Soil Diazotrophic Community Research Articles

Temporal dynamics of the diazotrophic community during corpse decomposition

Corpse decomposition affects soil organisms through the formation of “cadaver decomposition islands.” Soil diazotrophic microbes possess essential ecological functions on nitrogen input and nutrient cycling in the terrestrial ecosystem. However, our knowledge about how soil diazotrophic communities respond to corpse decomposition is lacking. In this study, we focused on the succession patterns and biological interaction of nitrogen-fixing microorganisms during animal (Ochotona curzoniae) corpse decomposition in terrestrial ecosystems by targeting nifH gene with high-throughput sequencing. Our results revealed that corpse decomposition of pikas reduced the α diversity and significantly impacted the β diversity of diazotrophic community across different decomposition stages. The divergent succession of diazotrophic community occurred under corpse pressure. Furthermore, the relative importance of stochasticity to the community assembly was improved by corpse decomposition, while the importance decreased over decomposition time. Cadaver decay also simplified the diazotrophic networks and weakened the biological interactions among diazotrophic populations. Notably, NH4-N was the most important factor affecting diazotrophic community, followed by time and total carbon. This work emphasized that corpse decomposition perhaps influences the process of biological nitrogen fixation by altering soil diazotrophic communities, which is of great significance for understanding the terrestrial ecosystems’ nitrogen cycle functions.Key points• Corpse decomposition reduced the α diversity of diazotrophic community.• Corpse decomposition improved the stochasticity of diazotrophic community assembly.• Corpse decomposition weakened the interactions among diazotrophic populations.

Responses of diazotrophic network structure and community diversity to alfalfa-maize intercropping are soil property-dependent.

Intercropping and soil properties both affect soil diazotrophic communities. However, the specific effects that alfalfa-maize intercropping has on diazotrophic networks and community diversity under different soil properties remain unclear. In this study, we investigated the soil diazotrophic communities of two crop systems, alfalfa monoculture (AA) and alfalfa-maize intercropping (A/M), in two sites with similar climates but different soil properties (poor vs. average). The diazotrophic network complexity and community diversity were higher at the site with poor soil than at the site with average soil (p < 0.05). Community structure also varied significantly between the sites with poor and average soil (p < 0.05). This divergence was mainly due to the differences in soil nitrogen, phosphorus, and organic carbon contents between the two sites. At the site with poor soil, the A/M system had lower diazotrophic diversity, lower network complexity and greater competition between diazotrophs than the AA system (p < 0.05) because intercropping intensified the soil phosphorus limitation under poor soil conditions. However, in the average soil, it was the A/M system that had an altered diazotrophic structure, with an increased abundance of 11 bacterial genera and a decreased abundance of three bacterial genera (p < 0.05). Our results indicated that the effects of alfalfa-maize intercropping on diazotrophic communities were soil property-dependent.

Endophytic diazotrophic communities from rice roots are diverse and weakly associated with soil diazotrophic community composition and soil properties.

Bacteria that promote plant growth, such as diazotrophs, are valuable tools for achieving a more sustainable production of important non-legume crops like rice. Different strategies have been used to discover new bacteria capable of promoting plant growth. This work evaluated the contribution of soil diazotrophs to the endophytic communities established in the roots of rice seedlings cultivated on seven representative soils from Uruguay. The soils were classified into two groups according to the C and clay content. qPCR, terminal restriction fragment length polymorphism (T-RFLP), and 454-pyrosequencing of the nifH gene were used for analyzing diazotrophs in soil and plantlets' roots grown from seeds of the same genotype for 25 days under controlled conditions. A similar nifH abundance was found among the seven soils, roots, or leaves. The distribution of diazotrophs was more uneven in roots than in soils, with dominance indices significantly higher than in soils (nifH T-RFLP). Dominant soils' diazotrophs were mainly affiliated to Alphaproteobacteria and Planctomycetota. Conversely, Alpha, Beta, Gammaproteobacteria, and Bacillota were predominant in different roots, though undetectable in soils. Almost no nifH sequences were shared between soils and roots. Root endophytic diazotrophs comprised a broader taxonomic range of microorganisms than diazotrophs found in soils from which the plantlets were grown and showed strong colonization patterns.

