Nitrogen Fixation Genes Research Articles

Correction: Isoptericola haloaureus sp. nov., a dimorphic actinobacterium isolated from mangrove sediments of southeast India, implicating biosaline agricultural significance through nitrogen fixation and salt tolerance genes

Correction: Isoptericola haloaureus sp. nov., a dimorphic actinobacterium isolated from mangrove sediments of southeast India, implicating biosaline agricultural significance through nitrogen fixation and salt tolerance genes

Grazing disturbance reduces biocrust-related cyanobacteria and N-fixer abundance but increases bacterial diversity: Implications for biocrust restoration in degraded drylands

Overgrazing is a major driver of dryland degradation, however, so far, there is limited understanding on how this process affects biocrust-related microbial community, and especially how the key groups respond to grazing disturbance. In this study, quantitative polymerase chain reaction (qPCR) and high throughout sequencing technologies were used to investigate the bacterial community abundance and diversity in the Horqin Sandland (China) experiencing different livestock grazing disturbances, in order to examine whether a shift in bacterial community (in particular the key biocrust components, cyanobacteria) was involved, and how this was related to biocrust development and altered soil carbon and nitrogen level. Our results revealed that a clear heterogeneous soil bacterial community was associated with grazing disturbance, which inhibited biocrust development. The decreased photosynthetic cyanobacterial abundance (81.81 % in relative abundance and 98.83 % in absolute abundance) and nitrogen-fixing genes (87.72 %), associated with the lower total carbon and nitrogen content (P<0.05), illustrated a low soil carbon and nitrogen-fixing capability and nutrient level in the grazing-disturbed soils. In particular, a switch of nitrogen-fixing dominance from cyanobacteria to proteobacteria was induced by the grazing disturbance. Compared to the grazing-disturbed soils, higher dominant cyanobacterial operational taxonomic units (OTUs) (especially the species of Nostoc and Scytonema) and lower bacterial diversity (e.g., Shannon, Ace, and Chao index) were observed in the undisturbed biocrust soils, highlighting that the dominants rather than diversity played more important roles in biocrust development. Collectively, our results demonstrate that cyanobacteria are very sensitive (even more than others, e.g., proteobacteria and actinobacteria) to grazing disturbance, therefore, we propose that cyanobacterial inoculation is likely an effective approach to supplement soil cyanobacterial abundance, which is expected to induce biocrust development and increase carbon and nitrogen level in the disturbed drylands.

Genomic insights into key mechanisms for carbon, nitrogen, and phosphate assimilation by the acidophilic, halotolerant genus Acidihalobacter members.

In-depth comparative genomic analysis was conducted to predict carbon, nitrogen, and phosphate assimilation pathways in the halotolerant, acidophilic genus Acidihalobacter. The study primarily aimed to understand how the metabolic capabilities of each species can determine their roles and effects on the microbial ecology of their unique saline and acidic environments, as well as in their potential application to saline water bioleaching systems. All four genomes encoded the genes for the complete tricarboxylic acid cycle, including 2-oxoglutarate dehydrogenase, a key enzyme absent in obligate chemolithotrophic acidophiles. Genes for a unique carboxysome shell protein, csoS1D, typically found in halotolerant bacteria but not in acidophiles, were identified. All genomes contained lactate and malate utilization genes, but only Ac. ferrooxydans DSM 14175T contained genes for the metabolism of propionate. Genes for phosphate assimilation were present, though organized differently across species. Only Ac. prosperus DSM 5130T and Ac. aeolianus DSM 14174T genomes contained nitrogen fixation genes, while Ac. ferrooxydans DSM 14175T and Ac. yilgarnensis DSM 105917T possessed genes for urease transporters and respiratory nitrate reductases, respectively. The findings suggest that all species can fix carbon dioxide but can also potentially utilize exogenous carbon sources and that the non-nitrogen-fixing species rely on alternative nitrogen assimilation mechanisms.

