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Comparative Responses of Invasive and Native Plant Species to Combined Cd and Microplastic Pollution.

The co-occurrence of heavy metal contamination and biodegradable microplastic (polylactic acid, PLA) pollution poses increasing risks to terrestrial plant communities and soil functioning, yet species-specific responses to combined stress remain poorly understood. Cd and microplastics frequently co-occur in agricultural soils, where microplastics can alter cadmium mobility, bioavailability, and transport pathways, potentially modifying metal toxicity and plant stress responses compared with single-pollutant exposure. We investigated the responses of the invasive Bidens pilosa and the native Solanum nigrum grown in monoculture and mixed culture under combined cadmium (Cd) and biodegradable microplastic (PLA) stress by integrating plant growth, photosynthetic performance, oxidative physiology, and rhizosphere biochemical processes. Combined Cd-MP exposure markedly reduced plant growth, chlorophyll content (SPAD), photosystem II efficiency (Fv/Fm), nitrogen accumulation, biomass production, and rhizosphere enzyme activities associated with carbon, nitrogen, and phosphorus cycling. However, B. pilosa maintained greater physiological stability under stress, characterized by higher antioxidant enzyme activities (SOD, CAT, POD), lower reactive oxygen species (H2O2, O2˙-) accumulation, and reduced lipid peroxidation (MDA), whereas S. nigrum exhibited stronger oxidative damage and functional impairment. Multivariate analyses further revealed that root antioxidant capacity was closely associated with rhizosphere microbial enzyme activity, suggesting a root-centered regulatory mechanism linking plant stress tolerance to soil functioning. Overall, the invasive species showed greater tolerance to combined contamination and maintained relatively higher rhizosphere functional activity than the native species, indicating that multi-pollutant stress may alter competitive interactions between invasive and native plants in contaminated environments.

Cadmium

Metagenomic analysis of microbial community dynamics in konjac rhizosphere during soft rot disease progression.

Amorphophallus konjac, the sole glucomannan-rich species in the Araceae family, faces significant yield and quality losses due to soft rot disease. Understanding the relationship between soil microbial communities and soft rot incidence is critical for sustainable konjac production. Metagenomic profiling was employed to systematically characterize the spatiotemporal dynamics of rhizosphere microbiomes during disease progression. Microbial alpha diversity (Chao1 index) exhibited a significant peak in the rhizosphere of diseased plants at the mature stage, contrasting with stable diversity patterns in healthy and latently infected groups, indicating dysbiosis-associated richness inflation during disease progression. Principal coordinate analysis (PCoA) revealed significant divergence in rhizosphere microbial structures between diseased and healthy/latently infected groups, with higher compositional variability observed in diseased samples. At the phylum level, Chloroflexi and Acidobacteria abundances in healthy mature plants exceeded those in diseased plants by 11.54% and 4.6%, respectively, while pathogenic Rhizopus arrhizus and Rhizopus microsporus were significantly enriched in diseased mature plants. Correlation analyses demonstrated predominantly negative associations between bacterial species and soil factors, contrasting with positive fungal correlations. KEGG pathway annotation identified carbohydrate metabolism and amino acid synthesis as core microbial functions in the konjac rhizosphere. Collectively, Chloroflexi and Acidobacteria were validated as putative biocontrol agents, while Rhizopus spp. emerged as key drivers of soft rot development. These findings provide mechanistic insights for designing microbiome-based biocontrol strategies to mitigate konjac soft rot, offering a sustainable alternative to conventional agrochemical reliance. KEY POINTS: • Diseased konjac microbial richness peaks; healthy plants enrich Chloroflexi/Acidobacteria. • Rhizopus pathogens drive soft rot; bacteria and fungi show opposing soil factor links. • Lays groundwork for microbiome approaches to cut agrochemicals in konjac rot control.

Rhizosphere

Two Saccharopolyspora isolates from archaeological excavation sites: polyphasic taxonomy, biosynthetic potential, bioactivity profiling and description of Saccharopolyspora antiqui sp. nov.

