Superiority of native soil core microbiomes in supporting plant growth.

Native core microbiomes represent a unique opportunity to support food provision and plant-based industries. Yet, these microbiomes are often neglected when developing synthetic communities (SynComs) to support plant health and growth. Here, we study the contribution of native core, native non-core and non-native microorganisms to support plant production. We construct four alternative SynComs based on the excellent growth promoting ability of individual stain and paired non-antagonistic action. One of microbiome based SynCom (SC2) shows a high niche breadth and low average variation degree in-vitro interaction. The promoting-growth effect of SC2 can be transferred to non-sterile environment, attributing to the colonization of native core microorganisms and the improvement of rhizosphere promoting-growth function including nitrogen fixation, IAA production, and dissolved phosphorus. Further, microbial fertilizer based on SC2 and composite carrier (rapeseed cake fertilizer + rice husk carbon) increase the net biomass of plant by 129%. Our results highlight the fundamental importance of native core microorganisms to boost plant production.

Synergistic interactions in core microbiome Rhizobiales accelerate 1,4-dioxane biodegradation.

Next-generation sequencing (NGS) has revolutionized taxa identification within contaminant-degrading communities. However, uncovering a core degrading microbiome in diverse polluted environments and understanding its associated microbial interactions remains challenging. In this study, we isolated two distinct microbial consortia, namely MA-S and Cl-G, from separate environmental samples using 1,4-dioxane as a target pollutant. Both consortia exhibited a persistent prevalence of the phylum Proteobacteria, especially within the order Rhizobiales. Extensive analysis confirmed that Rhizobiales as the dominant microbial population (> 90 %) across successive degradation cycles, constituting the core degrading microbiome. Co-occurrence network analysis highlighted synergistic interactions within Rhizobiales, especially within the Shinella and Xanthobacter genera, facilitating efficient 1,4-dioxane degradation. The enrichment of Rhizobiales correlated with an increased abundance of essential genes such as PobA, HpaB, ADH, and ALDH. Shinella yambaruensis emerged as a key degrader in both consortia, identified through whole-genome sequencing and RNA-seq analysis, revealing genes implicated in 1,4-dioxane degradation pathways, such as PobA and HpaB. Direct and indirect co-cultivation experiments confirmed synergistic interaction between Shinella sp. and Xanthobacter sp., enhancing the degradation of 1,4-dioxane within the core microbiome Rhizobiales. Our findings advocate for integrating the core microbiome concept into engineered consortia to optimize 1,4-dioxane bioremediation strategies.

Calmodulin-like protein MdCML15 interacts with MdBT2 to modulate iron homeostasis in apple.

BTB and TAZ domain proteins (BTs) function as specialized adaptors facilitating substrate recognition of the CUL3-RING ubiquitin ligase (CRL3) complex that targets proteins for ubiquitination in reaction to diverse pressures. Nonetheless, knowledge of the molecular mechanisms by which the apple scaffold protein MdBT2 responds to external and internal signals is limited. Here we demonstrate that a putative Ca 2+ sensor, calmodulin-like 15 (MdCML15), acts as an upstream regulator of MdBT2 to negatively modulate its functions in plasma membrane H+-ATPase regulation and iron deficiency tolerance. MdCML15 was identified to be substantially linked to MdBT2, and to result in the ubiquitination and degradation of the MdBT2 target protein MdbHLH104. Consequently, MdCML15 repressed the MdbHLH104 target, MdAHA8's expression, reducing levels of a specific membrane H+-ATPase. Finally, the phenotype of transgenic apple plantlets and calli demonstrated that MdCML15 modulates membrane H+-ATPase-produced rhizosphere pH lowering alongside iron homeostasis through an MdCML15-MdBT2-MdbHLH104-MdAHA8 pathway. Our results provide new insights into the relationship between Ca2+ signaling and iron homeostasis.