Depth-dependent Effects of Leguminous Crops on Soil Nitrogen-Fixing Microbial Communities Correlated with Carbon Degradation

Abstract Legumes play critical roles in agroecosystems by modulating nitrogen-fixing microorganisms to enhance soil fertility and promote crop productivity. Current research on the effects of legumes predominantly focuses on surface soil, lacking a comprehensive analysis of their overall impact across multiple soil layers and an in-depth understanding of associated microbial mechanisms. Here, the community structure of soil nitrogen-fixing microorganisms in three soil layers (0-20, 20-50, and 50-100 cm) under legume and non-legume cultivation was investigated through metagenomic sequencing. We found that only in topsoil (0-20 cm) legume treatment exhibited a significantly higher relative abundance of nitrogen-fixing genes than non-legume treatment. Under legume cultivation, the relative abundance of nitrogen-fixing genes was significantly higher in the topsoil layer than in deeper layers, whereas non-legume treatment displayed an inverse depth-dependent pattern. Combining soil physicochemical properties, the relative abundance of nitrogen-fixing genes correlated significantly with soil moisture, total carbon (TC), and dissolved organic carbon (DOC) content. Both TC and DOC were identified as key drivers of these genes. Subsequently, a similar depth-dependent pattern within the relative abundance of soil carbon degradation genes was found in response to the cultivation of both crops. The relative abundances of soil carbon degradation genes were negatively correlated with nitrogen-fixing genes under legume treatment individually, distinct from non-legume treatment. Our findings highlight the depth-dependent impact of legumes on nitrogen fixation and the critical interaction between soil carbon degradation and nitrogen fixation, providing insights into carbon management in legume cultivation practices to enhance nitrogen fixation in future agriculture.

SiO2 nanoparticles can enhance nitrogen retention and reduce copper resistance genes during aerobic composting of swine manure

SiO2 nanoparticles (SiO2 NPs) are low-cost, environmentally friendly materials with significant potential to remove pollutants from complex environments. In this study, SiO2 NPs were used for the first time as an additive in aerobic composting to enhance nitrogen retention and reduce the expression of copper resistance genes. The addition of 0.5 g kg−1 SiO2 NPs effectively reduced nitrogen loss by 72.33 % by decreasing denitrification genes (nosZ, nirK, and napA) and increasing nitrogen fixation gene (nifH). The dominant factors affecting nitrification and denitrification genes were Firmicutes and C/N ratio. Additionally, SiO2 NPs decreased copper resistance genes by 28.96 % − 37.52 % in compost products. Copper resistance genes decreased most in the treatment with 0.5 g kg−1 SiO2 NPs. In summary, 0.5 g kg−1 SiO2 NPs have the potential to reduce copper resistance genes and enhance nitrogen retention during aerobic composting, which may be used to improve compost quality.

IAA-producing bacteria from the rhizosphere of chickpea (Cicer arietinum L.): Isolation, characterization, and their effects on plant growth performance

Indole-3-acetic acid (IAA), a crucial plant hormone, regulates diverse physiological processes. This study aimed to isolate and characterize IAA-producing bacteria from the chickpea (Cicer arietinum L.) rhizosphere and evaluate their effects on plant growth. From 54 rhizosphere samples, 118 bacteria (designated as GAC) were isolated and screened for IAA production using a Salkowski colorimetric assay, and Bergey's manual was used for biochemical identification. Isolates were grown under various conditions and in vitro screened for their growth promotion traits. A PCR investigation was performed for IAA and nitrogen-fixing genes, and evaluated for greenhouse conditions. Among them, 27 isolates produced IAA, with eight high producers selected. Morphological and biochemical identification classified the six isolates as Pseudomonas and the other two as Bacillus. Optimal conditions for IAA production were observed at 500 μg/ml tryptophan, 35 °C, and pH 7.0. A 48-h incubation was ideal for IAA production, except for GAC-34 and GAC-73, which required 72 h. All the isolates achieved optimal IAA levels with tryptone and sucrose as nitrogen and carbon sources, respectively. Moreover, all isolates showed nitrogen fixation ability, and the six isolates exhibited phosphate solubilization. PCR confirmed the amplification of nifH (300 bp), nifK (360 bp), and ipdC (1170 bp) genes. Greenhouse experiments demonstrated that eight selected isolates significantly enhanced chickpea growth parameters (p < 0.001). These findings suggest that these IAA-producing bacteria have the potential to be used as biofertilizers to improve crop productivity, although further molecular identification and field studies are required.