Archaeological excavation sites represent underexplored microbial habitats with the potential to recover taxonomically and biotechnologically valuable actinomycetes. In this study, two Saccharopolyspora strains, 5N708T and 5N102, were isolated from soil samples collected from the Gaziantep-Doliche-Dülük and Bitlis-Ahlat-Selçuklu Cemetery archaeological excavation sites in Türkiye. A polyphasic taxonomic approach, including 16S rRNA gene sequencing, phylogenetic and phylogenomic analyses, average nucleotide identity, digital DNA-DNA hybridization, phenotypic characterization, and chemotaxonomic analyses, showed that strain 5N708T represents a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora antiqui sp. nov. is proposed, whereas strain 5N102 was assigned to Saccharopolyspora elongata. Both isolates were further evaluated for their antimicrobial, antioxidant, and cytotoxic activities, and their biosynthetic potential was investigated by genome mining. Both strains showed activity against Staphylococcus aureus, with strain 5N708T producing the larger inhibition zone. Strain 5N102 exhibited markedly stronger antioxidant activity than strain 5N708T in radical scavenging, ferric reducing antioxidant power, and reducing power assays. In contrast, strain 5N708T showed more promising cytotoxic activity, with relative selectivity toward MIA PaCa-2 pancreatic cancer cells compared with HEK293 cells after prolonged incubation. Genome mining revealed multiple biosynthetic gene clusters in both isolates, supporting their capacity to produce secondary metabolites. These findings indicate that archaeological soils are promising reservoirs of taxonomically novel and biologically active Saccharopolyspora strains.

Saccharopolyspora

Predictions of rhizosphere microbiome dynamics with a genome-informed and trait-based energy budget model.

Soil microbiomes are highly diverse, and to improve their representation in biogeochemical models, microbial genome data can be leveraged to infer key functional traits. By integrating genome-inferred traits into a theory-based hierarchical framework, emergent behaviour arising from interactions of individual traits can be predicted. Here we combine theory-driven predictions of substrate uptake kinetics with a genome-informed trait-based dynamic energy budget model to predict emergent life-history traits and trade-offs in soil bacteria. When applied to a plant microbiome system, the model accurately predicted distinct substrate-acquisition strategies that aligned with observations, uncovering resource-dependent trade-offs between microbial growth rate and efficiency. For instance, inherently slower-growing microorganisms, favoured by organic acid exudation at later plant growth stages, exhibited enhanced carbon use efficiency (yield) without sacrificing growth rate (power). This insight has implications for retaining plant root-derived carbon in soils and highlights the power of data-driven, trait-based approaches for improving microbial representation in biogeochemical models.

Rhizosphere

Balancing nutrient remobilization and photosynthesis: proteomic insights into the dual role of lupin cotyledons after germination.

Efficient nutrient mobilization from seed storage tissues is essential for seedling establishment, particularly in legumes such as Lupinus albus (white lupin), which thrive in nutrient-poor soils. This study investigates the role of cotyledons in nitrogen (N) and mineral remobilization after germination during their transition from storage organs to photosynthetically active tissues, including the metabolic challenges posed by the coexistence of these two functions in epigeal germination. We cultivated white lupin seedlings under nitrogen-deficient conditions, analyzing cotyledon composition and function over 28 days. Our results indicate that 60% of cotyledon-stored proteins are degraded within the first 8 days, with free amino acids transiently accumulating before being redistributed to support growth. The progressive depletion of cotyledon reserves was accompanied by structural and metabolic changes, including an increase in photosynthetic proteins. However, cotyledon photosynthetic capacity remained lower than that of true leaves, suggesting a transient role in energy metabolism. The loss of cotyledons before day 12 significantly impaired seedling development, emphasizing their critical contribution to nitrogen, phosphate, and micronutrient supply during early growth. Comparative proteomic analysis revealed dynamic shifts in nutrient transport, amino acid metabolism, and stress response pathways following cotyledon removal. These findings underscore the significance of cotyledon nutrient remobilization in legume adaptation to low-fertility soils and highlight potential targets for breeding strategies aimed at improving nutrient use efficiency. By optimizing cotyledon nutrient composition and function, future breeding efforts could enhance seedling vigor, reduce fertilizer dependency, and improve the nutritional value of lupin-based foods.

Lupinus

Isolation and characterization of two novel species Neorhizobium fuzhouense sp. nov. and Neotabrizicola paludis sp. nov.