Long-term conservation tillage changes the diversity, assembly and network stability of the diazotrophic community

Diazotrophs make important contributions to nitrogen (N) inputs in agricultural ecosystems. However, strong evidence on the effects of conservation tillage (CT) on the coexistence and assembly of diazotrophic communities and related mechanisms is lacking. Here, we studied the impacts of long-term CT on the coexistence and assembly patterns of diazotrophic communities. Compared to traditional tillage (CK), CT significantly reduced both the N fixation rate (0–10 cm) and the alpha diversity of the diazotrophic community while increasing the density of the diazotrophic and overall bacterial communities. CT also reduced the competitive relationships within the diazotrophic community and enhanced network stability. Furthermore, diazotroph assembly was dominated by deterministic processes (68.63%) under CK, and stochastic processes (58.82%) under CT. Soil depth and total N (TN) were identified as crucial predictors shaping the assembly processes of diazotrophic communities under different tillage practices. The relative influence of stochastic processes on the diazotrophic community under CT varied more significantly with increasing soil depth. Overall, tillage practices and soil depth had significant influences on the coexistence and assembly processes of the soil diazotrophic community. Moreover, long-term CT may impact the selection of N fixation agents and the specific taxa associated with N fixers. Our results indicated that in CT systems, relatively sufficient nutrient availability leads to a reduction in interspecies competition, an increase in network stability, and a greater influence of stochastic processes on community assembly. These findings may help us better understand biological N fixation in sustainable agricultural systems.

Non-targeted effects of nitrification inhibitors on soil free-living nitrogen fixation modified with weed management

Biological nitrogen fixation and nitrification inhibitor applications contribute to improving soil nitrogen (N) availability, however, free-living N fixation affected by nitrification inhibitors has not been effectively evaluated in soils under different weed management methods. In this study, the effects of the nitrification inhibitors dicyandiamide (DCD) and 3, 4-dimethylpyrazole phosphate (DMPP) on the nitrogenase, nifH gene，and diazotrophic communities in soils under different weed management methods (AMB, weeds growth without mowing or glyphosate spraying; GS, glyphosate spraying; MSG, mowing and removing weeds and glyphosate spraying; and WM, mowing aboveground weeds) were investigated. Compared to the control counterparts, the DCD application decreased soil nitrogenase activity and nifH gene abundance by 4.5 % and 37.9 %, respectively, under the GS management method, and the DMPP application reduced soil nitrogenase activity by 20.4 % and reduced the nifH gene abundance by 83.4 % under the MSG management method. The application of nitrification inhibitors significantly elevated soil NH4+-N contents but decreased NO3−-N contents, which had adverse impacts on soil nifH gene abundance and nitrogenase activity. The nifH gene abundances were also negatively impacted by dissolved organic N and Geobacter but were positively affected by available phosphorus and diazotrophic community structures. Nitrification inhibitors significantly inhibited Methylocella but stimulated Rhizobiales and affected soil diazotrophic communities. The nitrification inhibitors DCD and DMPP significantly altered soil diazotrophic community structures, but weed management outweighed nitrification inhibitors in reshaping soil diazotrophic community structures. The non-targeted effects of the nitrification inhibitors DMPP and DCD on soil free-living N fixation were substantially influenced by the weed management methods.