Responses of soil microbial metabolism, function and soil quality to long-term addition of organic materials with different carbon sources

Biochar and green manure have been widely applied in agricultural production and are important means to achieve sustainable agriculture. However, there is limited research systematically and comprehensively exploring the response of soil microbiota and the changes in soil metabolomics after the addition of two different carbon source amendments to the soil, and the differential mechanisms of soil metabolomics between them remain unclear. In this study, a long-term field experiment (initiated in 2019) was conducted to investigate the effects of biochar and green manure application on soil nutrients and soil functions driven by soil microbes. Compared to the pure fertilizer treatment, biochar increased soil total carbon by 14.54% to 27.04% and soil available potassium by 4.67% to 27.46%. Ryegrass significantly increased soil available phosphorus and organic matter. Under different fertilization regimes, the ecological niches of soil microbes changed significantly. Network analysis revealed that long-term ryegrass returning reduced the complexity of soil microbial networks. Ryegrass and biochar increased dispersal limitation in fungal assemblages (reaching 93.33% and 86.67%, respectively), with biochar particularly enhancing variable selection in bacterial assemblages (accounting for 53.33%). Variation partitioning analysis based on redundancy analysis indicated that humic substances had the highest explanatory power for microbial community variation, with humic substances explaining 38.49% of bacteria and 52.19% of fungi variation. The ryegrass treatment mainly changed the abundance of carbohydrates (CH), amines (AM), c (AH), and lipids (LP), while the BC treatment mainly altered the abundance of organic acids (AC), amines (AM), and carbohydrates (CH). Meanwhile, both treatments significantly reduced the bisphenol A, one of the soil pollutants. Ryegrass incorporation significantly increased the abundance of genes related to soil C, N, P, and S cycling, especially genes involved in carbon decomposition, while biochar significantly enhanced the abundance of nitrogen fixation genes nifH and Hao in soil. Random forest model results indicated that carbohydrates, alcohols, aromatics (AR), and ester (ES) were the main categories of metabolites in soil influenced by differential microbes, and Finegoldia served as a common important metabolic driving species. In summary, this study reveals the processes of soil function, microbial community succession, and metabolism driven by ryegrass and biochar, providing important insights for optimizing soil management and improving soil quality.Graphical

Microbiome of Supraglacial Systems on the Aldegonda and Bertel Glaciers (Western Spitsbergen Island)