Two novel aerobic bacterial strains, designated SGZ-38T and sgz301269T, were isolated from the root of Pennisetum sp. and paddy soil, respectively. Strain SGZ-38T grew at 10-40 ℃ (optimum 30 °C) and pH 5.0-12.0 (optimum 6.5) and tolerated up to 1.0% NaCl (w/v), whereas strain sgz301269T grew at 15-37 °C (optimum 30 °C), pH 5.0-9.5 (optimum 7.0) and 0-2% NaCl (optimum 0%). Phylogenetic trees based on the 16S rRNA gene and genomes placed both strains into distinct lineages, forming separated clades from their closest relatives. Strain SGZ-381T exhibited the highest 16S rRNA gene similarities to "Neorhizobium deserti" ACCC 61627T (97.4%), and strain sgz301269T had the highest 16S rRNA gene sequence similarity to Neotabrizicola shimadae N10T (97.6%). The respiratory quinone in both strains was ubiquinone-10. The main fatty acids of SGZ-381T were Summed feature 8, Summed feature 2 and C16:0, whereas strain sgz301269T included C10:0 3OH, C18:0 3OH and Summed feature 8. The DNA G+C content of SGZ-381T and sgz301269T was 62.1% and 65.5%, respectively. The average nucleotide identity and digital DNA-DNA hybridization values between each strain and their respective closest species were 74.6% and 20.1%, 75.3% and 17.4% respectively, below the thresholds for species delineation. Based on the comprehensive chemotaxonomic, phylogenetic, and phenotypic evidence, proposed names of the novel strains are Neorhizobium fuzhouense sp. nov. (type strain SGZ-381T=GDMCC1.4207T=JCM 36770T), Neotabrizicola paludis sp. nov. (type strain sgz301269T=MCCC 1K09178T=KCTC 8856T).

Bacterial Typing Techniques

Proposal of three novel species of the family Xanthobacteraceae: Xanthobacter pollutisoli sp. nov., Xanthobacter luteus sp. nov. and Aquabacter albus sp. nov., isolated from oil-contaminated soils.

Three Gram-stain-negative bacterial strains, KR7-65T, KR7-225T and CN5-332T isolated from oil-contaminated soil in Korea and China were identified. Phylogenetic analysis based on 16S rRNA gene sequences placed the strains within the family Xanthobacteraceae, with KR7-65T and KR7-225T affiliated with the genus Xanthobacter and CN5-332T with the genus Aquabacter. Sequence similarities to type strains of validly published species were below 98.5%. Core genome phylogeny showed that the four strains formed distinct clusters occupying different positions in the phylogenetic tree and exhibited different closest relatives. Average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization (dDDH) values between KR7-65T and KR7-225T and members of Xanthobacter were 78.7-87.0%, 74.8-87.8% and 22.7-32.3%, respectively, whereas those between CN5-332T and members of Aquabacter were 80.2-80.7%, 79.6-80.5% and 23.4-23.9%, supporting their assignment as novel species. The DNA G+C contents were 68.0, 69.9 and 66.5 mol% for KR7-65T, KR7-225T and CN5-332T, respectively. Strains KR7-65T and KR7-225T contained phosphatidylcholine, phosphatidylglycerol, phosphatidyl monomethyl ethanolamine, diphosphatidylglycerol (DPG) and an unidentified glycolipid as major polar lipids, whereas DPG was absent in strain CN5-332T. The primary fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), cyclo C19 : 0ω8c and C16 : 0. On the basis of phylogenetic, genomic and phenotypic evidence, strains KR7-65T and KR7-225T represent two novel species of the genus Xanthobacter, for which the names Xanthobacter pollutisoli sp. nov. (type strain KR7-65T=KACC 23453T=NBRC 116939T) and Xanthobacter luteus sp. nov. (type strain KR7-225T=KACC 23282T=NBRC 116940T) are proposed. Strain CN5-332T represents a novel species of the genus Aquabacter, for which the name Aquabacter albus sp. nov. (type strain CN5-332T=KACC 23276T=CCTCC AB 2024343T) is proposed.

Phylogeny

Neobacillus driksii sp. nov. isolated from a Mars 2020 spacecraft assembly facility and genomic potential for lasso peptide production in Neobacillus.

