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Abstract Introduction Protists are unicellular eukaryotes including important predators, parasites and phototrophs, and play pivotal roles in organic matter decomposition, biogeochemical nutrient cycling and various ecosystem functions. Unravelling the impact of climate warming on soil protists is paramount in predicting how these microorganisms will continue to provide essential ecosystem services in the face of changing climatic conditions. Materials and Methods We examined the effects of 5‐year simulated climate warming scenario, with temperatures increased by 4°C above ambient levels, on the diversity and community composition of soil protists, as well as their interactions with other microorganisms in both natural and plantation forest ecosystems during three seasons: summer, autumn and winter. Results We found a season‐dependent response of protists to climate warming, with a significant decrease in protist diversity during winter in natural forests. Furthermore, we identified significant alterations in the community compositions of protists during summer in both natural and plantation forests, as well as during winter in both forest types, under warming. Our analysis pinpointed specific functional protist taxa, such as consumers, parasites and phototrophs, which exhibited significant shifts in their relative abundances under warming. Additionally, we found that warming facilitated trophic interactions between protists and bacteria, while also strengthening interactions between bacterial and fungal communities. Warming could induce direct modifications in protist community compositions or indirectly affect them by modifying bacterial and fungal communities, as revealed by structural equation modelling. Conclusion These findings demonstrate the substantial impact of warming on the taxonomic and functional compositions of protists in forest ecosystems, with the magnitude of these effects varying across seasons. Our study implicates that ongoing climate warming could have significant consequences for the profiles of soil protists, as well as their trophic interactions with bacteria and fungi, highlighting the importance of considering these effects for the sustainable provision of ecosystem functions.

ABSTRACTPlant-associated diazotrophs strongly relate to plant nitrogen (N) supply and growth. However, our knowledge of diazotrophic community assembly and microbial N metabolism in plant microbiomes is largely limited. Here we examined the assembly and temporal dynamics of diazotrophic communities across multiple compartments (soils, epiphytic and endophytic niches of root and leaf, and grain) of three cereal crops (maize, wheat, and barley) and identified the potential N-cycling pathways in phylloplane microbiomes. Our results demonstrated that the microbial species pool, influenced by site-specific environmental factors (e.g., edaphic factors), had a stronger effect than host selection (i.e., plant species and developmental stage) in shaping diazotrophic communities across the soil–plant continuum. Crop diazotrophic communities were dominated by a few taxa (~0.7% of diazotrophic phylotypes) which were mainly affiliated with Methylobacterium, Azospirillum, Bradyrhizobium, and Rhizobium. Furthermore, eight dominant taxa belonging to Azospirillum and Methylobacterium were identified as keystone diazotrophic taxa for three crops and were potentially associated with microbial network stability and crop yields. Metagenomic binning recovered 58 metagenome-assembled genomes (MAGs) from the phylloplane, and the majority of them were identified as novel species (37 MAGs) and harbored genes potentially related to multiple N metabolism processes (e.g., nitrate reduction). Notably, for the first time, a high-quality MAG harboring genes involved in the complete denitrification process was recovered in the phylloplane and showed high identity to Pseudomonas mendocina. Overall, these findings significantly expand our understanding of ecological drivers of crop diazotrophs and provide new insights into the potential microbial N metabolism in the phyllosphere.IMPORTANCEPlants harbor diverse nitrogen-fixing microorganisms (i.e., diazotrophic communities) in both belowground and aboveground tissues, which play a vital role in plant nitrogen supply and growth promotion. Understanding the assembly and temporal dynamics of crop diazotrophic communities is a prerequisite for harnessing them to promote plant growth. In this study, we show that the site-specific microbial species pool largely shapes the structure of diazotrophic communities in the leaves and roots of three cereal crops. We further identify keystone diazotrophic taxa in crop microbiomes and characterize potential microbial N metabolism pathways in the phyllosphere, which provides essential information for developing microbiome-based tools in future sustainable agricultural production.

Abstract Predatory protists are widely recognized as critical biotic forces driving soil microbial communities, but their top‐down controls on ammonia‐oxidizing microorganisms (AOMs), the major players in nitrification, are largely unresolved. Here, we investigated the communities of predatory protists and their associations with AOMs using high‐throughput sequencing and network analysis in soil aggregates following various long‐term organic substitutions. We found that organic substitutions increased while soil aggregation decreased the alpha diversity of predatory protists. Predatory protistan communities were significantly associated with AOMs. Variosea, an important group of Amoebozoa, were the keystone predatory protists associated with the AOMs. Collectively, our findings highlight the importance of predatory protists, especially Variosea, in regulating the communities of AOMs in an acidic Ultsisol, with implications for managing nitrification by predatory protists in agricultural soils.