The positive effects of mineral-solubilizing microbial inoculants on asymbiotic nitrogen fixation of abandoned mine soils are driven by keystone phylotype

Toward the restoration of the increasing numbers of abandoned mines across China, external-soil spray seeding technologies have become more extensively utilized. However, considerable challenges remain that seriously hamper the effectiveness of these technologies, such as inadequate nutrient availability for plants. Previous studies have shown that mineral-solubilizing microbial inoculants can increase the nodules of legumes. However, their effects on symbiotic nitrogen fixation (SNF), asymbiotic nitrogen fixation (ANF), and diazotrophic communities remain unknown. Further, research into the application of functional microorganisms for the restoration of abandoned mines has been conducted either in greenhouses, or their application in the field has been too brief. Thus, we established a four-year field experiment in an abandoned mine and quantified the SNF, ANF, and diazotrophic communities. To the best of our knowledge, this study is the first to describe the long-term application of specific functional microorganisms for the remediation of abandoned mine sites in the field. We revealed that mineral-solubilizing microbial inoculants significantly increased the soil ANF rate and SNF content. There was no significant correlation between the diazotrophic alpha diversity and soil ANF rate; however, there were strong positive associations between the relative abundance and biodiversity of keystone phylotype (module #5) within ecological clusters and the ANF rate. Molecular ecological networks indicated that microbial inoculants increased network complexity and stability. Moreover, the inoculants significantly enhanced the deterministic ratio of diazotrophic communities. Furthermore, homogeneous selection predominantly mediated the assembly of soil diazotrophic communities. It was concluded that mineral-solubilizing microorganisms played a critical role in maintaining and enhancing nitrogen, which offers a new solution with great potential for the restoration of ecosystems at abandoned mine sites.

Control factors of soil diazotrophic community assembly and nitrogen fixation rate across eastern China

Biological nitrogen (N) fixation is an essential N input into terrestrial ecosystems and plays a critical role in global N cycling. However, the driving factors of the diazotrophic community composition and soil N fixation rate at the regional scale remain largely unclear, especially in different land-use types. Here, we explored the environmental adaptation of diazotrophic communities and controlling factors of N fixation in maize, rice, and forest soils across eastern China. Results showed that the diversity, composition, and co-occurrence network differed significantly among these habitats, with a relatively higher alpha diversity and network complexity in rice fields. The mean annual precipitation plays a predominant role in shaping diazotrophic community composition. Dispersal limitation, belonging to stochastic processes, contributed most to diazotrophic community assembly in terrestrial ecosystems. Habitat niche breadth analysis indicated that the environmental adaptation of diazotrophic communities was highest in rice fields, followed by forest and maize fields. The overall N fixation rate was higher in rice fields, followed by maize and forest soils. Structural equation modeling revealed that the N fixation rate was mainly affected by soil properties in maize and forest soils. In contrast, the N fixation rate was more influenced by diazotrophic community composition and climatic factors in rice habitats. Our results revealed that the environmental adaptation of the diazotrophic communities and controlling factors of the N fixation function were different among maize, rice, and forest ecosystems. These findings will help us understand and predict the N bioavailability in different ecosystem types under climate changes.

Effects of copper oxide nanoparticles on soil diazotrophic communities in maize rhizosphere

Effects of copper oxide nanoparticles on soil diazotrophic communities in maize rhizosphere

Enhancement of nitrogen fixation and diazotrophs by long-term polychlorinated biphenyl contamination in paddy soil

Biological nitrogen fixation (BNF) driven by diazotrophs is a major means of increasing available nitrogen (N) in paddy soil, in addition to anthropogenic fertilization. However, the influence of long-term polychlorinated biphenyl (PCB) contamination on the diazotrophic community and nitrogen fixation in paddy soil is poorly understood. In this study, samples were collected from paddy soil subjected to > 30 years of PCB contamination, and the soil diazotrophic community and N2 fixation rate were evaluated by Illumina MiSeq sequencing and acetylene reduction assays, respectively. The results indicated that high PCB contamination increased diazotrophic abundance and the N2 fixation rate, and altered diazotrophic community structure in the paddy soil. The random forest model demonstrated that the β-diversity of the diazotrophic community was the most significant predictor of the N2 fixation rate. Structure equation modeling identified a specialized keystone diazotrophic ecological cluster, predominated by Bradyrhizobium, Desulfomonile, and Cyanobacteria, as the key driver of N2 fixation. Overall, our findings indicated that long-term PCB contamination enhanced the N2 fixation rate by altering diazotrophic community abundance and structure, which may deepen our understanding of the ecological function of diazotrophs in organic-contaminated soil.