A wild melon reference genome provides novel insights into the domestication of a key gene responsible for melon fruit acidity.

KEY MESSAGE: A wild melon reference genome elucidates the genomic basis of fruit acidity domestication. Structural variants (SVs) have been reported to impose major effects on agronomic traits, representing a significant contributor to crop domestication. However, the landscape of SVs between wild and cultivated melons is elusive and how SVs have contributed to melon domestication remains largely unexplored. Here, we report a 379-Mb chromosome-scale genome of a wild progenitor melon accession "P84", with a contig N50 of 14.9 Mb. Genome comparison identifies 10,589 SVs between P84 and four cultivated melons with 6937 not characterized in previously analysis of 25 melon genome sequences. Furthermore, the population-scale genotyping of these SVs was determined in 1175 accessions, and 18 GWAS signals including fruit acidity, fruit length, fruit weight, fruit color and sex determination were detected. Based on these genotyped SVs, we identified 3317 highly diverged SVs between wild and cultivated melons, which could be the potential SVs associated with domestication-related traits. Furthermore, we identify novel SVs affecting fruit acidity and proposed the diverged evolutionary trajectories of CmPH, a key regulator of melon fruit acidity, during domestication and selection of different populations. These results will offer valuable resources for genomic studies and genetic improvement in melon.

Fe-ZSM-5 zeolite catalyst for heterogeneous Fenton oxidation of 1,4-dioxane: effect of Si/Al ratios and contributions of reactive oxygen species.

Heterogeneous Fenton oxidation using traditional catalysts with H2O2 for the degradation of 1,4-dioxane (1,4-DX) still presents challenge. In this study, we explored the potential of Fe-ZSM-5 zeolites (Fe-zeolite) with three Si/Al ratios (25, 100, 300) as heterogeneous Fenton catalysts for the removal of 1,4-DX from aqueous solution. Fe2O3 or ZSM-5 alone provided ineffective in degrading 1,4-DX when combined with H2O2. However, the efficient removal of 1,4-DX using H2O2 was observed when Fe2O3 was loaded on ZSM-5. Notably, the Brønsted acid sites of Fe-zeolite played a crucial role during the degradation of 1,4-DX. Fe-zeolites, in combination with H2O2, effectively removed 1,4-DX via a combination of adsorption and oxidation. Initially, Fe-zeolites demonstrated excellent affinity for 1,4-DX, achieving adsorption equilibrium rapidly in about 10 min, followed by effective catalytic oxidative degradation. Among the Fe-ZSM-5 catalysts, Fe-ZSM-5 (25) exhibited the highest catalytic activity and degraded 1,4-DX the fastest. We identified hydroxyl radicals (·OH) and singlet oxygen (1O2) as the primary reactive oxygen species (ROS) responsible for 1,4-DX degradation, with superoxide anions (HO2·/O2·-) mainly converting into 1O2 and ·OH. The degradation primarily occurred at the Fe-zeolite interface, with the degradation rate constants proportional to the amount of Brønsted acid sites on the Fe-zeolite. Fe-zeolites were effective over a wide working pH range, with alkaline pH conditions favoring 1,4-DX degradation. Overall, our study provides valuable insights into the selection of suitable catalysts for effective removal of 1,4-DX using a heterogeneous Fenton technology.

Self-assembling a 1,4-dioxane-degrading consortium and identifying the key role of Shinella sp. through dilution-to-extinction and reculturing.

IMPORTANCE: Assembling a functional microbial consortium and identifying key degraders involved in the degradation of 1,4-dioxane are crucial for the design of synergistic consortia used in enhancing the bioremediation of 1,4-dioxane-contaminated sites. However, due to the vast diversity of microbes, assembling a functional consortium and identifying novel degraders through a simple method remain a challenge. In this study, we reassembled 1,4-dioxane-degrading microbial consortia using a simple and easy-to-operate method by combining dilution-to-extinction and reculture techniques. We combined differential analysis of community structure and metabolic function and confirmed that Shinella species have a stronger 1,4-dioxane degradation ability than Xanthobacter species in the enriched consortium. In addition, a new dioxane-degrading bacterium was isolated, Shinella yambaruensis, which verified our findings. These results demonstrate that DTE and reculture techniques can be used beyond diversity reduction to assemble functional microbial communities, particularly to identify key degraders in contaminant-degrading consortia.

