Search

With increasing land resource constraints, wetlands, as ecological hotspots, are expected to enhance biogeochemical processes to mitigate nitrogen (N) pollution, particularly nitrate-nitrogen (NO 3 -N). However, the interactions among bacteria, algae, and macrophytes in wetlands, which are crucial for N removal, remain largely unknown. This study explored how macrophyte coverage influences bacterial-algal interactions, shifting from mutualism to inhibition, thereby affecting N removal. Moderate coverage enhanced NO - -N). However, the interactions among bacteria, algae, and macrophytes in wetlands, which are crucial for N removal, remain largely unknown. This study explored how macrophyte coverage influences bacterial-algal interactions, shifting from mutualism to inhibition, thereby affecting N removal. Moderate coverage enhanced NO 3 - -N and total nitrogen (TN) removal (P < 0.05), which was correlated with increased microbial abundance (P < 0.05). This may have resulted from moderate algal photosynthesis, reduced physiological stress, and the expansion of ecological niches for microbes. Insufficient coverage promoted algal growth (chlorophyll-a > 31.8 μg·L -1 ), leading to increased competition for substrates and elevated pH, which further inhibited bacterial activity. Excessive coverage also inhibited bacterial activity by reducing illumination and oxidation-reduction potential. Consequently, insufficient and excessive coverage decreased N removal efficiencies by 2.7-15.7 % (NO 3 - -N) and 3.7-11.1 % (TN) while increasing methane emission potential by 1.4-6.9 times compared with moderate coverage. These findings offer insights into solving NO 3 - -N contamination using near-natural methods and balancing the ecological and practical considerations for small wetlands., 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.)

Background: The assembly of the rhizosphere community, even the diazotroph community, is mainly shaped by soil environmental factors (including soil climate and physiochemical characteristics) and plant selection. To better understand the driving forces on the active overall and nitrogen-fixing bacterial community compositions, we characterized the communities of tobacco rhizosphere soil collected from three sampling sites with a large geographic scale (> 600 km)., Results: The results indicate that the diversity and community composition of the overall bacterial and diazotroph communities are obviously differed according to the sampling sites. Still, no significant difference is found between the communities in rootzone and rhizosphere samples. Climate variables including mean annual precipitation (MAP) and mean annual temperature (MAT), soil physiochemical characteristics including available nitrogen (AN), available potassium (AK) and pH are main factors that affect the bacterial and diazotroph community structures in the three sampling sites. Furthermore, MAP and MAT, AN and available phosphorus (AP), total nitrogen (TN) and organic carbon (OC), AK and electrical conductivity (EC) showed similar effects, but pH showed independent effect on the composition of the overall bacteria and diazotroph communities. However, the alpha diversity indices of active overall and nitrogen-fixing bacteria in the rhizosphere are obviously higher than in the rootzone samples, and no significant differences are observed among different sampling sites. Proteobacteria is the predominant active phylum of all samples for overall and nitrogen-fixing bacteria. Escherichia-Shigella, Achromobacter, Streptomyces and Sphingomonas are the dominant active bacterial genera, and Bradyrhizobium, Skermanella and Extensimonas are dominant active nitrogen-fixing bacteria genera in rhizosphere. Furthermore, the high active abundance of Escherichia-Shigella but low abundance of Ralstonia in all three sampling sites indicate high root-knot nematode infection and low wilt disease endemic risk., Conclusion: These results indicate that soil environmental factors contribute more to the tobacco rhizosphere bacterial community assemblage, but the rhizosphere contributes more to the diversity of active overall bacteria and nitrogen-fixing bacteria in the community. Our study provides novel knowledge for the assemble of rhizosphere bacterial and active bacteria communities across a large geographical scale., (© 2024. The Author(s).)

The rhizosphere soil properties and microbial communities of Lilium tsingtauense , an endangered wild plant, have not been examined in previous studies. Here, we characterized spatial variation in soil properties and microbial communities in the rhizosphere of L. tsingtauense . We measured the abundance of L. tsingtauense at different altitudes and collected rhizosphere and bulk soils at three representative altitudes. The results showed that L. tsingtauense was more abundant, and the rhizosphere soil was richer in nitrogen, phosphorus, potassium, water content, and organic matter and more acidic at high altitudes than at lower altitudes. The diversity and richness of rhizosphere bacteria and fungi increased with altitude and were higher in rhizosphere soil than in bulk soil. In addition, ectomycorrhizal fungi, endophytic fungi, and nitrogen-fixing bacteria were more abundant, and plant-pathogenic fungi were less abundant at high altitudes. Co-occurrence network analysis identified four key phyla (Bacteroidota, Proteobacteria, Ascomycota, and Basidiomycota) in the microbial communities. We identified a series of microbial taxa (Acidobacteriales, Xanthobacteraceae, and Chaetomiaceae) and rhizosphere soil metabolites (phosphatidylcholine and phosphatidylserine) that are crucial for the survival of L. tsingtauense . Correlation analysis and random forest analysis showed that some environmental factors were closely related to the rhizosphere soil microbial community and played an important role in predicting the distribution and growth status of L. tsingtauense . In sum, the results of this study revealed altitudinal variation in the rhizosphere microbial communities of L. tsingtauense and the factors driving this variation. Our findings also have implications for habitat restoration and the conservation of this species., Importance: Our study highlighted the importance of the rhizosphere microbial community of the endangered plant L. tsingtauense . We found that soil pH plays an important role in the survival of L. tsingtauense . Our results demonstrated that a series of microbial taxa (Acidobacteriales, Xanthobacteraceae, Aspergillaceae, and Chaetomiaceae) and soil metabolites (phosphatidylcholine and phosphatidylserine) could be essential indicators for L. tsingtauense habitat. We also found that some environmental factors play an important role in shaping rhizosphere microbial community structure. Collectively, these results provided new insights into the altitudinal distribution of L. tsingtauense and highlight the importance of microbial communities in their growth., Competing Interests: The authors declare no conflict of interest.

Wastewater discharge is a global threat to freshwater resources. Streams, in particular, are receiving waterbodies that are directly impacted chemically and biologically due to effluent discharge. However, it is largely unknown how wastewater serves as a subsidy or a stressor to aquatic biodiversity, particularly microbiota, over space. Nutrient-diffusing substrata (NDS) were deployed; NDS release nutrients through diffusion into the water column into a wastewater-dependent stream across three reaches. We used N, P, and N + P treatments for the measurement of single nutrient and co-nutrient limitation, and a no-nutrient control. Both algal and total biofilm biomass was measured and the 16S ribosomal RNA genes via targeted amplicon sequencing was used to assess bacterial/archaeal community diversity. Data indicated that total organic matter in biofilms differs spatially with the greatest organic matter (OM) concentrations in the confluence downstream of wastewater inputs. Biofilm OM concentrations were greatest in P and N + P treatments in the confluence site relative to control or N-only treatments. This indicates heterotrophic microbial communities-likely bacteria that dominate stream biofilms-are P-limited in this ecosystem even with upstream wastewater inputs. In conjunction, bacteria/archaeal communities differed the greatest among nutrient treatments versus spatially and had several indicator taxa belonging to Flavobacterium spp. in N treatments relative to controls. Collectively with historical water quality data, we conclude that this wastewater-fed stream is primarily N-enriched but potentially P-limited, which results in significant shifts in biofilm bacterial communities and likely their overall biomass in this urban watershed., Importance: Streams in arid and semi-arid biomes are often dependent on their flow from municipal sources, such as wastewater effluent. However, wastewater has been shown to contain high concentrations of nutrients and chemical pollutants that can potentially harm aquatic ecosystems and their biota. Understanding if and the type of microorganisms that respond to pollution sources, specifically effluent from wastewater treatment facilities, in regions where flow is predominantly from treatment facilities, is critical for developing a predictive monitoring approach for eutrophication or other ecological degradation states for freshwaters., Competing Interests: The authors declare no conflict of interest.