Fungal plant pathogens threaten crop production and sustainable agricultural development. However, the environmental factors driving their diversity and nationwide biogeographic model remain elusive, impacting our capacity to predict their changes under future climate scenarios. Here, we analyzed potential fungal plant pathogens from 563 samples collected from 57 agricultural fields across China. Over 28.0% of fungal taxa in the phyllosphere were identified as potential plant pathogens, compared to 22.3% in the rhizosphere. Dominant fungal plant pathogen groups were Cladosporium (in the phyllosphere) and Fusarium (in the rhizosphere), with higher diversity observed in the phyllosphere than in rhizosphere soil. Deterministic processes played an important role in shaping the potential fungal plant pathogen community assembly in both habitats. Mean annual precipitation and temperature were the most important factor influencing phyllosphere fungal plant pathogen richness. Significantly negative relationships were found between fungal pathogen diversity and sorghum yield. Notably, compared to the rhizosphere, the phyllosphere fungal plant pathogen diversity played a more crucial role in sorghum yield. Together, our work provides novel insights into the factors governing the spatial patterns of fungal plant pathogens in the crop microbiome, and highlights the potential significance of aboveground phyllosphere fungal plant pathogens in crop productivity.

Abstract Introduction Shelterbelts, which provide protection for livestock and crops against severe weather conditions, have been recognised as important contributors to increased crop yields. Soil microorganisms play important roles in nutrient cycling, soil health and plant performance, thereby exerting significant influence on ecological services in terrestrial ecosystems. However, impacts of shelterbelts on soil microbial communities in pasture ecosystems remain poorly studied. Materials and Methods Here, we assessed the effect of shelterbelts (composed of Eucalyptus and native shrubs) on the pasture and nutrient productivity as well as soil bacterial and fungal communities at four distances from the shelterbelts with two different heights in two fields during spring and summer by applying amplicon sequencing, physicochemical and nutritional value analyses. Results The results showed that the shelterbelt height, distances from shelterbelts, seasons and their interactive effects substantially affected pasture yield, neutral detergent fibre, metabolisable energy and crude protein yields, but did not increase the pasture productivity in the rows adjacent to the shelterbelts. We also identified that the alpha diversity of bacteria and fungi did not significantly change across distances from the shelterbelts in both fields, but the community structure of soil bacteria and fungi was significantly influenced by distances from the shelterbelts. Interestingly, soil calcium (Ca) and shelterbelts were identified as top predictors of fungal community while soil pH and Ca was key drivers of bacterial community. Conclusion Our study provides novel knowledge in the impact of shelterbelts on pasture and nutritional production and soil microbial communities, which contributes to appropriate application and strategic management of the windbreak systems in agriculture production.

Abstract Hydrogen‐oxidising bacteria play a key role in maintaining the composition of gases within the atmosphere and are ubiquitous in agricultural soils. While studies have shown that hydrogen accumulates in soil surrounding legume nodules and the soil surface, soils as a whole act as a net sink for hydrogen, raising questions about how hydrogen is internally recycled by soils. Can the energy derived from hydrogen oxidation be directly funnelled into plants to promote their growth or does it only act as a booster for other plant‐growth promoting bacteria? Moreover, while the fertilisation effect of hydrogen on plants has previously been shown to be beneficial, questions remain about the upper limit of hydrogen uptake by plants before it becomes detrimental. Agricultural practices such as fertilisation may impact the balance of hydrogen‐oxidisers and hydrogen‐producers in these ecosystems, potentially having detrimental effects on not only agricultural land but also global biogeochemical cycles. In this perspectives piece, we highlight the importance of understanding the contribution of hydrogen to agricultural soils and the effects of agricultural practices on the ability for bacteria to cycle hydrogen in agricultural soils. We propose a framework to gain better insights into microbial hydrogen cycling within agroecosystems, which could contribute to the development of new agricultural biotechnologies.

Widespread forest fires pose significant challenges to the diverse communities of soil-dwelling organisms and the multiple ecosystem functions they support. However, whether the biodiversity and interactions of various taxonomic groups respond to fire consistently in terms of direction and extent, and their relative role in regulating post-fire soil functioning, remains largely unexplored. In this study, we investigated whether the diversity and co-occurrence networks of soil organisms across various trophic levels (including bacteria, fungi, protists, and invertebrates) in subtropical forests exhibit consistent responses to fire. Furthermore, we investigated their contribution to regulating soil multifunctionality, which is measured by a range of soil extracellular enzyme activities, available nutrients and reduced potential fungal plant pathogens. Our findings revealed that fire led to a decline in the richness of fungi, protists, and invertebrates, without significantly impacting bacterial richness. Fire also simplified the microbial co-occurrence networks while complexifying the invertebrate networks. Interestingly, soil multifunctionality tended to decrease with the richness of lower-trophic communities (i.e., bacteria), whereas it increased with that of high-trophic communities (i.e., protists and invertebrates). Moreover, fire indirectly influenced soil multifunctionality by altering biodiversity and network complexity, particularly pronounced in high-trophic communities. Overall, our results underscored the divergent vulnerability of biodiversity and networks to fires across taxa groups, highlighting the crucial role of biodiversity and interactions of higher trophic taxa groups in shaping the recovery and functionality of fire-affected soils.

