Changes in Transformation of Soil Organic C and Functional Diversity of Soil Microbial Community Under Different Land Uses

Changes in soil biological and biochemical properties under different land uses in the subtropical region of China were investigated in order to develop rational cultivation and fertilization management. A small watershed of subtropical region of China was selected for this study. Land uses covered paddy fields, vegetable farming, fruit trees, upland crops, bamboo stands, and forestry. Soil biological and biochemical properties included soil organic C and nutrient contents, mineralization of soil organic C, and soil microbial biomass and community functional diversity. Soil organic C and total N contents, microbial biomass C and N, and respiration intensity under different land uses were changed in the following order： paddy fields （and vegetable farming） 〉 bamboo stands 〉 fruit trccs （and upland）. The top surface （0-15 cm） paddy fields （and vegetable farming） were 76.4 and 80.8% higher in soil organic C and total N contents than fruit trees （and upland） soils, respectively. Subsurface paddy soils （15-30 cm） were 59.8 and 67.3% higher in organic C and total N than upland soils, respectively. Soil microbial C, N and respiration intensity in paddy soils （0-15 cm） were 6.36, 3.63 and 3.20 times those in fruit tree （and upland） soils respectively. Soil microbial metabolic quotient was in the order： fruit trees （and upland） 〉 forestry 〉 paddy fields. Metabolic quotient in paddy soils was only 47.7% of that in fruit tree （and upland） soils. Rates of soil organic C mineralization during incubation changed in the order： paddy fields 〉 bamboo stands 〉 fruit trees （and upland） and soil bacteria population： paddy fields 〉 fruit trees （and upland） 〉 forestry. No significant difference was found for fungi and actinomycetes populations. BIOLOG analysis indicated a changing order of paddy fields 〉 fruit trees （and upland） 〉 forestry in values of the average well cell development （AWCD） and functional diversity indexes of microbial community. Results also showed that the conversion from paddy fields to vegetable farming for 5 years resulted in a dramatic increase in soil available phosphorus content while insignificant changes in soil organic C and total N content due to a large inputs of phosphate fertilizers. This conversion caused 53, 41.5, and 41.3% decreases in soil microbial biomass C, N, and respiration intensity, respectively, while 23.6% increase in metabolic quotient and a decrease in soil organic C mineralization rate. Moreover, soil bacteria and actinomycetes populations were increased slightly, while fungi population increased dramatically. Functional diversity indexes of soil microbial community decreased significantly. It was concluded that land uses in the subtropical region of China strongly affected soil biological and biochemical properties. Soil organic C and nutrient contents, mineralization of organic C and functional diversity of microbial community in paddy fields were higher than those in upland and forestry. Overuse of chemical fertilizers in paddy fields with high fertility might degrade soil biological properties and biochemical function, resulting in deterioration of soil biological quality.

Microbial community structure and functional metabolic diversity are associated with organic carbon availability in an agricultural soil

Exploration of soil environmental characteristics governing soil microbial community structure and activity may improve our understanding of biogeochemical processes and soil quality. The impact of soil environmental characteristics especially organic carbon availability after 15-yr different organic and inorganic fertilizer inputs on soil bacterial community structure and functional metabolic diversity of soil microbial communities were evaluated in a 15-yr fertilizer experiment in Changping County, Beijing, China. The experiment was a wheat-maize rotation system which was established in 1991 including four different fertilizer treatments. These treatments included： a non-amended control（CK）, a commonly used application rate of inorganic fertilizer treatment（NPK）; a commonly used application rate of inorganic fertilizer with swine manure incorporated treatment（NPKM）, and a commonly used application rate of inorganic fertilizer with maize straw incorporated treatment（NPKS）. Denaturing gradient gel electrophoresis（DGGE） of the 16 S r RNA gene was used to determine the bacterial community structure and single carbon source utilization profiles were determined to characterize the microbial community functional metabolic diversity of different fertilizer treatments using Biolog Eco plates. The results indicated that long-term fertilized treatments significantly increased soil bacterial community structure compared to CK. The use of inorganic fertilizer with organic amendments incorporated for long term（NPKM, NPKS） significantly promoted soil bacterial structure than the application of inorganic fertilizer only（NPK）, and NPKM treatment was the most important driver for increases in the soil microbial community richness（S） and structural diversity（H）. Overall utilization of carbon sources by soil microbial communities（average well color development, AWCD） and microbial substrate utilization diversity and evenness indices（H＇ and E） indicated that long-term inorganic fertilizer with organic amendments incorporated（NPKM, NPKS） could significantly stimulate soil microbial metabolic activity and functional diversity relative to CK, while no differences of them were found between NPKS and NPK treatments. Principal component analysis（PCA） based on carbon source utilization profiles also showed significant separation of soil microbial community under long-term fertilization regimes and NPKM treatment was significantly separated from the other three treatments primarily according to the higher microbial utilization of carbohydrates, carboxylic acids, polymers, phenolic compounds, and amino acid, while higher utilization of amines/amides differed soil microbial community in NPKS treatment from those in the other three treatments. Redundancy analysis（RDA） indicated that soil organic carbon（SOC） availability, especially soil microbial biomass carbon（Cmic） and Cmic/SOC ratio are the key factors of soil environmental characteristics contributing to the increase of both soil microbial community structure and functional metabolic diversity in the long-term fertilization trial. Our results showed that long-term inorganic fertilizer and swine manure application could significantly improve soil bacterial community structure and soil microbial metabolic activity through the increases in SOC availability, which could provide insights into the sustainable management of China＇s soil resource.