Background: Relay intercropping of maize and soybean can improve land productivity. However, the mechanism behind N 2 O emissions in this practice remains unclear. A two-factor randomized block field trial was conducted to reveal the mechanism of N 2 O emissions in a full additive maize-soybean relay intercropping. Factor A was three cropping systems - that is, monoculture maize (Zea mays L.), monoculture soybean (Glycine max L. Merr.) and maize-soybean relay intercropping. Factor B was different N supply, containing no N, reduced N and conventional N. Differences in N 2 O emissions, soil properties, rhizosphere bacterial communities and yield advantage were evaluated., Results: The land equivalent ratio was 1.55-2.44, and the cumulative N 2 O emission ( C E N 2 O ) was notably lower by 60.2% in intercropping than in monoculture, respectively. Reduced N declined C E N 2 O without penalty on the yield advantages. The relay intercropping shifted soil properties - for example, soil organic matter, total N, NH 4 + and protease activity - and improved the soil microorganism community - for example, Proteobacteria and Acidobacteria. Intercropping reduced C E N 2 O by directly suppressing nirS- and amoA-regulated N 2 O generation during soil N cycling, or nirS- and amoA-mediated soil properties shifted to reduce C E N 2 O indirectly. Reduced N directly reduced C E N 2 O by decreasing soil N content and reducing soil microorganism activities to alleviate N 2 O produced in soil N cycling., Conclusion: Conducting a full additive maize-soybean relay intercropping with reduced nitrogen supply provides a way to alleviate N 2 O emissions without the penalty on the yield advantage by changing rhizosphere bacterial communities and soil N cycling. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Established as a plant macronutrient, potassium (K) substantially bestows plant growth and thus, global food production. It is absorbed by plants as potassium cation (K + ) from soil solution, which is enriched through slow-release from soil minerals or addition of soluble fertilizers. Contribution of bioavailable K + from soil is usually insignificant (< 2 %), although the earth's crust is rich in K-bearing minerals. However, K is fixed largely in interlayer spaces of K-bearing minerals, which can be released by K-solubilizing bacteria (KSB) such as Bacillus, Pseudomonas, Enterobacter, and Acidithiobacillus. The underlying mechanisms of K dissolution by KSB include acidolysis, ion exchange reactions, chelation, complexolysis, and release of various organic and inorganic acids such as citric, oxalic, acetic, gluconic, and tartaric acids. These acids cause disintegration of K-bearing minerals and bring K + into soil solution that becomes available to the plants. Current literature review updates the scientific information about microbial species, factors, and mechanisms governing the bio-intrusion of K-bearing minerals. Moreover, it explores the potential of KSB not only for K-solubilization but also to enhance bioavailability of phosphorus, nitrogen, and micronutrients, as well as its other beneficial impact on plant growth. Thus, in the context of sustainable agricultural production and global food security, utilization of KSB may facilitate plant nutrient availability, conserve natural resources, and reduce environmental impacts caused by chemical fertilizers., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Microorganisms are significant drivers of organic matter mineralization and are essential in marine biogeochemical cycles. However, the variations and influencing factors in prokaryotic communities from cold-seep sediments to the water column and the specific role of these microorganisms in biogeochemical cycles in the water column above cold seep remain unclear. Here, we investigated prokaryotic communities and their roles in nitrogen/sulfur cycling processes and conducted in situ dissolved organic matter (DOM) enrichment experiments to explore the effects of diverse sources of DOM on prokaryotic communities. Field investigations showed that the prokaryotic communities in the near-bottom water were more similar to those in the deep layer of the euphotic zone (44.60%) and at a depth of 400 m (50.89%) than those in the sediment (18.00%). DOM enrichment experiments revealed that adding dissolved organic nitrogen (DON) and phosphorus DOP caused a notable increase in the relative abundances of Rhodobacterales and Vibrionales, respectively. A remarkable increase was observed in the relative abundance of Alteromonadales and Pseudomonadales after the addition of dissolved organic sulfur (DOS). The metagenomic results revealed that Proteobacteria served as the keystone taxa in mediating the biogeochemical cycles of nitrogen, phosphorus, and sulfur in the Haima cold seep. This study highlights the responses of prokaryotes to DOM with different components and the microbially driven elemental cycles in cold seeps, providing a foundational reference for further studies on material energy metabolism and the coupled cycling of essential elements mediated by deep-sea microorganisms., Importance: Deep-sea cold seeps are among the most productive ecosystems, sustaining unique fauna and microbial communities through the release of methane and other hydrocarbons. Our study revealed that the influence of seepage fluid on the prokaryotic community in the water column is surprisingly limited, which challenges conventional views regarding the impact of seepage fluids. In addition, we identified that different DOM compositions play a crucial role in shaping the prokaryotic community composition, providing new insights into the factors driving microbial diversity in cold seeps. Furthermore, the study highlighted Proteobacteria as key and multifaceted drivers of biogeochemical cycles in cold seeps, emphasizing their significant contribution to complex interactions and processes. These findings offer a fresh perspective on the dynamics of cold-seep environments and their microbial communities, advancing our understanding of the biogeochemical functions in deep-sea environments., Competing Interests: The authors declare no conflict of interest.

The adverse effects of intensified cropland practices on soil quality and biodiversity become especially evident in India, where nearly 60% of land is dedicated to cultivation and almost 30% of soil is already degraded. Intensive agricultural practice significantly contributes to soil degradation, highlighting the crucial need for effective countermeasures to support sustainable development goals. A long-term experiment, established in the semi-arid Nimar Valley (India) in 2007, monitors the effect of organic and conventional management on the plant-soil system in a Vertisol. The focus of our study was to assess how organic and conventional farming systems affect biological and chemical soil quality indicators. Additionally, we followed the community structure of the soil microbiome throughout the vegetation phase under soya or cotton cultivation in the year 2019. We found that organic farming enhanced soil organic carbon and nitrogen content, increased microbial abundance and activity, and fostered distinct microbial communities associated with traits in nutrient mineralization. In contrast, conventional farming enhanced the abundance of bacteria involved in ammonium oxidation suggesting high nitrification and subsequent nitrogen losses with regular mineral fertilization. Our findings underscore the value of adopting organic farming approaches in semi-arid subtropical regions to rectify soil quality and minimize nitrogen losses., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Climate change and anthropogenic activities have significantly contributed to the degradation of wet meadows on the Qinghai-Tibet Plateau (QTP). Soil nitrogen (N) availability is a crucial determinant of the productivity of wet meadow vegetation. Furthermore, soil microbial nitrogen functional genes (NFGs) are critical in the transformation of soil N. Nevertheless, the dynamics of NFGs in response to vegetation degradation, as well as the underlying drivers, remain poorly understood. In this study, wet meadows at varying levels of vegetation degradation on the QTP, categorized as non-degraded (ND), slightly degraded (SD), moderately degraded (MD), and heavily degraded (HD), were examined. Soil samples from depths of 0 to 10 cm and 10 to 20 cm were collected during different growth cycles (June 2020, August 2020, and May 2021). The analysis focused on NFGs involved in organic nitrogen fixation ( nifH ), archaeal and bacterial ammonia oxidation ( amoA-AOA and amoA-AOB , respectively), and nitrite reduction ( nirK ), utilizing real-time fluorescence quantitative PCR. Our findings indicate a significant decline in the abundance of NFGs with intensified vegetation degradation, exhibiting notable spatial and temporal fluctuations. Specifically, the relative NFGs followed the pattern: nirK > amoA- AOA > amoA- AOB > nifH . Redundancy analysis revealed that vegetation cover was the primary regulator of NFGs abundance, accounting for 56.1%-57% of the variation. Additionally, soil total nitrogen, pH, and total phosphorus content were responsible for 38.5%, 28.2%, and 7% of the variability in NFGs, respectively. The ( amoA-AOA + amoA-AOB + nirK ) ratios associated with effective N transformation indicated that the vegetation degradation process moderately increased the nitrification potential., Importance: Our research investigates how the degradation of meadows affects the tiny organisms in soil that help plants use nitrogen, which is essential for their growth. In the Qinghai-Tibet Plateau, a region known for its unique ecosystems, we found that as meadows deteriorate-due to climate change and human activities-the number of these beneficial organisms significantly decreases. This decline could reduce soil fertility, impacting plant life and the overall health of the ecosystem. Understanding these changes helps us grasp how environmental pressures influence soil and plant health. Such knowledge is crucial for developing strategies to preserve these vulnerable ecosystems and ensure they continue to sustain biodiversity and provide resources for local communities., Competing Interests: The authors declare no conflict of interest.

