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The inefficient nitrogen removal in constructed wetlands (CWs) can be attributed to insufficient carbon sources for low carbon-to-nitrogen (C/N) ratio wastewater. In this study, sugarcane bagasse fermentation liquid (SBFL) was used as a supplemental carbon source in intermittently aerated CWs to enhance nitrogen removal. The impact of different regulated influent C/N ratios on nitrogen removal and greenhouse gas (GHG) emissions was investigated. Results demonstrated that SBFL addition significantly enhanced the denitrification capacity, resulting in faster NO 3 - -N removal compared to sucrose. Moreover, intermittently aerated CWs significantly improved NH 4 + -N removal efficiency compared to non-aerated CWs. The highest total nitrogen removal efficiency (98.3 %) was achieved at an influent C/N ratio of 5 in intermittently aerated CWs with SBFL addition. The addition of SBFL resulted in a reduction of N 2 O emissions by 17.8 %-43.7 % compared to sucrose. All CWs exhibited low CH 4 ) resulting in lower emissions compared to sucrose. Additionally, higher abundance of denitrification (nirK, nirS and nosZ) genes as well as more abundant denitrifying bacteria were shown in CWs of SBFL inputs. The results of this study provide a feasible strategy for applying SBFL as a carbon source to improve nitrogen removal efficiency and mitigate GHG emissions in CWs.-2 h -1 ) resulting in lower emissions compared to sucrose. Additionally, higher abundance of denitrification (nirK, nirS and nosZ) genes as well as more abundant denitrifying bacteria were shown in CWs of SBFL inputs. The results of this study provide a feasible strategy for applying SBFL as a carbon source to improve nitrogen removal efficiency and mitigate GHG emissions in CWs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Perchlorate (ClO 4 - ) mainly exists in the form of ammonium perchlorate in industrial production. However, the degradation mechanisms of different concentrations of ammonium nitrogen (NH 4 mixed pollutants in the environment are not well understood. This study aims to explore the potential of different types of carbon sources for ClO + -N) and ClO 4 -N biodegradation. Experimental results showed that the concentration and type of carbon sources are decisive to simultaneous removal of NH - mixed pollutants in the environment are not well understood. This study aims to explore the potential of different types of carbon sources for ClO 4 - and NH 4 + -N biodegradation. Experimental results showed that the concentration and type of carbon sources are decisive to simultaneous removal of NH 4 + -N and ClO 4 - . Under condition of C(COD)/C(ClO 4 - ) ratio of 21.15 ± 4.40, the simultaneously removal efficiency of ClO 4 - and NH 4 degrading bacteria (such as Proteobacteria, Chloroflexi, and Firmicutes) significantly increased by 528.57 % in Glu-C. It can be inferred that the concentration of carbon source and NH + -N in acetate (Ace) was relatively higher than that in methanol (Met). C(NH 4 + -N)/C(ClO 4 - ) ratio of 9.66 ± 0.51 and C(COD)/C(ClO 4 - ) ratio of 2.51 ± 0.87 promoted ClO 4 - reduction in glucose-C (Glu-C). However, high concentration of Glu could cause pH decrease (from 7.57 to 4.59), thereby inhibiting ClO 4 - reduction. High-throughput sequencing results indicated that Proteobacteria and Bacteroidetes have made a major contribution to the simultaneous removal of NH 4 + -N and ClO 4 - . They are two representative bacterial phyla for participating in both ClO 4 - reduction and denitrification. Notably, the abundance of main ClO 4 - degrading bacteria (such as Proteobacteria, Chloroflexi, and Firmicutes) significantly increased by 528.57 % in Glu-C. It can be inferred that the concentration of carbon source and NH 4 + -N were the most important factors determining the removal efficiency of ClO 4 - by influencing changes in the core microbial community. This study will provide new techniques and mechanistic insights for the simultaneous removal of mixed ClO 4 - and nitrogen pollutants, which can also provide theoretical support for innovation in future biological treatment processes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study investigated the ability of lactic acid bacteria (LAB) isolated from oranges to use fish by-products (FB) and chicken by-products (CB) as nitrogen sources alternative to yeast extract for lactic acid (LA) production in a papaya by-product medium as a carbon source. Once the fermentation agents had been isolated, they were subjected to biochemical and molecular characterization. Inexpensive nitrogen sources, precisely CB and FB, were prepared, freeze-dried, and yield evaluated. Also, before to the fermentation experiments, the Total Kjehdahl Nitrogen (TKN) of these by-products and that of the yeast extract were determined. Then, three production media differing in terms of nitrogen source were formulated from these nitrogen sources. From the 22 LAB isolated from orange, two isolates of interest (NGO25 and NGO23) were obtained; all belonging to the Lactiplantibacillus plantarum species based on 16S rRNA gene sequencing. Furthermore, the production yield powder obtained after lyophilization of 1 L of CB and FB surpernatant were, respectively, 16.6 g and 12.933 g. The TKN of different nitrogen sources powder were 71.4 ± 0.000% DM (FB), 86.145 ± 0.001% DM (CB), and 87.5 ± 0.99% DM (yeast extract). The best kinetic parameters of LA production (LA (g/L): 31.945 ± 0.078; volumetric productivity (g/L.h): 1.331 ± 0.003; LA yield (mg/g) 63.89 ± 0.156; biomass (g/L) 7.925 ± 0.035; cell growth rate (g/L.h): 0.330 ± 0.001) were recorded by Lactiplantibacillus plantarum NGO25 after 24 h of fermentation. The latter data were obtained in the production medium containing CB as nitrogen sources. In addition, this production medium cost only $0.152 to formulate, compared to yeast extract which required $1.692 to formulate. Thus, freeze-dried CB can be used as an alternative to yeast extract in large-scale production of LA., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2024 Romial Joel Ngouénam et al.)

The main challenge in removing nutrients from municipal wastewater in China is the lack of available carbon sources. While hydrolysis acidification tanks can improve wastewater biodegradability by effectively utilizing internal carbon sources, high sludge concentrations are difficult to control in traditional tank variants. In this study, an innovative anaerobic filter (AnF) hydrolysis acidification reactor composed of a continuously stirred tank reactor (CSTR) and cloth media filter was designed to regulate and maintain high sludge concentrations in the hydrolysis acidifier. The reactor was used as a pretreatment unit for the anaerobic/anoxic/oxic (AAO) units and combined into an AnF-AAO system to explore the effectiveness of internal carbon source utilization in wastewater. The results indicate that as the sludge concentration in the hydrolysis acidifier increased, the hydrolysis and acidification processes became more efficient. The optimal sludge concentration was 40 g/L, which significantly increased the production of soluble chemical oxygen demand and volatile fatty acids. Above this concentration, the efficiency decreased. Compared to traditional AAO processes, the AnF-AAO system achieved superior total nitrogen and phosphorus removal with shorter hydraulic retention times and reduced sludge production by a significant amount of 35%. Due to its capacity for enhancing internal carbon source utilization, the AnF-AAO system constitutes a promising approach for sustainable urban wastewater treatment., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Hongchen Wang reports financial support was provided by Renmin University of China School of Environment and Natural Resource. Hongchen Wang reports a relationship with Renmin University of China School of Environment and Natural Resource that includes: employment., (Copyright © 2024. Published by Elsevier Ltd.)