Acidification suppresses the natural capacity of soil microbiome to fight pathogenic Fusarium infections.

Soil-borne pathogens pose a major threat to food production worldwide, particularly under global change and with growing populations. Yet, we still know very little about how the soil microbiome regulates the abundance of soil pathogens and their impact on plant health. Here we combined field surveys with experiments to investigate the relationships of soil properties and the structure and function of the soil microbiome with contrasting plant health outcomes. We find that soil acidification largely impacts bacterial communities and reduces the capacity of soils to combat fungal pathogens. In vitro assays with microbiomes from acidified soils further highlight a declined ability to suppress Fusarium, a globally important plant pathogen. Similarly, when we inoculate healthy plants with an acidified soil microbiome, we show a greatly reduced capacity to prevent pathogen invasion. Finally, metagenome sequencing of the soil microbiome and untargeted metabolomics reveals a down regulation of genes associated with the synthesis of sulfur compounds and reduction of key traits related to sulfur metabolism in acidic soils. Our findings suggest that changes in the soil microbiome and disruption of specific microbial processes induced by soil acidification can play a critical role for plant health.

Root Exudates of Ginger Induced by Ralstonia solanacearum Infection Could Inhibit Bacterial Wilt.

Bacterial wilt caused by Ralstonia solanacearum (Rs) is one of the most important diseases found in ginger; however, the disease resistance mechanisms dependent on root bacteria and exudates are unclear. In the present study, we analyzed the changes in the composition of rhizobacteria, endobacteria, and root exudates during the pathogenesis of bacterial wilt using high-throughput sequencing and gas chromatography-mass spectrometry (GC-MS). Rs caused bacterial wilt in ginger with an incidence of 50.00% and changed the bacterial community composition in both endosphere and rhizosphere. It significantly reduced bacterial α-diversity but increased the abundance of beneficial and stress-tolerant bacteria, such as Lysobacter, Ramlibacter, Pseudomonas, and Azospirillum. Moreover, the change in rhizobacterial composition induced the changes in endobacterial and root exudate compositions. Moreover, the upregulated exudates inhibited ginger bacterial wilt, with the initial disease index (77.50%) being reduced to 40.00%, suggesting that ginger secretes antibacterial compounds for defense against bacterial pathogens.

Treatment with organic manure inoculated with a biocontrol agent induces soil bacterial communities to inhibit tomato Fusarium wilt disease.

Introduction: Organic manure, plant growth-promoting microorganisms, and biocontrol agents are widely used to sustainably control soil-borne diseases. However, how and whether organic manure inoculated with biocontrol agents alters soil microbiota and reduces disease severity is poorly understood. Methods: Here, we examined changes to the soil microbial community, soil properties, and incidence of Fusarium wilt disease in response to several fertilization regimes. Specifically, we studied the effects of inorganic chemical fertilization (CF), organic manure fertilization (OF), and Erythrobacter sp. YH-07-inoculated organic manure fertilization (BF) on the incidence of Fusarium wilt in tomato across three seasons. Results: BF-treated soils showed increased microbial abundance, richness, and diversity compared to other treatments, and this trend was stable across seasons. BF-treated soils also exhibited a significantly altered microbial community composition, including increased abundances of Bacillus, Altererythrobacter, Cryptococcus, and Saprospiraceae, and decreased abundances of Chryseolinea and Fusarium. Importantly, BF treatment significantly suppressed the incidence of Fusarium wilt in tomato, likely due to direct suppression by Erythrobacter sp. YH-07 and indirect suppression through changes to the microbial community composition and soil properties. Discussion: Taken together, these results suggest that Erythrobacter sp. YH-07-inoculated organic manure is a stable and sustainable soil amendment for the suppression of Fusarium wilt diseases.