Background: The ecosystems of marine ranching have enhanced marine biodiversity and ecological balance and have promoted the natural recovery and enhancement of fishery resources. The microbial communities of these ecosystems, including bacteria, fungi, protists, and viruses, are the drivers of biogeochemical cycles. Although seasonal changes in microbial communities are critical for ecosystem functioning, the current understanding of microbial-driven metabolic properties and their viral communities in marine sediments remains limited. Here, we employed amplicon (16S and 18S) and metagenomic approaches aiming to reveal the seasonal patterns of microbial communities, bacterial-eukaryotic interactions, whole metabolic potential, and their coupling mechanisms with carbon (C), nitrogen (N), and sulfur (S) cycling in marine ranching sediments. Additionally, the characterization and diversity of viral communities in different seasons were explored in marine ranching sediments., Results: The current study demonstrated that seasonal variations dramatically affected the diversity of microbial communities in marine ranching sediments and the bacterial-eukaryotic interkingdom co-occurrence networks. Metabolic reconstruction of the 113 medium to high-quality metagenome-assembled genomes (MAGs) was conducted, and a total of 8 MAGs involved in key metabolic genes and pathways (methane oxidation - denitrification - S oxidation), suggesting a possible coupling effect between the C, N, and S cycles. In total, 338 viral operational taxonomic units (vOTUs) were identified, all possessing specific ecological characteristics in different seasons and primarily belonging to Caudoviricetes, revealing their widespread distribution and variety in marine sediment ecosystems. In addition, predicted virus-host linkages showed that high host specificity was observed, with few viruses associated with specific hosts., Conclusions: This finding deepens our knowledge of element cycling and viral diversity in fisheries enrichment ecosystems, providing insights into microbial-virus interactions in marine sediments and their effects on biogeochemical cycling. These findings have potential applications in marine ranching management and ecological conservation. Video Abstract., (© 2024. The Author(s).)

Seasonal dynamics of bacterial composition and functions were demonstrated for the biological fluidized-bed bioreactors combined in the anoxic/aerobic1/aerobic2 (AOO) coking wastewater (CWW) treatment sequences. The bacterial composition and functions in the CWW activated sludge samples were revealed by 16S rRNA genes amplicon sequencing. Thiobacillus, Cloacibacterium, Alkaliphilus and Pseudomonas were determined as core genera with seasonal changes. Mutable microbial community composition fluctuated in different seasons in same bioreactor. Distributions of predicted KEGG pathways along four seasons consistently demonstrated enrichment in biodegradation of carbon- and nitrogen-containing compounds. The major contaminants were removed from CWW by biochemical pathway of xenobiotics biodegradation and metabolism. This Level 2 pathway mainly owned the Level 3 pathways of benzoate degradation, drug metabolism-other enzymes, drug metabolism-cytochrome P450, metabolism of xenobiotics by cytochrome P450, and aminobenzoate degradation. The RDA results showed that dissolved oxygen with seasonal fluctuation was the main parameter shaping the microbial community. The observed dynamics within the microbial community composition, coupled with the maintained stability of CWW treatment efficiencies and a consistent profile of microbial functional pathways, underscore the presence of functional redundancy in the AOO bioreactors. The study underscored stable and effective operational performances of bioreactors in the AOO sequences, contributing the knowledge of microbiological basics to the advancement of CWW biological treatment. KEY POINTS: • Seasonal fluctuations of bacterial composition described for the AOO system. • Seasonal distributions of metabolic functions focused on carbon and nitrogen removal. • Functional redundancy was revealed in the AOO microbial community., (© 2024. The Author(s).)

The biotechnological potential for agricultural applications in the soil in the thawing process on Whalers Bay, Deception Island, Antarctica was evaluated using a metagenomic approach through high-throughput sequencing. Approximately 22.70% of the sequences were affiliated to the phyla of the Bacteria dominion, followed by 0.26% to the Eukarya. Proteobacteria (Bacteria) and Ascomycota (Fungi) were the most abundant phyla. Thirty-two and thirty-six bacterial and fungal genera associated with agricultural biotechnological applications were observed. Streptomyces and Pythium were the most abundant genera related to the Bacteria and Oomycota, respectively. The main agricultural application associated with bacteria was nitrogen affixation; in contrast for fungi, was associated with phytopathogenic capabilities. The present study showed the need to use metagenomic technology to understand the dynamics and possible metabolic pathways associated with the microbial communities present in the soil sample in the process of thawing recovered from the Antarctic continent, which presented potential application in processes of agro-industrial interest., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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

The ammonia oxidation process driven by microorganisms is a dominant source for nitrous oxide （N 2 O） emissions. Here, we examined the influence of greenhouse vegetable planting ages on soil ammonia-oxidizing archaea （AOA） and ammonia-oxidizing bacteria （AOB）, which is of great significance for assessing the soil quality status and greenhouse gas transformations. A field study was conducted at different times （1, 5, 10, and 20 a） in greenhouse vegetable soils of Gaoyi, Hebei Province. Chemical analysis and Illumina NovaSeq high-throughput sequencing were used to analyze the soil physicochemical properties and community structures and diversity of AOA and AOB. The variation in AOA and AOB communities and the driving factors in greenhouse soils at different ages were also investigated. The results showed that the contents of total nitrogen, organic matter, alkali-hydrolyzable nitrogen, available phosphorus, and available potassium first increased and then decreased with the prolongation of growth. The contents of nitrate nitrogen, ammonium nitrogen, and electrical conductivity first decreased and then increased with the prolongation of growth. The pH value of soils decreased with the prolongation of growth. The abundance and diversity index of AOA and AOB first decreased and then increased with the prolongation of growth. Nitrososphaeria, unclassified Thaumarchaeota, and Candidatus Nitrosocaldus were the dominant species of AOA, while Betaproteobacteria and Nitrosospira were the dominant species of AOB. The composition of the soil AOA community varied greatly compared to that of AOB with the prolongation of growth. Correlation analysis showed that the changes in soil nutrient factors had a significant correlation with AOA and AOB communities. Redundancy analysis indicated that ammonium nitrogen, alkali-hydrolyzable nitrogen, and nitrate nitrogen were key factors of AOA communities, while electrical conductivity, available potassium, and nitrate nitrogen were key factors for AOB. In summary, long-term planting of greenhouse vegetables significantly affected the abundance and composition of soil AOA and AOB communities. Our results provide a theoretical basis for further studies on the greenhouse gas transformation and microbial mechanisms of the nitrogen cycle in greenhouse soils.

Cordyceps militaris infects insects and forms sclerotia within the insect remains, establishing insect-microbe complexes. Here, C . militaris sclerotia samples from a single location in China over a 5-year period were subjected to high-throughput DNA sequencing, and the core microbes (which were stably enriched in the sclerotia over the 5 years) were identified. Next, seven bacterial strains were isolated from the C. militaris sclerotia, their biochemical characteristics were assessed, and they were co-cultured with C. militaris to study their effects on C. militaris metabolite production and biomass. Furthermore, the effects of NH 4 , NO 3 , and peptone media on C. militaris were compared. The results showed that Rhodococcus , Phyllobacterium , Pseudomonas , Achromobacter , Ensifer , Stenotrophomonas , Sphingobacterium , Variovorax , and Acinetobacter were the core microbes. Although co-culture of C. militaris with the seven bacterial strains isolated from the sclerotia did not directly increase the cordycepin level, they all had NO 3 reduction ability, and four had urea decomposition ability. Meanwhile, C. militaris in NH 4 medium had an increased cordycepin level compared to C. militaris in the other two media. From this, we inferred that bacteria in the sclerotia can convert NO 3 to NH 4 , and then cordycepin is produced using NH 4 , which was confirmed by RNA-seq and real-time fluorescence quantitative PCR. Thus, bacteria in the sclerotia may indirectly affect the C. militaris metabolite production by regulating nitrogen metabolism. In summary, there are stable core microbes in the C. militaris sclerotia, and they may directly and indirectly affect the growth and metabolite production of C. militaris ., Importance: The model Cordyceps species Cordyceps militaris is rich in therapeutic compounds. It has recently been demonstrated that symbiotic microbes in sclerotia affect Cordyceps' growth, development, and secondary metabolite production. In this study, core microbes were identified based on C. militaris sclerotia samples obtained from the same site over 5 years. Additionally, bacterial strains isolated from C. militaris sclerotia were found to affect metabolite production and nitrogen utilization, based on functional tests. Moreover, based on the bacterial nitrogen metabolism capacity in the sclerotia and its influence on C. militaris metabolite production, we deduced that bacteria in the sclerotia can indirectly affect C. militaris metabolite production by regulating nitrogen metabolism. This is the first report on how bacteria in the sclerotia affect C. militaris metabolite production from the perspective of the nitrogen cycle. The results increase our understanding of microbial functions in C. militaris sclerotia., Competing Interests: The authors declare no conflict of interest.