Potassium (K) and magnesium (Mg) play analogous roles in regulating plant photosynthesis and carbon and nitrogen (C-N) metabolism. Based on this consensus, we hypothesize that appropriate Mg supplementation may alleviate growth inhibition under low K stress. We monitored morphological, physiological, and molecular changes in G935 apple plants under different K (0.1 and 6 mmol L -1 ) and Mg supply (3 and 6 mmol L -1 ) conditions. Low K stress caused changes in root and leaf structure, inhibited photosynthesis, and limited the root growth of the apple rootstock. Further study on Mg supplementation showed that it could promote the uptake of K + and NO 3 - by upregulating the expression of K + transporter proteins such as Arabidopsis K + transporter 1 (MdAKT1), high-affinity K + transporter 1 (MdHKT1), and potassium transporter 5 (MdPT5) and nitrate transporters such as nitrate transporter 1.1/1.2/2.1/2.4 (MdNRT 1.1/1.2/2.1/2.4). Mg promoted the translocation of 15 N from roots to leaves and enhanced photosynthetic N utilization efficiency (PNUE) by increasing the proportion of photosynthetic N and alleviating photosynthetic restrictions. Furthermore, Mg supplementation improved the synthesis of photosynthates by enhancing the activities of sugar-metabolizing enzymes (Rubisco, SS, SPS, S6PDH). Mg also facilitated the transport of sucrose and sorbitol from leaves to roots by upregulating the expression of sucrose transporter 1.1/1.2/4.1/4.2 (MdSUT 1.1/1.2/4.1/4.2) and sorbitol transporter 1.1/1.2 (MdSOT 1.1/1.2). Overall, Mg effectively alleviated growth inhibition in apple rootstock plants under low K stress by facilitating the uptake of N and K uptake, optimizing nitrogen partitioning, enhancing nitrogen use efficiency (NUE) and PNUE, and promoting the photosynthate synthesis and translocation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Immobilized microalgae biotechnologies can conserve water and space by low-carbon wastewater treatment and resource recovery in a recirculating aquaculture system (RAS). However, technical process parameters have been unoptimized considering the mutual interaction between factors. In this study, machine learning optimized the parameters of alginate-immobilized Chlorella vulgaris (C. vulgaris), that is, 474 μmol/(m 2 ·s) of light intensity, 23 × 10 6 cells/mL for initial cell number, and 2.07 mm particle size. Importantly, under continuous illumination, the immobilized C. vulgaris and microalgal-bacterial consortium improved water purification and biomass reutilization. Transcriptomics of C. vulgaris showed enhanced nitrogen removal by increasing pyridine nucleotide and lipid accumulation via enhanced triacylglycerol synthesis. Symbiotic bacteria upregulated genes for nitrate reduction and organic matter degradation, which stimulated biomass accumulation through CO 2 fixation and starch synthesis. The recoverable microalgae (1.94 g/L biomass, 47 % protein, 26.23 % lipids), struvite (64.79 % phosphorus), and alginate (79.52 %) every two weeks demonstrates a low-carbon resource recovery in RAS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The efficient use of livestock and poultry manure waste has become a global challenge, with microorganisms playing an important role. To investigate the impact of novel ammonifying microorganism cultures (NAMC) on microbial community dynamics and carbon and nitrogen metabolism, five treatments [5% (v/w) sterilized distilled water, Amm-1, Amm-2, Amm-3, and Amm-4] were applied to cow manure compost. Inoculation with NAMC improved the structure of bacterial and fungal communities, enriched the populations of the functional microorganisms, enhanced the role of specific microorganisms, and promoted the formation of tight modularity within the microbial network. Further functional predictions indicated a significant increase in both carbon metabolism (CMB) and nitrogen metabolism (NMB). During the thermophilic phase, inoculated NAMC treatments boosted carbon metabolism annotation by 10.55%-33.87% and nitrogen metabolism annotation by 26.69%-63.11. Structural equation modeling supported the NAMC-mediated enhancement of NMB and CMB. In conclusion, NAMC inoculation, particularly with Amm-4, enhanced the synergistic interaction between bacteria and fungi. This collaboration promoted enzymatic catabolic and synthetic processes, resultng in positive feedback loops with the endogenous microbial community. Understanding these mechanisms not only unravels how ammonifying microorganisms influence microbial communities but also paves the way for the development of the composting industry and global waste management practices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Despite the growing catalogue of studies detailing the taxonomic and functional composition of soil bacterial communities, the life history traits of those communities remain largely unknown. This study analyzes a global dataset of soil metagenomes to explore environmental drivers of growth potential, a fundamental aspect of bacterial life history. We find that growth potential, estimated from codon usage statistics, was highest in forested biomes and lowest in arid latitudes. This indicates that bacterial productivity generally reflects ecosystem productivity globally. Accordingly, the strongest environmental predictors of growth potential were productivity indicators, such as distance to the equator, and soil properties that vary along productivity gradients, such as pH and carbon to nitrogen ratios. We also observe that growth potential was negatively correlated with the relative abundances of genes involved in carbohydrate metabolism, demonstrating tradeoffs between growth and resource acquisition in soil bacteria. Overall, we identify macroecological patterns in bacterial growth potential and link growth rates to soil carbon cycling., (© 2024. The Author(s).)

Nanoparticle (NP) pollution has negative impacts and is a major global environmental problem. However, the molecular response of alfalfa (Medicago sativa L.) to titanium dioxide nanoparticles (TiO 2 NPs) is limited. Herein, the dual effects of TiO 2 NPs (0-1000 mg L -1 ) on carbon (C) and nitrogen (N) metabolisms in alfalfa were investigated. The results showed that 500 mg L -1 TiO 2 NPs (Ti-500) had the highest phytotoxicity in the C/N metabolizing enzymes; and it significantly increased total soluble sugar, starch, sucrose, and sucrose-phosphate synthase. Furthermore, obvious photosynthesis responses were found in alfalfa exposed to Ti-500. By contrast, 100 mg L -1 TiO 2 NPs (Ti-100) enhanced N metabolizing enzymes. RNA-seq analyses showed 4265 and 2121 differentially expressed genes (DEGs) in Ti-100 and Ti-500, respectively. A total of 904 and 844 differentially expressed proteins (DEPs) were identified in Ti-100 and Ti-500, respectively. Through the physiological, transcriptional, and proteomic analyses, the DEGs and DEPs related to C/N metabolism, photosynthesis, chlorophyll synthesis, starch and sucrose metabolism, and C fixation in photosynthetic organisms were observed. Overall, TiO 2 NPs at low doses improve photosynthesis and C/N regulation, but high doses can cause toxicity. It is valuable for the safe application of NPs in agriculture., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Tree-mycorrhizal associations are associated with patterns in nitrogen (N) availability and soil organic matter storage; however, we still lack a mechanistic understanding of what tree and fungal traits drive these patterns and how they will respond to global changes in soil N availability. To address this knowledge gap, we investigated how arbuscular mycorrhizal (AM)- and ectomycorrhizal (EcM)-associated seedlings alter rhizodeposition in response to increased seedling inorganic N acquisition. We grew four species each of EcM and AM seedlings from forests of the eastern United States in a continuously 13 C-labeled atmosphere within an environmentally controlled chamber and subjected to three levels of 15 N-labeled fertilizer. We traced seedling 15 N uptake from, and 13 C-labeled inputs (net rhizodeposition) into, root-excluded or -included soil over a 5-month growing season. N uptake by seedlings was positively related to rhizodeposition for EcM- but not AM-associated seedlings in root-included soils. Despite this contrast in rhizodeposition, there was no difference in soil C storage between mycorrhizal types over the course of the experiment. Instead root-inclusive soils lost C, while root-exclusive soils gained C. Our findings suggest that mycorrhizal associations mediate tree belowground C investment in response to inorganic N availability, but these differences do not affect C storage. Continued soil warming and N deposition under global change will increase soil inorganic N availability and our seedling results indicate this could lead to greater belowground C investment by EcM-associated trees. This potential for less efficient N uptake by EcM-trees could contribute to AM-tree success and a shift toward more AM-dominated temperate forests., (© 2024 John Wiley & Sons Ltd.)

The feasibility of inducing simultaneous nitrification and denitrification (SND) by S 0 for low carbon to nitrogen (C/N) ratio wastewater remediation was investigated. Compared with S 0 and/or organics absent systems (-3.4 %∼5.0 %), the higher nitrogen removal performance (18.2 %∼59.8 %) was achieved with C/N ratios and S 0 dosages increasing when S 0 and organics added simultaneously. The synergistic effect of S 0 and organics stimulated extracellular polymeric substances secretion and weakened intermolecular binding force of S 0 , facilitating S 0 bio-utilization and reducing the external organics requirement. It also promoted microbial metabolism (0.16 ∼ 0.24 μg O 2 /(g VSS·h)) and ammonia assimilation (5.9 %∼20.5 %), thereby enhancing the capture of organics and providing more electron donors for SND. Furthermore, aerobic denitrifiers (15.91 %∼27.45 %) and aerobic denitrifying (napA and nirS) and ammonia assimilating genes were accumulated by this synergistic effect. This study revealed the mechanism of SND induced by coordination of S 0 and organics and provided an innovative strategy for triggering efficient and stable SND., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigated the nutrient removal and microbial community succession in moving bed biofilm reactor under stable and three levels of influent carbon/nitrogen (C/N) ratio fluctuation (± 10%, ± 20%, and ± 30%). Under the conditions of influent C/N ratio fluctuation, the removal efficiency of COD and PO 4 3 - -P decreased 4.7-6.4% and 3.7-12.9%, respectively, while the nitrogen removal was almost unaffected. A sharp decrease in the content of culturable functional bacteria related to nitrogen and phosphorus removal including nitrite-oxidizing bacteria (NOB), aerobic denitrifying bacteria (DNB), and polyphosphate-accumulating organisms (PAOs) from the carrier biofilm was observed. Sequencing analysis revealed that the abundance of Candidatus Competibacter increased 10.3-25.9% and became the dominant genus responsible for denitrification, potentially indicating that nitrate was removed via endogenous denitrification under the influent C/N ratio fluctuation. The above results will provide basic data for the nutrient removal in decentralized wastewater treatment under highly variable influent conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Effective nitrate removal is a key challenge when treating low carbon-to-nitrogen ratio wastewater. How to select an effective inorganic electron donor to improve the autotrophic denitrification of nitrate nitrogen has become an area of intense research. In this study, the nitrate removal mechanism of three iron-based materials in the presence and absence of microorganisms was investigated with Fe 2+ /Fe 0 as an electron donor and nitrate as an electron acceptor, and the relationship between the iron materials and denitrifying microorganisms was explored. The results indicated that the nitrogen removal efficiency of each iron-based material coupled sludge systems was higher than that of iron-based material. Furthermore, compared with the sponge iron coupled sludge system (60.6%-70.4%) and magnetite coupled sludge (56.1%-65.3%), the pyrite coupled sludge system had the highest removal efficiency of TN, and the removal efficiency increased from 62.5% to 82.1% with time. The test results of scanning electron microscope, X-ray photoelectron spectroscopy and X-ray diffraction indicated that iron-based materials promoted the attachment of microorganisms and the chemical reduction of nitrate in three iron-based material coupled sludge systems. Furthermore, the pyrite coupled sludge system had the highest nitrite reductase activity and can induce microorganisms to secrete more extracellular polymer substances. Combined with high-throughput sequencing and PICRUSt2 functional predictive analysis software, the total relative abundance of the dominant bacterial in pyrite coupled sludge system was the highest (72.06%) compared with the other iron-based material systems, and the abundance of Blastocatellaceae was relatively high. Overall, these results suggest that the pyrite coupled sludge system was more conducive to long-term stable nitrate removal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Researches on the advanced nitrogen (N) removal of municipal tailwater always overlooked the value of refractory degradable dissolved organic matter (R-DOM). In this study, a novel electric field coupled iron‑carbon biofilter (E-ICBF) was utilized to explore the performance and microbial changes with polyethylene glycol (PEG) as the representative R-DOM. Results demonstrated that the removal efficiencies of E-ICBF for nitrate nitrogen (NO 3 - -N), ammonia nitrogen (NH 4 + -N), and total nitrogen (TN) improved by 28.76 %, 12.96 %, and 28.45 %, compared to quartz sand biofilter (SBF). Moreover, removal efficiencies of NO 3 - -N and TN in E-ICBF with R-DOM went up by 12.11 % and 14.02 % compared to methanol. Additionally, both PEG and the electric field reduced the microbial richness and diversity. However, PEG promoted the increase of denitrifying bacteria abundance including unclassified_f_Comamonadaceae, Thauera, and unclassified_f_Gallionellaceae. The electric field improved the abundances of genes related to N removal (hao, nasC, nasA, nifH, nifD, nifK) and PEG further enhanced the effect. The abundances of key enzymes [EC:1.7.5.1], [EC:1.7.2.1], [EC:1.7.2.4], and [EC:1.7.2.5] decreased due to the addition of PEG and the electric field mitigated the negative influence. Additionally, the electric field changed relationships between microorganisms and pollutant removal, and improved interspecific relationships between denitrifying bacterial genera and other genera in E-ICBF., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this original manuscript., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Poly-γ-glutamic acid (γ-PGA) is a promising biopolymer for various applications. In this study, we isolated a novel γ-PGA-producing strain, Bacillus halotolerans F29. The one-factor-at-a-time method was used to investigate the influence of carbon sources, nitrogen sources, and culture parameters on γ-PGA production. The optimal carbon and nitrogen sources were sucrose and (NH 4 ) 2 SO 4 , respectively. The optimal culture conditions for γ-PGA production were determined to be 37 °C and a pH of 5.5. Response surface methodology was used to determine the optimum medium components: 77.6 g/L sucrose, 43.0 g/L monosodium glutamate, and 2.2 g/L K 2 HPO 4 . The γ-PGA titer increased significantly from 8.5 ± 0.3 g/L to 20.7 ± 0.7 g/L when strain F29 was cultivated in the optimized medium. Furthermore, the γ-PGA titer reached 50.9 ± 1.5 g/L with a productivity of 1.33 g/L/h and a yield of 2.23 g of γ-PGA/g of L-glutamic acid with the optimized medium in fed-batch fermentation. The maximum γ-PGA titer reached 45.3 ± 1.1 g/L, with a productivity of 1.06 g/L/h when molasses was used as a carbon source. It should be noted that the γ-PGA yield in this study was the highest of all reported studies, indicating great potential for the industrial production of γ-PGA., (© 2024. The Author(s), under exclusive licence to Microbiological Society of Korea.)