UNLABELLED: During microbial surveillance of the Mars 2020 spacecraft assembly facility, two novel bacterial strains, potentially capable of producing lasso peptides, were identified. Characterization using a polyphasic taxonomic approach, whole-genome sequencing and phylogenomic analyses revealed a close genetic relationship among two strains from Mars 2020 cleanroom floors (179-C4-2-HS, 179-J1A1-HS), one strain from the Agave plant (AT2.8), and another strain from wheat-associated soil (V4I25). All four strains exhibited high 16S rRNA gene sequence similarity (>99.2%) and low average nucleotide identity (ANI) with Neobacillus niacini NBRC 15566T, delineating new phylogenetic branches within the genus. Detailed molecular analyses, including gyrB (90.2%), ANI (86.4%), average amino acid identity (87.8%) phylogenies, digital DNA-DNA hybridization (32.6%), and percentage of conserved proteins (77.7%) indicated significant divergence from N. niacini NBRC 15566T. Consequently, these strains have been designated Neobacillus driksii sp. nov., with the type strain 179-C4-2-HST (DSM 115941T = NRRL B-65665T). N. driksii grew at 4°C to 45°C, pH range of 6.0 to 9.5, and 0.5% to 5% NaCl. The major cellular fatty acids are iso-C15:0 and anteiso-C15:0. The dominant polar lipids include diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified aminolipid. Metagenomic analysis within NASA cleanrooms revealed that N. driksii is scarce (17 out of 236 samples). Genes encoding the biosynthesis pathway for lasso peptides were identified in all N. driksii strains and are not commonly found in other Neobacillus species, except in 7 out of 26 recognized species. This study highlights the unique metabolic capabilities of N. driksii, underscoring their potential in antimicrobial research and biotechnology. IMPORTANCE: The microbial surveillance of the Mars 2020 assembly cleanroom led to the isolation of novel N. driksii with potential applications in cleanroom environments, such as hospitals, pharmaceuticals, semiconductors, and aeronautical industries. N. driksii genomes were found to possess genes responsible for producing lasso peptides, which are crucial for antimicrobial defense, communication, and enzyme inhibition. Isolation of N. driksii from cleanrooms, Agave plants, and dryland wheat soils, suggested niche-specific ecology and resilience under various environmentally challenging conditions. The discovery of potent antimicrobial agents from novel N. driksii underscores the importance of genome mining and the isolation of rare microorganisms. Bioactive gene clusters potentially producing nicotianamine-like siderophores were found in N. driksii genomes. These siderophores can be used for bioremediation to remove heavy metals from contaminated environments, promote plant growth by aiding iron uptake in agriculture, and treat iron overload conditions in medical applications.

Phylogeny

Genome-based predictions of metabolic preferences and substrate phenotypes in psychrotrophic bacteria from permafrost environments.

Genomes reveal vast functional potential, but harbor genomic noise that obscures prediction of metabolic and environmental preferences. Genomic databases are skewed towards clinically relevant and easily cultivated bacteria, limiting predictions for diverse and underrepresented environmental taxa. Psychrotrophic bacteria, which can survive and grow in cold, nutrient-limited, dry, and saline environments, are especially underrepresented despite their relevance for understanding microbial responses to changing cold environments and potential biotechnological value given growth at low temperatures. Assembling complete genomes of 48 isolates from Alaskan permafrost, seasonally frozen active layer soils, and terrestrial ice, we used Kyoto Encyclopedia of Genes and Genomes (KEGG) ortholog annotations to evaluate the predictability of metabolic resource-use traits observed using phenotypic tests. Genome-predicted values for glycolytic versus gluconeogenic catabolic preference index, or sugar-acid preference (SAP), explained over 50% of the variance in empirically observed SAP. SAP was inversely correlated to genomic GC content, which follows phylum-level trends, indicating that coarse metabolic preference covaries with phylogeny. Regularized elastic net models offered a more granular view, linking KEGG genes to specific substrate utilization and sensitivity phenotypes and yielding moderate but reproducible accuracy (AUC 0.70-0.79) for 11 substrates, demonstrating that specific substrate responses may be predictable from relatively small subsets of KO genes. These results extend recent advances, such as the SAP metric, and highlight associations among genomic GC content, phylum, and broad metabolic strategy. Linking genomic content to phenotype using isolates is a necessary step toward predictive models of microbial function in environmental communities, and this work can be used for hypothesis generation, with applications towards more expansive data sets.IMPORTANCECold region soils and ice host psychrotrophic bacteria with metabolic traits and adaptations that enable persistence in harsh, resource-limited environments. However, these taxa are underrepresented in genomic reference databases dominated by well-studied, mesophilic organisms. This gap limits inference of ecological strategies and our ability to predict how these microbes may influence the large, thaw-vulnerable carbon reservoirs in permafrost. Here, we show that genomic GC content is associated with the sugar-versus-acid catabolic preference (SAP) of isolates across major phyla, suggesting that broad genomic features may provide a coarse signal of metabolic strategy. We demonstrate that a modified SAP metric, using binary (positive/negative) substrate utilization rather than detailed growth rate measurements, is moderately predictive, thus extending its application to slow-growing or difficult-to-culture taxa. Together, these advances broaden the toolkit for linking genome content to resource-use traits (phenotype) in poorly characterized, cold-adapted bacteria and offer a tractable entry point to broad prediction and hypothesis generation.