A large amount of greenhouse gas nitrous oxide (N 2 O) will be produced during the biological nitrogen removal process for organic wastewater of low C/N ratio. One of the effective methods to solve this problem is to incorporate inexpensive carbon source. In this study, raw wastewater (RW) from pig farm, that was not anaerobically digested, was utilized as exogenous carbon in both A/O and SBR aerobic reactor to treat liquid digestate with high ammonia nitrogen and low C/N ratio. The results showed that N 2 O emission in SBR was higher than that of A/O process under the same nitrogen load. The N 2 O conversion in the biological nitrogen removal process was investigated by the strategy of integrating stable isotope method and metagenomics. The δO 18 -N 2 O, δN 15 -N 2 O, and SP values of the SBR were closer to the denitrification values of Ammonia-Oxidizing Bacteria (AOB) than those of A/O. The abundance of AOB in the SBR reactor was higher than that in the A/O reactor, while the abundance of denitrifying bacteria was lower. The amoA/B/C gene abundance in the SBR was greater than that in the A/O, and the NOS gene abundance was the opposite. The results indicated that both AOB denitrification and bacterial denitrification led to the increase of N 2 O emissions of the SBR., 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.)

Climate and edaphic properties drive the biogeographic distribution of dominant soil microbial phylotypes in terrestrial ecosystems. However, the impact of plant species and their root nutritional traits on microbial distribution in coastal wetlands remains unclear. Here, we investigated the nutritional traits of 100 halophyte root samples and the bacterial communities in the corresponding soil samples from coastal wetlands across eastern China. This study spans 22° of latitude, covering over 2500 km from north to south. We found that 1% of soil bacterial phylotypes accounted for nearly 30% of the soil bacterial community abundance, suggesting that a few bacterial phylotypes dominated the coastal wetlands. These dominated phylotypes could be grouped into three ecological clusters as per their preference over climatic (temperature and precipitation), edaphic (soil carbon and nitrogen), and plant factors (halophyte vegetation, root carbon, and nitrogen). We further provide novel evidence that plant root nutritional traits, especially root C and N, can strongly influence the distribution of these ecological clusters. Taken together, our study provides solid evidence of revealing the dominance of specific bacterial phylotypes and the complex interactions with their environment, highlighting the importance of plant root nutritional traits on biogeographic distribution of soil microbiome in coastal wetland ecosystems., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

The complex niche of fish gut is often characterized by the associated microorganisms that have implications in fish gut-health nexus. Although efforts to distinguish the microbial communities have highlighted their disparate structure along the gut length, remarkably little information is available about their distinct structural and functional profiles in different gut compartments in different fish species. Here, we performed comparative taxonomic and predictive functional analyses of the foregut and hindgut microbiota in an omnivorous freshwater fish species, Cyprinus carpio var. specularis, commonly known as mirror carp. Our analyses showed that the hindgut microbiota could be distinguished from foregut based on the abundance of ammonia-oxidizing, denitrifying, and nitrogen-fixing commensals of families such as Rhodospirillaceae, Oxalobacteraceae, Nitrosomonadaceae, and Nitrospiraceae. Functionally, unique metabolic pathways such as degradation of lignin, 2-nitrobenzoate, vanillin, vanillate, and toluene predicted within hindgut also hinted at the ability of hindgut microbiota for assimilation of nitrogen and detoxification of ammonia. The study highlights a major role of hindgut microbiota in assimilating nitrogen, which remains to be one of the limiting nutrients within the gut of mirror carp., (© 2024. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

The combination of microalgal culture and wastewater treatment is an emerging topic. This study investigated the use of different microalgae to treat different types of dairy farm wastewater. The results showed that the removal of ammonia nitrogen and total phosphorus by mixed microalgae was over 99% and 80%, respectively. The highest production of protein in biomass and extracellular polymeric substances was observed in high-concentration wastewater. In the phycosphere, the abundance of Proteobacteria and Cyanobacteria increased, while that of Bacteroidota decreased. Phycosphere bacteria were strongly correlated with microalgal growth and the composition of extracellular polymeric substances, especially with bound extracellular polymeric substances relative to soluble extracellular polymeric substances. Genes associated with photosynthesis and respiration in phycosphere bacteria were upregulated, contributing to the material exchange capacity in the microalgal-bacterial systems. The interaction between microalgae and phycosphere bacteria thus represents the core of the binary cultivation system-based wastewater treatment and requires further investigation., 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.)

Antagonistic bacterial strains from Bacillus spp. have been widely studied and utilized in the biocontrol of phytopathogens and the promotion of plant growth, but their impacts on the rhizosphere microecology when applied to crop plants are unclear. Herein, the effects of applying the antagonistic bacterium Bacillus subtilis S1 as a biofertilizer on the rhizosphere microecology of cucumbers were investigated. In a pot experiment on cucumber seedlings inoculated with S1, 3124 bacterial operational taxonomic units (OTUs) were obtained from the rhizosphere soils using high-throughput sequencing of 16S rRNA gene amplicons, and the most abundant phylum was Proteobacteria that accounted for 49.48% in the bacterial community. S1 treatment significantly reduced the abundances of soil bacterial taxa during a period of approximately 30 days but did not affect bacterial diversity in the rhizosphere soils of cucumbers. The enzymatic activities of soil nitrite reductase (S-Nir) and dehydrogenase (S-DHA) were significantly increased after S1 fertilization. However, the activities of soil urease (S-UE), cellulase (S-CL), and sucrase (S-SC) were significantly reduced compared to the control group. Additionally, the ammonium- and nitrate-nitrogen contents of S1-treated soil samples were significantly lower than those of the control group. S1 fertilization reshaped the rhizosphere soil bacterial community of cucumber plants. The S-CL activity and nitrate-nitrogen content in rhizosphere soil affected by S1 inoculation play important roles in altering the abundance of rhizosphere soil microbiota., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Human decomposition in terrestrial ecosystems is a dynamic process creating localized hot spots of soil microbial activity. Longer-term (beyond a few months) impacts on decomposer microbial communities are poorly characterized and do not typically connect microbial communities to biogeochemistry, limiting our understanding of decomposer communities and their functions. We performed separate year-long human decomposition trials, one starting in spring, another in winter, integrating bacterial and fungal community structure and abundances with soil physicochemistry and biogeochemistry to identify key drivers of microbial community change. In both trials, soil acidification, elevated microbial respiration, and reduced soil oxygen concentrations occurred. Changes in soil oxygen concentrations were the primary driver of microbial succession and nitrogen transformation patterns, while fungal community diversity and abundance was related to soil pH. Relative abundance of facultative anaerobic taxa (Firmicutes and Saccharomycetes) increased during the period of reduced soil oxygen. The magnitude and timing of the decomposition responses were amplified during the spring trial relative to the winter, even when corrected for thermal inputs (accumulated degree days). Further, soil chemical parameters, microbial community structure, and fungal gene abundances remained altered at the end of 1 year, suggesting longer-term impacts on soil ecosystems beyond the initial pulse of decomposition products., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Background: Oligotrophy and hypereutrophy represent the two extremes of lake trophic states, and understanding the distribution of bacterial communities across these contrasting conditions is crucial for advancing aquatic microbial research. Despite the significance of these extreme trophic states, bacterial community characteristics and co-occurrence patterns in such environments have been scarcely interpreted. To bridge this knowledge gap, we collected 60 water samples from Lake Fuxian (oligotrophic) and Lake Xingyun (hypereutrophic) during different hydrological periods., Results: Employing 16S rRNA gene sequencing, our findings revealed distinct community structures and metabolic potentials in bacterial communities of hypereutrophic and oligotrophic lake ecosystems. The hypereutrophic ecosystem exhibited higher bacterial α- and β-diversity compared to the oligotrophic ecosystem. Actinobacteria dominated the oligotrophic Lake Fuxian, while Cyanobacteria, Proteobacteria, and Bacteroidetes were more prevalent in the hypereutrophic Lake Xingyun. Functions associated with methanol oxidation, methylotrophy, fermentation, aromatic compound degradation, nitrogen/nitrate respiration, and nitrogen/nitrate denitrification were enriched in the oligotrophic lake, underscoring the vital role of bacteria in carbon and nitrogen cycling. In contrast, functions related to ureolysis, human pathogens, animal parasites or symbionts, and phototrophy were enriched in the hypereutrophic lake, highlighting human activity-related disturbances and potential pathogenic risks. Co-occurrence network analysis unveiled a more complex and stable bacterial network in the hypereutrophic lake compared to the oligotrophic lake., Conclusion: Our study provides insights into the intricate relationships between trophic states and bacterial community structure, emphasizing significant differences in diversity, community composition, and network characteristics between extreme states of oligotrophy and hypereutrophy. Additionally, it explores the nuanced responses of bacterial communities to environmental conditions in these two contrasting trophic states., (© 2024. The Author(s).)