Microbial biomass, diversity of cultivated bacteria and micromycetes, as well as the number of functional nitrogen cycle genes in the supraglacial systems of the Aldegonde and Bertel glaciers were studied. Biomass of microorganisms varied from 2.54 to 722 µg/g of substrate. It has been shown for the first time that the majority (78.7–99.8%) of the microbial biomass of supraglacial objects is represented by fungi rather than prokaryotes. Main part (from 70 to 90%) of the fungal biomass was mycelium, the length of which varied from 6.70 to 537.51 m/g of substrate. The number of prokaryotes varied from 2.4 × 108 to 1.95 × 109 cells/g of substrate. The length of actinomycete mycelium varied from 2.6 to 62.61 m/g of substrate. The abundance of cultivated bacteria and actinomycetes varied from 3.3 × 104 to 1.2 × 106 CFU/g of substrate, and that of micromycetes varied from 2.2 × 101 to 1.7 × 104 CFU/g of substrate. Bacteria of the genera Arthrobacter, Bacillus, Rhodococcus, and Streptomyces, as well as micromycetes of the genera Antarctomyces, Cadophora, Hyphozyma, Teberdinia and Thelebolus dominated. Micromycetes Antarctomyces psychrotrophicus, Hyphozyma variabilis and Teberdinia hygrophila were found in Svalbard for the first time. The number of amoA genes in ammonium-oxidizing bacteria varied from 5.33×106 to 4.86 × 109; nitrogen fixation genes nifH, from 9.89 × 107 to 9.81 × 1010; nirK denitrification genes, from 4.82 × 107 to 3.34 × 1010 gene copies/g of substrate. The results obtained indirectly indicate the leading role of fungi in the microbiome of the supraglacial objects of Svalbard and the significant contribution of prokaryotes to the emission of greenhouse gases from them.

Enhancing bioremediation of petroleum-contaminated soil by sophorolipids-modified biochar: Combined metagenomic and metabolomic analyses

In this study, sophorolipids (SLs)-modified biochar (BC-SLs) was used to enhance the bioremediation of petroleum hydrocarbons (PHs) contaminated soil. The biodegradation rate of petroleum hydrocarbons (PHs) by BC-SLs and BC treatments were 62.86 % and 52.64 % after 60 days of remediation experiments, respectively, higher than non-biochar treatment group (24.09 %). The metagenomic analysis showed that the abundance of petroleum-degrading bacteria Actinobacteria and Proteobacteria were increased by 3.8 % and 5.3 %, respectively in BC-SLs treatment, and the abundance of functional genes for PHs degradation, such as alkB, nidA and pcaG, were significantly increased by 12.85 %, 30.08 % and 21.01 %, respectively. The metabolomic analysis showed that BC-SLs facilitated the metabolic process of PHs, the microbial metabolism of petroleum hydrocarbons (PHs) became more active. Fatty acid degradation and polycyclic aromatic hydrocarbons (PAHs) degradation were up-regulated, indicating the promoting effect of the BC-SLs for PHs metabolism. The combined metagenomic and metabolomic analysis demonstrated the strong positive correlations between PHs metabolites and PHs-degrading bacteria, such as lauric acid vs. Actinobacteria, benzoic vs. Proteobacteria. The strong positive correlations between PHs metabolites and PHs-degrading genes were also observed, such as o-ehyltoluene vs. nahD, 4-isopropylbenzoic acid vs. etbAa. The modification of biochar with SLs increased the oxygen-containing functional groups on the surface of biochar. Meanwhile, the emulsification and solubilization of SLs promoted the bioavailability of PHs. The effects of BC-SLs on the nitrogen cycle during PHs remediation showed that it facilitated the accumulation of nitrogen-fixing genes, promoted nitrification but inhibited denitrification process. This study confirms that the application of BC-SLs is an effective remediation of PHs contamination and a sustainable method for controlling agricultural waste resources.

Repercussions of Prolonged Pesticide Use on Natural Soil Microbiome Dynamics Using Metagenomics Approach.