Response of soil bacterial community to biochar application in a boreal pine forest

Boreal forests commonly suff er from nitrogen defi ciency due to low rate of nitrogen mineralization.Biochar may promote soil organic matter decomposition and accelerate nitrogen mineralization.In this study,Illumina NovaSeq sequencing combined with functional annotation of prokaryotic taxa(FAPROTAX)analysis was used to investigate the eff ect of biochar pyrolysis temperatures,the amount of applied biochar,and the period since the biochar application(2-and 3-year)on soil bacterial communities.The results show that biochar pyrolysis temperatures(500℃ and 650℃)and the amount of applied biochar(0.5 kg m^(−2)and 1.0 kg m^(−2))did not change soil properties.Nevertheless,the interaction of biochar pyrolysis temperature and the amount had signifi cant eff ects on bacterial species richness and evenness(P<0.05).The application of biochar produced at 500℃ had a lower abundance of Actinobacteria and Verrucomicrobia,while that produced at 650℃ had a higher abundance of Conexibacter and Phenylobacterium.When biochar produced at 650℃ was applied,applying 0.5 kg m^(−2)had a higher abundance of Cyanobacteria,Conexibacter,and Phenylobacterium than that of 1.0 kg m^(−2)(P<0.05).Functionally,the abundance of the aromatic compound degradation group increased with the extension of application time and increase of pyrolysis temperature.The time since application played an important role in the formation of soil bacterial communities and their functional structure.Long-term studies are necessary to understand the consequence of biochar on bacterial communities in boreal forests.

Microbial Community Composition and Function in Sediments from the Pearl River Mouth Basin

This study was conducted to characterize the diversity and function of microbial communities in marine sediments of the Pearl River Mouth Basin(PRMB)in the South China Sea.The results showed that the bacterial and archaeal communities varied greatly with depth.Proteobacteria in bacterial communities and Nitrososphaeria and Woesearchaeota in archaeal communities were dominant in the shallow sediments(1-40 cm),while Chloroflexi in bacterial communities and Bathyarchaeia in archaeal communities were dominant in the deep sediments(50-200 cm).Regarding ecological functions based on the metatranscriptomic data,genes involved in various pathways of nitrogen metabolism and sulfur metabolism were observed in the tested sediment samples.Metagenomic analysis revealed that Proteobacteria contribute the most to nearly all genes involved in nitrogen and sulfur metabolism.Moreover,Thaumarchaeota contribute the most to certain genes involved in nitrification,denitrification and assimilatory sulfate reduction pathways.The most abundant bacterial genus,Candidatus Scalindua,is crucial for nitrification,dissimilatory nitrate reduction,denitrification and assimilatory sulfate reduction pathways.