Soil diazotrophic abundance, diversity, and community assembly mechanisms significantly differ between glacier riparian wetlands and their adjacent alpine meadows.

Global warming can trigger dramatic glacier area shrinkage and change the flux of glacial runoff, leading to the expansion and subsequent retreat of riparian wetlands. This elicits the interconversion of riparian wetlands and their adjacent ecosystems (e.g., alpine meadows), probably significantly impacting ecosystem nitrogen input by changing soil diazotrophic communities. However, the soil diazotrophic community differences between glacial riparian wetlands and their adjacent ecosystems remain largely unexplored. Here, soils were collected from riparian wetlands and their adjacent alpine meadows at six locations from glacier foreland to lake mouth along a typical Tibetan glacial river in the Namtso watershed. The abundance and diversity of soil diazotrophs were determined by real-time PCR and amplicon sequencing based on nifH gene. The soil diazotrophic community assembly mechanisms were analyzed via iCAMP, a recently developed null model-based method. The results showed that compared with the riparian wetlands, the abundance and diversity of the diazotrophs in the alpine meadow soils significantly decreased. The soil diazotrophic community profiles also significantly differed between the riparian wetlands and alpine meadows. For example, compared with the alpine meadows, the relative abundance of chemoheterotrophic and sulfate-respiration diazotrophs was significantly higher in the riparian wetland soils. In contrast, the diazotrophs related to ureolysis, photoautotrophy, and denitrification were significantly enriched in the alpine meadow soils. The iCAMP analysis showed that the assembly of soil diazotrophic community was mainly controlled by drift and dispersal limitation. Compared with the riparian wetlands, the assembly of the alpine meadow soil diazotrophic community was more affected by dispersal limitation and homogeneous selection. These findings suggest that the conversion of riparian wetlands and alpine meadows can significantly alter soil diazotrophic community and probably the ecosystem nitrogen input mechanisms, highlighting the enormous effects of climate change on alpine ecosystems.

Changes in community assembly processes and co-occurrence networks of soil diazotrophs along an elevational gradient in Tibetan alpine meadows

Shifts in soil microbial community composition along elevational gradients have been extensively studied; yet, the assembly process and co-occurrence network of soil microbes underlying these shifts remain to be explored, especially for diazotrophs that contribute to the most nitrogen input in natural ecosystems. In this study, we examined how assembly processes and co-occurrence networks of soil diazotrophs changed along an elevational gradient with four elevation levels ranging from 3200 m to 3800 m using high-throughput sequencing of the nifH gene in Tibetan alpine meadows. Our analyses showed that α-diversity of soil diazotrophs was lower at low elevations (i.e., 3200 m and 3400 m) relative to high elevations (i.e., 3600 m and 3800 m). Moreover, dominant diazotrophic phyla shifted from Actinobacteria at low elevations to Proteobacteria at high elevations. The change in diazotrophic community composition along the elevation gradient was related to the differences in soil temperature, moisture, NH4+-N concentration and pH. Species rank-abundance distribution models showed that soil diazotrophic communities were mainly assembled by deterministic processes at 3200 m, 3600 m and 3800 m elevations, and by stochastic processes at the 3400 elevation. Additionally, the complexity of diazotrophic co-occurrence networks increased with increasing elevations. Collectively, our findings highlight the change in soil diazotrophic communities along the elevation gradient and the high environmental sensitivity of soil diazotrophs, and suggest that global change could have consequences for soil diazotrophic community composition and structure in montane ecosystems.