The ecological function of biological crusts in arid and semi-arid areas is of great importance. Bacteria, as a crucial microbial group in biological crusts, play a key role in the formation, nutrient cycling, and regulation of these crusts. However, the succession of biological crusts and the diversity of bacterial communities, along with key environmental factors in the Loess Plateau's hilly and gully areas, remain unclear. This study investigated soil bacterial abundance and diversity in bare soil (BS), alga-lichen mixed crust (MC), and alga-lichen mixed crust subsoil (MCS) using high-throughput sequencing methods. It explored the relationship between the bacterial community in biological crusts and key environmental factors. The results indicated that the Chao1, Shannon index, and phylogenetic diversity of bacteria significantly increased with the succession of biological crusts. There were notable differences in the community composition and structure of bacteria at different stages of crust development, with Rubrobacteria and Cyanobacteriia dominating in MCS. Effective phosphorus, available potassium, nitrogen, pH, and total organic carbon were identified as key environmental factors affecting soil bacterial communities. In summary, the succession of biological crusts alters soil physicochemical characteristics and creates different ecological niches for bacterial communities. Soil nutrients and pH play a crucial role in the selection of bacterial species and the shaping of bacterial communities in the Loess Plateau's hilly and gully areas., (© 2024. The Author(s).)

Anammox bacteria (AnAOB) can be easily enriched under high temperatures and high substrate concentrations, while the application of the mainstream anammox process in low substrate municipal sewage is still relatively uncommon. Therefore, this study investigated the enrichment of AnAOB under conditions of low ammonia nitrogen and nitrite concentration at 25 °C. Results showed that using inoculated aerobic sludge, four ASBRs (R1, R2, R3 and R4) were successfully initiated with different influent substrate (NO 2 - -N/NH 4 + -N) ratios of 1.2, 1.32, 1.4 and 1.5, respectively, with reactor start-up times were 162, 150, 120 and 134 days, respectively. The values of ΔNO 2 - -N/ΔNH 4 + -N in reactors were stable at 1.17, 1.32, 1.43 and 1.53 respectively. The increase in influent substrate ratios resulted in improved TN removal rates and accelerated consumption of chemical oxygen demand (COD) during the initial start-up stage. The maximum TN removal rates achieved in the four reactors were 76.09%, 79.24%, 82.82% and 82.63%, respectively. The color of sludge gradually changes from yellowish-brown to reddish-brown. Furthermore, the surface of sludge exhibited a porous mineral structure, with crater-like cavities. The dominant anammox species in the system was identified as Candida Brocadia (3.04%). According to qPCR, the abundance of hzsB in the system is 1.65 × 10 12 copies/g VSS, confirming the effective enrichment of AnAOB., 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.)

Guanidine is a chemically stable nitrogen compound that is excreted in human urine and is widely used in manufacturing of plastics, as a flame retardant and as a component of propellants, and is well known as a protein denaturant in biochemistry 1-3 . Guanidine occurs widely in nature and is used by several microorganisms as a nitrogen source, but microorganisms growing on guanidine as the only substrate have not yet been identified. Here we show that the complete ammonia oxidizer (comammox) Nitrospira inopinata and probably most other comammox microorganisms can grow on guanidine as the sole source of energy, reductant and nitrogen. Proteomics, enzyme kinetics and the crystal structure of a N. inopinata guanidinase homologue demonstrated that it is a bona fide guanidinase. Incubation experiments with comammox-containing agricultural soil and wastewater treatment plant microbiomes suggested that guanidine serves as substrate for nitrification in the environment. The identification of guanidine as a growth substrate for comammox shows an unexpected niche of these globally important nitrifiers and offers opportunities for their isolation., (© 2024. The Author(s).)

Heterotrophic nitrification (HN) bacteria use organic carbon sources to remove ammonia nitrogen (NH 4 -N); however, the mechanisms of carbon and nitrogen metabolism are unknown. To understand this mechanism, HN functional microbial communities named MG and MA were enriched with glucose and sodium acetate, respectively. The NH + -N removal efficiencies were 98.87 % and 98.91 %, with 88.06 % and 69.77 % nitrogen assimilation for MG and MA at 22 h and 10 h, respectively. Fungi (52.86 %) were more competitive in MG, and bacteria (99.99 %) were dominant in MA. Metagenomic and metabolomic analyses indicated that HN might be a signaling molecule (NO) in the production and detoxification processes when MG metabolizes glucose (amo, hao, and nosZ were not detected). MA metabolizes sodium acetate to produce less energy and promotes nitrogen oxidation reduction; however, genes (hao, hox, and NOS2) were not detected. These results suggest that NO and energy requirements induce microbial HN.4 + -N removal efficiencies were 98.87 % and 98.91 %, with 88.06 % and 69.77 % nitrogen assimilation for MG and MA at 22 h and 10 h, respectively. Fungi (52.86 %) were more competitive in MG, and bacteria (99.99 %) were dominant in MA. Metagenomic and metabolomic analyses indicated that HN might be a signaling molecule (NO) in the production and detoxification processes when MG metabolizes glucose (amo, hao, and nosZ were not detected). MA metabolizes sodium acetate to produce less energy and promotes nitrogen oxidation reduction; however, genes (hao, hox, and NOS2) were not detected. These results suggest that NO and energy requirements induce microbial HN., 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 work unraveled discrepant ecological patterns between biofilms and flocs in a deteriorated bioreactor inoculated with mature partial nitrification-anammox (PN/A) sludge. Based on 16S rRNA analysis, a comprehensive evaluation of neutral and null models, along with niche width, delineated that the bacterial community assembly in biofilms and flocs was dominantly driven by the stochastic process, and dispersal limitation critically shaped the community assembly. Co-occurrence network analysis revealed that environmental stress caused decentralized and fragmented bacterial colonies, and anammox bacteria were mainly peripheral in biofilms network and less involved in interspecific interactions. Simultaneous PN/A and partial denitrification-anammox (PD/A) processes were identified, whereas PN and PD process primarily occurred in the biofilms and flocs, respectively, as evidenced by metagenomics. Collectively, these outcomes are expected to deepen the basic understanding of complex microbial community and nitrogen metabolism under environmental disturbance, thereby better characterizing and serving the artificial 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. Published by Elsevier Ltd.)

