GHs) predominate in carbon metabolism across all samples. In nitrogen metabolic pathway, genes related to organic degradation and synthesis are more abundant in the Yellow River Estuary than the other two habitats. In sulfur metabolic pathway, genes involved in assimilatory sulfate reduction are significantly enriched. Assimilatory sulfate reduction might be crucial for sulfur metabolism in coastal regions, with a full assimilatory nitrate reduction pathway found in Desulfobacterota. This research offers insights into the compositional diversity, metabolic functions, and biogeographic distribution patterns of bacterial communities in sediments from typical marine areas of northern China.

Morpho-Molecular and Genomic Characterization of Penicillium mexicanum Isolates Retrieved from a Forsaken Gold Mine

During the ongoing studies designed to examine the fungal diversity present within the abandoned and flooded Escádia Grande gold mine (Góis, Portugal), we repeatedly isolated several specimens belonging to a Penicillium species. Molecular phylogenetic analysis, coupled with morphological observations, positioned this fungus within subgen. Penicillium sect. Paradoxa, series Atramentosa, pinpointing its identity as Penicillium mexicanum (the first record for mining soils and the country). Given the limited research conducted on Penicillia isolated from similar environments, the species genome was sequenced utilizing the Oxford Nanopore® MinION™ methodology and studied through bioinformatic analysis. The obtained genome has a size of 29.62 Mb, containing a 47.72% GC content, 10,156 genes, with 44 rRNAs and 178 tRNAs/tmRNAs, providing the first genomic resource for this microorganism. Bioinformatic analysis allowed us to identify multiple genomic traits that can contribute towards this species survival in these extreme environments, including the presence of high levels of major facilitator transporters (MFS), Zn (2)-C6 fungal-type DNA-binding domains, P-loop containing nucleoside triphosphate hydrolases, specific fungal transcription factors and sugar transporters. Furthermore, putative advantageous metabolic traits, such as methylotrophy, assimilatory nitrate and sulfate reduction abilities, were also detected. In addition, the results also highlighted a strong genomic and metabolic organization and investment towards arsenic detoxification (transport and oxidation). Lastly, thirty-two putative biosynthetic gene clusters were predicted, including some with high similarity values to monascorubrin, nidulanin A, histidyltryptophanyldiketopiperazine/dehydrohistidyltryptophanyldiketopiperazine/roquefortine D/roquefortine C/glandicoline A/glandicoline B/meleagrine, YWA1 and choline. Overall, this study expands the current Penicillia knowledge from mining environments while also enhancing our understanding regarding fungal arsenic resistance.

Rhodobacteraceae are Prevalent and Ecologically Crucial Bacterial Members in Marine Biofloc Aquaculture.

Bioflocs are microbial aggregates primarily composed of heterotrophic bacteria that play essential ecological roles in maintaining animal health, gut microbiota, and water quality in biofloc aquaculture systems. Despite the global adoption of biofloc aquaculture for shrimp and fish cultivation, our understanding of biofloc microbiota-particularly the dominant bacterial members and their ecological functions-remains limited. In this study, we employed integrated metataxonomic and metagenomic approaches to demonstrate that the family Rhodobacteraceae of Alphaproteobacteria consistently dominates the biofloc microbiota and plays essential ecological roles. We first analyzed a comprehensive metataxonomic dataset consisting of 200 16S rRNA gene amplicons collected across three Asian countries: South Korea, China, and Vietnam. Taxonomic investigation identified Rhodobacteraceae as the dominant and consistent bacterial members across the datasets. The predominance of this taxon was further validated through metagenomics approaches, including read taxonomy and read recruitment analyses. To explore the ecological roles of Rhodobacteraceae, we applied genome-centric metagenomics, reconstructing 45 metagenome-assembled genomes. Functional annotation of these genomes revealed that dominant Rhodobacteraceae genera, such as Marivita, Ruegeria, Dinoroseobacter, and Aliiroseovarius, are involved in vital ecological processes, including complex carbohydrate degradation, aerobic denitrification, assimilatory nitrate reduction, ammonium assimilation, and sulfur oxidation. Overall, our study reveals that the common practice of carbohydrate addition in biofloc aquaculture systems fosters the growth of specific heterotrophic bacterial communities, particularly Rhodobacteraceae. These bacteria contribute to maintaining water quality by removing toxic nitrogen and sulfur compounds and enhance animal health by colonizing gut microbiota and exerting probiotic effects.