Genome, Bacterial

Description of two novel Marinobacter species isolated from saline-alkali soil: Marinobacter alkalisoli sp. nov. and Marinobacter shunpengi sp. nov.

Four Gram-staining negative, non-motile, rod-shape bacteria, named strains GN3S48T, HN1S83, LN3S78T, and M1N3S26, were isolated from the bulk saline soils, in Baotou, China. Among them, strains GN3S48T and HN1S83 could degrade 100 mg l-1n-hexadecane as sole carbon and energy source for their growth. Phylogenetic analyses showed that the four strains always formed two distinct clades: Strain LN3S78T clustered with strain M1N3S26, and strain GN3S48T clustered with strain HN1S83. Nonetheless, all four strains tightly clustered and shared the highest 16S rRNA gene similarities with Marinobacter species. Specifically, clade of strains LN3S78T and M1N3S26 cluster with Marinobacter lipolyticus CGMCC 1.7282T, while clade of strains GN3S48T and HN1S83 clustered with Marinobacter zhanjiangensis CCTCC AB 208029T. The ANIb and AAI values between strains GN3S48T and HN1S83 were 96.4% and 94.6%, respectively, while those between strains LN3S78T and M1N3S26 were 99.3% and 99.1%, respectively. All ANI and AAI values between the four strains and their closest relatives were below the 95.0% species delineation threshold. The predominant respiratory quinone of the four strains was Q-9. Based on this polyphasic result, the two clades should be identified as two novel species within the genus Marinobacter. Thus, Marinobacter alkalisoli sp. nov. (type strain GN3S48T = CGMCC 1.62232T = KCTC 8701T = JCM 37359 T) and Marinobacter shunpengi sp. nov. (type strain LN3S78T = CGMCC 1.62233T = KCTC 8702T = JCM 37360T) are proposed. The metagenomic analysis revealed that the two new species are globally distributed in high-salt habitats. In addition, comparative genomic analysis confirmed that alkane-degrading genes are ubiquitous in Marinobacter strains.

Marinobacter

Motile and non-motile Listeria species adopt distinct ecological and evolutionary strategies to achieve broad geographic ranges across soil ecosystems.

Broad geographic ranges often reflect ecological versatility and are associated with lower extinction risk. Motility is a key physiological and ecological trait in bacteria. However, how some motile and non-motile bacteria achieve broad geographic ranges remains poorly understood. Here, we analyzed the genomes of 141 Listeria welshimeri and 90 Listeria booriae isolates systematically obtained from soils, representing widespread motile and non-motile species, respectively. We show that L. welshimeri lacks clear phylogeographic structure, suggesting minimal geographic barriers to dispersal. Its wide distribution is likely associated with enhanced motility and effective host colonization that facilitate wildlife-driven dispersal, particularly by regional-terrestrial birds. This pattern is supported by positive selection on flagellar and chemotaxis genes, strong associations with wildlife movement patterns, and close genomic relatedness between soil and wild bird isolates. In contrast, L. booriae displays clade endemism and a strong distance-decay relationship, suggesting dispersal limitation. Despite lacking a dispersal advantage, L. booriae's wide distribution appears to be linked to genomic flexibility and metabolic versatility that support adaptation to diverse environmental conditions, especially those shaped by iron concentration and precipitation. This is evidenced by its large, open pangenome characterized by abundant and diverse metabolic pathways and broad substrates utilization capacity; pronounced positive selection on genes involved in inorganic ion, amino acid, and coenzyme transport and metabolism; and strong associations between gene richness and abiotic factors as well as bacterial community composition. These findings suggest distinct genomic foundations and ecological and evolutionary mechanisms underlying the success of motile and non-motile cosmopolitan bacteria in soil ecosystems.