The limited availability of carbon sources in low carbon source wastewater has always hindered nitrogen removal efficiency. The residual slurry liquid after anaerobic digestion has the potential to be used as a carbon source. This study investigated the optimal parameters of dissolved oxygen (DO) for enhancing the treatment of low carbon source wastewater using slurry, and revealed the characteristics of carbon metabolism gene enrichment and carbon fixation potential driven by DO. The results indicated that treating wastewater under high DO concentrations (3-4 mg/L) conditions could meet the emission standards set by wastewater treatment plants in China. However, the lower-cost DO concentration of 3 mg/L is considered a more cost-effective parameter, effectively removing 85.68% of chemical oxygen demand and 91.56% of total nitrogen. Mechanistic analysis suggested that reducing DO concentration increased the diversity of microbial communities. Regulating DO concentration reshaped the co-metabolic network of microorganisms with different DO sensitivities by influencing Hydrogenophaga and Chlorobium. This ultimately led to the reconstruction of heterotrophic microbial communities dominated by Sphaerotilus and Acidovorax under high DO conditions, and heterotrophic-autotrophic co-enriched microbial communities dominated by Chlorobium under low DO conditions (1-2 mg/L). Additionally, under high DO conditions, high microbial mass transfer efficiency and the enrichment of functional genes were crucial for achieving high nitrogen removal performance. Further, the microbial carbon fixation potential was relatively high under the DO 3 mg/L condition, helping to reduce the consumption of additional carbon sources. This study provided innovative ideas for the sustainable and low-carbon development of wastewater treatment technology., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In constructed wetlands (CWs), carbon source availability profoundly affected microbial metabolic activities engaged in both iron cycle and nitrogen metabolism. However, research gaps existed in understanding the biotransformation of nitrogen and iron in response to fluctuations in organic carbon content under day-night alterations. Results demonstrated increased removal efficiency of NO 3 - -N (75.70 %) under light conditions, attributed to increased total organic carbon (TOC). This enhancement promoted the relative abundance of bacteria involved in nitrogen and iron processes, establishing a more stable microbial network. Elevated TOC content also upregulated genes for iron metabolism and glycolysis, facilitating denitrification. Spearman correlation analysis supported the synergistic mechanisms between FeS 4 -based autotrophic denitrification and TOC-mediated heterotrophic denitrification under light conditions. The significant impact of carbon sources on microbial activities underscores the critical role of organic carbon availability in enhancing nitrogen removal efficiency, providing valuable insights for optimizing FeS + -N (75.70 %) under light conditions, attributed to increased total organic carbon (TOC). This enhancement promoted the relative abundance of bacteria involved in nitrogen and iron processes, establishing a more stable microbial network. Elevated TOC content also upregulated genes for iron metabolism and glycolysis, facilitating denitrification. Spearman correlation analysis supported the synergistic mechanisms between FeS 2 -based autotrophic denitrification and TOC-mediated heterotrophic denitrification under light conditions. The significant impact of carbon sources on microbial activities underscores the critical role of organic carbon availability in enhancing nitrogen removal efficiency, providing valuable insights for optimizing FeS 2 -based CWs design and operation strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Pseudomonas sp. ZHL02, removing nitrogen via ammonia nitrogen (NH 4 + ) → hydroxylamine (HN 2 OH) → nitrite (NO 2 - ) → nitrate (NO 3 O) pathway was employed for getting in-depth information on the heterotrophic nitrification-aerobic denitrification (HNAD) pathway from carbon oxidation, nitrogen conversion, electron transport process, enzyme activity, as well as gene expression while sodium succinate, sodium citrate, and sodium acetate were utilized as the carbon sources. The nitrogen balance analysis results demonstrated that ZHL02 mainly removed NH - -N through assimilation. The carbon source metabolism resulted in the discrepancies in electron transport chain and nitrogen removal between different HNAD bacteria. Moreover, the prokaryotic strand-specific transcriptome method showed that, amo and hao were absent in ZHL02, and unknown genes may be involved in ZHL02 during the HNAD process. As a fascinating process for removing nitrogen, the HNAD process is still puzzling, and the relationship between carbon metabolism and nitrogen metabolism among different HNAD pathways should be studied further.2 - → nitric oxide (NO) → nitrous oxide (N 2 O) pathway was employed for getting in-depth information on the heterotrophic nitrification-aerobic denitrification (HNAD) pathway from carbon oxidation, nitrogen conversion, electron transport process, enzyme activity, as well as gene expression while sodium succinate, sodium citrate, and sodium acetate were utilized as the carbon sources. The nitrogen balance analysis results demonstrated that ZHL02 mainly removed NH 4 + -N through assimilation. The carbon source metabolism resulted in the discrepancies in electron transport chain and nitrogen removal between different HNAD bacteria. Moreover, the prokaryotic strand-specific transcriptome method showed that, amo and hao were absent in ZHL02, and unknown genes may be involved in ZHL02 during the HNAD process. As a fascinating process for removing nitrogen, the HNAD process is still puzzling, and the relationship between carbon metabolism and nitrogen metabolism among different HNAD pathways should be studied further., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Thioredoxins play an essential role in regulating enzyme activity in response to environmental changes, especially in photosynthetic organisms. They are crucial for metabolic regulation in cyanobacteria, but the key redox-regulated central processes remain to be determined. Physiological, metabolic, and transcriptomic characterization of a conditional mutant of the essential Synechocystis sp. PCC 6803 thioredoxin trxA gene (STXA2) revealed that decreased TrxA levels alter cell morphology and induce a dormant-like state. Furthermore, TrxA depletion in the STXA2 strain inhibited protein synthesis and led to changes in amino acid pools and nitrogen/carbon reserve polymers, accompanied by oxidation of the elongation factor-Tu. Transcriptomic analysis of TrxA depletion in STXA2 revealed a robust transcriptional response. Downregulated genes formed a large cluster directly related to photosynthesis, ATP synthesis, and CO2 fixation. In contrast, upregulated genes were grouped into different clusters related to respiratory electron transport, carotenoid biosynthesis, amino acid metabolism, and protein degradation, among others. These findings highlight the complex regulatory mechanisms that govern cyanobacterial metabolism, where TrxA acts as a critical regulator that orchestrates the transition from anabolic to maintenance metabolism and regulates carbon and nitrogen balance., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Microbial communities are crucial for water quality and biogeochemical cycling in freshwaters. Microbes secrete extracellular enzymes to decompose organic matter for their needs of nutrients and scarce elements. Yet, there is a lack of knowledge on microbial metabolic limitations in freshwaters, especially in lake sediments. Here, we examined the carbon, nitrogen, and phosphorus-acquiring extracellular enzyme activities and the bacterial and fungal communities of 30 sediments across Xingkai Lake, the largest freshwater lake in Northeast Asia. We further analyzed the microbial metabolic limitations via extracellular enzyme stoichiometry and explored the direct and indirect effects of abiotic and biotic factors on the limitations. We found that microbial metabolisms were primarily limited by phosphorus in Xingkai Lake. For instance, microbial carbon and phosphorus limitations were closely correlated to abiotic factors like water depth, total dissolved solids, sediment total carbon, and conductivity. The metabolic limitations were also affected by biotic factors, such as showing positive relationships with the alpha and beta diversity of bacteria, and with the beta diversity of fungi. In addition, community compositions of bacteria and fungi were mainly correlated to abiotic factors such as total carbon and dissolved organic carbon, respectively. Collectively, microbial metabolic limitations were affected directly or indirectly by abiotic factors and microbial communities. Our findings indicate that microbial metabolic limitations are not only driven by bacteria and fungi but also by abiotic factors such as water depth and total nitrogen, and thus provide empirical evidence for effective management of freshwater lakes under climate warming and intensified human activities., (© 2024. The Author(s).)