Soil metagenomics reveals reduced tillage improves soil functional profiles of carbon, nitrogen, and phosphorus cycling in bulk and rhizosphere soils

Conservation tillage practices (CAT) are known to benefit soil health and soil ecosystem functions relative to conventional tillage (CVT); however, much uncertainty remains concerning microbial functional traits and their subsequent effects on soil nutrients under different tillage practices. We analyzed the functional profiles of the C, N, and P cycles in response to CAT of no-tillage (NT), reduced tillage (RT), and CVT of moldboard plowing (MP) in bulk and rhizosphere soils using shotgun sequencing. CAT induced distinct microbial functional patterns relative to CVT, and these differences were generally more evident in rhizosphere soils than in bulk soils. CAT promotes multiple metabolic pathways such as C and N decomposition, fermentation, CO oxidation, N fixation, nitrate reduction and inorganic-P and organic-P transformations in bulk and/or rhizosphere soils. Variations in these metabolic pathways were mainly driven by Bradyrhizobium, Mesorhizobium, Nitrososphaera, Phenylobacterium, Rhizobium which are affiliated with Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidota and Thaumarchaeota. Furthermore, 24 high-quality metagenome-assembled genomes (MAGs) were reconstructed, of which three novel MAGs (TL100, TL46, and TL57) harbored functional genes regulating all metabolic pathways. In particular, NT-enriched MAGs (such as Sphingomonas) promote fermentation, resulting in the reduction of soil total carbon (TC) relative to MP in bulk soils. RT retained the contents of soil TC and total nitrogen (TN) well and up-regulated the phoR gene carried by Streptomyces, which promoted the regulation of P-starvation concomitantly with the increase in the contents of total phosphorous (TP) and available phosphorous (AP) in bulk soil. Additionally, assimilatory nitrate reduction coupled with organic-P mineralization was facilitated by CAT in rhizosphere soil, leading to the mitigation of N loss and the activation of soil organic-P for crop uptake. Overall, our results revealed that CAT significantly accelerated multiple metabolic potentials, and RT could sustain soil nutrient contents better than NT.

Revealing the metabolic potential and environmental adaptation of nematophagous fungus, Purpureocillium lilacinum, derived from hadal sediment.

The extreme environment shapes fungi in deep-sea sediments with novel metabolic capabilities. The ubiquity of fungi in deep-sea habitats supports their significant roles in these ecosystems. However, there is limited research on the metabolic activities and adaptive mechanisms of filamentous fungi in deep-sea ecosystems. In this study, we investigated the biological activities, including antibacterial, antitumor and nematicidal activity of Purpureocillium lilacinum FDZ8Y1, isolated from sediments of the Mariana Trench. A key feature of P. lilacinum FDZ8Y1 was its tolerance to high hydrostatic pressure (HHP), up to 110 MPa. We showed that HHP affected its vegetative growth, development, and production of secondary metabolites, indicating the potential for discovering novel natural products from hadal fungi. Whole-genome sequencing of P. lilacinum FDZ8Y1 revealed the metabolic potential of this piezotolerant fungus in carbon (carbohydrate metabolism), nitrogen (assimilatory nitrate reduction and protein degradation) and sulfur cycling processes (assimilatory sulfate reduction). Transcriptomic analysis under elevated HHP showed that P. lilacinum FDZ8Y1 may activate several metabolic pathways and stress proteins to cope with HHP, including fatty acid metabolism, the antioxidant defense system, the biosynthetic pathway for secondary metabolites, extracellular enzymes and membrane transporters. This study provides valuable insights into the metabolic potential and adaptation mechanisms of hadal fungi to the challenging conditions of the hadal environment.

