Belowground Bacterial Communities Research Articles

Above and below-ground bacterial communities shift in seagrass beds with warmer temperatures

Current rates of ocean warming are predicted to exacerbate ongoing declines in seagrass populations. Above-ground responses of seagrass to increasing temperatures have been studied from a direct physiological perspective while indirect effects, including changes to microbially-mediated below-ground processes, remain poorly understood. To test potential effects of increased temperature on seagrass growth and associated microbial communities, we sampled seagrass beds experiencing ambient and elevated water temperatures at Lake Macquarie, Australia. Sites with warmer water were associated with a plume from a power station discharge channel with temperatures analogous to conditions predicted by 2100 under current rates of ocean warming (+3°C). The microbial community composition in both sediments and leaf tissues varied significantly between warm and ambient water temperatures with higher relative abundances of putative sulphate-reducing bacteria such as Desulfocapsaceae, Desulfobulbaceae and Desulfosarcinaceae in sedimentary communities in warm water. Above-ground biomass and seagrass growth rates were greater at warm sites while below-ground biomass and detrital decomposition rates showed no difference suggesting potential buffering of temperature effects below-ground. These findings suggest a 3°C rise in temperate regions is unlikely to induce mortality in seagrass however, it may shift microbial communities towards more homogenous structure and composition.

The Influence of the Genotype and Planting Density on the Structure and Composition of Root and Rhizosphere Microbial Communities in Maize.

Maize has the largest cultivation area of any crop in the world and plays an important role in ensuring food security. High-density planting is essential for maintaining high maize yields in modern intensive agriculture. Nonetheless, how high-density planting and the tolerance of individual genotypes to such planting shape the root-associated microbiome of maize is still unknown. In this study, we analyzed the root and rhizosphere bacterial communities of two maize accessions with contrasting shoot architectures grown under high- and low-density planting conditions. Our results suggested that maize hosted specific, distinct bacterial communities in the root endocompartment and that the maize genotype had a significant effect on the selection of specific microbes from the rhizosphere. High-density planting also had significant effects on root-associated bacterial communities. Specifically, genotype and high-density planting coordinated to shape the structure, composition, and function of root and rhizosphere bacterial communities. Taken together, our results provide insights into how aboveground plant architecture and density may alter the belowground bacterial community in root-associated compartments of maize.

Pedogenesis of typical zonal soil drives belowground bacterial communities of arable land in the Northeast China Plain

Belowground bacterial communities play essential roles in maintaining ecosystem multifunction, while our understanding of how and why their distribution patterns and community compositions may change with the distinct pedogenetic conditions of different soil types is still limited. Here, we evaluated the roles of soil physiochemical properties and biotic interactions in driving belowground bacterial community composition across three typical zonal soil types, including black calcium soil (QS), typical black soil (HL) and dark brown soil (BQL), with distinct pedogenesis on the Northeast China Plain. Changes in soil bacterial diversity and community composition in these three zonal soil types were strongly correlated with soil pedogenetic features. SOC concentrations in HL were higher than in QS and BQL, but bacterial diversity was low, and the network structure revealed greater stability and connectivity. The composition of the bacterial community correlated significantly with soil pH in QS but with soil texture in BQL. The bacterial co-occurrence network of HL had higher density and clustering coefficients but lower edges, and different keystone species of networks were also detected. This work provides a basic understanding of the driving mechanisms responsible for belowground bacterial biodiversity and distribution patterns over different pedogenetic conditions in agroecosystems.

Belowground bacterial communities and carbon components contribute to soil respiration in a subtropical forest

Belowground bacterial communities and carbon components contribute to soil respiration in a subtropical forest

Immediate impacts of soybean cover crop on bacterial community composition and diversity in soil under long-term Saccharum monoculture.