The denitrifying sulfur (S) conversion-associated enhanced biological phosphorus removal (DS-EBPR) process for treating saline wastewater is characterized by its unique microbial ecology that integrates carbon (C), nitrogen (N), phosphorus (P), and S biotransformation. However, operational instability arises due to the numerous parameters and intricates bacterial interactions. This study introduces a two-stage interpretable machine learning approach to predict S conversion-driven P removal efficiency and optimize DS-EBPR process. Stage one utilized the XGBoost regression model, achieving an R 2 value of 0.948 for predicting sulfate reduction (SR) intensity from anaerobic parameters with feature engineering. Stage two involved the CatBoost classification and regression model integrating anoxic parameters with the predicted SR values for predicting P removal, reaching an accuracy of 94% and an R 2 value of 0.93, respectively. This study identified key environmental factors, including SR intensity (20-45 mg S/L), influent P concentration (<9.0 mg P/L), mixed liquor volatile suspended solids (MLVSS)/mixed liquor suspended solids (MLSS) ratio (0.55-0.72), influent C/S ratio (0.5-1.0), anoxic reaction time (5-6 h), and MLSS concentration (>6.50 g/L). A user-friendly graphic interface was developed to facilitate easier optimization and control. This approach streamlines the determination of optimal conditions for enhancing P removal in the DS-EBPR process.

Mixotrophy is an important trophic strategy for bacterial survival in the ocean. However, the global relevance and identity of the major mixotrophic taxa remain largely elusive. Here, we combined phylogenetic, metagenomic, and metatranscriptomic analyses to characterize ubiquitous Arcobacteraceae based on our deep-sea in situ incubations and the global data. The phylogenomic tree of Arcobacteraceae is divided into three large clades, among which members of clades A and B are almost all from terrestrial environments, while those of clade C are widely distributed in various marine habitats in addition to some terrestrial origins. All clades harbor genes putatively involved in chitin degradation, sulfide oxidation, hydrogen oxidation, thiosulfate oxidation, denitrification, dissimilatory nitrate reduction to ammonium, microaerophilic respiration, and metal (iron/manganese) reduction. Additionally, in clade C, more unique pathways were retrieved, including thiosulfate disproportionation, ethanol fermentation, methane oxidation, fatty acid oxidation, cobalamin synthesis, and dissimilatory reductions of sulfate, perchlorate, and arsenate. Within this clade, two mixotrophic Candidatus genera represented by UBA6211 and CAIJNA01 harbor genes putatively involved in the reverse tricarboxylic acid pathway for carbon fixation. Moreover, the metatranscriptomic data in deep-sea in situ incubations indicated that the latter genus is a mixotroph that conducts carbon fixation by coupling sulfur oxidation and denitrification and metabolizing organic matter. Furthermore, global metatranscriptomic data confirmed the ubiquitous distribution and global relevance of Arcobacteraceae in the expression of those corresponding genes across all oceanic regions and depths. Overall, these results highlight the contribution of previously unrecognized Arcobacteraceae to carbon, nitrogen, and sulfur cycling in global oceans.IMPORTANCEMarine microorganisms exert a profound influence on global carbon cycling and ecological relationships. Mixotrophy, characterized by the simultaneous utilization of both autotrophic and heterotrophic nutrition, has a significant impact on the global carbon cycling. This report characterizes a group of uncultivated bacteria Arcobacteraceae that thrived on the "hot time" of bulky particulate organic matter and exhibited mixotrophic strategy during the in situ organic mineralization. Compared with clades A and B, more unique metabolic pathways were retrieved in clade C, including the reverse tricarboxylic acid pathway for carbon fixation, thiosulfate disproportionation, methane oxidation, and fatty acid oxidation. Global metatranscriptomic data from the Tara Oceans expeditions confirmed the ubiquitous distribution and extensive transcriptional activity of Arcobacteraceae with the expression of genes putatively involved in carbon fixation, methane oxidation, multiple sulfur compound oxidation, and denitrification across all oceanic regions and depths., Competing Interests: The authors declare no conflict of interest.

To understand the effects of nitrogen deposition on element cycling and nutrient limitation status in forest ecosystems, we examined the effects of nitrogen deposition on the stoichiometric characteristics of forest soil-microbial-extracellular enzymes in Pinus yunnanensis forest. We conducted a field experiment with control (CK, 0 g N·m -2 ·a -1 ), low nitrogen (LN, 10 g N·m -2 ·a -1 ), medium nitrogen (MN, 20 g N·m -2 ·a -1 ) and high nitrogen (HN, 25 g N·m -2 ·a -1 ) since 2019. We collected soil samples (0-5 cm, 5-10 cm and 10-20 cm) at September 2022, and measured the contents of soil organic, total nitrogen, total phosphorus, microbial biomass carbon, nitrogen and phosphorus (MBC, MBN, MBP) and the activities of C, N, and P acquisition enzymes. The results showed that nitrogen deposition significantly reduced soil organic content, C:N and C:P by 6.9%-29.8%, 7.6%-45.2% and 6.5%-28.6%, and increased soil total N content and N:P by 10.0%-45.0% and 19.0%-46.0%, respectively. Nitrogen addition did not affect soil total P content. Except for soil C:N and C:P, soil nutrient content and stoichiometric ratio were highest in 0-5 cm soil layer. MN and HN treatments significantly decreased MBN by 11.0%-12.7%. MBC, MBP, and their stoichiometry did not change significantly under nitrogen deposition. Soil microbial nutrient content in 0-5 cm soil layer was significantly higher than that in other soil layers. Nitrogen deposition significantly decreased the activities of cellobiose hydrolase and leucine aminopeptidase (decreased by 14.5%-16.2% and 48.7%-66.3%). HN treatment promoted β-1,4-glucosidase activity (increased by 68.0%), but inhibited soil enzyme stoichiometric carbon to nitrogen ratio and nitrogen to phosphorus ratio (decreased by 95.4% and 88.4%). LN and MN treatment promoted β-1,4-N-acetylglucosaminidase activity (increased by 68.3%-116.6%), but inhibited enzyme stoichiometric carbon to phosphorus ratio (decreased by 14.9%-29.4%). Alkaline phosphatase activity had no significant change. Soil enzyme activities were significantly decreased with increasing soil depth. Soil total N and total P and microbial nutrients were negatively correlated with vector angle (representing microbial nitrogen or phosphorus limitation), while vector length (representing microbial carbon limitation) was consistently significantly positively correlated with vector angle, suggesting the synergistic promotion between microbial carbon limitation and phosphorus limitation. Nitrogen deposition gradually shifted to phosphorus limitation while alleviating microbial nitrogen limitation in P. yunnanensis forest. In addition, microbial activities in this region was limited by C availability, and the relationship between microbial C and P limitation was proportional.

Graphene oxide (GO) is a novel nanomaterial being applied in different fields, but was less used as foliar fertilizer in agriculture. We conducted a pot experiment to analyze the effects of foliar spraying GO from 0 (control), 50 (T 1 ), 100 (T 2 ), 150 (T 3 ) and 200 mg·L -1 (T 4 ) on the morphogenesis and carbon and nitrogen metabolism of kidney bean plants during the initiation of flowering to clarify the physiological effects of foliar spraying GO. The results showed that dry matter accumulation, the content of photosynthetic pigments, soluble sugars of T 1 to T 4 treatments, were significantly increased by 40.7%-43.4%, 10.4%-80.7%, 6.4%-9.1% in kidney bean plants compared with CK treatment, respectively. T 3 treatment performed the best. Meanwhile, the activities of sucrose phosphate synthase, acid converting enzyme and neutral converting enzyme of T 3 and T 4 treatments were increased by 25.7%-45.5%, 17.4%-28.6%, and 14.7%-20.1%, and the activities of nitrate reductase, glutamine synthetase, and glutamate synthetase of T 2 and T 3 treatments were increased by 8.1%-15.2%, 11.5%-25.0%, and 89.7%-93.1%, respectively. In conclusion, foliar spraying of appropriate GO in early flowering stage of kidney bean could increase the content of photosynthetic pigments, improve the level of photosynthetic carbon and nitrogen metabolism, and increase dry matter accumulation. T 3 treatment (150 mg·L -1 ) was the most effective in this study.