This study investigated the abundance of comammox bacteria and canonical ammonia-oxidizing bacteria (AOB) and archaea (AOA), and their relative contribution to nitrification along a chronosequence of deglaciated forelands. The results showed that nitrification related gene abundance tended to increase with glacier retreat, with comammox bacteria and AOA appearing to be the most critical drivers for soil nitrification rates. These findings provide new evidence for the presence of comammox bacteria in glacier forelands and enhance our understanding of the niche differentiation of canonical nitrifier and comammox bacteria.

Abstract Soil contamination is one of the main threats to ecosystem health and sustainability. Yet little is known about the extent to which soil contaminants differ between urban greenspaces and natural ecosystems. Here we show that urban greenspaces and adjacent natural areas (i.e., natural/semi-natural ecosystems) shared similar levels of multiple soil contaminants (metal(loid)s, pesticides, microplastics, and antibiotic resistance genes) across the globe. We reveal that human influence explained many forms of soil contamination worldwide. Socio-economic factors were integral to explaining the occurrence of soil contaminants worldwide. We further show that increased levels of multiple soil contaminants were linked with changes in microbial traits including genes associated with environmental stress resistance, nutrient cycling, and pathogenesis. Taken together, our work demonstrates that human-driven soil contamination in nearby natural areas mirrors that in urban greenspaces globally, and highlights that soil contaminants have the potential to cause dire consequences for ecosystem sustainability and human wellbeing.

With the pressures of a changing global climate and ever-growing population, the need for sustainable agricultural practices that increase crop yields while decreasing greenhouse gas emissions are critical. Currently used practices to increase yields can often be problematic due to low nitrogen use efficiency or a potential overreliance on agrichemicals that can alter the community composition of a given ecosystem, although this is typically system and situation dependent. As such, the next generation of enhanced efficiency fertilisers that combine chemical, materials engineering and biological components are likely to be a game changer. Integral to their success is a better understanding of how plant–soil microbiomes interact with the new enhanced efficiency fertilisers, and how we can best tailor the fertilisers to suit different plant–soil combinations. In particular, the biodegradation properties of new fertiliser coatings must be given careful consideration so as to not further burden agricultural soils with microplastics or cause ecotoxicity problems. This perspective proposes novel, interdisciplinary strategies to generate highly efficient, biodegradable fertiliser coatings for use in the agricultural sector.

Abstract Background Little is known about the global distribution and environmental drivers of key microbial functional traits such as antibiotic resistance genes (ARGs). Soils are one of Earth’s largest reservoirs of ARGs, which are integral for soil microbial competition, and have potential implications for plant and human health. Yet, their diversity and global patterns remain poorly described. Here, we analyzed 285 ARGs in soils from 1012 sites across all continents and created the first global atlas with the distributions of topsoil ARGs. Results We show that ARGs peaked in high latitude cold and boreal forests. Climatic seasonality and mobile genetic elements, associated with the transmission of antibiotic resistance, were also key drivers of their global distribution. Dominant ARGs were mainly related to multidrug resistance genes and efflux pump machineries. We further pinpointed the global hotspots of the diversity and proportions of soil ARGs. Conclusions Together, our work provides the foundation for a better understanding of the ecology and global distribution of the environmental soil antibiotic resistome. Video Abstract

Abstract Soil pollution is an important stressor affecting biodiversity and ecosystem functioning. However, we lack a holistic understanding of how soil microbial communities respond to heavy metal pollution in agricultural ecosystems. Here, we explored the distribution patterns and inter-kingdom interactions of entire soil microbiome (including bacteria, fungi, and protists) in 47 paired paddy and upland fields along a gradient of legacy mercury (Hg) pollution. We found that the richness and composition of protistan community had stronger responses to Hg pollution than those of bacterial and fungal communities in both paddy and upland soils. Mercury polluted soils harbored less protistan phototrophs but more protistan consumers. We further revealed that long-term Hg pollution greatly increased network complexity of protistan community than that of bacterial and fungal communities, as well as intensified the interactions between protists and the other microorganisms. Moreover, our results consistently indicated that protistan communities had stronger responses to long-term Hg pollution than bacterial and fungal communities in agricultural soils based on structural equation models and random forest analyses. Our study highlights that soil protists can be used as bioindicators of Hg pollution, with important implications for the assessment of contaminated farmlands and the sustainable management of agricultural ecosystems.