Community Profile and Drivers of Predatory Myxobacteria under Different Compost Manures.

Myxobacteria are unique predatory microorganisms with a distinctive social lifestyle. These taxa play key roles in the microbial food webs in different ecosystems and regulate the community structures of soil microbial communities. Compared with conditions under conventional management, myxobacteria abundance increases in the organic soil, which could be related to the presence of abundant myxobacteria in the applied compost manure during organic conditions. In the present study,16S rRNA genes sequencing technology was used to investigate the community profile and drivers of predatory myxobacteria in four common compost manures. According to the results, there was a significant difference in predatory myxobacteria community structure among different compost manure treatments (p < 0.05). The alpha-diversity indices of myxobacteria community under swine manure compost were the lowest (Observed OTU richness = 13.25, Chao1 = 14.83, Shannon = 0.61), and those under wormcast were the highest (Observed OTU richness = 30.25, Chao1 = 31.65, Shannon = 2.62). Bacterial community diversity and Mg2+ and Ca2+ concentrations were the major factors influencing the myxobacteria community under different compost manure treatments. In addition, organic carbon, pH, and total nitrogen influenced the community profile of myxobacteria in compost manure. The interaction between myxobacteria and specific bacterial taxa (Micrococcales) in compost manure may explain the influence of bacteria on myxobacteria community structure. Further investigations on the in-situ community profile of predatory myxobacteria and the key microorganism influencing their community would advance our understanding of the community profile and functions of predatory microorganisms in the microbial world.

The inhibitory effects of different types of Brassica seed meals on the virulence of Ralstonia solanacearum.

BACKGROUND: Understanding the specific inhibitory effects of different Brassica seed meals (BSMs) on soilborne pathogens is important for their application as biocontrol agents for controlling plant disease. In this study, the seed meals of Brassica napus L. (BnSM), Brassica campestris L. (BcSM), and Brassica juncea L. (BjSM), and the combined seed meal of BcSM and BjSM (CSM, 1:1), were selected for investigation. The inhibitory effects of these seed meals on the plant pathogen Ralstonia solanacearum (Smith) and tomato bacterial wilt, were assessed and compared. RESULTS: All the BSMs significantly inhibited the growth of R. solanacearum in vitro. Furthermore, the BSMs could effectively suppress R. solanacearum virulence traits, including motility, exopolysaccharide production, dehydrogenase activity, virulence-related gene expression, and colonization in the soil. Among them, BjSM showed the best inhibiting effects, and CSM displayed synergic toxicity against R. solanacearum. In addition, the predominant antibacterial compounds in BcSM and BjSM were identified as the volatile compounds, 3-butenyl isothiocyanate and allyl isothiocyanate, respectively. Finally, pot experiment verified that the control effects of BjSM and CSM on tomato wilt reached more than 90%. CONCLUSION: This is the first study to report on the ability of different kinds of BSMs to suppress the virulence of R. solanacearum and biocontrol efficiencies against bacterial wilt in tomato plants. Furtherly, the main antibacterial compounds in the BSMs were identified. The results demonstrated that CSM may possess potential for controlling bacterial wilt caused by R. solanacearum. The results provide a fresh perspective for comprehending the mechanism underlying BSM suppression of pathogens and plant disease.

Profiling of Plant Growth-Promoting Metabolites by Phosphate-Solubilizing Bacteria in Maize Rhizosphere.