Microbial Inoculants Modify the Functions of Resident Soil Microbes to Expedite the Field Restoration of the Abandoned Mine

ABSTRACTGlobal‐scale mining activities have had significant deleterious impacts on local ecosystems and the overall environment, which will necessitate robust restoration efforts. A practical approach includes combining microbial inoculants with the technology of external soil spray seeding. This approach holds the potential for sustainable abandoned mine site restoration by enhancing plant growth through the modulation of soil nutrients and microbial communities. Nonetheless, the detailed effects of microbial inoculants on specific aspects of soil microbial community functions and their complex interactions with plant growth remain underexplored, particularly in the context of restoration efforts. To bridge this gap, we performed a four‐year field study at an abandoned carbonate mine location, using metagenomic sequencing to evaluate the influence of microbial inoculants on soil microbial functionality. Our research revealed that introducing microbial inoculants greatly enhanced essential soil parameters and notably increased plant biomass. Additionally, these inoculants altered the functional gene makeup of the microbial community, significantly boosting the relative abundance of processes such as nitrogen fixation, nitrification, denitrification, assimilatory nitrate reduction (ANRA), dissimilatory nitrate reduction (DNRA), and organic phosphorus mineralization. Conversely, there was a decrease in the relative abundance of carbon degradation, phosphorus regulation, and transport processes. We observed strong correlations between the abundance of nitrogen and phosphorus cycles and plant biomass. Crucially, microbial inoculants affect plant biomass by initially altering soil properties and subsequently coordinating nitrogen and phosphorus cycles. These findings provide valuable insights into the role of microbial inoculants in mine site restoration and offer a theoretical foundation for their broader practical application.

Adaptive denitrification adjustment by marimo for enhanced nitrate nitrogen removal in lake restoration

This work investigated the effectiveness of marimo in restoring lake ecosystems, with a focus on its capacity for adaptive denitrification. Observations revealed a significant increase in granule size, benefiting its settling capability and thereby facilitating substantial reductions in nitrate levels and overall lake water purification. Notably, nitrate nitrogen was identified as pivotal in the nitrogen cycle, exhibiting a strong coupling relationship with denitrification processes. Marimo autonomously adjusted its denitrification in response to environmental nitrate levels, enhancing its nitrogen removal efficiency. Furthermore, Rhodobacteraceae was found to play an important role in nitrogen metabolism, with competitive interactions between Proteobacteria and Bacteroidota, influencing the overall efficiency of nitrogen removal. It was found that at first, the accumulation of nitrate enhanced denitrification, while afterwards, the decrease in nitrate concentration weakened denitrification. In addition, assimilatory nitrate reduction showed an upward trend throughout the entire process. Analysis of metabolic pathways further supported the adaptive denitrification adjustment mechanism of marimo. These findings highlight the potential of marimo as a natural solution for water quality improvement in lake ecosystems.

Functional Genes and Metabolic Pathways of Nitrogen Metabolism Microorganisms in Lake Sediments：A Case Study of Hongfeng Lake, Guizhou Province