Saccharum yield decline results from long-term monoculture practices. Changes in cropping management can improve soil health and productivity. Below-ground bacterial community diversity and composition across soybean (Glycine max (L.) Merr) cover crop, Saccharum monoculture (30+ year) and fallowed soil were determined. Near full length (~1,400 base pairs) of 16S rRNA gene sequences were extracted from the rhizospheres of sugarcane and soybean and fallowed soil were compared. Higher soil bacterial diversity was observed in the soybean cover crop than sugarcane monoculture across all measured indices (observed operationational taxonomic units, Chao1, Shannon, reciprocal Simpson and Jackknife). Acidocateria, Proteobacteria, Bacteroidetes and Planctomycetes were the most abundant bacterial phyla across the treatments. Indicator species analysis identified nine indicator phyla. Planctomycetes, Armatimonadetes and candidate phylum FBP were associated with soybean; Proteobacteria and Firmicutes were linked with sugarcane and Gemmatimonadetes, Nitrospirae, Rokubacteria and unclassified bacteria were associated with fallowed soil. Non-metric multidimensional scaling analysis showed distinct groupings of bacterial operational taxonomic units (97% identity) according to management system (soybean, sugarcane or fallow) indicating compositional differences among treatments. This is confirmed by the results of the multi-response permutation procedures (A = 0.541, p = 0.00045716). No correlation between soil parameters and bacterial community structure was observed according to Mantel test (r = 211865, p = 0.14). Use of soybean cover-crop fostered bacterial diversity and altered community structure. This indicates cover crops could have a restorative effect and potentially promote sustainability in long-term Saccharum production systems.

Water pH, not soil pH, alters bacterial community structural pattern and nitrogen cycling pathways in date palm (Phoenix dactylifera L.) roots and bulk soil under freshwater irrigation regime

Date palms are widely cultivated in arid agroecosystems, where knowledge of irrigation water effect on their soil and root-associated bacterial communities is limited. Using 16S rDNA metabarcoding, we studied soil and root-associated bacterial communities of date palms growing in United Arab Emirates. Overall, 12.18% of bacterial operational taxonomic units (OTUs) were unique to roots, whereas 41.55% were specific to soil. The bacterial diversity was lower in root and community patterns were distinct between compartments, wherein irrigation water pH was a key structuring factor in both compartments, while salinity (electrical conductivity) was important only in the soil. Co-occurrence network analysis revealed a decrease in complexity in the soil–root continuum, and specific taxa/modules also varied with water pH. We observed a higher abundance of endophyte–saprotroph (Bacillus, Streptomyces and Dongia) dual-role OTUs in both compartments, possibly involved in nutrient mobilisation and plant growth. Based on PICRUSt and trait-based analyses, we showed that these OTUs are putatively involved in the nitrogen cycle (nitrification, denitrification, and assimilatory nitrate reduction). Taken together, we suggest that irrigation water pH, not soil pH, transiently affects belowground bacterial communities and selects bacteria in specific pH ranges, which may be important for nutrient cycling (i.e. nitrogen cycle) in arid agroecosystems.

Rhizosphere and bulk soil bacterial community succession is influenced more by changes in soil properties than in rhizosphere metabolites across a maize growing season

The rhizosphere microbiome composition shifts across plant developmental stages. In addition, the bulk soil microbiome also changes during the growing season. However, the factors governing the rhizosphere and bulk soil microbiome succession across plant development are still not well identified. In this study we analyzed the changes in rhizosphere and bulk soil bacterial community structure through 16S rRNA amplicon sequencing across three maize growth stages (vegetative leaf 5 [V5], vegetative leaf 10 [V10] and reproductive stage 2 [R2]), as well as their correlations with temporal shifts in soil physicochemical properties and in root metabolites quantified in rhizosphere soils by LC/GC–MS. Twelve maize genotypes were used in a replicated field experiment for a comprehensive study. Results indicated significant correlations between the rhizosphere and bulk soil bacterial community succession with the temporal changes in rhizosphere metabolites and soil physicochemical properties. The changes in soil properties (mainly soil N and moisture) across stages explained more of the variation in the rhizosphere and bulk soil bacterial communities than the rhizosphere metabolites. However, shifts in rhizosphere metabolites (mainly sugars and gibberellins) across stages also strongly affected the rhizosphere bacterial community succession, but this effect was weaker on the bulk soil bacterial community. In addition to the changes in microbiome composition and diversity, we also detected changes in soil microbial respiration between plant developmental stages, with greater microbial activity at the V10 stage. In sum, our study showed that changes in soil properties have a great influence on both rhizosphere and bulk soil bacterial community succession across a growing season. The changes in plant root metabolites across the cropping season also affected the succession of the belowground bacterial communities, but the impact was much larger on the rhizosphere than on the bulk soil microbiome. Understanding the ecological processes shaping the belowground microbiomes through time is pivotal to develop future strategies for the most appropriate crop management across the whole growing season.