Excessive nitrogen release during composting poses significant challenges to both the environment and compost quality. Biological enhancement of humification and nitrogen conservation is an environmentally friendly and cost-effective approach to composting. The aim of this study was to develop a psychrophilic and thermophilic nitrifying bacterial consortium (CNB) and investigate its role in nitrogen transformation and humification during cow manure composting. Analysis revealed that CNB inoculation promoted microbial proliferation and metabolism, significantly increased the number of nitrifying bacteria (p < 0.05), and elevated the activity of nitrite oxidoreductase and nxrA gene abundance. Compared to the control, CNB inoculation promoted the formation of NO 3 - -N (77.87-82.35 %), while reducing NH 3 O (20.05 %) emissions, and increased humus content (16.22 %). Mantel analysis showed that the higher abundance of nitrifying bacteria and nxrA facilitated the nitrification of NH 2 O (20.05 %) emissions, and increased humus content (16.22 %). Mantel analysis showed that the higher abundance of nitrifying bacteria and nxrA facilitated the nitrification of NH 4 + -N. The improvement in nitrite oxidoreductase activity promoted NO 3 - -N formation, leading to increased humus content and enhanced compost safety., 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 investigates the performance, resilience and microbial community dynamics of two anaerobic processes, i.e. pure anammox (R1) and partial denitrification/anammox (PD/A) (R2), following a 30-day starvation period. The tolerance to starvation was assessed by comparing nitrogen removal efficiency and microbial activity across both reactors. Results show that the PD/A process recovery to pre-starvation performance levels within just one day, as compared to the pure anammox process. Notably, although the activity of anammox bacteria decreased in both processes during starvation, the decay rate in R1 was 69.59 % higher than in R2, potentially explaining the quicker recovery of R2. Furthermore, enhanced secretion of extracellular polymeric substance (EPS) during starvation served as a protective mechanism. The potential functions and genes in microorganisms, as well as the pathway of nitrogen cycling, were demonstrated through analyses using the KEGG database. This research reveals essential mechanistic insights and strategic guidance for the effective implementation of anammox-based biological nitrogen removal 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 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In the past few decades, the pressure of higher food production to satisfy the demand of ever rising population has inevitably increased the use synthetic agrochemicals which have deterioration effects. Biostimulants containing beneficial microbes (single inoculants and microbial consortium) were found as an ideal substitute of synthetic chemical fertilizers. In recent years, microbial consortium is known as a better bioinoculant in comparison to single inoculant bioformulation because of multifarious plant growth-promoting advantages. Looking at the advantageous effect of consortium, in present investigation, different bacteria were isolated from rhizospheric soil and plant samples collected from the Himalayan mountains on the green slopes of the Shivaliks, Himachal Pradesh. The isolated bacteria were screened for nitrogen (N) fixation, phosphorus (P) solubilization and potassium (K) solubilization plant growth promoting attributes, and efficient strains were identified through 16S rRNA gene sequencing and BLASTn analysis. The bacteria showing a positive effect in NPK uptake were developed as bacterial consortium for the growth promotion of eggplant crop. A total of 188 rhizospheric and endophytic bacteria were sorted out, among which 13 were exhibiting nitrogenase activity, whereas 43 and 31 were exhibiting P and K solubilization traits, respectively. The selected three efficient and potential bacterial strains were identified using 16S rRNA gene sequencing as Enterobacter ludwigii EU-BEN-22 (N-fixer; 35.68 ± 00.9 nmol C 2 H 4 per mg protein per h), Micrococcus indicus EU-BRP-6 (P-solubilizer; 201 ± 0.004 mg/L), and Pseudomonas gessardii EU-BRK-55 (K-solubilizer; 51.3 ± 1.7 mg/mL), and they were used to develop a bacterial consortium. The bacterial consortium evaluation on eggplant resulted in the improvement of growth (root/shoot length and biomass) and physiological parameters (chlorophyll, carotenoids, total soluble sugar, and phenolic content) of the plants with respect to single culture inoculation, chemical fertilizer, and untreated control. A bacterial consortium having potential to promote plant growth could be used as bioinoculant for horticulture crops growing in hilly regions., (© 2024. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

The quality of tobacco is directly affected by macromolecular content, fermentation is an effective method to improve biochemical properties. In this study, we utilized CBHA (cellobiohydrolase A) glycosylase, which was expressed by Pichia pastoris , as an additive for fermentation. The contents of main chemical components of tobacco leaves after fermentation were determined, and the changes of microbial community structure and abundance in tobacco leaves during fermentation were analyzed. The relationship between chemical composition and changes in microbial composition was investigated, and the function of bacteria and fungi in fermentation was predicted to identify possible metabolic pathways. After 48 h of CBHA fermentation, the contents of starch, cellulose and total nitrogen in tobacco leaf decreased by 17.60%, 28.91% and 16.05%, respectively. The microbial community structure changed significantly, with Aspergillus abundance decreasing significantly, while Filobasidum , Cladosporium , Bullera , Komagataella , etc., increased in CBHA treated group. Soluble sugar was most affected by microbial community in tobacco leaves, which was negatively correlated with starch, cellulose and total nitrogen. During the fermentation process, the relative abundance of metabolism-related functional genes increased, and the expressions of cellulase and endopeptidase also increased. The results showed that the changes of bacterial community and dominant microbial community on tobacco leaves affected the content of chemical components in tobacco leaves, and adding CBHA for fermentation had a positive effect on improving the quality of tobacco leaves.

The microbial processes occurring in constructed wetlands (CWs) are difficult to understand owing to the complex interactions occurring between a variety of substrates, microorganisms, and plants under the given physicochemical conditions. This frequently leads to very large unexplained nitrogen losses in these systems. In continuation of our findings on Anammox contributions, our research on full-scale field CWs has suggested the significant involvement of the sulfur cycle in the conventional C-N cycle occurring in wetlands, which might closely explain the nitrogen losses in these systems. This paper explored the possibility of the sulfur-driven autotrophic denitrification (SDAD) pathway in different types of CWs, shallow and deep and passive and aerated systems, by analyzing the metagenomic bacterial communities present within these CWs. The results indicate a higher abundance of SDAD bacteria (Paracoccus and Arcobacter) in deep passive systems compared to shallow systems and presence of a large number of SDAD genera (Paracoccus, Thiobacillus, Beggiatoa, Sulfurimonas, Arcobacter, and Sulfuricurvum) in aerated CWs. The bacteria belonging to the functional category of dark oxidation of sulfur compounds were found to be enriched in deep and aerated CWs hinting at the possible role of the SDAD pathway in total nitrogen removal in these systems. As a case study, the percentage nitrogen removal through SDAD pathway was calculated to be 15-20% in aerated wetlands. The presence of autotrophic pathways for nitrogen removal can prove highly beneficial in terms of reducing sludge generation and hence reducing clogging, making aerated CWs a sustainable wastewater treatment solution., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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).)

Soil microorganisms are the most active participants in terrestrial ecosystems, and have key roles in biogeochemical cycles and ecosystem functions. Despite the extensive research on soil pH as a key predictor of microbial community and composition, a limitation of these studies lies in determining whether bacterial and/or fungal communities are directly or indirectly influenced by pH. We conducted a controlled laboratory experiment to investigate the effects of soil pH amendment (+/- 1-2 units) with six levels on soil microbial communities in two contrasting Chinese agricultural soils (pH 8.43 in Dezhou, located in the North China Plain, Shandong vs pH 6.17 in Wuxi, located in the Taihu Lake region, Jiangsu, east China). Results showed that the fungal diversity and composition were related to soil pH, but the effects were much lower than the effects of soil pH on bacterial community in two soils. The diversity and composition of bacterial communities were more closely associated with soil pH in Wuxi soils compared to Dezhou soils. The alpha diversity of bacterial communities peaked near in situ pH levels in both soils, displaying a quadratic fitting pattern. Redundancy analysis and variation partition analysis indicated that soil pH affected bacterial community and composition by directly imposing a physiological constraint on soil bacteria and indirectly altering soil characteristics (e.g., nutrient availability). The study also examined complete curves of taxa relative abundances at the phylum and family levels in response to soil pH, with most relationships conforming to a quadratic fitting pattern, indicating soil pH is a reliable predictor. Furthermore, soil pH amendment affected the transformation of nitrogen and the abundances of functional genes involved in the nitrogen cycle, and methane production and consumption. Overall, results from this study would enhance our comprehension of how soil microorganisms in contrasting farmlands will respond to soil pH changes, and would contribute to more effective soil management and conservation strategies., Importance: This study delves into the impact of soil pH on microbial communities, investigating whether pH directly or indirectly influences bacterial and fungal communities. The research involved two contrasting soils subjected to a 1-2 pH unit amendment. Results indicate bacterial community composition was shaped by soil pH through physiological constraints and nutrient limitations. We found that most taxa relative abundances at the phylum and family levels responded to pH with a quadratic fitting pattern, indicating that soil pH is a reliable predictor. Additionally, soil pH was found to significantly influence the predicted abundance of functional genes involved in the nitrogen cycle as well as in methane production and consumption processes. These insights can contribute to develop more effective soil management and conservation strategies., Competing Interests: The authors declare no conflict of interest.

Global patterns in soil microbiomes are driven by non-linear environmental thresholds. Fertilization is known to shape the soil microbiome of terrestrial ecosystems worldwide. Yet, whether fertilization influences global thresholds in soil microbiomes remains virtually unknown. Here, utilizing optimized machine learning models with Shapley additive explanations on a dataset of 10,907 soil samples from 24 countries, we discovered that the microbial community response to fertilization is highly dependent on environmental contexts. Furthermore, the interactions among nitrogen (N) addition, pH, and mean annual temperature contribute to non-linear patterns in soil bacterial diversity. Specifically, we observed positive responses within a soil pH range of 5.2-6.6, with the influence of higher temperature (>15°C) on bacterial diversity being positive within this pH range but reversed in more acidic or alkaline soils. Additionally, we revealed the threshold effect of soil organic carbon and total nitrogen, demonstrating how temperature and N addition amount interacted with microbial communities within specific edaphic concentration ranges. Our findings underscore how complex environmental interactions control soil bacterial diversity under fertilization., (© 2024 John Wiley & Sons Ltd.)