The nitrogen cycle is of great importance for material circulation and energy flow in lake ecosystems. It is driven by microorganisms in lake sediments and can contribute to balancing lake ecosystems. In this study, physical and chemical properties of the sediments sampled from Hongfeng Lake in Guizhou Province were assayed and analyzed using metagenomics to reveal relevant microorganisms, functional genes, metabolic pathways, and their relationships throughout nitrogen metabolism. The results showed that bacteria were dominant, and the top three relative abundant genera were Thiobacillus （16.64%）, Rubrivivax（9.43%）, and Nitrospira （7.09%）. Only six pathways, including nitrogen fixation, nitrification, denitrification, assimilatory nitrate reduction, dissimilatory nitrate reduction, and complete nitrification, were detected in total, of which denitrification and dissimilatory nitrate reduction were the primary processes, but anaerobic ammonia oxidation was not detected. Bacteria and archaea participated in these six pathways, while eukaryotes only functioned in dissimilatory nitrate reduction, denitrification, and complete nitrification. Ammonia nitrogen, nitrate nitrogen, and total phosphorus, as main environmental factors affecting the distribution of functional genes for nitrogen metabolism, differentiated with each other in their respective real-world conditions. A positive correlation （95.04%） was observed between the functional genes and microorganisms, and narG, narZ, and nxrA possessed the highest abundance and the highest host genes. On this basis, these findings are expected to further elucidate the nitrogen cycle of typical karst lakes in Guizhou Province.

Boreal moss-microbe interactions are revealed through metagenome assembly of novel bacterial species

Moss-microbe interactions contribute to ecosystem processes in boreal forests. Yet, how host-specific characteristics and the environment drive the composition and metabolic potential of moss microbiomes is still poorly understood. In this study, we use shotgun metagenomics to identify the taxonomy and metabolic potential of the bacteria of four moss species of the boreal forests of Northern Québec, Canada. To characterize moss bacterial community composition and diversity, we assembled the genomes of 110 potentially novel bacterial species. Our results highlight that moss genus, species, gametophyte section, and to a lesser extent soil pH and soil temperature, drive moss-associated bacterial community composition and diversity. In the brown gametophyte section, two Stigonema spp. showed partial pathway completeness for photosynthesis and nitrogen fixation, while all brown-associated Hyphomicrobiales had complete assimilatory nitrate reduction pathways and many nearly complete carbon fixation pathways. Several brown-associated species showed partial to complete pathways for coenzyme M and F420 biosynthesis, important for methane metabolism. In addition, green-associated Hyphomicrobiales (Methylobacteria spp.) displayed potential for the anoxygenic photosystem II pathway. Overall, our findings demonstrate how host-specific characteristics and environmental factors shape the composition and metabolic potential of moss bacteria, highlighting their roles in carbon fixation, nitrogen cycling, and methane metabolism in boreal forests.

Antarctic Soils Select Copiotroph-Dominated Bacteria.

The life strategies of bacterial communities determine their structure and function and are an important driver of biogeochemical cycling. However, the variations in these strategies under different soil resource conditions remain largely unknown. We explored the bacterial life strategies and changes in structure and functions between Antarctic soils and forest (temperate, subtropical, and tropical) soils. The results showed that the weighted mean rRNA operon copy number in temperate soils was 19.5% lower than that in Antarctic soils, whereas no significant differences were observed among Antarctic, subtropical, and tropical soils. An unexpected result was that bacterial communities in Antarctic soils tended to be copiotrophs, such as Actinobacteriota and Bacteroidota, whereas those in temperate soils tended to be oligotrophs, such as Acidobacteriota and Chloroflexi. Functional predictions showed that in comparison to copiotrophs in Antarctic soils, temperate-inhabiting oligotrophic bacteria exhibited an 84.2-91.1% lower abundance of labile C decomposition genes (hemicellulose, cellulose, monosaccharides, and disaccharides), whereas a 74.4% higher abundance of stable C decomposition (lignin). Genes involved in N cycling (nitrogen fixation, assimilatory nitrate reduction, and denitrification) were 24.3-64.4% lower in temperate soils than in Antarctic soils. Collectively, our study provides a framework for describing the life strategies of soil bacteria, which are crucial to global biogeochemical cycles.

Short-Term Effects of Cenchrus fungigraminus/Potato or Broad Bean Interplanting on Rhizosphere Soil Fertility, Microbial Diversity, and Greenhouse Gas Sequestration in Southeast China.