Abstract Introduction Nitrification inhibitors and straw incorporation are used to reduce microbial mediated nitrogen (N) losses, thereby increasing N use efficiency (NUE) in cropland soil, but their effects on nitrous oxide (N2O) emission across different soil types and the underlying molecular mechanisms remain less understood. Materials and Methods In this study, we investigated how nitrification inhibitor nitrapyrin and straw incorporation affected soil N balance, N2O emission, nitrifiers and denitrifiers in three cropland soils (black, fluvo‐aquic and red soils). Results Compared with fertilization‐only treatment (N), nitrapyrin addition increased ammonium, while it decreased nitrate concentration in all soils; however, the performance of straw incorporation was less significant. Nitrapyrin decreased N2O emission only in the fluvo‐aquic soil, while straw incorporation strongly increased N2O emission in the red soil. Terminal restriction fragment length polymorphism analysis revealed that the denitrifier community was distinct across the three soils, but not strongly structured by nitrapyrin or straw incorporation. Compared with the N treatment, straw incorporation and nitrapyrin increased nosZ gene abundance only in the black soil. Structural equational modelling further confirmed that, after accounting for different soil properties, straw incorporation significantly increased N2O emission from black and red soils, and a decrease in nosZ gene abundance was the main biological factor for increased N2O emission in red soil. Conclusion Taken together, our work provides new knowledge that the performance of nitrification inhibitor and straw incorporation on N2O emission is soil type‐dependent, and management practices should be used as per soil type to balance NUE and N2O emission.

Summary It is a grand challenge to ensure the food security for a predicted world population of exceeding 9.7 billion by 2050, especially in an era of global climate change, land degradation and biodiversity loss. Current agricultural productions are mainly relying on synthetic chemical fertilisers to boost plant productivity but have undesirable effects on the environment and soil biodiversity. A promising direction in sustainable agriculture is to harness naturally occurring processes of beneficial plant‐associated microbiomes to ensure sustained crop production and global food security. Despite the significant progress made in the development of beneficial microbes as inoculants to enhance plant performance, challenges remain with the translation of knowledge of plant and soil microbiomes to successful microbial products in the agricultural sector. Here, we highlight how fertilizer technology should be renovated by harnessing microbiome‐based innovations to promote plant productivity and contribute to the end of hunger.

Abstract Introduction Fungal plant pathogens are an emerging threat to economically important crop production worldwide and a significant risk to global food security. However, we have limited knowledge of how agricultural management practices drive the emergence and spread of pathogens within crop microbiomes and the underlying ecological mechanisms. Materials and Methods We characterized the profiles of potential fungal plant pathogens, as well as bacterial and protistan communities, in sorghum phyllosphere, root endosphere, and rhizosphere and bulk soils collected from a long‐term experiment with multiple inorganic and organic fertilization regimes. Results We found contrasting patterns of fungal plant pathogens across the four sorghum–soil compartments and that organic fertilization regime significantly reduced the diversity and proportions of fungal plant pathogens in rhizosphere and bulk soils. We further found that the changes in fungal plant pathogens were driven more by resident bacterial and protistan communities than by soil physicochemical parameters. There was a significantly negative relationship between the diversity of fungal plant pathogens in the rhizosphere and bulk soils with sorghum yield and protein contents. Structural equation modeling revealed that long‐term organic fertilization regimes contributed to the suppression of fungal plant pathogens mainly through modulating the resident bacterial and protistan communities. Conclusion These findings advance our understanding of the responses of fungal plant pathogens in crop microbiomes to fertilization regimes, with implications for more targeted strategies to manage the impacts of fungal pathogens on plant health and economic losses.