Atmospheric nitrogen (N) deposition has been substantially reduced due to declines in the reactive N emission in major regions of the world. Nevertheless, the impact of reduced N deposition on soil microbial communities and the mechanisms by which they are regulated remain largely unknown. Here, we examined the effects of N addition and cessation of N addition on plant and soil microbial communities through a 17-year field experiment in a temperate grassland. We found that extreme N input did not irreversibly disrupt the ecosystem, but ceasing high levels of N addition led to greater resilience in bacterial and fungal communities. Fungi exhibited diminished resilience compared to bacteria due to their heightened reliance on changes in plant communities. Neither bacterial nor fungal diversity fully recovered to their original states. Their sensitivity and resilience were mainly steered by toxic metal ions and soil pH differentially regulating on functional taxa. Specifically, beneficial symbiotic microbes such as N-fixing bacteria and arbuscular mycorrhizal fungi experienced detrimental effects from toxic metal ions and lower pH, hindering their recovery. The bacterial functional groups involved in carbon decomposition, and ericoid mycorrhizal and saprotrophic fungi were positively influenced by soil metals, and demonstrated gradual recovery. These findings could advance our mechanistic understanding of microbial community dynamics under ongoing global changes, thereby informing management strategies to mitigate the adverse effects of N enrichment on soil function., (© 2024 John Wiley & Sons Ltd.)

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 -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.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.)

Understanding the microbial community structure of sludge is crucial for improving the design, operation and optimisation of full-scale wastewater treatment plants (WWTPs). This study aimed to have a comprehensive comparison of microbial communities between aerobic granular sludge and flocculent sludge from two full-scale sequential batch reactors-based WWTPs with nutrient removal for the first time. To better understand key functional bacteria such as polyphosphate accumulating bacteria (PAOs), competitive bacteria such as glycogen accumulating bacteria (GAOs) and nitrifying bacteria for both nitrogen and phosphorus removal, another two full-scale WWTPs with only carbon (C) removal and C and nitrogen (N) removal were compared too. It was found that the richness and diversity of the microbial population in sludge increased with pollutant removal from only C, C and N, to C,N, P removal. For C, N P removal, granule structure led to a more diverse and rich microbial community structure than flocculent structure. Although more abundant nitrifying bacteria were enriched in granular sludge than flocculent sludge, the abundance of total putative PAOs was equivalent. However, the most typical putative PAOs such as Tetrasphaera and Candidatus Accumulibacter seemed to be more correlated with biological phosphorus removal performance, which might be more proper to be used as an indication for P removal potential. The higher abundance of GAOs in flocculent sludge with better phosphorus removal performance might suggest that further investigation is needed to understand the functions of GAOs. In addition, the equivalent abundances of PAOs in the WWTPs with only C removal and with C, N, and P removal, respectively, indicate that many newly reported putative PAOs might not contribute to P removal. This study provides insight into the microbial communities and functional bacteria in aerobic granular sludge and flocculent sludge in full-scale SBRs, which can provide microbes-informed optimisation of reactor operation for better nutrient removal., Competing Interests: Declaration of competing interest There is no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study, two bioprocess models were first constructed with the newly-discovered comammox process described as one-step and two-step nitrification and evaluated against relevant experimental data. The validated models were then applied to reveal the potential effect of comammox bacteria on the granular bioreactor particularly suitable for undertaking partial nitritation/anammox (PN/A) under different operating conditions of bulk dissolved oxygen (DO) and influent NH 4 . The results showed although comammox bacteria-based PN/A could achieve > 80.0 % total nitrogen (TN) removal over a relatively wider range of bulk DO and influent NH + . The results showed although comammox bacteria-based PN/A could achieve > 80.0 % total nitrogen (TN) removal over a relatively wider range of bulk DO and influent NH 4 + (i.e., 0.25-0.40 g-O 2 /m 3 and 470-870 g-N/m 3 to avoid the undesired full nitrification by comammox bacteria. Comparatively, conventional ammonium-oxidizing bacteria (AOB)-based PN/A not only required higher bulk DO to achieve > 80.0 % TN removal but also suffered from 1.7 %∼2.8 % N 2 O) production (< 0.1 %), the bulk DO should be finely controlled based on the influent NH 4 + to avoid the undesired full nitrification by comammox bacteria. Comparatively, conventional ammonium-oxidizing bacteria (AOB)-based PN/A not only required higher bulk DO to achieve > 80.0 % TN removal but also suffered from 1.7 %∼2.8 % N 2 O production., 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 order to explore the operation performance, kinetic characteristics and bacterial community of the short-cut nitrification and denitrification (SND) system, the SND system with pre-cultured short cut nitrification and denitrification sludge was established and operated under different ferrous ion (Fe (II)) conditions. Experimental results showed that the average NH 4 + -N removal efficiency (ARE) of SND system was 97.3% on Day 5 and maintained a high level of 94.9% ± 1.3% for a long operation period. When the influent Fe(II) concentration increased from 2.3 to 7.3 mg L -1 , the sedimentation performance, sludge concentration and organic matter removal performance were improved. However, higher Fe(II) of 12.3 mg L -1 decreased the removal of nitrogen and COD Cr with the relative abundance (RA) of Proteobacteria and Bacteroidetes decreased to 30.28% and 19.41%, respectively. Proteobacteria, Bacteroidetes and Firmicutes were the dominant phyla in SND system. Higher Fe(II) level of 12.3 mg L -1 increase the RA of denitrifying genus Trichococcus (33.93%), and the denitrifying genus Thauera and Tolumonas dominant at Fe(II) level of no more than 7.3 mg L -1 ., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Soil extracellular enzyme plays a vital role in changing soil nitrogen (N) mineralization of rice field. However, the effects of soil extracellular enzyme activities (EEA) and microbial community composition response to N mineralization of rice field under short-term tillage treatment needed to be further explored. In this study, we investigated the impact of short-term (8-year) tillage practices on rhizosphere soil N transformation rate, soil enzyme activities, soil microbial community structure, and the N mineralization function gene abundances in double-cropping rice field in southern China. The experiment consisted of four tillage treatments: rotary tillage with crop straw input (RT), conventional tillage with crop straw input (CT), no-tillage with crop straw retention (NT), and rotary tillage with all crop straw removed as a control (RTO). The results indicated that the rhizosphere soil N transformation rate in paddy field under the NT and RTO treatments was significantly decreased compared to RT and CT treatments. In comparison to the NT and RTO treatments, soil protease, urease, β -glucosaminidase, and arginase activities were significantly improved by the CT treatment, as were abundances of soil sub , npr , and chiA with CT and RT treatments. Moreover, the overall diversity of soil bacterial communities in NT and RTO treatments was significantly lower than that in RT and CT treatments. Soil chitinolytic and bacterial ureolytic communities were also obviously changed under a combination of tillage and crop straw input practices.

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).)

Leveraging comammox Nitrospira and anammox bacteria for shortcut nitrogen removal can drastically lower the carbon footprint of wastewater treatment facilities by decreasing aeration energy, carbon, alkalinity, and tank volume requirements while also potentially reducing nitrous oxide emissions. However, their co-occurrence as dominant nitrifying bacteria is rarely reported in full-scale wastewater treatment. As a result, there is a poor understanding of how operational parameters, in particular, dissolved oxygen, impact their activity and synergistic behavior. Here, we report the impact of dissolved oxygen concentration (DO = 2, 4, 6 mg/L) on the microbial community's transcriptomic expression in a full-scale integrated fixed film activated sludge (IFAS) municipal wastewater treatment facility where nitrogen removal is predominantly performed by comammox Nitrospira and anammox bacterial populations. 16S rRNA transcript compositions revealed anammox bacteria and Nitrospira were significantly more active in IFAS biofilms compared to suspended sludge biomass. In IFAS biofilms, anammox bacteria significantly increased hzo expression at lower dissolved oxygen concentrations and this increase was highly correlated with the amoA expression levels of comammox bacteria. Interestingly, the genes involved in nitrite oxidation by comammox bacteria were significantly more upregulated, relative to the genes involved in ammonia oxidation with decreasing dissolved oxygen concentrations. Ultimately, our findings suggest that comammox Nitrospira supplies anammox bacteria with nitrite via ammonia oxidation and that this synergistic behavior is dependent on dissolved oxygen concentrations.