Cenchrus fungigraminus is a new species and is largely used as forage and mushroom substrate. However, it can usually not be planted on farmland on account of local agricultural land policy. Interplanting Cenchrus fungigraminus with other crops annually (short-term) is an innovative strategy to promote the sustainable development of the grass industry in southern China. To further investigate this, C. fungigraminus mono-planting (MC), C. fungigraminus-potato interplanting (CIP) and C. fungigraminus-broad bean interplanting (CIB) were performed. Compared to MC, soil microbial biomass carbon (SMBC), soil organic matter (SOM), ammoniacal nitrogen (AMN), pH and soil amino sugars had a positive effect on the rhizosphere soil of CIP and CIB, as well as enhancing soil nitrogenase, nitrite reductase, and peroxidase activities (p < 0.05). Moreover, CIP improved the root vitality (2.08 times) and crude protein (1.11 times). In addition, CIB enhanced the crude fiber of C. fungigraminus seedlings. These two interplanting models also improved the microbial composition and diversity (Actinobacteria, Firmicutes, and Bacteroidota, etc.) in the rhizosphere soil of C. fungigraminus seedlings. Among all the samples, 189 and 59 genes were involved in methane cycling and nitrogen cycling, respectively, which improved the presence of the serine cycle, ribulose monophosphate, assimilatory nitrate reduction, methane absorption, and glutamate synthesis and inhibited denitrification. Through correlation analysis and the Mantel test, the putative functional genes, encoding functions in both nitrogen and methane cycling, were shown to have a significant positive effect on pH, moisture, AMN, SOM, SMBC, and soil peroxidase activity, while not displaying a significant effect on soil nitrogenase activity and total amino sugar (p < 0.05). The short-term influence of the interplanting model was shown to improve land use efficiency and economic profitability per unit land area, and the models could provide sustainable agricultural production for rural revitalization.

Nitrogen removal pathways in lake restoration using microalgal-bacterial granular sludge: Unraveling influence of organics and carbon to nitrogen ratio

This study investigated the performance of microalgal-bacterial granular sludge (MBGS) in the restoration of Qingling Lake and Huangjia Lake, focusing on nitrogen removal under varying water quality conditions. Significant color changes in MBGS and differences in granule characteristics were observed, with Qingling Lake demonstrating superior removal efficiencies for ammonia nitrogen, nitrate nitrogen, and total nitrogen compared to Huangjia Lake. Stoichiometric analysis revealed that when the chemical oxygen demand (COD) and carbon-to-nitrogen (C/N) ratios were less than 20 mg/L and 20, respectively, assimilatory nitrate reduction was positively correlated with both, whereas denitrification was negatively correlated. Gene function analysis showed that Qingling Lake had a more active microbial community supporting efficient nitrogen metabolism. The findings highlighted the enormous potential of MBGS in lake restoration, demonstrating its ability to adapt to different COD concentrations and C/N ratios by altering its nitrogen removal pathways.

Differential regulation of belowground rhizospheric ecosystem by biological and chemical nitrogen supplies: implications for maize yield enhancement mechanisms.

Nitrogen (N) content affects aboveground maize growth and nutrient absorption by altering the belowground rhizospheric ecosystem, impacting both yield and quality. However, the mechanisms through which different N supply methods (chemical and biological N supplies) regulate the belowground rhizospheric ecosystem to enhance maize yield remain unclear. To address this issue, we conducted a field experiment in northeast China, comprising three treatments: maize monocropping without N fertilizer application (MM), maize/alfalfa intercropping without N fertilizer application (BNF), and maize monocropping with N fertilizer application (CNS). The MM treatment represents the control, while the BNF treatment represents the biological N supply form, and CNS treatment represents the chemical N supply form. In the autumn of 2019, samples of maize and rhizospheric soil were collected to assess parameters including yield, rhizospheric soil characteristics, and microbial indicators. Both BNF and MM significantly increased maize yield and different yield components compared with MM, with no statistically significant difference in total yield between BNF and CNS. Furthermore, BNF significantly improved N by 12.61% and available N (AN) by 13.20% compared with MM. Furthermore, BNF treatment also significantly increased the Shannon index by 1.90%, while the CNS treatment significantly increased the Chao1 index by 28.1% and ACE index by 29.49%, with no significant difference between CNS and BNF. However, CNS had a more pronounced impact on structure of the rhizosphere soil bacterial community compared to BNF, inducing more significant fluctuations within the microbial network (modularity index and negative cohesion index). Regarding N transformation pathways predicted by bacterial functions, BNF significantly increased the N fixation pathway, while CNS significantly increased assimilatory nitrate reduction. In CNS, AN, NO3-N, NH4-N, assimilatory nitrate reduction, and community structure contributed significantly to maize yield, whereas in BNF, N fixation, community structure, community stability, NO3-N, and NH4-N played significant roles in enhancing maize yield. While CNS and BNF can achieve comparable maize yields in practical agricultural production, they have significantly different impacts on the belowground rhizosphere ecosystem, leading to different mechanisms of yield enhancement.