Abstract Introduction Phyllosphere microbiome is intrinsically linked to plant performance and ecosystem functioning. However, our knowledge about the role of ecological processes in shaping the biogeography of different phyllosphere microbial communities is scarce. Materials and Methods Here, we compared the biogeographic patterns of bacterial and fungal communities in phyllosphere samples of plants belonging to the genus of Eucalyptus of the Myrtaceae family collected from an over 4000 km transect. We investigated the relative importance of two major ecosystem processes (stochastic vs. deterministic) in shaping phyllosphere microbial community assemblies. Results Our results demonstrated that the neutral community model, which can quantify the degree of a community assembly determined by stochastic processes, explained 64.2% of the variations in bacterial community assembly, which had a normalized stochasticity ratio of 71.8%. These results suggest that the dominant role of stochastic processes in shaping bacterial community assembly. In contrast, phyllosphere fungal community assembly was mainly shaped by deterministic processes as revealed by a relatively small explanation rate of the neutral community model (48.7%) and a normalized stochasticity ratio of 25.1%. Variation partitioning analysis and random forest modelling results indicated that climatic factors, particularly mean annual precipitation and aridity index, were important in driving both bacterial and fungal biogeographic patterns in the phyllosphere. Edaphic factors, such as soil organic and mineral nitrogen content, were more closely related to fungal community assembly than to bacterial community assembly. The differential responses of bacterial and fungal communities to environmental factors could be attributed to the different traits of bacteria and fungi, that is, the higher potential dispersal rate and wider habitat niche of bacteria than fungi. Conclusion Our findings demonstrated that phyllosphere bacterial and fungal communities followed distinct community assembly processes, which is supported by the ‘size plasticity’ hypothesis that smaller organisms (bacteria) are less influenced by environmental conditions and are more homogeneous across space than larger organisms (fungi). These findings provide new insights into the microbial ecology of plant phyllosphere microbiomes.

Rice-crayfish co-culture (RC) has been widely and rapidly promoted as a sustainable agricultural system in many countries. The accumulation of crayfish residues could enhance soil organic matters; however, impacts of this integrated farming model on the dissemination and pathogenicity of resistance and virulence genes remain poorly understood. Here, we characterized antibiotic resistance genes (ARGs), biocide resistance genes (BRGs), metal resistance genes (MRGs) and virulence factor genes (VFGs) using metagenomic methods in paired RC and rice monoculture (RM) systems across China. The RC model did not increase the abundance of soil ARGs, BRGs, MRGs, or VFGs in comparison to the RM model, but selectively enriched 35 subtypes of these potential resistance and virulence genes. Network analysis revealed that resistance and virulence genes had a higher number of connections with mobile genetic elements (MGEs) in the RC system than that in the RM system, suggesting a higher horizontal transfer potential of these genes. Moreover, the RC model had a higher abundance of human opportunistic pathogens such as Salmonella enterica, Vibrio cholerae, and Shigella dysenteriae which were potential hosts of VFGs such as phoP, fleS, and gspE, suggesting a potential threat to human health. We further unraveled that stochastic process was the main driver of the assembly of resistance and virulence genes in the RC system. The abundance of ARGs and VFGs were primarily associated with microbial community compositions, while the abundance of BRGs and MRGs were mainly associated with that of MGEs. Taken together, our results suggest that the RC model has potential to cause the dissemination and pathogenicity of resistance and virulence genes, which has important implications for the control of soil-borne biological risks and the strategic management of sustainable agriculture.

The excessive usage of nitrogen (N) fertilizers can accelerate the tendency of global climate change. Biological N fixation by diazotrophs contributes substantially to N input and is a viable solution to sustainable agriculture via reducing inorganic N fertilization. However, how manure application influences the abundance, community structure and assembly process of diazotrophs in soil aggregates is not fully understood. Here, we investigated the effect of manure amendment on diazotrophic communities in soil aggregates of an arable soil. Manure application increased soil aggregation, crop yield and the abundance of nifH genes. The abundance of nifH genes increased with aggregate sizes, indicating that diazotrophs prefer to live in larger aggregates. The abundance of nifH genes in large macroaggregates, rather than in microaggregates and silt and clay, was positively associated with plant biomass and crop yield. Both manure application and aggregate size did not alter the Shannon diversity of diazotrophs but significantly changed the diazotrophic community structure. The variation of diazotrophic community structure explained by manure application was greater than that by aggregate size. Manure application promoted the relative abundance of Firmicutes but reduced that of α-Proteobacteria. Stochastic processes played a dominant role in the assembly of diazotrophs in the control treatment. Low-rate manure (9 Mg ha−1) application, rather than medium-rate (18 Mg ha−1) and high-rate (27 Mg ha−1) manure, significantly increased the relative importance of deterministic processes in diazotrophic community assembly. Taken together, our findings demonstrated that long-term manure application increased nifH gene abundance and altered the community structure and assembly process of diazotrophs in soil aggregates, which advanced our understanding of the ecophysiology and functionality of diazotrophs in acidic Ultisols.