Alpine ecosystems are important terrestrial carbon (C) pools, and microbial decomposers play a key role in litter decomposition. Microbial metabolic limitations in these ecosystems, however, remain unclear. The objectives of this study aim to elucidate the characteristics of microbial nutrient limitation and their C use efficiency (CUE), and to evaluate their response to environmental factors. Five ecological indicators were utilized to assess and compare the degree of microbial elemental homeostasis and the nutrient limitations of the microbial communities among varying stages of litter decomposition (L, F, and H horizon) along an altitudinal gradient (2800, 3000, 3250, and 3500 m) under uniform vegetation (Abies fabri) on Gongga Mountain, eastern Tibetan Plateau. In this study, microorganisms in the litter reached a strictly homeostatic of C content exclusively during the middle stage of litter decomposition (F horizon). Based on the stoichiometry of soil enzymes, we observed that microbial N- and P-limitation increased during litter degradation, but that P-limitation was stronger than N-limitation at the late stages of degradation (H horizon). Furthermore, an increase in microbial CUE corresponded with a reduction in microbial C-limitation. Additionally, redundancy analysis (RDA) based on forward selection further showed that microbial biomass C (MBC) is closely associated with the enzyme activities and their ratios, and MBC was also an important factor in characterizing changes in microbial nutrient limitation and CUE. Our findings suggest that variations in MBC, rather than N- and P-related components, predominantly influence microbial metabolic processes during litter decomposition on Gongga Mountain, eastern Tibetan Plateau., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Liamocins are molecules with a polyol lipid structure produced by rare strains of Aureobasidium pullulans. In recent years, liamocins have attracted attention due to their antibacterial, anticancer and surface-active properties, and promising potential applications have been identified in the food, agriculture, medical and pharmaceutical industries. This study is the first to investigate the effects of different carbon and nitrogen sources on the growth and liamocin production kinetics of A. pullulans NBRC 100716 strain. This strain was selected among six different A. pullulans strains whose liamocin productions were tested by us for the first time. In fermentations carried out in shaking water baths, the carbon source that most supported the liamocin production of this strain was fructose, and the nitrogen source was peptone-yeast extract combination. In the medium containing fructose and the peptone-yeast extract mixture, A. pullulans NBRC 100716 produced 4.26 g liamocin L -1 . The specific liamocin production rate (qp) of the strain in this medium was 0.0090 g liamocin/g mo.h. This study is also the first to produce liamocin with a fructophilic A. pullulans strain. Present findings in this research also demonstrated the excellent biosurfactant capacity of the liamocin produced by this strain. The obtained liamocin reduced the water surface tension to a degree that can compete with synthetic surfactants. Furthermore, this is the first report to reveal that the fatty acid profile of liamocin obtained from A. pullulans NBRC 100716 contains an appreciable amount of unsaturated fatty acids and is similar to the composition of vegetable oil., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The phosphorus (P) concentration is increasing in parts of the Baltic Sea following the spring bloom. The fate of this excess P-pool is an open question, and here we investigate the role of microbial degradation processes in the excess P assimilation phase. During a 17-day-long mesocosm experiment in the southwest Finnish archipelago, we examined nitrogen, phosphorus, and carbon acquiring extracellular enzyme activities in three size fractions (<0.2, 0.2-3, and >3 µm), bacterial abundance, production, community composition, and its predicted metabolic functions. The mesocosms received carbon (C) and nitrogen (N) amendments individually and in combination (NC) to distinguish between heterotrophic and autotrophic processes. Alkaline phosphatase activity occurred mainly in the dissolved form and likely contributed to the excess phosphate conditions together with grazing. At the beginning of the experiment, peptidolytic and glycolytic enzymes were mostly produced by free-living bacteria. However, by the end of the experiment, the NC-treatment induced a shift in peptidolytic and glycolytic activities and degradation of phosphomonoesters toward the particle-associated fraction, likely as a consequence of higher substrate availability. This would potentially promote retention of nutrients in the surface as opposed to sedimentation, but direct sedimentation measurements are needed to verify this hypothesis., (The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Carbon metabolism and nutrient removal are crucial for biological wastewater treatment. This study focuses on analyzing carbon allocation and utilization by heterotrophic bacteria in response to increasing COD concentration in the influent. The study also assesses the effect of denitrification and biological phosphorus removal, particularly in combination with anaerobic ammonia oxidation (anammox). The experiment was conducted in a SBR operating under anaerobic/anoxic/oxic conditions. As COD concentration in the influent increased from 100 to 275 mg/L, intracellular COD accounted for 95.72 % of the COD removed. By regulating the NO 3 concentration in the anoxic stage from 10 to 30 mg/L, the nitrite accumulation rate reached 69.46 %, which could serve as an electron acceptor for anammox. Most genes related to the tricarboxylic acid (TCA) cycle declined, while the genes involved in the glyoxylate cycle, gluconeogenesis, PHA synthesis increased. This suggests that glycogen accumulation and carbon storage, rather than direct carbon oxidation, was the dominant pathway for carbon metabolism. However, the genes responsible for the reduction of NO - concentration in the anoxic stage from 10 to 30 mg/L, the nitrite accumulation rate reached 69.46 %, which could serve as an electron acceptor for anammox. Most genes related to the tricarboxylic acid (TCA) cycle declined, while the genes involved in the glyoxylate cycle, gluconeogenesis, PHA synthesis increased. This suggests that glycogen accumulation and carbon storage, rather than direct carbon oxidation, was the dominant pathway for carbon metabolism. However, the genes responsible for the reduction of NO 2 - -N (nirK) and NO (nosB) decreased, contributing to NO 2 - accumulation. The study also employed metagenomic analysis to reveal microbial interactions. The enrichment of specific bacterial species, including Dechloromonas sp. (D2.bin.10), Ca. Competibacteraceae bacterium (D9.bin.8), Ca. Desulfobacillus denitrificans (D6.bin.17), and Ignavibacteriae bacterium (D3.bin.9), played a collaborative role in facilitating nutrient removal and promoting the combination with anammox., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The glutamine synthetase/glutamic acid synthetase (GS/GOGAT) cycle plays important roles in N metabolism, growth, development, and stress resistance in plants. Excess ammonium (NH4+) restricts growth, but GS can help to alleviate its toxicity. In this study, the 84K model clone of hybrid poplar (Populus alba × P. tremula var. glandulosa), which has reduced biomass accumulation and leaf chlorosis under high-NH4+ stress, showed less severe symptoms in transgenic lines overexpressing GLUTAMINE SYNTHETASE 1;2 (GS1;2-OE), and more severe symptoms in RNAi lines (GS1;2-RNAi). Compared with the wild type, the GS1;2-OE lines had increased GS and GOGAT activities and higher contents of free amino acids, soluble proteins, total N, and chlorophyll under high-NH4+ stress, whilst the antioxidant and NH4+ assimilation capacities of the GS1;2-RNAi lines were decreased. The total C content and C/N ratio in roots and leaves of the overexpression lines were higher under stress, and there were increased contents of various amino acids and sugar alcohols, and reduced contents of carbohydrates in the roots. Under high-NH4+ stress, genes related to amino acid biosynthesis, sucrose and starch degradation, galactose metabolism, and the antioxidant system were significantly up-regulated in the roots of the overexpression lines. Thus, overexpression of GS1;2 affected the carbon and amino acid metabolism pathways under high-NH4+ stress to help maintain the balance between C and N metabolism and alleviate the symptoms of toxicity. Modification of the GS/GOGAT cycle by genetic engineering is therefore a potential strategy for improving the NH4+ tolerance of cultivated trees., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The simulations and predictions obtained from mathematical models of bioprocesses conducted by microorganisms are not overvalued. Mechanistic models are bringing a better process understanding and the possibility of simulating unmeasurable variables. The Dynamic Energy Budget (DEB) model is an energy balance that can be formulated for any living organism and can be classified as a structured model. In this study, the DEB model was used to describe E. coli growth in a batch reactor in carbon and nitrogen substrate limitation conditions. The DEB model provides a possibility to follow the changes in the microbes' cells including their elemental composition and content of some important cell ingredients in different growth phases in substrate limitation conditions which makes it more informative compared to Monod's model. The model can be used as an optimal choice between Monod-like models and flux-based approaches. KEY POINTS: • The DEB model can be used to catch changes in elemental composition of E. coli • Bacteria batch culture growth phases can be explained by the DEB model • The DEB model is more informative compared to Monod's based models., (© 2024. The Author(s).)

Plant acclimation to an ever-changing environment is decisive for growth, reproduction, and survival. Light availability limits biomass production on both ends of the intensity spectrum. Therefore, the adjustment of plant metabolism is central to high-light (HL) acclimation, and the accumulation of photoprotective anthocyanins is commonly observed. However, mechanisms and factors regulating the HL acclimation response are less clear. Two Arabidopsis mutants of spliceosome components exhibiting a pronounced anthocyanin overaccumulation in HL were isolated from a forward genetic screen for new factors crucial for plant acclimation. Time-resolved physiological, transcriptome, and metabolome analysis revealed a vital function of the spliceosome components for rapidly adjusting gene expression and metabolism. Deficiency of INCREASED LEVEL OF POLYPLOIDY1 (ILP1), NTC-RELATED PROTEIN1 (NTR1), and PLEIOTROPIC REGULATORY LOCUS1 (PRL1) resulted in a marked overaccumulation of carbohydrates and strongly diminished amino acid biosynthesis in HL. While not generally limited in N-assimilation, ilp1, ntr1, and prl1 showed higher glutamate levels and reduced amino acid biosynthesis in HL. The comprehensive analysis reveals a function of the spliceosome components in the conditional regulation of the carbon:nitrogen balance and the accumulation of anthocyanins during HL acclimation. The importance of gene expression, metabolic regulation, and re-direction of carbon towards anthocyanin biosynthesis for HL acclimation are discussed., (© 2024 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Atmospheric nitrogen (N) deposition in forests can affect soil microbial growth and turnover directly through increasing N availability and indirectly through altering plant-derived carbon (C) availability for microbes. This impacts microbial residues (i.e., amino sugars), a major component of soil organic carbon (SOC). Previous studies in forests have so far focused on the impact of understory N addition on microbes and microbial residues, but the effect of N deposition through plant canopy, the major pathway of N deposition in nature, has not been explicitly explored. In this study, we investigated whether and how the quantities (25 and 50 kg N ha -1  year -1 ) and modes (canopy and understory) of N addition affect soil microbial residues in a temperate broadleaf forest under 10-year N additions. Our results showed that N addition enhanced the concentrations of soil amino sugars and microbial residual C (MRC) but not their relative contributions to SOC, and this effect on amino sugars and MRC was closely related to the quantities and modes of N addition. In the topsoil, high-N addition significantly increased the concentrations of amino sugars and MRC, regardless of the N addition mode. In the subsoil, only canopy N addition positively affected amino sugars and MRC, implying that the indirect pathway via plants plays a more important role. Neither canopy nor understory N addition significantly affected soil microbial biomass (as represented by phospholipid fatty acids), community composition and activity, suggesting that enhanced microbial residues under N deposition likely stem from increased microbial turnover. These findings indicate that understory N addition may underestimate the impact of N deposition on microbial residues and SOC, highlighting that the processes of canopy N uptake and plant-derived C availability to microbes should be taken into consideration when predicting the impact of N deposition on the C sequestration in temperate forests., (© 2024 John Wiley & Sons Ltd.)