Soil Microbiology

Description of two nitrogen-fixing bacteria, Azospirillum mesophilum sp. nov. and Azospirillum terrae sp. nov., isolated from paddy soils.

Two novel aerobic, rod-shaped, motile bacterial strains, designated as sgz302134T and sgz301742T, were isolated from paddy soil in Fujian Province. Strains sgz302134T and sgz301742T shared the highest 16S rRNA gene sequence similarities with the type strains Azospirillum isscasi C340-1T (98.2%) and Azospirillum thiophilum DSM 21654T (97.4%), respectively. The phylogenetic tree based on 16S rRNA gene sequences showed that two strains clustered with members of the genus Azospirillum. Growth of strains sgz302134T and sgz301742T was observed at 10-45 °C, pH 5.0-9.5 and 0-0.5% (w/v) NaCl and 15-37 °C, pH 6.0-9.0 and 0-1.0% (w/v) NaCl, respectively. Strains sgz302134T and sgz301742T contained Q-10 as the main quinone. The main fatty acids (>10%) of both strains were summed feature 2 (C12 : 0 aldehyde), summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and C16 : 0. The genomic DNA G+C content of strains sgz302134T and sgz301742T was 68.4 and 68.3%, respectively. The digital DNA-DNA hybridization and average nucleotide identity values between the two strains and their related reference strains were 27.8 and 87.4% and 22.0 and 84.3%, respectively. Both strains possessed nif genes nifBDEHKN. Based on the above results, these two strains represent two novel species of the genus Azospirillum, for which the names Azospirillum mesophilum sp. nov. and Azospirillum terrae sp. nov. are proposed. The type strains are sgz302134T (=MCCC 1K09520T=KCTC 8840T) and sgz301742T (=MCCC 1K09804T=KCTC 18149T), respectively.

Soil Microbiology

Whole genome-based reclassification of the genus Metabacillus: Proposal for five novel genera, Chryseobacillus gen. nov., Cohnibacillus gen. nov., Salimetabacillus gen. nov., Pantoeobacillus gen. nov., and Lutimetabacillus gen. nov. and the description of one novel bacterial species, Chryseobacillus diguaensis sp. nov. isolated from soil in the Digua reservoir.

Comprehensive phylogenomic and comparative genomic analyses were conducted to clarify the taxonomic boundaries of the genus Metabacillus. Phylogenetic trees reconstructed from a set of single-copy orthologous proteins (SCOPs) revealed that the genus, as currently defined, is polyphyletic. The type species of the genus Metabacillus and its closest relatives formed a consistent clade, herein designated as Metabacillus sensu stricto. The remaining species were grouped into three well-supported clades: Kandeliae, Indicus, and Mangrovi, and two single-taxon lineages: M. arenae and M. lacus. The phylogenomic delineation found in these divergent taxa was corroborated by either inconsistent distribution patterns or the absence of previously defined conserved signature indels (CSIs) specific to Metabacillus. Genomic metrics, including Average Nucleotide Identity (ANI), Average Amino acid Identity (AAI), and digital DNA-DNA hybridization (dDDH) further supported the taxonomic delineation proposed here. The observed genomic divergence was mirrored by phenotypic differences, including variations in GC content ranges. Based on this polyphasic evidence, we propose the reclassification of the genus Metabacillus taxa into five novel genera: Chryseobacillus gen. nov. (encompassing the Kandeliae clade), Cohnibacillus gen. nov. (M. lacus), Salimetabacillus gen. nov. (M. arenae), Pantoeobacillus gen. nov. (Indicus clade), and Lutimetabacillus gen. nov. (Mangrovi clade). The core lineage is retained as Metabacillus sensu stricto, for which an emended description of the genus Metabacillus is also provided. A novel bacterial strain, designated as MAU-250T, was isolated from a soil sample collected on the shore of an artificial reservoir in the Andean foothills of the Maule Region in central Chile. Public metagenome screening supported a low-abundance taxon with broad ecological adaptability, preferentially associated with soil habitats. A polyphasic analysis based on phenotypic traits and genomic distances (78.0% ANIb and 19.8% dDDH against its closest relative) also supported its designation as a novel species, for which the name Chryseobacillus diguaensis sp. nov. is proposed. The type strain is MAU-250T (=RGM 3146T = IMI 507634T).