ABSTRACT Deciphering the relationships between microbes and their host plants is critical for a better understanding of microbial diversity maintenance and community stability. Here, we investigated fungal diversity and community assembly in the phyllosphere and rhizosphere of 13 tree species in a subtropical common-garden experiment. The results showed that fungal community structures significantly differed across compartments (leaf, root, and soil) and different tree species. Higher α-diversity was observed in the phyllosphere than in the roots and rhizospheric soil. Fungal community composition (β-diversity) was significantly affected by both compartment and species identity. The fungal community compositions were significantly correlated with soil pH in the roots and the soils as well as with soil nitrate and leaf total phosphorus in the leaves. We found that fungal community assemblies were mainly driven by deterministic processes, regardless of compartments. Moreover, host preference analyses indicated that stronger plant/fungus preferences occurred in leaves than in roots and soils. Our results highlight the differences in tree mycobiome between aboveground and belowground compartments and have important implications for the promotion of biodiversity conservation and management sustainability for the subtropical forest. IMPORTANCE Subtropical mountain forests are widely distributed in Southern China and are characterized by high biodiversity. The interactions between plants and fungi play pivotal roles in biodiversity maintenance and community stability. Nevertheless, knowledge of fungal diversity and of the community assembly patterns of woody plants is scarce. Here, we investigated fungal diversity and community assembly in the phyllosphere and rhizosphere of 13 tree species in a common-garden experiment. We found that both compartment and plant identity influenced fungal diversity, community, and guild compositions, while deterministic processes mainly governed the fungal community assembly, especially in the rhizospheric fungal communities. Our results demonstrate that tree leaves represent stronger host/fungi preferences than do roots and soils. Together, our findings enhance the understanding of the roles of compartment and plant identity in structuring fungal communities as well as promote fungal diversity maintenance in subtropical mountain forest ecosystems.

ABSTRACT Livestock wastes contain high levels of antibiotic resistance genes (ARGs) and a variety of human-related pathogens. Bioconversion of livestock manure using larvae of the beetle Protaetia brevitarsis is an effective technique for waste reduction and value creation; however, the fate of manure ARGs during gut passage and interaction with the gut microbiome of P. brevitarsis remains unclear. To investigate this, we fed P. brevitarsis with dry chicken manure for 6 days and measured bacterial community dynamics and ARG abundance and diversity along the P. brevitarsis gut tract using high-throughput quantitative PCR and metagenomics approaches. The diversity of ARGs was significantly lower in larval midgut, hindgut, and frass than in raw chicken manure, and around 80% of pathogenicity-related genes (PRGs) exhibited reduced abundance. Network analysis demonstrated that Bacteroidetes and Firmicutes were the key bacterial phyla associated with ARG reduction. Metagenomic analysis further indicated that ARGs, mobile genetic elements (MGEs), and PRGs were simultaneously attenuated in the hindgut, implicating a decreased likelihood for horizontal gene transfer (HGT) of ARGs among bacteria and pathogens during manure bioconversion. Our findings demonstrated that the attenuation of ARGs is strongly associated with the variation of the gut microbiome of P. brevitarsis, providing insights into mechanisms of risk mitigation of ARG dissemination during manure bioconversion. IMPORTANCE Saprophagous fauna like the oriental edible beetle (P. brevitarsis) plays a fundamental role in converting organic wastes into biofertilizer. Accumulating evidence has shown that soil fauna can reduce the abundance of ARGs, although the underlying mechanism of ARG reduction is still unclear. In our previous research, we found a large reduction of ARGs in vegetable roots and leaves from frass compared with raw manure, providing a promising biofertilizer for soil-vegetable systems. Therefore, in this study, temporal dynamic changes in the microbiomes of the donor (chicken manure) and host (P. brevitarsis) were investigated, and we found a close association between the gut microbiome and the alteration of ARGs. These results shed new light on how the insect gut microbiome can mitigate manure-borne ARGs and provide insights into the bioconversion process via a typical member of the saprophagous fauna, P. brevitarsis.

Field application of organic fertilizers may promote the spread of antibiotic resistance genes (ARGs) in farmland ecosystems, which represents a major threat to public health. However, impacts of multi-year organic fertilization on the spread of soil and phyllosphere ARGs and the possible transmission pathways remain elusive. Here, we investigated the patterns of ARGs and structures of bacterial and fungal communities in soil and wheat crop phyllosphere, under long-term fertilization of swine manure or sewage sludge using high-throughput quantitative PCR and amplicon sequencing. It was found that application of swine manure and sewage sludge had more influence on the prevalence of ARGs in phyllosphere than in soil. SourceTracker analysis revealed that a few bacterial taxa from swine manure and sewage sludge could survive in soil, but hardly in phyllosphere. Mantel test and variation partitioning analysis further revealed that shift in the microbial community compositions was an important factor affecting soil ARGs, while horizontal gene transfer mediated by mobile genetic elements might contribute to the spread of phyllosphere ARGs. Overall, these results demonstrated that long-term application of swine manure and sewage sludge differently impacts the ARGs in soil and phyllosphere, which have implications for sustainable agricultural management.