The residual pesticides in soil can affect the natural microbiome composition and genetic profile that drive nutrient cycling and soil fertility. In the present study, metagenomic approach was leveraged to determine modulations in nutrient cycling and microbial composition along with connected nexus of pesticide, antibiotic, and heavy metal resistance in selected crop and fallow soils having history of consistent pesticide applications. GC-MS analysis estimated residuals of chlorpyrifos, hexachlorbenzene, and dieldrin showing persistent nature of pesticides that pose selective pressure for microbial adaptation. Taxonomic profiling showed increased abundance of pesticide degrading Streptomyces, Xanthomonas, Cupriavidus, and Pseudomonas across the selected soils. Genes encoding for pesticide degrading cytochrome p450, organophosphorus hydrolase, aldehyde dehydrogenase, and oxidase were predominant and positively correlated with Bacillus, Sphingobium, and Burkholderia. Nitrogen-fixing genes (nifH, narB, and nir) were relatively less abundant in crop soils, correlating to the decrease in nitrogen-fixing bacteria (Anabaena, Pantoea, and Azotobacter). Microbial enzymes involved in carbon (pfkA, gap, pgi, and tpiA) and phosphorus cycle (gmbh and phnJ) were significantly higher in crop soils indicating extensive utilization of pesticide residuals as a nutrient source by the indigenous soil microbiota. Additionally, presence of antibiotic and heavy metal resistance genes suggested potential cross-resistance under pressure from pesticide residues. The results implied selective increase in pesticide degrading microbes with decrease in beneficial bacteria that resulted in reduced soil health and fertility. The assessment of agricultural soil microbial profile will provide a framework to develop sustainable agriculture practices to conserve soil health and fertility.

Metagenomics Analysis of the Impact of Protein-Degrading Functional Microbial Agents on Composting of Chicken Manure from Cereal Hulls

In this study, four highly efficient protein-degrading bacteria (Siccibactercolletis, Bacillus thuringiensis, Bacillus cereus, and Bacillus sp. (in: Firmicutes)) were screened from soil and fermentation beds and prepared into a mixed microbial agent in a ratio of 1:1:1:1. The effects of inoculation with protein-degrading functional bacteria on nitrogen transformation rate, microbial community, and functional genes during chicken manure–rice husk composting were studied. With the addition of functional agents, the nitrogen loss in chicken manure composting was reduced to 17.05%, and ammonia emissions were also reduced. Firmicutes, Proteobacteria, Bacteroidetes, Cocci, and Actinobacteria became the dominant bacterial communities, accounting for 85.41%~98.52% of the overall bacterial community in the compost; it promoted the growth of microorganisms such as Pseudogracilibacillus and Lachnospiraceae in the compost. Metagenomic analysis revealed that the addition of functional bacterial agents enhanced the expression of nitrogen fixation genes (nifK, nifH, and glnA) during the high-temperature phase, increased the diversity of bacteria associated with the nitrogen cycle in the compost, and improved the absorption and fixation of nitrogen source elements by microorganisms. Additionally, it strengthened the correlation between microbial communities, the composting environment, and functional genes. This study provides a theoretical basis for the efficient application of microbial agents and the reduction of pollution in chicken manure hull composting.

Improvement of the nitrogenase activity in Escherichia coli that expresses the nitrogen fixation-related genes from Azotobacter vinelandii

The transfer of nitrogen fixation (nif) genes from diazotrophs to non-diazotrophic hosts is of increasing interest for engineering biological nitrogen fixation. A recombinant Escherichia coli strain expressing Azotobacter vinelandii 18 nif genes (nifHDKBUSVQENXYWZMF, nifiscA, and nafU) were previously constructed and showed nitrogenase activity. In the present study, we constructed several E. coli strain derivatives in which all or some of the 18 nif genes were additionally integrated into the fliK locus of the chromosome in various combinations. E. coli derivatives with the chromosomal integration of nifiscA, nifU, and nifS, which are involved in the biosynthesis of the [4Fe–4S] cluster of dinitrogenase reductase, exhibited enhanced nitrogenase activity. We also revealed that overexpression of E. coli fldA and ydbK, which encode flavodoxin and flavodoxin-reducing enzyme, respectively, enhanced nitrogenase activity, likely by facilitating electron transfer to dinitrogenase reductase. The additional expression of nifM, putatively involved in maturation of dinitrogenase reductase, further enhanced nitrogenase activity and the amount of soluble NifH. By combining these factors, we successfully improved nitrogenase activity 10-fold.