Is foliar spectrum predictive of belowground bacterial diversity? A case study in a peach orchard.

Rhizosphere bacteria can have wide-ranging effects on their host plants, influencing plant biochemical and structural characteristics, and overall productivity. The implications of plant-microbe interactions provides an opportunity to interfere agriculture ecosystem with exogenous regulation of soil microbial community. Therefore, how to efficiently predict soil bacterial community at low cost is becoming a practical demand. Here, we hypothesize that foliar spectral traits can predict the diversity of bacterial community in orchard ecosystem. We tested this hypothesis by studying the ecological linkages between foliar spectral traits and soil bacterial community in a peach orchard in Yanqing, Beijing in 2020. Foliar spectral indexes were strongly correlated with alpha bacterial diversity and abundant genera that can promote soil nutrient conversion and utilization, such as Blastococcus, Solirubrobacter, and Sphingomonas at fruit mature stage. Certain unidentified or relative abundance <1% genera were also associated with foliar spectral traits. We selected specific indicators (photochemical reflectance index, normalized difference vegetable index, greenness index, and optimized soil-adjusted vegetation index) of foliar spectral indexes, alpha and beta diversities of bacterial community, and quantified the relations between foliar spectral traits and belowground bacterial community via SEM. The results of this study indicated that foliar spectral traits could powerfully predict belowground bacterial diversity. Characterizing plant attributes with easy-accessed foliar spectral indexes provides a new thinking in untangling the complex plant-microbe relationship, which could better cope with the decreased functional attributes (physiological, ecological, and productive traits) in orchard ecosystem.

Aridity modulates belowground bacterial community dynamics in olive tree.

SummaryAridity negatively affects the diversity and abundance of edaphic microbial communities and their multiple ecosystem services, ultimately impacting vegetation productivity and biotic interactions. Investigation about how plant‐associated microbial communities respond to increasing aridity is of particular importance, especially in light of the global climate change predictions. To assess the effect of aridity on plant associated bacterial communities, we investigated the diversity and co‐occurrence of bacteria associated with the bulk soil and the root system of olive trees cultivated in orchards located in higher, middle and lower arid regions of Tunisia. The results indicated that the selective process mediated by the plant root system is amplified with the increment of aridity, defining distinct bacterial communities, dominated by aridity‐winner and aridity‐loser bacteria negatively and positively correlated with increasing annual rainfall, respectively. Aridity regulated also the co‐occurrence interactions among bacteria by determining specific modules enriched with one of the two categories (aridity‐winners or aridity‐losers), which included bacteria with multiple PGP functions against aridity. Our findings provide new insights into the process of bacterial assembly and interactions with the host plant in response to aridity, contributing to understand how the increasing aridity predicted by climate changes may affect the resilience of the plant holobiont.

A microbiota\u2013root\u2013shoot circuit favours Arabidopsis growth over defence under suboptimal light

Bidirectional root–shoot signalling is probably key in orchestrating stress responses and ensuring plant survival. Here, we show that Arabidopsis thaliana responses to microbial root commensals and light are interconnected along a microbiota–root–shoot axis. Microbiota and light manipulation experiments in a gnotobiotic plant system reveal that low photosynthetically active radiation perceived by leaves induces long-distance modulation of root bacterial communities but not fungal or oomycete communities. Reciprocally, microbial commensals alleviate plant growth deficiency under low photosynthetically active radiation. This growth rescue was associated with reduced microbiota-induced aboveground defence responses and altered resistance to foliar pathogens compared with the control light condition. Inspection of a set of A. thaliana mutants reveals that this microbiota- and light-dependent growth–defence trade-off is directly explained by belowground bacterial community composition and requires the host transcriptional regulator MYC2. Our work indicates that aboveground stress responses in plants can be modulated by signals from microbial root commensals.

Evaluating domestication and ploidy effects on the assembly of the wheat bacterial microbiome.