ABSTRACT Soil-borne fungal phytopathogens are important threats to soil and crop health. However, their community composition and environmental determinants remain unclear. Here, we explored the effects of agricultural fertilization regime (i.e., organic material application) on soil fungal phytopathogens, using data sets from a combination of field survey and long-term experiment. We found that soil organic carbon was the key factor that affected the diversity and relative abundance of fungal phytopathogens in agricultural soils. The dominant genera of phytopathogens including Monographella was also strongly associated with soil organic carbon. In addition, the elevated soil organic carbon enhanced the node proportion of phytopathogens and the positive interactions within the fungal community in the network. Results of the long-term experiment revealed that applications of crop straw and fresh livestock manure significantly increased the proportion of phytopathogens, which were associated with the elevated soil organic carbon. This work offers new insights into the occurrence and environmental factors of fungal phytopathogens in agricultural soils, which are fundamental to control their impacts on the soil and crop systems. IMPORTANCE Fungal phytopathogens are important threats to soil and crop health, but their community composition and environmental determinants remain unclear. We found that soil organic carbon is the key factor of the prevalence of fungal phytopathogens through a field survey, which is also supported by our long-term (6-year) experiment showing the applications of crop straw and fresh livestock manure significantly increased the proportion of fungal phytopathogens. These findings advance our understanding of the occurrence and environmental drivers of soil-borne fungal phytopathogens under agricultural fertilization regime and have important implications for the control of soil-borne pathogens.

Soil age is thought to be an important driver of ecosystem development. Here, the authors perform a global survey of soil chronosequences and meta-analysis to show that, contrary to expectations, soil age is a relatively minor ecosystem driver at the biome scale once other drivers such as parent material, climate, and vegetation type are accounted for.

Fungi regulate nutrient cycling, decomposition, symbiosis, and pathogenicity in cropland soils. However, the relative importance of generalist and specialist taxa in structuring soil fungal community remains largely unresolved. We hypothesized that generalist fungi, which are adaptable to various environmental conditions, could potentially dominate the community and become the basis for fungal coexisting networks in cropping systems. In this study, we identified the generalist and habitat specialist fungi in cropland soils across a 2,200 kms environmental gradient, including three bioclimatic regions (subtropical, warm temperate, and temperate). A few fungal taxa in our database were classified as generalist taxa (~1%). These generalists accounted for >35% of the relative abundance of all fungal populations, and most of them are Ascomycota and potentially pathotrophic. Compared to the specialist taxa (5–17% of all phylotypes in three regions), generalists had a higher degree of connectivity and were often identified as hub within the network. Structural equation modeling provided further evidence that after accounting for spatial and climatic/edaphic factors, generalists had larger contributions to the fungal coexistence pattern than habitat specialists. Taken together, our study provided evidence that generalist taxa are crucial components for fungal community structure. The knowledge of generalists can provide important implication for understanding the ecological preference of fungal groups in cropland systems.

Soil microbial communities play a central role in driving multiple ecosystem functions and ecological processes that are key to maintaining the plant productivity. However, we lack sound evidence for the linkage between soil microbial diversity and plant productivity, which hinders our ability to predict the consequences of microbial diversity loss for food security under the context of global environmental change. Here, we used the dilution-to-extinction approach to test the consequences of soil microbial diversity loss for the aboveground plant biomass in a glasshouse experiment. Compared with original soils, the bacterial alpha-diversity (Observed operational taxonomic units and Shannon index) significantly decreased in treatments with serially diluted inoculum. Principal coordinates analysis showed that the overall bacterial community compositions (beta-diversity) in original soils were clearly separated from the treatments with serially diluted inoculum. The aboveground biomass of lettuce harvested from the original soils was significantly higher than that from the sterilized soils regardless of the inoculation. The ordinary least squares regression model showed a significant linear relationship between the plant biomass and bacterial alpha-diversity, indicating that reduction in soil microbial diversity could result in a significant decline in the biomass of lettuce. No significant correlation was observed between plant biomass and soil processes including soil basal respiration and denitrification rates. Structural equation models suggested that the effects of soil microbial diversity on the plant biomass were maintained even when simultaneously accounting for other drivers (soil properties and biological processes). Our study provides experimental evidence that soil microbial diversity is important to the maintenance of the plant productivity and suggests that the functional redundancy in soil microbial communities may be overestimated especially in the agroecological system.