Fertilization practices affect biological nitrogen fixation by modulating diazotrophic communities in an acidic soil in southern China

Biological nitrogen (N) fixation (BNF) driven by diazotrophs is an important pathway for N input in agricultural ecosystems. However, free-living BNF and its associated diazotrophic communities under different fertilization practices in acidic soils are poorly studied. Here, we conducted a long-term fertilization experiment (29 years), including an unfertilized control (CK), chemical N, P and K fertilizer (NPK), NPK plus lime (NPKL), NPK plus straw (NPKS), NPK plus straw and lime (NPKSL) and NPK plus manure (NPKM), and explored how fertilization practices affect free-living BNF by changing biotic and abiotic variables. Compared with CK (1.51 nmol C2H2 d−1 g−1), BNF rates were significantly increased in NPKM (1.99 nmol C2H2 d−1 g−1) but reduced in NPK (0.55 nmol C2H2 d−1 g−1), NPKL (0.61 nmol C2H2 d−1 g−1) and NPKS (0.69 nmol C2H2 d−1 g−1). Similarly, chemical fertilization, regardless of the addition of lime or straw, reduced the abundance (0.71-1.18 × 108 gene copies g−1) and α-diversity (1.11-2.43) of the diazotrophic community, while manure application had a positive effect on diazotrophic abundance (3.23 × 108 gene copies g−1) and α-diversity (3.36). Non-parametric multivariate analysis of variance (PERMANOVA) suggested that manure application (R2 = 0.212, p = 0.001) showed the stronger influence on diazotrophic community composition than additions of lime (R2 = 0.115, p = 0.019) and straw (R2 = 0.064, p = 0.161). Random forest model analysis revealed that BNF rates can be significantly (p < 0.05) explained by soil pH (9.9%), diazotrophic community attributes (composition, 8.5%; Chao1, 8.1%; abundance, 6.0%; Shannon, 5.7%), and soil total carbon (5.1%). Partial least squares path modeling (PLS-PM) suggested that diazotrophic community and soil properties mainly provided direct and indirect contributions to the variations of BNF rates, respectively. The dominant genera Pelomonas, Azospirillum and Dechloromonas were positively associated with BNF rates, with their members observed as keystone species in the community network. Together, chemical fertilizer combined with manure is an effective practice for improving BNF in acidic soil by affecting soil diazotrophic community.

Global Grassland Diazotrophic Communities Are Structured by Combined Abiotic, Biotic, and Spatial Distance Factors but Resilient to Fertilization

Grassland ecosystems cover around 37% of the ice-free land surface on Earth and have critical socioeconomic importance globally. As in many terrestrial ecosystems, biological dinitrogen (N2) fixation represents an essential natural source of nitrogen (N). The ability to fix atmospheric N2 is limited to diazotrophs, a diverse guild of bacteria and archaea. To elucidate the abiotic (climatic, edaphic), biotic (vegetation), and spatial factors that govern diazotrophic community composition in global grassland soils, amplicon sequencing of the dinitrogenase reductase gene—nifH—was performed on samples from a replicated standardized nutrient [N, phosphorus (P)] addition experiment in 23 grassland sites spanning four continents. Sites harbored distinct and diverse diazotrophic communities, with most of reads assigned to diazotrophic taxa within the Alphaproteobacteria (e.g., Rhizobiales), Cyanobacteria (e.g., Nostocales), and Deltaproteobacteria (e.g., Desulforomonadales) groups. Likely because of the wide range of climatic and edaphic conditions and spatial distance among sampling sites, only a few of the taxa were present at all sites. The best model describing the variation among soil diazotrophic communities at the OTU level combined climate seasonality (temperature in the wettest quarter and precipitation in the warmest quarter) with edaphic (C:N ratio, soil texture) and vegetation factors (various perennial plant covers). Additionally, spatial variables (geographic distance) correlated with diazotrophic community variation, suggesting an interplay of environmental variables and spatial distance. The diazotrophic communities appeared to be resilient to elevated nutrient levels, as 2–4 years of chronic N and P additions had little effect on the community composition. However, it remains to be seen, whether changes in the community composition occur after exposure to long-term, chronic fertilization regimes.