Exploring the potential of biochar for the remediation of microbial communities and element cycling in microplastic-contaminated soil

The detrimental effects of microplastics (MPs) on soil microbial and elemental raise significant environmental concerns. The potential of remediation with biochar to mitigate these negative impacts remains an open question. The remediation effects of biochar derived from corn and cotton straw on MPs concerning soil microorganisms and element cycling were investigated. Specifically, biochar induced substantial remediations in microbial community structure following MP exposure, restoring and fortifying the symbiotic network while exerting dominance over microbial community changes. A combined treatment of biochar and MPs exhibited a noteworthy increase in the abundance of NH4+, NO3−, and available phosphorous by 0.46–2.1 times, reversing the declining trend of dissolved organic carbon, showing a remarkable increase by 0.36 times. This combined treatment also led to a reduction in the abundance of the nitrogen fixation gene nifH by 0.46 times, while significantly increasing the expression of nitrification genes (amoA and amoB) and denitrification genes (nirS and nirK) by 22.5 times and 1.7 times, respectively. Additionally, the carbon cycle cbbLG gene showed a 2.3-fold increase, and the phosphorus cycle gene phoD increased by 0.1-fold. The mixed treatment enriched element-cycling microorganisms by 4.8–9.6 times. In summary, the addition of biochar repaired the negative effects of MPs in terms of microbial community dynamics, element content, gene expression, and functional microbiota. These findings underscore the crucial role of biochar in alleviating the adverse effects of MPs on microbial communities and elemental cycling, providing valuable insights into sustainable environmental remediation strategies.

Nodulating Aeschynomene indica without Nod Factor Synthesis Genes: In Silico Analysis of Evolutionary Relationship

Aeschynomene indica rhizobia (AIRs) are special classes of bacteria capable of nodulating without nodulation factors and have photosynthetic capacity. With an aim to characterize the structural variations in Bradyrhizobium genomes during its evolution, the genomes of AIRs and the reference Bradyrhizobium strains were compared utilizing molecular biology, bioinformatics, and biochemistry techniques. The presence of symbiotic nitrogen fixation (nif) genes and photosynthetic genes, as well as components of the T3SS (Type III secretion system) and T3CP (Type III chaperone) in the genome of AIRs, was also assessed. Additionally, the origin, evolutionary history, and genes associated with the NF-independent nodulation pattern in AIRs were explored. The results indicate that horizontal gene transfer events have occurred in AIRs, and three distinct origins of AIRs were estimated: early differentiated AIRs, non-symbiotic Bradyrhizobium, and non-AIRs. In contrast to the significant genetic transformations observed in the second and third groups, the first group of AIRs displays a rich evolutionary history, exhibits high species diversity, and primarily relies on vertical transmission of nitrogen fixation and photosynthetic genes. Overall, the findings provide a fundamental theoretical foundation for gaining a deeper understanding of the phylogeny and genealogy of AIRs.

Isoptericola haloaureus sp. nov., a dimorphic actinobacterium isolated from mangrove sediments of southeast India, implicating biosaline agricultural significance through nitrogen fixation and salt tolerance genes.