The Mariana Trench (MT) is the deepest part of the ocean on Earth. Previous studies have described the microbial community structures and functional potential in the seawater and surface sediment of MT. Still, the metabolic features and adaptation strategies of the microorganisms involved in nitrogen cycling processes are poorly understood. In this study, comparative metagenomic approaches were used to study microbial nitrogen cycling in three MT habitats, including hadal seawater [9,600-10,500 m below sea level (mbsl)], surface sediments [0-46 cm below seafloor (cmbsf) at a water depth between 7,143 and 8,638 mbsl], and deep sediments (200-306 cmbsf at a water depth of 8,300 mbsl). We identified five new nitrite-oxidizing bacteria (NOB) lineages that had adapted to the oligotrophic MT slope sediment, via their CO 2 fixation capability through the reductive tricarboxylic acid (rTCA) or Calvin-Benson-Bassham (CBB) cycle; an anammox bacterium might perform aerobic respiration and utilize sedimentary carbohydrates for energy generation because it contains genes encoding type A cytochrome c oxidase and complete glycolysis pathway. In seawater, abundant alkane-oxidizing Ketobacter species can fix inert N 2 released from other denitrifying and/or anammox bacteria. This study further expands our understanding of microbial life in the largely unexplored deepest part of the ocean., Importance: The metabolic features and adaptation strategies of the nitrogen cycling microorganisms in the deepest part of the ocean are largely unknown. This study revealed that anammox bacteria might perform aerobic respiration in response to nutrient limitation or O 2 fluctuations in the Mariana Trench sediments. Meanwhile, an abundant alkane-oxidizing Ketobacter species could fix N 2 in hadal seawater. This study provides new insights into the roles of hadal microorganisms in global nitrogen biogeochemical cycles. It substantially expands our understanding of the microbial life in the largely unexplored deepest part of the ocean., Competing Interests: The authors declare no conflict of interest.

Human activities have caused an imbalance in the input nitrogen and phosphorus (N/P) in the biosphere. The imbalance of N/P is one of the characteristics of water eutrophication, which is the fundamental factor responsible for the blooms. The effects of the N/P imbalance on diatom and phycospheric bacteria in blooms are poorly understood. In this study, the N/P molar ratio in real water (14:1) and the predicted N/P molar ratio in future water (65:1) were simulated to analyze the response of Cyclotella sp. and phycospheric bacteria to the N/P imbalance. The results showed that the N/P imbalance inhibited the growth of Cyclotella sp., but prolonged diatom bloom duration. The resistance of Cyclotella sp. to the N/P imbalance is related to phycospheric bacteria, and there are dynamic regulatory mechanisms within the phycospheric bacteria community to resist the N/P imbalance: (1) the increase of HNA bacterial density, the decrease of LNA bacterial density, (2) the increase of phycospheric bacterial diversity and eutrophic bacteria abundance, and the change of denitrifying bacteria abundance, (3) the activity of nitrogen and phosphorus metabolism of HNA bacteria enhanced, while that of LNA bacteria decreased. And the gene hosts of nitrogen and phosphorus metabolism were most enriched in Proteobacteria, indicating that Proteobacteria played an important role in maintaining the stability of phycospheric bacteria and was the dominant phylum resistant to the N/P imbalance. This study clarified that the algal-bacteria system was resistant to the N/P imbalance and implied that the N/P imbalance had little effect on the occurrence of diatom bloom events due to the presence of phycospheric bacteria., 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.)

Microbially-driven soil formation process is an emerging technology for the ecological rehabilitation of alkaline tailings. However, the dominant microorganisms and their specific roles in soil formation processes remain unknown. Herein, a 1-year field-scale experiment was applied to demonstrate the effect of nitrogen input on the structure and function of the microbiome in alkaline bauxite residue. Results showed that the contents of nutrient components were increased with Penicillium oxalicum (P. oxalicum) incorporation, as indicated by the increasing of carbon and nitrogen mineralization and enzyme metabolic efficiency. Specifically, the increasing enzyme metabolic efficiency was associated with nitrogen input, which shaped the microbial nutrient acquisition strategy. Subsequently, we evidenced that P. oxalicum played a significant role in shaping the assemblages of core bacterial taxa and influencing ecological functioning through intra- and cross-kingdom network analysis. Furthermore, a recruitment experiment indicated that nitrogen enhanced the enrichment of core microbiota (Nitrosomonas, Bacillus, Pseudomonas, and Saccharomyces) and may provide benefits to fungal community bio-diversity and microbial network stability. Collectively, these results demonstrated nitrogen-based coexistence patterns among P. oxalicum and microbiome and revealed P. oxalicum-mediated nutrient dynamics and ecophysiological adaptations in alkaline microhabitats. It will aid in promoting soil formation and ecological rehabilitation of bauxite residue. ENVIRONMENT IMPLICATION: Bauxite residue is a highly alkaline solid waste generated during the Bayer process for producing alumina. Attempting to transform bauxite residue into a stable soil-like substrate using low-cost microbial resources is a highly promising engineering. However, the dominant microorganisms and their specific roles in soil formation processes remain unknown. In this study, we evidenced the nitrogen-based coexistence patterns among Penicillium oxalicum and microbiome and revealed Penicillium oxalicum-mediated nutrient dynamics and ecophysiological adaptations in alkaline microhabitats. This study can improve the understanding of core microbes' assemblies that affect the microbiome physiological traits in soil formation 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.)

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.)

The estuarine plastisphere, a novel ecological habitat in the Anthropocene, has garnered global concerns. Recent geochemical evidence has pointed out its potential role in influencing nitrogen biogeochemistry. However, the biogeochemical significance of the plastisphere and its mechanisms regulating nitrogen cycling remain elusive. Using 15 N- and 13 C-labelling coupled with metagenomics and metatranscriptomics, here we unveil that the plastisphere likely acts as an underappreciated nitrifying niche in estuarine ecosystems, exhibiting a 0.9 ~ 12-fold higher activity of bacteria-mediated nitrification compared to surrounding seawater and other biofilms (stone, wood and glass biofilms). The shift of active nitrifiers from O 2 -sensitive nitrifiers in the seawater to nitrifiers with versatile metabolisms in the plastisphere, combined with the potential interspecific cooperation of nitrifying substrate exchange observed among the plastisphere nitrifiers, collectively results in the unique nitrifying niche. Our findings highlight the plastisphere as an emerging nitrifying niche in estuarine environment, and deepen the mechanistic understanding of its contribution to marine biogeochemistry., (© 2024. The Author(s).)

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 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.)

Long-term application of green manure (GM) and nitrogen (N) fertilizers markedly improved soil fertility and boosted rice yield in ecologically fragile karst paddy fields. However, the precise response mechanisms of the soil bacterial community to varying amounts of green manure alone and in combination with N fertilizer in such environments remain poorly elucidated. In this study, we investigated the soil bacterial communities, keystone taxa, and their relationship with soil environmental variables across eight fertilization treatments. These treatments included group without N addition (N 0 M 0 , no N fertilizer and no GM; N 0 M 22.5 , 22.5 t/ha GM; N 0 M 45 , 45 t/ha GM, N 0 M 67.5 , 67.5 t/ha GM) and group with N addition (NM 0 , N fertilizer and no GM; NM 22.5 , N fertilizer and 22.5 t/ha GM; NM 45 , N fertilizer and 45 t/ha GM; NM 67.5 , N fertilizer and 67.5 t/ha GM). The results revealed that increasing green manure input significantly boosted rice yield by 15.51-22.08% and 21.84-35% in both the group without and with N addition, respectively, compared to N 0 M 0 treatment. Moreover, with escalating green manure input, soil TN, AN, AK, and AP showed an increasing trend in the group without N addition. However, following the addition of N fertilizer, TN and AN content initially rose, followed by a decline due to the enhanced nutrient availability for rice. Furthermore, the application of a large amount of N fertilizer decreased the C: N ratio in the soil, resulting in significant changes in both the soil microbial community and its function. Particularly noteworthy was the transition of keystone taxa from their original roles as N-fixing and carbon-degrading groups (oligotrophs) to roles in carbon degradation (copiotrophs), nitrification, and denitrification. This shift in soil community and function might serve as a primary factor contributing to enhanced nutrient utilization efficiency in rice, thus significantly promoting rice yield., (© 2024. The Author(s).)