This study successfully established Iron Sulfide-Mediated mixotrophic Partial Denitrification/Anammox system, achieving nitrogen and phosphorus removal efficiency of 97.26% and 78.12%, respectively, with COD/NO 3 - -N, creating more niche space for autotrophic bacteria and other heterotrophic denitrifiers. The prediction of functional genes illustrated that iron Sulfide upregulated genes related to carbon metabolism, denitrification, anammox and sulfur oxidation-reduction, facilitating the establishment of carbon-nitrogen-sulfur cycle. Furthermore, this cycle primarily produced electrons via nicotinamide adenine dinucleotide and sulfur oxidation-reduction processes, subsequently utilized within the electron transfer chain. In summary, the Partial Denitrification/Anammox system under the influence of iron sulfide achieved effient nitrogen removal by expediting electron transfer through the carbon-nitrogen-sulfur cycle.3 - -N, creating more niche space for autotrophic bacteria and other heterotrophic denitrifiers. The prediction of functional genes illustrated that iron Sulfide upregulated genes related to carbon metabolism, denitrification, anammox and sulfur oxidation-reduction, facilitating the establishment of carbon-nitrogen-sulfur cycle. Furthermore, this cycle primarily produced electrons via nicotinamide adenine dinucleotide and sulfur oxidation-reduction processes, subsequently utilized within the electron transfer chain. In summary, the Partial Denitrification/Anammox system under the influence of iron sulfide achieved effient nitrogen removal by expediting electron transfer through the carbon-nitrogen-sulfur cycle., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Elevated CO 2 concentrations may inhibit photosynthesis due to nitrogen deficiency, but legumes may be able to overcome this limitation and continue to grow. Our study confirms this conjecture well. First, we placed the two-year-old potted saplings of Ormosia hosiei (O. hosiei) (a leguminous tree species) in the open-top chamber (OTC) with three CO 2 concentrations of 400 (CK), 600 (E1), and 800 μmol·mol -1 (E2) to simulate the elevated CO 2 concentration environment. After 146 days, the light saturation point (LSP), light compensation point (LCP), apparent quantum efficiency (AQE), and dark respiration rate (Rd) of O. hosiei were increased under increasing CO 2 concentration and obtain the maximum ribulose diphosphate (RuBP) carboxylation rate (V c max ) and RuBP regenerated photosynthetic electron transfer rate (J max ) were also significantly increased under E2 treatment (P < 0.05). This results in a significant increase of the maximum assimilation rate (A max ) under elevated CO 2 concentrations. Sucrose phosphate synthase (SPS) activity in sucrose metabolism increased in the leaves, more soluble sugars, starches, and sucrose was produced, but sucrose content only in leaves increased at E2, and more carbon flows to the roots. The activity of the NH 4 + assimilating enzymes glutamine synthetase (GS), glutamate synthetase (GOGAT), and glutamate dehydrogenase (GDH) in the leaves of O. hosiei increases under elevated CO 2 concentrations to promote nitrogen synthesis that reduces the content of ammonium nitrogen and increases the content of nitrate nitrogen. In addition, under E1 conditions, sucrose synthase (SS), direction of synthesis activity was highest and sucrose invertase (INV) activity was lowest, this means that the balance of C and N metabolism is maintained. While under E2 conditions SS activity decreased and INV activity increased, this increased C/N and nitrogen use efficiency. So, the elevated CO 2 concentration promotes the accumulation of O. hosiei biomass, especially in the aboveground part, but did not have a significant effect on the accumulation of root biomass. This means that O. hosiei is able to cope under the elevated CO 2 concentration without showing photosynthetic adaptation during the experimental period., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Microbial communities are fundamental components driving critical biogeochemical carbon (C), nitrogen (N) and sulfur (S) cycles in groundwater ecosystems. The reduction-oxidation (redox) potential is one important environmental factor influencing the microbial community composition. Here, we developed a bio-trap method using in-situ sediment as a matrix to collect aquifer sediment samples and evaluate the response of microbial composition and C/N/S cycling functions to redox variations created by providing sole O 2 , joint O 2 and H 2 , and sole H 2 to three wells. Illumina sequencing analyses showed that the microbial communities in the bio-trap sediment could respond quickly to redox changes in the wells, demonstrating that this bio-trap method is promising for detecting microbial variation in the aquifer sediment. The microbial metabolic functions related to C, N and S cyclings and organic pollutants degradation were predicted by the Kyoto Encyclopedia of Genes and Genomes (KEGG) approach. It was found that the joint O 2 and H 2 injection produced medium oxidation-reduction potential (ORP -346 and -614 mV) and enhanced more microbial functions than sole O 2 or H 2, which mainly include oxidative phosphorylation, most carbon source metabolism, various pollutants degradation, and nitrogen and sulfur metabolism. Moreover, the functional genes encoding phenol monooxygenase, dioxygenase, nitrogen fixation, nitrification, aerobic and anaerobic nitrate reductase, nitrite reductase, nitric oxide reductase, and sulfur oxidation increased. These findings tell us the contaminant bioremediation and N, S metabolism can be promoted by adjusting ORP realised by injecting joint O 2 and H 2 .

Anthropogenic activities have raised nitrogen (N) input worldwide with profound implications for soil carbon (C) cycling in ecosystems. The specific impacts of N input on soil organic matter (SOM) pools differing in microbial availability remain debatable. For the first time, we used a much-improved approach by effectively combining the 13 C natural abundance in SOM with 21 years of C 3 -C 4 vegetation conversion and long-term incubation. This allows to distinguish the impact of N input on SOM pools with various turnover times. We found that N input reduced the mineralization of all SOM pools, with labile pools having greater sensitivity to N than stable ones. The suppression in SOM mineralization was notably higher in the very labile pool (18%-52%) than the labile and stable (11%-47%) and the very stable pool (3%-21%) compared to that in the unfertilized control soil. The very labile C pool made a strong contribution (up to 60%) to total CO 2 release and also contributed to 74%-96% of suppressed CO 2 with N input. This suppression of SOM mineralization by N was initially attributed to the decreased microbial biomass and soil functions. Over the long-term, the shift in bacterial community toward Proteobacteria and reduction in functional genes for labile C degradation were the primary drivers. In conclusion, the higher the availability of the SOM pools, the stronger the suppression of their mineralization by N input. Labile SOM pools are highly sensitive to N availability and may hold a greater potential for C sequestration under N input at global scale., (© 2024 The Author(s). Global Change Biology published by John Wiley & Sons Ltd.)

Moving bed bioreactors are efficient for the removal of nitrogen effluents by simultaneous nitrification and denitrification (SND), which is particularly challenging for effluents with low C/N ratio due to insufficient carbon available for denitrification. The addition of external organic carbon in such cases increases treatment costs. Endogenous degradation of biomass offers a cost-effective and persistent source of carbon. Internal carbon leads to low denitrification rates and optimizing the biomass content leads to a sustainable solution for treating such effluents. The present study aims to optimize the process of SND using endogenous carbon by investigating the effect of operational parameters like dissolved oxygen, influent ammonium (NH 4 -N) loading, hydraulic retention time (HRT) and carrier filling ratio (CFR). The synthetic wastewater used in the study does not inherently contain any organic carbon, thus indicating the responsible carbon for denitrification to be of internal origin. Using a novel carrier for multispecies biofilm development, initial NH 4 -N concentration variation showed an insignificant effect on SND efficiency. At higher loading, nitrite accumulation due to incomplete denitrification was observed. The effect of CFR was investigated in a range of 20-35%, beyond which non-homogeneous mixing was caused in the reactor. Under controlled DO and temperature of about 5 mg/L and 28 ± 2°C, respectively, an optimum filling ratio of 35% was finalised, resulting in an SND efficiency of 97.9% at 22 h HRT for high influent NH 4 -N concentration of 500 mg/L. Bacteriological species identification revealed the coexistence of nitrogen-fixing, aerobic nitrifying and anaerobic denitrifying bacteria in the biofilm growing on the biocarrier.Mechanism of SND occurring in aerobic moving bed bioreactors: Laboratory-scale aerobic Moving Bed Bioreactor setup for a continuous study evaluating the influence of operational parameters on SNDOccurrence of aerobic and anoxic zones in biofilm grown on the surface of novel carriers due to diffusional gradient of dissolved oxygen.