Microbial treatment has recently been attracting attention as a sustainable agricultural strategy addressing the current problems caused by unreasonable agricultural practices. However, the mechanism through which microbial inoculants promote plant growth is not well understood. In this study, two phosphate-solubilizing bacteria (PSB) were screened, and their growth-promoting abilities were explored. At day 7 (D7), the lengths of the root and sprout with three microbial treatments, M16, M44, and the combination of M16 and M44 (Com), were significantly greater than those with the non-microbial control, with mean values of 9.08 and 4.73, 7.15 and 4.83, and 13.98 and 5.68 cm, respectively. At day 14 (D14), M16, M44, and Com significantly increased not only the length of the root and sprout but also the underground and aboveground biomass. Differential metabolites were identified, and various amino acids, amino acid derivatives, and other plant growth-regulating molecules were significantly enhanced by the three microbial treatments. The profiling of key metabolites associated with plant growth in different microbial treatments showed consistent results with their performances in the germination experiment, which revealed the metabolic mechanism of plant growth-promoting processes mediated by screened PSB. This study provides a theoretical basis for the application of PSB in sustainable agriculture.

Variations in bacterial taxonomic profiles and potential functions in response to the gut transit of earthworms (Eisenia fetida) feeding on cow manure.

Earthworms play an important role in the organic matter decomposition in terrestrial ecosystems. Earthworms interact directly with the microorganisms to affect the organic matter decomposition via gut transit, i.e., the digestion and assimilation of organic matter in the foregut and midgut and its excretion by the hindgut. However, how the microbial community ingested by earthworms respond to the transit processes in different gut segments of earthworms is not clear. We used composted cow manure to feed earthworms and sampled vermicompost and the contents of foregut, midgut and hindgut for bacterial 16S rRNA gene sequencing analysis. We observed that earthworm gut transit decreased the abundances of the dominant phyla Proteobacteria and Bacteroidetes but increased Actinobacteria, Chloroflexi and Acidobacteria. The alpha diversity of bacterial community in midgut was the lowest of the different gut segments, and the bacterial community structure of the foregut was significantly different from the midgut and hindgut. The enrichment analysis results revealed different selective stimulatory and inhibitory effects on the ingested bacterial community in the different gut segments, which extended to vermicompost. The FAPROTAX data indicated that C and N metabolic microbes were enriched in the earthworm gut. Microbes involved in fermentation and methanogenesis were enriched in the hindgut, and denitrification microbes were enriched in the foregut. The N metabolism microbes in vermicompost were significantly enriched after the stimulation of earthworm gut transit (P < 0.05), and the pathogenic microbes of animals and plants were inhibited. Combined with the results of subsequent correlation and biochemical analyses, earthworm gut transit significantly altered the structure and function of the bacterial community to accelerate the degradation and mineralization of organic matter and the enrichment of phosphorus and potassium. Our study suggests that the gut transit process of earthworms plays an important role in regulating organic matter dynamics in terrestrial ecosystems.

Biological characterization and regulation of soil health.

How to determine the soil health status effectively is the basic issue to realize the agriculture green development. In the existing soil health assessment system, the importance of soil organi-sms in the maintenance of soil health is rarely considered. From the perspective of soil biological health, we discussed the connotation of soil health, and summarized the biological indicators of soil health, including soil microorganisms, soil enzyme activity, soil micro-food web and earthworm. Based on the above-mentioned indicators, the regulation approaches were elaborated from the aspects of crop and soil management practices. In addition, the future research on soil biological health was prospected. The main aim of this study is to enhance the understanding of scientists and decision makers on the maintenance of soil biological health, and to give full consideration of the important role of soil organisms in ecosystem services.

Identification of novel 1,4-dioxane degraders and related genes from activated sludge by taxonomic and functional gene sequence analysis.