While numerous studies implicate the microbiome in host fitness, contributions of host evolution to microbial recruitment remain largely uncharacterized. Past work has shown that plant polyploidy and domestication can influence plant biotic and abiotic interactions, yet impacts on broader microbiome assembly are still unknown for many crop species. In this study, we utilized three approaches—two field studies and one greenhouse-based experiment—to determine the degree to which patterns in bacterial community assembly in wheat (Triticum sp.) roots and rhizospheres are attributable to the host factors of ploidy level (2n, 4n, 6n) and domestication status (cultivated vs. wild). Profiling belowground bacterial communities with 16S rRNA gene amplicon sequencing, we analyzed patterns in diversity and composition. From our initial analyses of a subsetted dataset, we observed that host ploidy level was statistically significant in explaining variation in alpha and beta diversity for rhizosphere microbiomes, as well as correlated with distinct phylum-level shifts in composition, in the field. Using a reduced complexity field soil inoculum and controlled greenhouse conditions, we found some evidence suggesting that genomic lineage and ploidy level influence root alpha and beta diversity (p-value<0.05). However, in a follow-up field experiment using an expanded set of Triticum genomes that included both wild and domesticated varieties, we did not find a strong signal for either diploid genome lineages, domestication status, or ploidy level in shaping rhizosphere bacterial communities. Taken together, these results suggest that while host ploidy and domestication may have some minor influence on microbial assembly, these impacts are subtle and difficult to assess in belowground compartments for wheat varieties. By improving our understanding of the degree to which host ploidy and cultivation factors shape the plant microbiome, this research informs perspectives on what key driving forces may underlie microbiome structuring, as well as where future efforts may be best directed towards fortifying plant growth by microbial means. The greatest influence of the host on the wheat microbiome appeared to occur in the rhizosphere compartment, and we suggest that future work focuses on this environment to further characterize how host genomic and phenotypic changes influence plant-microbe communications.

Soil bacterial communities vary more by season than with over two decades of experimental warming in Arctic tussock tundra

High latitude ecosystems are characterized by cold soils and long winters, with much of their biogeochemistry directly or indirectly controlled by temperature. Climate warming has led to an expansion of shrubby plant communities across tussock tundra, but whether these clear aboveground shifts correspond to changes in the microbial community belowground remains less certain. Using bromodeoxyuridine to label growing cells, we evaluated how total and actively growing bacterial communities varied throughout a year and following 22 years of passive summer warming. We found that changes in total and actively growing bacterial community structures were correlated with edaphic factors and time point sampled, but were unaffected by warming. The aboveground plant community had become more shrub-dominated with warming at this site, and so our results indicate that belowground bacterial communities did not track changes in the aboveground plant community. As such, studies that have used space-for-time methods to predict how increased shrub cover has altered bacterial communities may not be representative of how the microbial community will be affected by in situ changes in the plant community as the Arctic continues to warm.

Heat waves intensify the effects of drought on bacterial diversity but not community composition in Solanum lycopersicum soil

Droughts and heat waves are expected to occur in combination more frequently under climate change; however, the interactive effect of these extreme events on soil microorganisms remains poorly understood. Here, we investigated the effects of drought, heat wave, and rehydration on bacterial diversity and community composition in Solanum lycopersicum grown soil. A growth chamber experiment with two factors (drought and heat wave) was conducted. Seedlings of Solanum lycopersicum were grown in well-watered or moderate drought stress at ambient temperature prior to a 7-day heat wave treatment. Rewatering from drought was conducted after the heat wave treatment. Soils were sampled at three time points, and bacterial 16S ribosomal RNA genes extracted from total soil DNA were amplified by PCR. Results showed that heat wave significantly affected bacterial diversity and community composition, while drought only significantly affected community composition. Their combination had significant interactive effect on diversity but not community composition. Moderate drought slightly increased bacterial diversity, but heat wave plus drought caused a decline in diversity. Heat wave stress suppressed the recovery of bacterial diversity from drought stress. Our results suggest that heat wave can aggravate the effect of drought on soil bacterial diversity, and the combination of heat wave and drought stress causes a lower recovery potential for bacterial diversity, indicating that extreme weather events would strongly affect belowground bacterial community composition in future climatic scenarios.