Strain MP-1014T, an obligate halophilic actinobacterium, was isolated from the mangrove soil of Thandavarayancholanganpettai, Tamil Nadu, India. A polyphasic approach was utilized to explore its phylogenetic position completely. The isolate was Gram-positive, filamentous, non-motile, and coccoid in older cultures. Ideal growth conditions were seen at 30°C and pH 7.0, with 5% NaCl (W/V), and the DNA G + C content was 73.3%. The phylogenic analysis of this strain based upon 16S rRNA gene sequence revealed 97-99.8% similarity to the recognized species of the genus Isoptericola. Strain MP-1014T exhibits the highest similarity to I. sediminis JC619T (99.7%), I. chiayiensis KCTC19740T (98.9%), and subsequently to I. halotolerans KCTC19646T (98.6%), when compared with other members within the Isoptericola genus (< 98%). ANI scores of strain MP-1014T are 86.4%, 84.2%, and 81.5% and dDDH values are 59.7%, 53.6%, and 34.8% with I. sediminis JC619T, I. chiayiensis KCTC19740T and I. halotolerans KCTC19646T respectively. The major polar lipids of the strain MP-1014T were phosphatidylinositol, phosphatidylglycerol, diphosphotidylglycerol, two unknown phospholipids, and glycolipids. The predominant respiratory menaquinones were MK9 (H4) and MK9 (H2). The major fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C14:0, C15:0, and C16:0. Also, initial genome analysis of the organism suggests it as a biostimulant for enhancing agriculture in saline environments. Based on phenotypic and genetic distinctiveness, the strain MP-1014 T represents the novel species of the genus Isoptericola assigned Isoptericola haloaureus sp. nov., is addressed by the strain MP-1014 T, given its phenotypic, phylogenetic, and hereditary uniqueness. The type strain is MP-1014T [(NCBI = OP672482.1 = GCA_036689775.1) ATCC = BAA 2646T; DSMZ = 29325T; MTCC = 13246T].

Employing Genomic Tools to Explore the Molecular Mechanisms behind the Enhancement of Plant Growth and Stress Resilience Facilitated by a Burkholderia Rhizobacterial Strain.

The rhizobacterial strain BJ3 showed 16S rDNA sequence similarity to species within the Burkholderia genus. Its complete genome sequence revealed a 97% match with Burkholderia contaminans and uncovered gene clusters essential for plant-growth-promoting traits (PGPTs). These clusters include genes responsible for producing indole acetic acid (IAA), osmolytes, non-ribosomal peptides (NRPS), volatile organic compounds (VOCs), siderophores, lipopolysaccharides, hydrolytic enzymes, and spermidine. Additionally, the genome contains genes for nitrogen fixation and phosphate solubilization, as well as a gene encoding 1-aminocyclopropane-1-carboxylate (ACC) deaminase. The treatment with BJ3 enhanced root architecture, boosted vegetative growth, and accelerated early flowering in Arabidopsis. Treated seedlings also showed increased lignin production and antioxidant capabilities, as well as notably increased tolerance to water deficit and high salinity. An RNA-seq transcriptome analysis indicated that BJ3 treatment significantly activated genes related to immunity induction, hormone signaling, and vegetative growth. It specifically activated genes involved in the production of auxin, ethylene, and salicylic acid (SA), as well as genes involved in the synthesis of defense compounds like glucosinolates, camalexin, and terpenoids. The expression of AP2/ERF transcription factors was markedly increased. These findings highlight BJ3's potential to produce various bioactive metabolites and its ability to activate auxin, ethylene, and SA signaling in Arabidopsis, positioning it as a new Burkholderia strain that could significantly improve plant growth, stress resilience, and immune function.

Construction of high-quality genomes and gene catalogue for culturable microbes of sugarcane (Saccharum spp.)

Microbes living inside or around sugarcane (Saccharum spp.) are crucial for their resistance to abiotic and biotic stress, growth, and development. Sequences of microbial genomes and genes are helpful to understand the function of these microbes. However, there is currently a lack of such knowledge in sugarcane. Here, we combined Nanopore and Illumina sequencing technologies to successfully construct the first high-quality metagenome-assembled genomes (MAGs) and gene catalogues of sugarcane culturable microbes (GCSCMs), which contained 175 species-level genome bins (SGBs), and 7,771,501 non-redundant genes. The SGBs included 79 novel culturable bacteria genomes, and 3 bacterial genomes with nitrogen-fixing gene clusters. Four single scaffold near-complete circular MAGs (cMAGs) with 0% contamination were obtained from Nanopore sequencing data. In conclusion, we have filled a research gap in the genomes and gene catalogues of culturable microbes of sugarcane, providing a vital data resource for further understanding the genetic basis and functions of these microbes. In addition, our methodology and results can provide guidance and reference for other plant microbial genome and gene catalogue studies.