The microbial community structure and nitrogen cycle of high-altitude pristine saline lakes on the Qinghai-Tibetan plateau.

The nitrogen (N) cycle is the foundation of the biogeochemistry on Earth and plays a crucial role in global climate stability. It is one of the most important nutrient cycles in high-altitude lakes. The biogeochemistry of nitrogen is almost entirely dependent on redox reactions mediated by microorganisms. However, the nitrogen cycling of microbial communities in the high-altitude saline lakes of the Qinghai-Tibet Plateau (QTP), the world's "third pole" has not been investigated extensively. In this study, we used a metagenomic approach to investigate the microbial communities in four high-altitude pristine saline lakes in the Altun mountain on the QTP. We observed that Proteobacteria, Bacteroidota, and Actinobacteriota were dominant in these lakes. We reconstructed 1,593 bacterial MAGs and 8 archaeal MAGs, 1,060 of which were found to contain nitrogen cycle related genes. Our analysis revealed that nitrite reduction, nitrogen fixation, and assimilatory nitrate reduction processes might be active in the lakes. Denitrification might be a major mechanism driving the potential nitrogen loss, while nitrification might be inactive. A wide variety of microorganisms in the lake, dominated by Proteobacteria, participate together in the nitrogen cycle. The prevalence of the dominant taxon Yoonia in these lakes may be attributed to its well-established nitrogen functions and the coupled proton dynamics. This study is the first to systematically investigate the structure and nitrogen function of the microbial community in the high-altitude pristine saline lakes in the Altun mountain on the QTP. As such, it contributes to a better comprehension of biogeochemistry of high-altitude saline lakes.

Different effects of seasonal impoundment and land use change on microbiome in a tributary sediment of the three gorgers reservoir

Anthropogenic activities significantly impact river ecosystem nutrient fluxes and microbial metabolism. Here, we examined the seasonal and spatial variation of sediments physicochemical parameters and the associated microbiome in the Pengxi river, a representative tributary of Three Gorges Reservoir, in response to seasonal impoundment and land use change by human activities. Results revealed that seasonal impoundment and land use change enhanced total organic carbon (TOC), total nitrogen (TN) and ammonium nitrogen (NH4+-N) concentration in the sediment, but have different effects on sediment microbiome. Sediment microbiota showed higher similarity during the seasonal high-water level (HWL) in consecutive two years. The abundant phyla Acidobacteria, Gemmatimonadetes, Cyanobacteria, Actinobacteria and Planctomycetes significantly increased as water level increased. Along the changes in bacterial taxa, we also observed changes in predicted carbon fixation functions and nitrogen-related functions, including the significantly higher levels of Calvin cycle, 4HB/3HP cycle, 3HP cycle and assimilatory nitrate reduction, while significantly lower level of denitrification. Though land use change significantly increased TOC, TN and NH4+-N concentration, its effects on spatial variation of bacterial community composition and predicted functions was not significant. The finding indicates that TGR hydrologic changes and land use change have different influences on the carbon and nitrogen fluxes and their associated microbiome in TGR sediments. A focus of future research will be on assessing on carbon and nitrogen flux balance and the associated carbon and nitrogen microbial cycling in TGR sediment.