Effect of pine wood nematode invasion on pine community functions in the Pinghu region, Zhejiang Province, eastern China

In order to investigate the effect of invastion by pine wood nematode(PWN), this study analyzed several functional indices, i.e., the increment in DBH and stand volume and biomass, in the damaged stands with various mixed percentages of Pinus massoniana and P. thunbergii and with different levels of damage. According to the results of rate of change in increment of DBH and stand volume, the forest ecosystem resistance against PWN increased with a reduction in the mixed ratio of pine. The resistance was highest with a mixed percentage of 50%. The invasion of PWN changed the corresponding relationship of increment between DBH and stand volume(pure stands > 7:3 conifer and broadleaf > 6:4 conifer and broadleaf > 5:5 conifer and broadleaf) among the P. thunbergii stands when there is no damage, but for P. massoniana stands this phenomenon did not occur. For the increment rate of DBH and stand volume, this significant change in P. thunbergii forest indicates that the resistance of pure P. thunbergii forest was higher than that of P. massoniana. The invasion of PWN accelerates the succession from pure stands to mixed stands and then to the broadleaf evergreen stands.

Partial function prediction of sulfate-reducing bacterial community from the rhizospheres of two typical coastal wetland plants in China

Sulfate-reducing bacteria(SRB)are ubiquitous anaerobic microorganisms that play signifi cant roles in the global biogeochemical cycle.Coastal wetlands,one of the major habitats of SRB,exhibit high sulfate-reducing activity and thus play signifi cant roles in organic carbon remineralization,benthic geochemical action,and plant-microbe interactions.Recent studies have provided credible evidence that the functional rather than the taxonomic composition of microbes responds more closely to environmental factors.Therefore,in this study,functional gene prediction based on PacBio single molecular real-time sequencing of 16S rDNA was applied to determine the sulfate-reducing and organic substrate-decomposing activities of SRB in the rhizospheres of two typical coastal wetland plants in North and South China:Zostera japonica and Scirpus mariqueter.To this end,some physicochemical characteristics of the sediments as well as the phylogenetic structure,community composition,diversity,and proportions of several functional genes of the SRB in the two plant rhizospheres were analyzed.The Z.japonic a meadow had a higher dissimilatory sulfate reduction capability than the S.mariqueter-comprising saltmarsh,owing to its larger proportion of SRB in the microbial community,larger proportions of functional genes involved in dissimilatory sulfate reduction,and the stronger ability of the SRB to degrade organic substrates completely.This study confi rmed the feasibility of applying microbial community function prediction in research on the metabolic features of SRB,which will be helpful for gaining new knowledge of the biogeochemical and ecological roles of these bacteria in coastal wetlands.

健康功能社区建设评价指标体系构建

目的 构建一套能指引健康功能社区建设并能较为客观地反映建设成效的评价指标体系。方法 通过文献回顾、小组讨论和专家会议法初步确定指标体系框架和指标池,在此基础上通过3轮专家咨询确定评价指标体系,采用层次分析法确定各指标权重。结果 健康功能社区建设评价指标包括4项基本条件,健康服务、健康环境、健康人群、健康文化、组织管理5项一级指标及14项二级指标、63项三级指标。3轮咨询的专家积极系数均为100%,专家平均权威系数为0.84,协调系数分别为0.179、0.232、0.309(P < 0.05)。层次分析的一级指标权向量为0.177 ~ 0.211,最大特征值为5.000 ~ 5.218;二级指标的权向量为0.035 ~ 0.105,最大特征值为2.000 ~ 4.201;三级指标的权向量为0.003 ~ 0.105,最大特征值为1.000 ~ 14.529。判断矩阵的一致性比率均低于0.100。结论 3轮咨询的专家代表性、积极性、权威程度和协调程度较高,判断矩阵的一致性比率符合要求,构建的指标体系可为推进健康功能社区建设提供指引,今后还需结合实证研究不断予以完善。

Long-term rice-rice-green manure rotation changing the microbial communities in typical red paddy soil in South China

On the basis of a long-term（30 years） field experiment that involved four rotation systems, rice-rice-winter fallow（RRF）, rice-rice-ryegrass（RRG）, rice-rice-rape（RRP）, and rice-rice-milk vetch（RRV）, this study described the effects of green manure on the microbial communities in the red paddy soils using 454 pyrosequencing for the 16 S r RNA gene. The Chao1 richness and non-parametric Shannon＇s index increased in all soil samples that received green manure treatments. The communities＇ structures with the green manure applications were significantly dissimilar from that under the winter fallow. Using Metastats tests, many genera in the RRG, RRP and RRV soils were significantly different from those in the RRF soil, including a number of genera that functioned in the nitrogen and sulfur cycles. Analyses of the genera with these functions revealed the shifts in microbial ecosystem functions after long-term green manuring. Changes in the microbial communities increased the ammonium supply and decreased the soil acidification in green-manure-amended soils. Together, these data suggested powerful effects of green manure on both the microbial communities and the biogeochemical cycle driven by the shifts in bacterial functional groups.