Phylogeny

Functional identification of the key gene Eh-fadB in nicosulfuron degradation by Enterobacter hormaechei ES1 based on multi-omics and enzymatic characterization.

Nicosulfuron is a sulfonylurea herbicide with residues that pose ecological risks in agricultural soils. Here we elucidated the degradation mechanism of Enterobacter hormaechei ES1 through whole-genome sequencing, transcriptomics, metabolomics, gene knockout, heterologous expression, and soil bioremediation assays. Under nicosulfuron stress, ES1 upregulated antioxidant enzymes including SOD, POD, and CAT, along with glutathione synthesis, to scavenge excess reactive oxygen species. HPLC-TOF-MS identified degradation intermediates such as ADMP and ASDM, indicating initial cleavage of the sulfonylurea bridge. Integrated multi-omics prioritized Eh-fadB, encoding a fatty acid β-oxidation multifunctional enzyme, as a novel degradative gene. Targeted knockout of Eh-fadB reduced nicosulfuron degradation from 87.6% to 37.04%, while genetic complementation restored nearly full activity. Purified Eh-FadB directly converted nicosulfuron, with optimal performance at 30 °C and pH 5-6; its activity was enhanced by Na+ and Pb2+ but inhibited by Fe3+. Molecular docking and dynamics identified His-450 and Asn-427 as key residues for substrate binding. In contaminated soil, inoculation with ES1 reduced nicosulfuron content within 21 days and promoted recovery of dehydrogenase and urease activities. This study provides the first genetic and biochemical evidence that a FadB-type enzyme participates in nicosulfuron catabolism, supporting sulfonylurea bridge cleavage and its potential for soil bioremediation.

Eh-fadB

Endophytic fungi isolated from coffee plants promote Arabidopsis thaliana growth and suppress soil-borne fungal pathogens.

Endophytic beneficial microorganisms are widely used in agriculture for promoting plant growth and enhancing plant defense mechanisms. This study aimed to characterize endophytic fungi isolated from the roots of coffee plants cultivated in organic agroforestry systems and evaluate their potential as biocontrol agents against fungal pathogens, as well as their ability to promote plant growth. Biocontrol activity was assessed using in vitro dual-culture assays on potato dextrose agar, measuring the inhibition of pathogen growth. Plant growth promotion was evaluated by co-cultivating Arabidopsis thaliana seedlings with fungal isolates on Murashige and Skoog medium. Isolates were further subjected to both qualitative and quantitative biochemical characterization. A total of 18 endophytic fungal strains were identified and classified in five genera: Colletotrichum, Fusarium, Simplicillium, Lasiodiplodia and Trichoderma. Among these, ten Trichoderma isolates demonstrated strong antagonistic activity against selected fungal pathogens and significantly enhanced the growth of Arabidopsis seedlings in vitro. These beneficial effects were associated with the production of siderophores and indole-3-acetic acid, as well as the apparent nitrogen availability -- likely mediated through interactions with nitrogen-fixing bacteria.

Arabidopsis

Rhodococcus folensis sp. nov., an orange-red-pigmented bacterium from mining soil.