Altitudinal niches of symbiotic, associative and free-living diazotrophs driven by soil moisture and temperature in the alpine meadow on the Tibetan Plateau

Legume-associated symbiotic diazotrophs contribute more to nitrogen (N) fixation than non-symbiotic diazotrophs in many terrestrial ecosystems. However, the percentage of legume biomass is low in alpine meadows on the Tibetan Plateau. Therefore, non-symbiotic diazotrophs may play important roles in N fixation in alpine meadow soils. Moreover, Tibetan alpine meadows are fragile and sensitive to global climate change, and the investigating of the key factor driving soil diazotrophic community still entails several challenges. To address these issues, we investigated diazotrophic spatial distribution and diversity along the elevational gradient between 3200 and 4200 m in the alpine meadow using amplicon sequencing of nifH gene. The result clearly showed that soil moisture and temperature were key factors driving soil diazotrophic community structures. Both altitude and soil depth significantly differentiated diazotrophic community composition. Alpha diversity indices of diazotrophic communities showed unimodal distribution along elevation gradient, strongly affected by soil moisture. Altitudinal niches were occupied by different diazotrophs. Soils at lower elevations were dominated by symbiotic diazotrophs and associative diazotrophs related to high biomass of plant hosts, while those at higher elevations were dominated by free-living psychrophiles such as Polaromonas. Furthermore, high moisture stimulated free-living anaerobes at middle elevations, such as Geobacter and Anaeromyxobacter, while suppressed legumes and symbiotic Mezorhizobium. Soil temperature not only directly affected temperature-sensitive diazotrophs, but also indirectly affected them through plants and soil properties such as pH and ammonium content. Our results suggest that climate change may strongly affect biological nitrogen fixation (BNF), and free-living diazotrophs may play important roles in BNF of alpine meadow system on the Tibetan Plateau.

Land use indirectly affects the cycling of multiple nutrients by altering the diazotrophic community in black soil.

Diazotrophic bacteria, as one of most important group of soil microorganisms, play critical roles in multiple ecosystem functions (i.e., multifunctionality). However, little information is available about the diazotrophic community in driving soil nutrient cycling and multifunctionality at different depths with distinct vegetation in the black soil region of northeastern China. To learn the interactions among land use, cycling of multiple nutrients and the diazotrophic community, we performed this study in grassland (GL), forested land and a cropland (CL) in soils at depths of 0-15 cm and 15-35 cm. The highest nifH gene abundances were found in the CL treatment, while the highest diazotrophic species richness and diversity were detected in the GL in both soil layers. The nifH gene abundance was directly/positively correlated with soil bulk density and negatively correlated with land use and soil depth. The index of multiple nutrient cycling was directly/negatively affected by soil depth and indirectly/positively affected by land use. Land use directly/negatively affected soil pH and thus indirectly affected the diazotrophic community composition and the nutrient cycling index. The diversity and community composition of the diazotrophs together accounted for 95% of the differences in the multiple nutrient cycling index. Soil diazotrophic communities undertake important roles in maintaining nutrient cycling and soil multifunctionality at depths of 0-15 cm and 15-35 cm layers with different land uses of the black soil region of China. © 2021 Society of Chemical Industry.

Black locust coppice stands homogenize soil diazotrophic communities by reducing soil net nitrogen mineralization