Microalgal technology holds great promise for both low C/N wastewater treatment and resource recovery simultaneously. Nevertheless, the advancement of microalgal technology is hindered by its reduced nitrogen removal efficiency in low C/N ratio wastewater. In this work, microalgae and waste oyster shells were combined to achieve a total inorganic nitrogen removal efficiency of 93.85% at a rate of 2.05 mg L -1  h -1 in low C/N wastewater. Notably, over four cycles of oyster shell reuse, the reactor achieved an average 85% ammonia nitrogen removal extent, with a wastewater treatment cost of only $0.092/ton. Moreover, microbial community analysis during the reuse of oyster shells revealed the critical importance of timely replacement in inhibiting the growth of non-functional bacteria (Poterioochromonas&#95;malhamensi). The work demonstrated that the oyster shell - microalgae system provides a time- and cost-saving, environmental approach for the resourceful treatment of harsh low C/N wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The efficient nitrogen removal from micro-polluted source water is an international challenge to be solved urgently. However, the inner denitrification mechanism of native aerobic denitrifying bacterial communities in response to carbon scarcity remains relatively unclear. Here, the bacterial community XT6, screened from an oligotrophic reservoir, exhibited aerobic denitrifying capacity under low-carbon environments. Up to 76.79-81.64 % of total organic carbon (TOC) and 51.48-67.60 % of NO 3 - -N were removed by XT6 within 48 h at C/N ratios of 2.0-3.0. Additionally, the nitrogen balance experiments further manifested that 26.27-38.13 % of NO 3 - -N was lost in gaseous form. As the C/N ratio decreased, XT6 tended to generate more extracellular polymeric substances (EPS), with the tightly bound EPS showing the largest increase. Pseudomonas and Variovorax were quite abundant in XT6, constituting 59.69 % and 28.65 % of the total sequences, respectively. Furthermore, metagenomics analysis evidenced that XT6 removed TOC and nitrate mainly through the tricarboxylic acid cycle and aerobic denitrification. Overall, the abovementioned results provide a deeper understanding of the nitrogen metabolic pathways of indigenous aerobic denitrifying bacterial communities with low C/N ratios and offer useful guidance for controlling nitrogen pollution in oligotrophic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Root exudation and its mediated nutrient cycling process driven by nitrogen (N) fertilizer can stimulate the plant availability of various soil nutrients, which is essential for microbial nutrient acquisition. However, the response of soil microbial resource limitations to long-term N fertilizer application rates in greenhouse vegetable systems has rarely been investigated. Therefore, we selected a 15-year greenhouse vegetable system, and investigated how N fertilizer application amount impacts on root carbon and nitrogen exudation rates, microbial resource limitations and microbial carbon use efficiency (CUE ST ). Four N treatments were determined: high (N3), medium (N2), low (N1), and a control without N fertilization (N0). Compared to the control (N0), the results showed that the root C exudation rates decreased significantly by 42.9 %, 57.3 % and 33.6 %, and the root N exudation rates decreased significantly by 29.7 %, 42.6 %, and 24.1 % under N1, N2, and N3 treatments, respectively. Interactions between fertilizer and plant roots altered microbial C, N, P limitations and CUE ST ; Microbial C and N/P limitations were positively correlated with root C and N exudation rates, negatively correlated with microbial CUE ST . Random Forest analysis revealed that the root C and N exudation rates were key factors for soil microbial resource limitations and microbial CUE ST . Through the structural equation model (SEM) analysis, soil NH 4 content had significant direct effects on the root exudation rates after long-term N fertilizer application. An increase in root exudation rates led to enhanced microbial resource limitations in the rhizosphere soils, potentially due to increased competition. This enhancement may reduce microbial carbon use efficiency (CUE), that is, microbial C turnover, thereby reducing soil C sequestration. Overall, this study highlights the critical role of root exudation rates in microbial resource limitations and CUE changes in plant-soil systems, and further improves our understanding of plant-microbial interactions.+ content had significant direct effects on the root exudation rates after long-term N fertilizer application. An increase in root exudation rates led to enhanced microbial resource limitations in the rhizosphere soils, potentially due to increased competition. This enhancement may reduce microbial carbon use efficiency (CUE), that is, microbial C turnover, thereby reducing soil C sequestration. Overall, this study highlights the critical role of root exudation rates in microbial resource limitations and CUE changes in plant-soil systems, and further improves our understanding of plant-microbial interactions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

In the face of 21st-century challenges driven by population growth and resource depletion, understanding the intricacies of climate change is crucial for environmental sustainability. This review systematically explores the interaction between rising atmospheric CO 2 concentrations and soil microbial populations, with possible feedback effects on climate change and terrestrial carbon (C) cycling through a meta-analytical approach. Furthermore, it investigates the enzymatic activities related to carbon acquisition, gene expression patterns governing carbon and nitrogen metabolism, and metagenomic and meta-transcriptomic dynamics in response to elevated CO 2 levels. The study reveals that elevated CO 2 levels substantially influence soil microbial communities, increasing microbial biomass C and respiration rate by 15 % and upregulating genes involved in carbon and nitrogen metabolism by 12 %. Despite a 14 % increase in C-acquiring enzyme activity, there is a 5 % decrease in N-acquiring enzyme activity, indicating complex microbial responses to CO 2 changes. Additionally, fungal marker ratios increase by 14 % compared to bacterial markers, indicating potential ecosystem changes. However, the current inadequacy of data on metagenomic and meta-transcriptomic processes underscores the need for further research. Understanding soil microbial feedback mechanisms is crucial for elucidating the role of rising CO 2 levels in carbon sequestration and climate regulation. Consequently, future research should prioritize a comprehensive elucidation of soil microbial carbon cycling, greenhouse gas emission dynamics, and their underlying drivers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Microbial necromass carbon (MNC) contributes significantly to the formation of soil organic carbon (SOC). However, the microbial carbon sequestration effect of biochar is often underestimated and influenced by nutrient availability. The mechanisms associated with the formation and stabilization of MNC remain unclear, especially under the combined application of biochar and nitrogen (N) fertilizer. Thus, in a long-term field experiment (11 years) based on biochar application, we utilized bacterial 16S rRNA gene sequencing, fungal ITS amplicon sequencing, metagenomics, and microbial biomarkers to examine the interactions between MNC accumulation and microbial metabolic strategies under combined treatment with biochar and N fertilizer. We aimed to identify the critical microbial modules and species involved, and to analyze the sites where MNC was immobilized from various components. Biochar application increased the MNC content by 13.9 %. Among the MNC components, fungal necromass contributed more to MNC, but bacteria were more readily enriched after biochar application. The microbial life-history strategies that affected MNC formation under the application of various amounts biochar were linked to the N application level. Under N added at 226.5 kg ha -1 , communities such as Actinobacteria and Bacteroidetes with high-growth yield strategies were prevalent and contributed to MNC production. By contrast, under N added at 113.25 kg ha -1 with high biochar application, Proteobacteria with strong resource acquisition strategies were dominant and MNC accumulation was lower. The mineral-associated organic carbon pool was rapidly saturated with the addition of biochar, so the contribution of fungal necromass carbon may have been reduced by reutilization, thereby resulting in the more rapid preservation of bacterial necromass carbon in the particulate organic carbon pool. Overall, our findings indicate that microbial life history traits are crucial for linking microbial metabolic processes to the accumulation and stabilization of MNC, thereby highlighting the their importance for SOC accumulation in farmland soils, and the need to tailor appropriate biochar and N fertilizer application strategies for agricultural soils., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