This study used integrated omics technologies to investigate the potential novel pathways and enzymes for 1,4-dioxane degradation by a consortium enriched from activated sludge of a domestic wastewater treatment plant. An unclassified genus belonging to Xanthobacteraceae increased significantly after magnetic nanoparticle-mediated isolation for 1,4-dioxane degraders. Species with relatively higher abundance (> 0.3%) were identified to present high metabolic activities in the biodegradation process through shotgun sequencing. The functional gene investigations revealed that Xanthobacter sp. 91, Xanthobacter sp. 126, and a Rhizobiales strain carried novel 1,4-dioxane-hydroxylating monooxygenase genes. Xanthobacter sp. 126 contained the genes coding for glycolate oxidase, which was the main enzyme responsible for utilization of 1,4-dioxane intermediates through the TCA cycle, and further proven by the specific glycolate oxidase inhibitor, α-hydroxy-2-pyridinemethanesulfonic acid. An expanded and detailed degradation pathway of 1,4-dioxane was proposed on the basis of the three major intermediates (2-hydroxy-1,4-dioxane, ethylene glycol, and oxalic acid) confirmed by metabolomics. These findings of microbial community and function as well as the novel pathway will be valuable in predicting natural attenuation or reconstruction of a bacterial consortium for enhanced remediation of 1,4-dioxane-contaminated sites as well as wastewater treatment.

Insights into enhanced biodegradation of sulfadimethoxine by catalyst: Transcriptomic responses and free radical interactions.

The occurrence of sulfonamides in the environment is a severe global threat to public health due to the increasing prevalence of antibiotic selection pressure that may lead to the development of antibiotic resistance. We report an enhanced biodegradation of sulfadimethoxine (SDM) by Phanerochaete chrysosporium (Pc) with lignocellulosic biomass (Lb) using Fe3O4-ZSM-5 as a catalyst (Pc/Fe3O4-ZSM-5/Lb). SDM was completely degraded within 4 days at pH 7.0 in the Pc/Fe3O4-ZSM-5/Lb system. Transcriptomic, metabolites and free radical analyses were performed to explore the detailed molecular mechanisms of SDM degradation. A total of 246 genes of Pc in the Pc/Fe3O4-ZSM-5/Lb system exhibited significant upregulation compared to that in Pc alone. Upregulated genes encoding cellulases, cytochrome P450, cellobiose quinone oxidoreductase, and cellobiose dehydrogenase were involved in SDM degradation in the Pc/Fe3O4-ZSM-5/Lb system. In addition, genes encoding glutathione S-transferase and cytochrome P450 genes related to oxidative stress and detoxification were all significantly upregulated (P < 0.01). Electron paramagnetic resonance revealed the generation of OH suggesting a free radical pathway could be catalyzed by Fe3O4-ZSM-5 and the enzymes. These findings of catalyst-assisted SDM biodegradation will be valuable for remediation of antibiotics from contaminated wastewater.

Biodegradation mechanisms of sulfonamides by Phanerochaete chrysosporium - Luffa fiber system revealed at the transcriptome level.

The overuse of antibiotics and subsequent enrichment of antibiotic resistant microbes in the natural and built environments is a severe threat to global public health. In this study, a Phanerochaete chrysosporium fungal-luffa fiber system was found to efficiently biodegrade two sulfonamides, sulfadimethoxine (SDM) and sulfadizine (SDZ), in cow urine wastewater. Biodegradation pathways were proposed on the basis of key metabolites identified using high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (HPLC-QqTOF-MS). Transcriptomic, metabolomic, and free radical analyses were performed to explore the functional groups and detailed molecular mechanisms of SDM and SDZ degradation. A total of 27 UniGene clusters showed significant differences between luffa fiber and luffa fiber-free systems, which were significantly correlated to cellulose catabolism, carbohydrate metabolism, and oxidoreductase activity. Carbohydrate-active enzymes and oxidoreductases appear to play particularly important roles in SDM and SDZ degradation. Electron paramagnetic resonance (EPR) spectroscopy revealed the generation and evolution of OH and R during the biodegradation of SDM and SDZ, suggesting that beyond enzymatic degradation, SDM and SDZ were also transformed through a free radical pathway. Luffa fiber also acts as a co-substrate to improve the activity of enzymes for the degradation of SDM and SDZ. This research provides a potential strategy for removing SDM and SDZ from agricultural and industrial wastewater using fungal-luffa fiber systems.