Black locust (BL, Robinia pseudoacacia) is considered a promising tree species for reforestation due to its great ability to fix nitrogen. However, after two or three coppice-harvesting rotations, the productivity of BL declines. Whether soil microbial communities are affected and how these groups correlate with the nitrogen mineralization process across multi-generation stands remains unclear. We investigated the composition and structure of free-living nitrogen-fixing microorganisms (diazotrophs) by sequencing the marker gene nifH and compared these results to levels of soil nitrogen mineralization in the bulk soil and rhizosphere in black locust plantations on Mount Tai, China. The results showed multi-generation BL coppice plantations decreased the total soil nitrogen (N), soil phosphorus (P), soil microbial biomass N (MBN), soil microbial biomass C (MBC), soil nitrification rate (Rn), soil ammonification rate (Ra), and net soil N mineralization rate (Rm), but significantly increased the concentration of soil NH4+-N to maintain sufficient NO3−-N. The dominant species in bulk soil and rhizosphere changed from Rhodopseudomonas (22.62% and 15.76%), unclassified_c_Alphaproteobacteria (22.37% and 29.28%), unclassified_o_Rhizobiales (15.40% and 13.31%), Bradyrhizobium (12.00% and 11.74%) in seedling plantations to Bradyrhizobium (45.95% and 47.86%) and Rhodopseudomonas (43.56% and 41.84%) in coppice plantations, respectively. Mantel test and Redundancy analysis (RDA) revealed that Rn, Ra, and Rm were the most important factors shaping the diazotrophic communities. Our results suggest that the multi-generation BL coppice plantation can homogenize soil diazotrophic communities, which is mainly regulated by the available N loss caused by nitrogen mineralization. Strengthening the management technology of coppice plantations will provide more beneficial external consumption.

Dynamic variability of soil diazotrophs in bulk‐rhizosphere and phenological stages under long‐term mulching in an eroded area in the Loess Plateau

AbstractFree‐living diazotrophs play a significant role in the process of soil biological nitrogen fixation. Long‐term field management has a cumulative effect on soil microbial communities. However, after long‐term mulching measures in erosion areas, the stability difference in diazotrophs in bulk‐rhizosphere soil and the temporal dynamic changes remain unclear. In this study, we analysed the dynamic variability of soil diazotrophic community structures using high‐throughput sequencing. The result of Chao1 diversity index showed that mulching effectively increased the richness of soil diazotrophs. Combining the community composition, it was found that the diversity level of the soil diazotrophic community under mulching decreased more drastically along the development stages compared with no‐tillage, and Bradyrhizobium played a major role in nitrogen fixation. In rhizosphere soil, mulching promoted the relative abundances of Azohydromonas, Bradyrhizobium, and Skermanella. Co‐occurrence network analysis indicated that mulching measures strengthened the connection between the dominant components to improve the aggregation of the network. The partial least squares‐path model showed that the richness of diazotrophs in rhizosphere soil had an indirect effect on mulching through the available carbon and nitrogen nutrients of bulk soil, thereby improving the response rate to environmental variables. Overall, our findings showed that under long‐term mulching, the richness and aggregation of dominant components of the soil diazotrophic community increased with the phenological period and distance from roots, revealing the importance of mulching in eroded areas to improve farmland soil nitrogen fixation capacity. This study provides a theoretical reference for microecology to effectively improve soil fertility in similar areas.

Expansion of Native Plant Stellera chamaejasme L. Alters the Structure of Soil Diazotrophic Community in a Salinized Meadow Grassland, Northeast China

The invasion of native plants has posed a serious risk to species diversity and ecosystem function. How they modify underground community and facilitate successful invasion remain unknown. Soil diazotrophs may play an important role in invasion by native plants. Stellera chamaejasme L. has expanded within around the heavily degraded Horqin Grassland in northeast China in recent decades. This study aims to detect the effect of the expansion of S. chamaejasme L. on soil diazotrophic community structure through high-throughput sequencing and examine the relationship between diazotrophic community structure and soil physicochemical properties. An extensive increase in S. chamaejasme population induced significant changes in soil diazotrophic community and marked shifts in the relative abundances of Bradyrhizobium and Desulfuromonas. Soil organic matter (SOM), total nitrogen, NO3−-N, and electrical conductivity (EC) increased, whereas NH4+-N and pH significantly decreased in soil invaded by S. chamaejasme. The diazotrophic community structure was correlated with SOM, nitrogen content, EC, and pH. The relative abundances of Bradyrhizobium and Desulfuromonas were significant negatively and positively correlated with soil EC, respectively. This study suggests that the interaction between S. chamaejasme and soil diazotrophic microbes and the durative increase in soil EC may facilitate invasion by this S. chamaejasme population.