An integrated process of sludge in-situ fermentation, biological phosphorus removal and endogenous denitrification (ISFPR-ED) was developed to treat low ratio of chemical oxygen demand to nitrogen (COD/N) wastewater and waste activated sludge (WAS) in a single reactor. Nutrient removal and WAS reduction were achieved due to Tetrasphaera-dominated sludge fermentation provided organic carbon in extending the anaerobic duration. The WAS reduction efficiency, effluent orthophosphate (PO 4 -P) and total inorganic nitrogen reached 28.1 %, less than 0.4 and 7.2 mg/L, respectively. While organic carbon was reduced by 67 %. Tetrasphaera, conventional polyphosphate accumulating organisms (PAOs) stored glycogen, amino acids, and PHA for nutrient removal. Excess energy from fermentation enhanced anaerobic PO 3- -P) and total inorganic nitrogen reached 28.1 %, less than 0.4 and 7.2 mg/L, respectively. While organic carbon was reduced by 67 %. Tetrasphaera, conventional polyphosphate accumulating organisms (PAOs) stored glycogen, amino acids, and PHA for nutrient removal. Excess energy from fermentation enhanced anaerobic PO 4 -P removal and fermentation bacteria, working synergistically with conventional PAOs and fermenting microorganisms. This integrated process improves nutrient removal efficiency and reduces operating costs for carbon addition and WAS disposal in wastewater treatment.3- -P uptake by Tetrasphaera. Tetrasphaera was the dominant PO 4 3- -P removal and fermentation bacteria, working synergistically with conventional PAOs and fermenting microorganisms. This integrated process improves nutrient removal efficiency and reduces operating costs for carbon addition and WAS disposal in wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Pitaya, a desert plant, has an underexplored flowering mechanism due to a lack of functional validation assays. This study reveals that the transition from vegetative to generative growth in pitaya is regulated by significant metabolic shift, underscoring the importance of understanding and address the challenging issue pitaya's phase change. Lateral buds from 6-years-old 'Guanhuahong' pitaya (Hylocereus monacanthus) plants were collected on April 8th, 18th, and 28th 2023, representing early, middle, and late stages of phase transition, respectively. Results showed diminished nitrogen levels concurrent with increased carbon levels and carbon-to-nitrogen (C/N) ratios during pitaya phase transition. Transcriptomic analysis identified batches of differentially expressed genes (DEGs) involved in downregulating nitrogen metabolism and upregulating carbon metabolism. These batches of genes play a central role in the metabolic shifts that predominantly regulate the transition to the generative phase in pitaya. This study unveils the intricate regulatory network involving 6 sugar synthesis and transport, 11 photoperiod (e.g., PHY, CRY, PIF) and 6 vernalization (e.g., VIN3) pathways, alongside 11 structural flowering genes (FCA, FLK, LFY, AGL) out of a vast array of potential candidates in pitaya phase change. These findings provide insights into the metabolic pathways involved in pitaya's phase transition, offering a theoretical framework for managing flowering, guiding breeding strategies to optimize flowering timing and improve crop yields under varied nitrogen conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Achieving mainstream short-cut nitrogen removal via nitrite has become a carbon and energy efficient way, but still remains challenging for low-strength municipal wastewaters. This study integrated sidestream enhanced biological phosphorus removal system in a pilot-scale adsorption/bio-oxidation (A-B) process (named A-B-S2EBPR system) and nitrite accumulation was successfully achieved for treating the municipal wastewater. Nitrite could accumulate to 5.5 ± 0.3 mg N/L in the intermittently aerated tanks of B-stage with the nitrite accumulation ratio (NAR) of 79.1 ± 6.5 %. The final effluent concentration and removal efficiency of total inorganic nitrogen (TIN) were 4.6 ± 1.8 mg N/L and 84.9 ± 5.6 %, respectively. In-situ process performance of nitrogen conversions, routine batch nitrification/denitrification activity tests and functional gene abundance of nitrifiers collectively suggested that the nitrite accumulation was mainly caused by partial denitrification rather than out-selection of nitrite oxidizing bacteria (NOB). Moreover, the single-cell Raman spectroscopy analysis first demonstrated that there was a specific microbial population that could utilize polyhydroxyalkanoates (PHA) as the potential internal carbon source during the partial denitrification process. The integration of S2EBPR brings unique features to the conventional A-B process, such as extended anaerobic retention time, lower oxidation-reduction potential (ORP), much higher and complex volatile fatty acids (VFAs) etc., which can largely reshape the microbial communities. The dominant genera were Acinetobacter and Comamonadaceae, which accounted for (17.8 ± 15.5)% and (6.7 ± 3.4)%, respectively, while the relative abundance of conventional nitrifiers was less than 0.2%. This study provides insights into phylogenetic and phenotypic shifts of microbial communities when incorporating S2EBPR into the sustainable A-B process to achieve mainstream short-cut nitrogen removal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Both autotrophic and heterotrophic denitrification are known as important bioprocesses of microbe-mediated nitrogen cycle in natural ecosystems. Actually, mixotrophic denitrification co-driven by organic matter and reduced sulfur substances are also common, especially in hypoxic environments such as estuarine sediments. However, carbon, nitrogen and sulfur co-metabolism during mixotrophic denitrification in natural water ecosystems has rarely been reported in detail. Therefore, this study investigated the co-metabolism of carbon, nitrogen and sulfur using samples collected from four distinct natural water ecosystems. Results demonstrated that samples from various sources all exhibited the ability for co-metabolism of carbon, nitrogen and sulfur. Microbial community analysis showed that Pseudomonas and Paracoccus were dominant bacteria ranging from 65.6% to 75.5% in mixotrophic environment. Enterobacter sp. HIT-SHJ4, a mixotrophic denitrifying strain which owned the capacity for co-metabolism of carbon, nitrogen and sulfur, was isolated and reported here for the first time. The strain preferred methanol as its carbon source and demonstrated remarkable efficiency for removing sulfide and nitrate with below 100 mg/L sulfide. Under weak acid conditions (pH 6.5-7.0), it exhibited enhanced capability in converting sulfide to elemental sulfur. Its bioactivity was evident within a temperature from 25 °C to 40 °C and C/N ratios from 0.75 to 3. This study confirmed the widespread presence of microbial-mediated synergistic carbon, nitrogen and sulfur metabolism in natural aquatic ecosystems. HIT-SHJ4 emerges as a novel strain, shedding light on carbon, nitrogen and sulfur co-metabolism in natural water bodies. Furthermore, it also serves as a promising candidate microorganism for in-situ ecological remediation, particularly in dealing with contamination posed by nitrate, sulfide, and organic matter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Recent evidence suggests that changes in carbon-degrading extracellular enzyme activities (C-EEAs) can help explain soil organic carbon (SOC) dynamics under nitrogen (N) addition. However, the factors controlling C-EEAs remain unclear, impeding the inclusion of microbial mechanisms in global C cycle models. Using meta-analysis, we show that the responses of C-EEAs to N addition were best explained by mycorrhizal association across a wide range of environmental and experimental factors. In ectomycorrhizal (ECM) dominated ecosystems, N addition suppressed C-EEAs targeting the decomposition of structurally complex macromolecules by 13.1 %, and increased SOC stocks by 5.2 %. In contrast, N addition did not affect C-EEAs and SOC stocks in arbuscular mycorrhizal (AM) dominated ecosystems. Our results indicate that earlier studies may have overestimated SOC changes under N addition in AM-dominated ecosystems and underestimated SOC changes in ECM-dominated ecosystems. Incorporating this mycorrhizal-dependent impact of EEAs on SOC dynamics into Earth system models could improve predictions of SOC dynamics under environmental changes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Cultivation of sloping land is a main cause for soil erosion. Conservation practices, such as soil and stone terraces, may reduce the impacts of erosion but their impacts on soil microbial diversity and functioning related to carbon (C) and nutrient metabolisms remain unclear. This study was conducted to evaluate the effects of slope gradients (5°, 8°, 15°, 25°) and conservation practices (cultivated, uncultivated, soil terrace, and stone terrace) on bacterial and fungal diversities, metagenomic and metabolomic functioning associated with basic soil properties. Our results showed that steep slopes at 25° significantly decreased soil pH, silt percentage, and bacterial and fungal abundances, but that soil and stone terraces increased soil organic C (SOC), silt and clay contents, and fungal abundance compared to sloping cultivated lands. In addition, soil and stone terraces increased both bacterial and fungal alpha diversities, and relative abundances of Crenarchaeota, Nitrospirota, and Latescibacterota, but reduced the proportions of Actinobacteriota and Patescibacteria, thus shifting microbial beta diversities, which were significantly associated with increased SOC and silt content. For metagenomics, soil and stone terraces greatly increased the relative abundance of functional genes related to Respiration, Virulence, disease and defense, Stress response, and nitrogen and potassium metabolisms, such as Denitrification and Potassium homeostasis. For soil metabolomics, a total of 22 soil metabolites was enriched by soil and stone terraces, such as Lipids and lipid-like molecules (Arachidonic acid, Gamma-Linolenic acid, and Pentadecanoic acid), and Organoheterocyclic compounds (Adenine, Laudanosine, Methylpyrazine, and Nicotinic acid). To sum up, soil and stone terraces could reduce some of the negative impacts of steep slope cultivation on soil microbial diversity as well as their metagenomic and metabolomic functioning related to C and nutrient metabolism useful for soil health improvement, potentially bolstering the impact of sustainable practices in erosion hotspots around the world., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Active iron oxides, especially poorly crystalline forms, benefit soil organic carbon (SOC) accumulation via directly bounding and indirectly promoting aggregation. However, it remains unclear on the impacts of active iron oxides on SOC accumulation in paddy and upland soils under long-term fertilization regimes. Here, we attempted to clarify the underlying mechanisms of amorphous (Fe O ) and organically complexed (Fe P ) iron oxides mediating SOC accumulation in paddy and upland soils based on two long-term fertilization experiments (both including no fertilization [CK]; chemical nitrogen, phosphorus and potassium [NPK] and NPK plus manure [NPKM] treatments). Results showed that compared to upland soil, Fe-bound organic carbon (Fe-bound OC) content in paddy soil, occupying 21-30% of SOC, was 77% higher on average, due to larger amounts of Fe O (+31%) and Fe P (+224%). The Fe O and Fe P were positively related to mean weight diameter (MWD) of soil aggregates across paddy and upland soils. Compared to NPK treatment, NPKM treatment strongly increased Fe O (+41%), Fe P (+60%) and associated Fe-bound OC (+19%) in paddy soil, and increased Fe O (+17%) and Fe P (+25%) while decreasing Fe-bound OC (-9%) in upland soil. These combined findings indicated the importance of poorly crystalline iron oxides facilitating Fe-bound OC formation and its contribution to SOC accumulation in paddy soil rather than upland soil. Moreover, long-term manure amendment could enhance SOC accumulation by increasing Fe-bound OC and aggregation stability in paddy soil and enhancing physical protection in upland soil, largely attributed to increased poorly crystalline iron oxides. Overall, these results highlight the potential mechanisms through which active iron oxides regulate SOC accumulation and guide fertilization management in paddy and upland soils., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aquaculture of bivalve shellfish and algae offers significant ecological benefits, yet the complex interactions between these organisms can substantially impact local carbon dynamics. This study investigated the effects of co-culturing four intertidal bivalve species Pacific oysters (Crassostrea gigas), Manila clams (Ruditapes philippinarum), Chinese clams (Cyclina sinensis), and hard clams (Mercenaria mercenaria) with microalgae (Isochrysis galbana) on specific water quality parameters, including total particulate matter (TPM), total organic matter (TOM), dissolved inorganic carbon (DIC), dissolved carbon dioxide (dCO 2 ), dissolved oxygen (DO), and ammonium (NH 4 + ), dissolved inorganic carbon (DIC), and CO 4 + ), dissolved inorganic carbon (DIC), and CO 2 in WP groups, indicating synergistic bioremediation with algae. M. mercenaria notably reduced TPM, highlighting its particle carbon sequestration potential. DO concentrations decreased in most WP or NP groups, reflecting respiration of the cultured bivalves or microalgae. NH 2 levels also declined for most species, indicating nitrogen assimilation by these creatures. Overall, the bivalve size significantly impacted carbon and nitrogen processing capacities. These findings reveal species-specific capabilities in regulating water carbon dynamics. Further research should explore integrating these bivalves in carbon-negative aquaculture systems to mitigate environmental impacts. This study provides valuable insights underlying local carbon dynamics in shallow marine ecosystems.2 in WP groups, indicating synergistic bioremediation with algae. M. mercenaria notably reduced TPM, highlighting its particle carbon sequestration potential. DO concentrations decreased in most WP or NP groups, reflecting respiration of the cultured bivalves or microalgae. NH 4 + levels also declined for most species, indicating nitrogen assimilation by these creatures. Overall, the bivalve size significantly impacted carbon and nitrogen processing capacities. These findings reveal species-specific capabilities in regulating water carbon dynamics. Further research should explore integrating these bivalves in carbon-negative aquaculture systems to mitigate environmental impacts. This study provides valuable insights underlying local carbon dynamics in shallow marine ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)