Soil Element Stoichiometry Drives Bacterial Community Composition Following Thinning in A Larix Plantation in the Subalpine Regions of Northern China

It is well established that forest thinning alters aboveground plant community composition and soil resource availability. However, how it regulates the composition and diversity of belowground microbial communities remains unclear. To quantify the effects of thinning on soil bacterial groups and the underlying mechanisms of these effects, this research was conducted in a Larix principis-rupprechtii Mayr. plantation with various thinning intensities, including a control (0% tree removal), a low-intensity treatment (15% tree removal), a medium-intensity treatment (35% tree removal), and a high-intensity treatment (50% tree removal). Compared to the control, the medium and high intensity thinning treatments significantly improved soil moisture, nutrient concentrations (including soil total carbon, nitrogen, phosphorus, and ammonium nitrogen), microbial biomass, and elemental stoichiometry ratios. The abundance and diversity of bacterial communities peaked in the medium-intensity treatment. Thinning also had strong effects on dominant bacterial groups at the phylum level. For instance, Bacteroidetes and Nitrospirae were significantly increased in the medium-intensity treatment (MIT), while the Gemmatimonadetes were significantly decreased in the low-intensity treatment (LIT). Combining Spearman correlation analysis and redundancy analysis demonstrated that thinning could facilitate the assembly of unique bacterial communities, and these shifts in microorganisms could probably be attributed to corresponding changes in soil resource stoichiometry. In conclusion, this study provides novel evidence that rational thinning could promote belowground bacterial community diversity and that elemental stoichiometry is an important indicator in shaping forest soil bacterial communities.

Meta-Analysis Reveals Consistent Bacterial Responses to Land Use Change Across the Tropics

Bacterial communities are a major component of global diversity and are intimately involved in most terrestrial biogeochemical processes. Despite their importance, we know far less about the response of bacteria to human-induced environmental change than we do about other organisms. Understanding the response of organisms to land use change is especially pressing for tropical rainforests, which are being altered at a higher rate than any other ecosystem. Here, we conduct a meta-analysis of studies performed in each of the major tropical rainforest regions to ask whether there are consistent responses of belowground bacterial communities to the conversion of tropical rainforest to agriculture. Remarkably, we find common responses despite wide variation across studies in the types of agriculture practiced and the research methodology used to study land use change. These responses include changes in the relative abundance of phyla, most notably decreases in Acidobacteria (-1.94±1.1 fold (average±95%CI)) and Proteobacteria (-1.38±1.0 fold) and increases in Actinobacteria (1.55±1.1 fold), Chloroflexi (3.47±1.2 fold) and Firmicutes (6.6±1.3 fold). We also find that alpha diversity (at the scale of single soil cores) consistently increases (1.17±1.0-fold) with ecosystem conversion. These consistent responses suggest that, while there is great diversity in agricultural practices across the tropics, common features such as the use of slash-and-burn tactics have the potential to alter bacterial community composition and diversity belowground.

Proximate grassland and shrub-encroached sites show dramatic restructuring of soil bacterial communities.

BackgroundChanges in aboveground community composition and diversity following shrub encroachment have been studied extensively. Recently, shrub encroachment was associated with differences in belowground bacterial communities relative to non-encroached grassland sites hundreds of meters away. This spatial distance between grassland and shrub sites left open the question of how soil bacterial communities associated with different vegetation types might differ within the same plot location.MethodsWe examined soil bacterial communities between shrub-encroached and adjacent (one m apart) grassland soils in Chinese Inner Mongolian, using high-throughput sequencing method (Illumina, San Diego, CA, USA).ResultsShrub-encroached sites were associated with dramatic restructuring of soil bacterial community composition and predicted metabolic function, with significant increase in bacterial alpha-diversity. Moreover, bacterial phylogenic structures showed clustering in both shrub-encroached and grassland soils, suggesting that each vegetation type was associated with a unique and defined bacterial community by niche filtering. Finally, soil organic carbon (SOC) was the primary driver varied with shifts in soil bacterial community composition. The encroachment was associated with elevated SOC, suggesting that shrub-mediated shifts in SOC might be responsible for changes in belowground bacterial community.DiscussionThis study demonstrated that shrub-encroached soils were associated with dramatic restructuring of bacterial communities, suggesting that belowground bacterial communities appear to be sensitive indicators of vegetation type. Our study indicates that the increased shrub-encroached intensity in Inner Mongolia will likely trigger large-scale disruptions in both aboveground plant and belowground bacterial communities across the region.