Oxidation degradation of 2,2',5-trichlorodiphenyl in a chelating agent enhanced Fenton reaction: Influencing factors, products, and pathways.

The sodium pyrophosphate (SP)-enhanced Fenton reaction has been proven to have promising potential in remediation of polychlorinated biphenyls in soils by keeping iron ions soluble at high pH and minimizing the useless decomposition of H2O2. However, little information can be obtained about the effect of environmental factors on its remediation performance. Thus, the effect of environmental factors on the degradation of 2,2',5-trichlorodiphenyl (PCB18), one of the main PCB congeners in Chinese sites, was investigated in this study. PCB18 degradation was sensitive to pH, which ranged from 39.8% to 99.5% as increased pH from 3.0 to 9.0. ·OH was responsible for PCB18 degradation at pH 5.0, while both ·OH and O2- resulted in PCB 18 degradation at pH 7.0 with the calculated reaction activation energy of 73.5 kJ mol-1. Bivalent cations and transition metal ions decreased PCB18 degradation markedly as their concentrations increased. The addition of humic acid had an inhibitory on PCB18 degradation, but no obvious inhibition of PCB18 removal was observed when the same concentration of fulvic acid was added. The addition of 1 and 10 µM model humic constituents (MHCs) promoted PCB18 degradation, but the addition of 100 µM MHCs decreased PCB18 removal. Biphenyl, two dichlorobiphenyl, and two hydroxy trichlorobiphenyl derivatives were identified as the major degradation products of PCB18 in the Fe2+/SP/H2O2 system at pH 7.0. Thus, an oxidative pathway contributed by OH and a reductive pathway induced by O2- were proposed as the main mechanisms for PCB18 degradation in the SP-enhanced Fenton reaction.

Long-term nitrogen application decreases the abundance and copy number of predatory myxobacteria and alters the myxobacterial community structure in the soil.

Myxobacteria are fascinating micro-predators due to their extraordinary social lifestyle, which is unique in the bacterial domain. These taxa are metabolically active in the soil microbial food web and control populations of soil microbes. However, the effects of fertilisation treatments on predatory myxobacteria in agricultural systems are often overlooked. Here, the high-throughput absolute abundance quantification (HAAQ) method was employed to investigate the abundance and cell density of myxobacteria in the Red Soil Experimental Station fields following 29 years of fertilisation. Using 16S rRNA gene amplicons, we detected a total of 419 myxobacterial operational taxonomic units (OTUs), accounting for 0.25-2.70% of the total bacterial abundance. Significantly different myxobacterial communities were found between nitrogen-fertilised (N_cluster) and manure-fertilised (M_cluster) samples by principal coordinate analysis (PCoA), analysis of similarities (ANOSIM), and Manhattan analysis (p < 0.05). N fertiliser treatments significantly decreased the myxobacterial abundance and copy number, species accumulation index (S), and Shannon index (p < 0.05). Furthermore, UpSet plots showed that the OTU number in the N fertiliser treatment was only 24.4% of that in the M treatment, as the application of N decreased the number of low-abundance myxobacterial OTUs. In addition, network analysis, redundancy analysis (RDA), and random forest (RF) analysis showed that myxobacterial abundance and copy number were the most important variables predicting the soil bacterial community and functional gene α- and ß-diversity (P < 0.05). Our findings imply that soil acidification caused by the application of nitrogen fertilisers is the most important driver of the decrease in the myxobacterial abundance and copy number in the soil. We suggest that the changes in the abundance and number of myxobacteria are strongly correlated with the overall bacterial α- and ß-diversity indices. In addition, such changes may be an important factor in the overall changes in microbial communities.