Long-term fertilization with high nitrogen rates decreased diversity and stability of diazotroph communities in soils of sweet potato

Sweet potato (Ipomoea batatas Lam) could produce acceptable root yield in low-nitrogen (N) soils, with substantial N uptake potentially attributed to supplies provided via biological N2-fixation by free-living diazotrophs. However, dynamics of diazotrophic communities as influenced by soil properties across the N fertilization gradients are still largely unclear. Long-term fertilization experiment under wheat-sweet potato rotation was established in an acid yellow brown soil since 2011. Soil samples were collected after sweet potato harvest (October 2018). The nitrogenase (nifH) gene real-time polymerase chain reaction (RT-PCR) and Hiseq high-throughput sequencing technologies were applied to soil samples from four N fertilizer treatments (0, 60, 120 and 180 kg ha−1). The results showed that long-term N fertilization significantly decreased abundance of the nifH gene, which was closely related to decreases in the content of available phosphorus (AP). The long-term high-N (120 and 180 kg ha−1) fertilization dramatically altered structure of soil diazotrophic community and lead to in decreased diversity of diazotrophs, whereas low-N (60 kg ha−1) fertilization maintained diazotrophic community diversity and stability. Compared with the low-N fertilizer inputs (0 and 60 kg ha−1), the high rates of N fertilization (180 kg ha−1) significantly decreased the relative abundance of nifH-harboring microorganisms, especially the phylum Cyanobacteria known as potential N2-fixers that could sustain fertility of sweet potato soils. There were negative correlations between N fertilization rates and the relative abundance of Proteobacteria, whereas the Bacteroidetes and Firmicutes showed a positive correlation. Moreover, the structural equation model (SEM) results suggested that the diazotrophic community diversity and structure were influenced mostly by soil pH rather than SOM and N forms (TN, NH4+-N and NO3−-N), and diazotrophs abundance mainly regulated by soil AP content. Our results implied that appropriate N fertilization is beneficial to sustain the stability and diversity of the diazotrophic community.

Nitrogen Fertilization and Native C4 Grass Species Alter Abundance, Activity, and Diversity of Soil Diazotrophic Communities.

Native C4 grasses have become the preferred species for native perennial pastures and bioenergy production due to their high productivity under low soil nitrogen (N) status. One reason for their low N requirement is that C4 grasses may benefit from soil diazotrophs and promote biological N fixation. Our objective was to evaluate the impact of N fertilization rates (0, 67, and 202 kg N ha–1) and grass species (switchgrass [Panicum virgatum] and big bluestem [Andropogon gerardii]) on the abundance, activity, diversity, and community composition of soil diazotrophs over three agricultural seasons (grass green-up, initial harvest, and second harvest) in a field experiment in East Tennessee, United States. Nitrogen fertilization rate had a stronger influence on diazotroph population size and activity (determined by nifH gene and transcript abundances) and community composition (determined by nifH gene amplicon sequencing) than agricultural season or grass species. Excessive fertilization (202 kg N ha–1) resulted in fewer nifH transcripts compared to moderate fertilization (67 kg N ha–1) and decreased both richness and evenness of diazotrophic community, reflecting an inhibitory effect of high N application rates on soil diazotrophic community. Overall, cluster I and cluster III diazotrophs were dominant in this native C4 grass system. Diazotroph population size and activity were directly related to soil water content (SWC) based on structural equation modeling. Soil pH, SWC, and C and N availability were related to the variability of diazotrophic community composition. Our results revealed relationships between soil diazotrophic community and associated soil properties, adding to our understanding of the response of soil diazotrophs to N fertilization and grass species in native C4 grass systems.