Shrub encroachment is associated with changes in soil bacterial community composition in a temperate grassland ecosystem

The effects of shrub encroachment on plant and soil properties have been well studied. However, little is known about how shrub encroachment influences soil bacterial communities. We investigated the effects of shrub encroachment on grassland soil bacterial communities along a soil depth gradient in the Inner Mongolian region of China. The belowground bacterial communities were examined using high-throughput sequencing of the 16S rRNA gene (V4-V5 region, Illumina MiSeq). Bacterial alpha-diversity was higher in shrub-encroached soils than in control grassland soils. Bacterial OTU richness was highest at 0–20 cm soil depths, while phylogenetic diversity was greatest at 10–20 cm soil depths. At each soil depth layer, shrub encroachment was associated with a significant shift in bacterial community composition. Change in soil pH was the factor most strongly related to change in bacterial community composition associated with shrub encroachment at all four depth horizons in the soils. Shrub encroachment appears to alter the distribution of bacterial life history strategies in the surface soil (i.e., showing an enrichment in copiotrophs and a depletion in oligotrophs) and shrubs are associated with an increase in nitrification potential in deeper soil horizons. Our results indicate that the influence of shrub encroachment on bacterial community composition extends deep into the soil. The intensity of shrub encroachment at this study site suggests that this ecosystem is undergoing dramatic succession towards shrub-dominance, which will likely trigger shifts in ecosystem function.

Plant-plant competition outcomes are modulated by plant effects on the soil bacterial community

Competition is a key process that determines plant community structure and dynamics, often mediated by nutrients and water availability. However, the role of soil microorganisms on plant competition, and the links between above- and belowground processes, are not well understood. Here we show that the effects of interspecific plant competition on plant performance are mediated by feedbacks between plants and soil bacterial communities. Each plant species selects a singular community of soil microorganisms in its rhizosphere with a specific species composition, abundance and activity. When two plant species interact, the resulting soil bacterial community matches that of the most competitive plant species, suggesting strong competitive interactions between soil bacterial communities as well. We propose a novel mechanism by which changes in belowground bacterial communities promoted by the most competitive plant species influence plant performance and competition outcome. These findings emphasise the strong links between plant and soil communities, paving the way to a better understanding of plant community dynamics and the effects of soil bacterial communities on ecosystem functioning and services.

Shifts in microbial communities in soil, rhizosphere and roots of two major crop systems under elevated CO2 and O3

Rising atmospheric concentrations of CO2 and O3 are key features of global environmental change. To investigate changes in the belowground bacterial community composition in response to elevated CO2 and O3 (eCO2 and eO3) the endosphere, rhizosphere and soil were sampled from soybeans under eCO2 and maize under eO3. The maize rhizosphere and endosphere α-diversity was higher than soybean, which may be due to a high relative abundance of Rhizobiales. Only the rhizosphere microbiome composition of the soybeans changed in response to eCO2, associated with an increased abundance of nitrogen fixing microbes. In maize, the microbiome composition was altered by the genotype and linked to differences in root exudate profiles. The eO3 treatment did not change the microbial communities in the rhizosphere, but altered the soil communities where hybrid maize was grown. In contrast to previous studies that focused exclusively on the soil, this study provides new insights into the effects of plant root exudates on the composition of the belowground microbiome in response to changing atmospheric conditions. Our results demonstrate that plant species and plant genotype were key factors driving the changes in the belowground bacterial community composition in agroecosystems that experience rising levels of atmospheric CO2 and O3.

Revegetation rewilds the soil bacterial microbiome of an old field.

Ecological restoration is a globally important and well-financed management intervention used to combat biodiversity declines and land degradation. Most restoration aims to increase biodiversity towards a reference state, but there are concerns that intended outcomes are not reached due to unsuccessful interventions and land-use legacy issues. Monitoring biodiversity recovery is essential to measure success; however, most projects remain insufficiently monitored. Current field-based methods are hard to standardize and are limited in their ability to assess important components of ecosystems, such as bacteria. High-throughput amplicon sequencing of environmental DNA (metabarcoding of eDNA) has been proposed as a cost-effective, scalable and uniform ecological monitoring solution, but its application in restoration remains largely untested. Here we show that metabarcoding of soil eDNA is effective at demonstrating the return of the native bacterial community in an old field following native plant revegetation. Bacterial composition shifted significantly after 8years of revegetation, where younger sites were more similar to cleared sites and older sites were more similar to remnant stands. Revegetation of the native plant community strongly impacted on the belowground bacterial community, despite the revegetated sites having a long and dramatically altered land-use history (i.e. >100years grazing). We demonstrate that metabarcoding of eDNA provides an effective way of monitoring changes in bacterial communities that would otherwise go unchecked with conventional monitoring of restoration projects. With further development, awareness of microbial diversity in restoration has significant scope for improving the efficacy of restoration interventions more broadly.