Unraveling the mechanism of assimilatory nitrate reduction and methane oxidation by Methylobacter sp. YHQ through dual N-O isotope analysis and kinetic modeling

Assimilatory nitrate reduction and methane (CH4) oxidation by bacteria play important roles in carbon (C) and nitrogen (N) biogeochemical cycles. Here, an investigation of enzymatic assimilatory nitrate reduction and CH4 oxidation by Methylobacter sp. YHQ from the wetlands is presented, specifically concentrating on N and oxygen (O) isotope fractionation with various initial nitrate and oxygen concentrations. The N enrichment factors (15εassimilation) increased from 4.2 ± 0.7‰ to 6.9 ±1.3‰ and the O isotope enrichment factors (18εassimilation) increased from 2.7 ± 0.9‰ to 4.7 ± 0.8‰ during nitrate assimilation when initial nitrate concentrations increased from 0.9 mM to 2 mM. Similar 18ε and 15ε values were observed at different oxygen concentrations. The values of 18ε and 15ε provided vital parameters for the assessment of assimilatory nitrate reduction via the Rayleigh equation approach. The ratios of O and N isotope enrichment factors (18ε:15ε)assimilation ranged from 0.64 ± 0.15 to 0.74 ± 0.18 during nitrate assimilation by Methylobacter sp. YHQ with Nas, which were different from (18ε:15ε)assimilation for assimilatory eukaryotic nitrate reductase (eukNR) from literature data. Thus, N and O isotope fractionation could be useful tools to distinguish eukNR from Nas during nitrate assimilation. Additionally, the rates of CH4 oxidation and nitrate reduction were evaluated with a reaction-based kinetic model, and it quantitatively described the enzymatic reactions of nitrate assimilation. Combining dual N-O isotope analysis with kinetic modeling provides new insights into the microbially driven C-N interactions.Graphical

Salinity affects microbial function genes related to nutrient cycling in arid regions.

Salinization damages soil system health and influences microbial communities structure and function. The response of microbial functions involved in the nutrient cycle to soil salinization is a valuable scientific question. However, our knowledge of the microbial metabolism functions in salinized soil and their response to salinity in arid desert environments is inadequate. Here, we applied metagenomics technology to investigate the response of microbial carbon (C), nitrogen (N), phosphorus (P), and sulfur (S) cycling and the key genes to salinity, and discuss the effects of edaphic variables on microbial functions. We found that carbon fixation dominated the carbon cycle. Nitrogen fixation, denitrification, assimilatory nitrate reduction (ANRA), and nitrogen degradation were commonly identified as the most abundant processes in the nitrogen cycle. Organic phosphorus dissolution and phosphorus absorption/transport were the most enriched P metabolic functions, while sulfur metabolism was dominated by assimilatory sulfate reduction (ASR), organic sulfur transformation, and linkages between inorganic and organic sulfur transformation. Increasing salinity inhibited carbon degradation, nitrogen fixation, nitrogen degradation, anammox, ANRA, phosphorus absorption and transport, and the majority of processes in sulfur metabolism. However, some of the metabolic pathway and key genes showed a positive response to salinization, such as carbon fixation (facA, pccA, korAB), denitrification (narG, nirK, norBC, nosZ), ANRA (nasA, nirA), and organic phosphorus dissolution processes (pstABCS, phnCD, ugpAB). High salinity reduced the network complexity in the soil communities. Even so, the saline microbial community presented highly cooperative interactions. The soil water content had significantly correlations with C metabolic genes. The SOC, N, and P contents were significantly correlated with C, N, P, and S network complexity and functional genes. AP, NH4+, and NO3- directly promote carbon fixation, denitrification, nitrogen degradation, organic P solubilization and mineralization, P uptake and transport, ASR, and organic sulfur transformation processes. Soil salinity in arid region inhibited multiple metabolic functions, but prompted the function of carbon fixation, denitrification, ANRA, and organic phosphorus dissolution. Soil salinity was the most important factor driving microbial functions, and nutrient availability also played important roles in regulating nutrient cycling.