Timely symbiosis: circadian control of legume-rhizobia symbiosis.

Legumes house nitrogen-fixing endosymbiotic rhizobia in specialised polyploid cells within root nodules. This results in a mutualistic relationship whereby the plant host receives fixed nitrogen from the bacteria in exchange for dicarboxylic acids. This plant-microbe interaction requires the regulation of multiple metabolic and physiological processes in both the host and symbiont in order to achieve highly efficient symbiosis. Recent studies have showed that the success of symbiosis is influenced by the circadian clock of the plant host. Medicago and soybean plants with altered clock mechanisms showed compromised nodulation and reduced plant growth. Furthermore, transcriptomic analyses revealed that multiple genes with key roles in recruitment of rhizobia to plant roots, infection and nodule development were under circadian control, suggesting that appropriate timing of expression of these genes may be important for nodulation. There is also evidence for rhythmic gene expression of key nitrogen fixation genes in the rhizobium symbiont, and temporal coordination between nitrogen fixation in the bacterial symbiont and nitrogen assimilation in the plant host may be important for successful symbiosis. Understanding of how circadian regulation impacts on nodule establishment and function will identify key plant-rhizobial connections and regulators that could be targeted to increase the efficiency of this relationship.

Deciphering the Nitrogen Fixation Gene Cluster in Vibrio natriegens: A Study on Optimized Expression and Application of Nitrogenase.

Microbial nitrogen fixation presents a viable alternative to chemical fertilizers, yet the limited colonization and specificity of naturally occurring nitrogen-fixing microorganisms present significant challenges to their widespread application. In this study, we identified a nitrogen fixation gene cluster (VNnif) in Vibrio natriegens (VN) and tested its nitrogenase activity through the acetylene reduction assay. We investigated the potential utilization of nitrogenase by incorporating the nitrogenase gene cluster from VN into plant growth-promoting rhizosphere bacteria Pseudomonas protegens CHA0 and enhancing its activity to 48.16 nmol C2H2/mg/h through promoter replacement and cluster rearrangement. The engineered strain CHA0-PVNnif was found to positively impact the growth of Arabidopsis thaliana col-0 and Triticum aestivum L. (wheat). This study expanded the role of plant growth-promoting rhizobacteria (PGPR) and provided a research foundation for enhancing nitrogenase activity.

Insights into the distribution characteristic of microbiota in alkaline–sandy soil with inorganic and organic fertiliser treatments: contrasting abundance, structure and functionality

ABSTRACT Fertiliser amendment is the most direct method for improving soil properties of ecologically vulnerable areas (e.g. desertification soil area), and then, understanding the response of microbial community activities and growth to different fertilisation treatments is of great significance in sandy soil restoration. Here, a field experiment with different fertiliser treatments in typical alkaline–sandy soil was conducted. In this study, the difference of physicochemical property, microbial communities and functions in different treatments of no fertiliser (CK), (ii) fly ash chemical fertilisers (CF), (iii) humic straw organic fertiliser (OF), (iv) functional microbial fertiliser (FMF), (v) sulphate-reducing bacterial (SRB) fertiliser (MF) and (vi) humic straw organic fertiliser + functional microbial fertiliser + SRB fertiliser (CMF) were evaluated. The result showed that the contents of soil total organic carbon, NO3-N, P and K in the FMF were highest. The abundance of keystone taxa (Gemmatimonadetes and Thaumarchaeota) increased significantly in the treatments of microbial fertiliser. Microbial fertiliser boosted genes for aerobic nitrite oxidation, nitrogen fixation, ammonia oxidation and aromatic compound degradation. The number of methylotrophy and dark sulphur oxidation genes was reduced. The bbove results indicated that the treatments with the inoculation of functional microbes are better than the others for the restoration of alkaline–sandy soil.