Urban green spaces are closely related to the activities and health of urban residents. Turf grass and soil are two major interfaces between the environmental and human microbiome, which represent potential pathways for the spread of antibiotic resistance genes (ARGs) from environmental to human microbiome through skin-surface contact. However, the information regarding the prevalence of ARGs in urban green spaces and drivers in shaping their distribution patterns remain unclear. Here, we profiled a wide spectrum of ARGs in grass phyllosphere and soils from 40 urban parks across Greater Melbourne, Australia, using high throughput quantitative PCR. A total of 217 and 218 unique ARGs and MGEs were detected in grass phyllosphere and soils, respectively, conferring resistance to almost all major classes of antibiotics commonly used in human and animals. The plant microbiome contained a core resistome, which occupied >84% of the total abundance of ARGs. In contrast, no core resistome was identified in the soil microbiome. The difference between plant and soil resistome composition was attributed to the difference in bacterial community structure and intensity of environmental and anthropogenic influence. Most importantly, the abundance of ARGs in urban green spaces was significantly positively related to industrial factors including total number of business, number of manufacturing, and number of electricity, gas, water and waste services in the region. Structural equation models further revealed that industrial distribution was a major factor shaping the ARG profiles in urban green spaces after accounting for multiple drivers. These findings have important implications for mitigation of the potential risks posed by ARGs to urban residents. Keywords: Resistome, Urban green spaces, Phyllosphere, Anthropogenic activities, Public health

The increasing antimicrobial resistance in manure-amended soil can potentially enter food chain, representing an important vehicle for antibiotic resistance genes (ARGs) transmission into human microbiome. However, the pathways for transmission of ARGs from soil to plant remain unclear. Here, we explored the impacts of poultry and cattle manure application on the patterns of resistome in soil and lettuce microbiome including rhizosphere, root endosphere, leaf endosphere and phyllosphere, to identify the potential transmission routes of ARGs in the soil-plant system. After 90 days of cultivation, a total of 144 ARGs were detected in all samples using high-throughput quantitative PCR. Rhizosphere soil samples harbored the most diverse ARGs compared with other components of lettuce. Cattle manure application increased the abundance of ARGs in root endophyte, while poultry manure application increased ARGs in rhizosphere, root endophyte and phyllosphere, suggesting that poultry manure may have a stronger impact on lettuce resistomes. The ARG profiles were significantly correlated with the bacterial community, and the enrichment of soil and plant resistomes was strongly affected by the bacterial taxa including Solibacteres, Chloroflexi, Acidobacteria, Gemm-1 and Gemmatimonadetes, as revealed by the network analyses. Moreover, the overlaps of ARGs between lettuce tissues and soil were identified, which indicated that plant and environmental resistomes are interconnected. Our findings provide insights into the transmission routes of ARGs from manured soil to vegetables, and highlight the potential risks of plant resistome migration to the human food chain. Keywords: Antibiotic resistance genes, Manure-amended soil, Plant resistome, Endophyte, Phyllosphere

Primary succession of plant and microbial communities in the glacier retreating foreland has been extensively studied, but shifts of antibiotic resistance genes (ARGs) with the glacier retreating due to global warming remain elusive. Unraveling the diversity and succession features of ARGs in pristine soil during glacier retreating could contribute to a mechanistic understanding of the evolution and development of soil resistome. In this study, we quantified the abundance and diversity of ARGs along a 50-year soil development chronosequence by using a high-throughput quantitative PCR (HT-qPCR) technique. A total of 24 ARGs and two mobile genetic elements (MGEs) were detected from all the glacier samples, and the numbers of detected ARGs showed a unimodal pattern with an increasing trend at the early stage (0∼8 years) but no significant change at later stages (17∼50 years). The oprJ and mexF genes encoding multidrug resistance were the only two ARGs that were detected across all the succession ages, and the mexF gene showed an increasing trend along the succession time. Structural equation models indicated the predominant role of the intI1 gene encoding the Class 1 integron-integrase in shaping the variation of ARG profiles. These findings suggested the presence of ARGs in pristine soils devoid of anthropogenic impacts, and horizontal gene transfer mediated by MGEs may contribute to the succession patterns of ARGs during the initial soil formation stage along the chronosequence.

Rising concern about the use of antibiotics in food production has resulted in many studies on the occurrence of antibiotic resistance genes (ARGs) in animal-associated bacterial communities. There are few baseline data on the abundance of ARGs on farms where chickens are intensively raised with little or no use of antibiotics. This study used a high-throughput quantitative PCR array to survey two antibiotic-free chicken farms for the occurrence of ARGs and mobile genetic elements known to enhance the spread of ARGs. No antibiotics had been used on the study farms for five years prior to this study. The results provide a baseline for the occurrence of resistance genes in the chicken production system without direct selective pressure.