Mining-impacted environments represent chemically complex ecosystems that may harbor metabolically versatile and pigment-producing microorganisms. During a survey of pigment-producing bacteria from abandoned mining soil in Trabzon, Türkiye, a red-pigmented strain, designated FMA22T, was isolated and characterized using a polyphasic taxonomic approach. 16 S rRNA gene sequence analysis placed the strain within the genus Rhodococcus, showing the highest similarity to R. corynebacterioides DSM 20,151T (99.57%), R. kroppenstedtii DSM 44908ᵀ (99.06%) and R. trifolii T8T (98.96%). The strain was Gram-stain-positive, aerobic and non-motile, and grew at 4-40 °C. Polar lipids included phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, phosphatidylcholine, five unidentified glycolipids, four unidentified lipids, one unidentified phospholipid and one unidentified phosphoglycolipid; MK-8(H2) was the major respiratory quinone. Major fatty acids were C18:1 ω9c, summed feature 3 (C16:1 ω7c/C16:1 ω6c) and C16:0. ANI and dDDH values with the closest relatives were below 76.8% and 20.5%, respectively. The draft genome (4.23 Mb; 67.2 mol% G + C; 4,106 CDSs) harbors a terpene-associated carotenoid cluster containing crtB, crtI and crtY. The orange-red pigment (λmax = 475 nm) showed antioxidant activity (DPPH SC₅₀ = 5.38 mg mL⁻¹; FRAP = 4.34 µmol TE g⁻¹) and weak but measurable HIV-1 reverse transcriptase inhibition (IC₅₀ = 22 mg mL⁻¹). These data support the proposal of Rhodococcus folensis sp. nov., with FMA22ᵀ (= LMG 34144ᵀ = DSM 120048ᵀ) as the type strain.

Soil Microbiology

Whole-Genome Analysis and Growth-Promoting Mechanism of Klebsiella pneumoniae YMK25 from Maize Rhizobacteria.

Plant growth-promoting rhizobacteria (PGPR) are microorganisms that enhance plant growth through various mechanisms. In the context of global agriculture, which faces fertilizer dependency and environmental pollution, developing eco-friendly microbial fertilizers has become crucial for enhancing agricultural sustainability. To identify highly effective PGPR, we isolated 102 bacterial strains from maize rhizosphere soil using the dilution plating method. The strains were screened for growth-promoting abilities using functional media, resulting in the selection of strain YMK25 for its exceptional capabilities in nitrogen fixation, solubilization of inorganic and organic phosphorus, indole-3-acetic acid (IAA) production, and siderophore production. Strain YMK25 produced IAA at a concentration of 80.49 ± 0.68 μg/mL and exhibited a relative siderophore expression level of 43.68%. Morphological analysis, 16S rDNA gene sequence analysis, and whole-genome sequencing confirmed that strain YMK25 is Klebsiella pneumoniae. Whole-genome analysis revealed a total genome length of 5,115,280 bp, a GC content of 57.61%, and it contained 4746 coding genes. Gene annotation results indicated genes involved in siderophore synthesis, phosphatase activity, and other plant growth-promoting functions, which align with the verified characteristics of strain YMK25. Furthermore, this strain exhibited significant metabolic capabilities. The pot experiment demonstrated that strain YMK25 promotes maize plant growth and assists in nutrient fixation in these plants. In conclusion, strain YMK25 is a high-quality PGPR with substantial potential for application in agricultural production, presenting promise for widespread use in sustainable agriculture.

Klebsiella pneumoniae

Untargeted metabolomics reveals anion and organ-specific metabolic responses of salinity tolerance in willow.

Willows can alleviate soil salinisation while generating sustainable feedstock for biorefinery, yet the metabolomic adaptations underlying their tolerance remain poorly understood. Salix miyabeana was treated with two environmentally abundant salts, NaCl and Na2SO4, in a 12-week pot trial. Willows tolerated salts across all treatments (up to 9.1 dS m-1 soil ECe), maintaining biomass while selectively partitioning ions, confining Na+ to roots and accumulating Cl- andin the canopy and adapting to osmotic stress via reduced stomatal conductance. Untargeted metabolomics captured >5000 putative compounds, including 278 core willow metabolome compounds constitutively produced across organs. Across all treatments, salinity drove widespread metabolic reprogramming, altering 28% of the overall metabolome, with organ-tailored strategies. Comparing salt forms at equimolar sodium, shared differentially abundant metabolites were limited to 3% of the metabolome, representing the generalised salinity response, predominantly in roots. Anion-specific metabolomic responses were extensive. NaCl reduced carbohydrates and tricarboxylic acid cycle intermediates, suggesting potential carbon and energy resource pressure, and accumulated root structuring compounds, antioxidant flavonoids, and fatty acids. Na2SO4 salinity triggered accumulation of sulphur-containing larger peptides, suggesting excess sulphate incorporation leverages ion toxicity to produce specialised salt-tolerance-associated metabolites. This high-depth picture of the willow metabolome underscores the importance of capturing plant adaptations to salt stress at organ scale and considering ion-specific contributions to soil salinity.

Salix