Co-expression analysis reveals distinct alliances around two carbon fixation pathways in hydrothermal vent symbionts

Most autotrophic organisms possess a single carbon fixation pathway. The chemoautotrophic symbionts of the hydrothermal vent tubeworm Riftia pachyptila, however, possess two functional pathways: the Calvin–Benson–Bassham (CBB) and the reductive tricarboxylic acid (rTCA) cycles. How these two pathways are coordinated is unknown. Here we measured net carbon fixation rates, transcriptional/metabolic responses and transcriptional co-expression patterns of Riftia pachyptila endosymbionts by incubating tubeworms collected from the East Pacific Rise at environmental pressures, temperature and geochemistry. Results showed that rTCA and CBB transcriptional patterns varied in response to different geochemical regimes and that each pathway is allied to specific metabolic processes; the rTCA is allied to hydrogenases and dissimilatory nitrate reduction, whereas the CBB is allied to sulfide oxidation and assimilatory nitrate reduction, suggesting distinctive yet complementary roles in metabolic function. Furthermore, our network analysis implicates the rTCA and a group 1e hydrogenase as key players in the physiological response to limitation of sulfide and oxygen. Net carbon fixation rates were also exemplary, and accordingly, we propose that co-activity of CBB and rTCA may be an adaptation for maintaining high carbon fixation rates, conferring a fitness advantage in dynamic vent environments.

Low Bacterial Diversity and Nitrate Levels in Cores from Deep Boreholes in Pristine Karst.

This study investigates the nitrate gradients within the deep biosphere of karst carbonate rocks and their resident microbiota. Samples were taken from borehole cores at depths down to 350 m below the surface, collected during geological site investigations for proposed railway tunnels and analysed using 16S rRNA amplicon sequencing. 16S rRNA amplicon sequencing analysis revealed relatively low microbial diversity, which can serve as a reliable indicator of the pristine nature of deep karst. However, some local hotspots of diversity are independent of depth. Pseudomonadota dominated the samples, with Gammaproteobacteria dominating at the class level. The low nitrate content in deep karst, in contrast to higher values closer to the surface, serves as an additional marker of its undisturbed and unpolluted status. Based on the prediction of functional profiles from 16S rRNA sequencing data, nitrates remain low due to indigenous microbial denitrification and assimilatory nitrate reduction. Pathways related to nitrogen fixation, ammonia assimilation, and nitrification were not confirmed. When elevated nitrate levels are observed in karst, they are most probably related to anthropogenic activities. Environmental factors other than depth and nitrate content play an important role in shaping bacterial communities.

Efficient nitrogen removal by microalgal-bacterial granular sludge-marimo coupling process

Current biological wastewater treatment processes usually have a drawback of insufficient nitrogen (N) removal, contributing to the ubiquitous eutrophication of aquatic ecosystems globally. To address such a challenging situation, this study explored an innovative microalgal-bacterial granular sludge-marimo (MBGS-MA) coupling process. The process removed 83.4 % of N with the effluent N concentration of 4.0 mg/L. With the growth of MBGS, there was a shift towards genes associated with nitrification and denitrification, and away from ammonia assimilation genes, revealing internal mechanism of the shift of N removal pathway. Contrarily, MA could use gaseous N2 with the N fixing genes in MA enriched, and the genes abundance related to assimilatory nitrate reduction were also raised under the mutualistic interactions between Proteobacteria and Cyanobacteria, which was beneficial to achieve efficient N removal. These findings may open a new horizon for developing innovative hybrid microalgal-bacterial processes aimed at high-efficiency N removal from wastewater.