Hydrological variability and long-term floristic-structural modifications in different habitats of a tropical semi-deciduous forest

Shifts in hydrological regimes alter river flow rates and flood pulses, decrease environmental heterogeneity and the floristic-structural complexity of associated plant communities. We tested the hypothesis that drought events affect plant community composition and structure at a small-scale within a riparian fragment towards a reduction in floristic-structural complexity. The tree community was sampled in three habitats (wet, transitional and dry) and monitored in seven inventories carried out between 1991 and 2018. Hydrological variations were evaluated through annual rainfalls, river flow rates and water level data. The species richness and the detrended correspondence analysis axes were used to characterise the temporal modifications in floristic composition. Community structure was described in terms of biomass: accumulated, growth of survivors, mortality and recruitment. Generalised linear mixed models were fitted to evaluate the effects of time and environment in community. It was concluded that the climate has become drier in recent years due to declining precipitation that has affected flow rates and water levels. The floristic-structural complexity of the study fragment was maintained during the monitoring period. However, prolonged and extreme drought events displayed the potential to impact floristic-structural patterns.

Urbanization reduces resource use efficiency of phytoplankton community by altering the environment and decreasing biodiversity

Urbanization often exerts multiple effects on aquatic and terrestrial organisms,including changes in biodiversity,species composition and ecosystem functions.However,the impacts of urbanization on river phytoplankton in subtropical urbanizing watersheds remain largely unknown.Here,we explored the effects of urbanization on phytoplankton community structure(i.e.,biomass,community composition and diversity)and function(i.e.,resource use efficiency)in a subtropical river at watershed scale in southeast China over 6 years.A total of 318 phytoplankton species belonging into 120 genera and 7 phyla were identified from 108 samples.Bacillariophyta biomass showed an increasing trend with increasing urbanization level.The phytoplankton community shifted from Chlorophyta dominance in rural upstream waters to Bacillariophyta dominance in urbanized downstream waters.Furthermore,phytoplankton diversity and resource use efficiency(RUE=phytoplankton biomass/total phosphorus)were significantly decreased with increasing urbanization level from upstream to downstream.Phytoplankton RUE exhibited a significant positive correlation with species richness,but a negative correlation with phytoplankton evenness.The variation in environmental factors(turbidity,total nitrogen,NH_(4)^(+)-N,total phosphorus,PO_(4)^(3-)-P and percentage urbanized area)was significantly correlated with phytoplankton diversity and RUE.Overall,our results revealed the influence of urbanization on phytoplankton community structure and ecosystem function was due to its altering the environmental conditions.Therefore,human-driven urbanization may play crucial roles in shaping the structure and function of phytoplankton communities in subtropical rivers,and the mechanism of this process can provide important information for freshwater sustainable uses,watershed management and conservation.

Impacts of produced water origin on bacterial community structures of activated sludge

The purpose of this study was to reveal how activated sludge communities respond to influent quality and indigenous communities by treating two produced waters from different origins in a batch reactor in succession. The community shift and compositions were investigated using Polymerase Chain Reaction–denaturing gradient gel electrophoresis（PCR–DGGE） and further16 S ribosomal DNA（r DNA） clone library analysis. The abundance of targeted genes for polycyclic aromatic hydrocarbon（PAH） degradation, nah Ac/phn Ac and C12O/C23 O, was tracked to define the metabolic ability of the in situ microbial community by Most Probable Number（MPN） PCR. The biosystem performed almost the same for treatment of both produced waters in terms of removals of chemical oxygen demand（COD） and PAHs. Sludge communities were closely associated with the respective influent bacterial communities（similarity 〉 60%）, while one sludge clone library was dominated by the Betaproteobacteria（38%） and Bacteriodetes（30%）and the other was dominated by Gammaproteobacteria（52%）. This suggested that different influent and water quality have an effect on sludge community compositions. In addition, the existence of catabolic genes in sludge was consistent with the potential for degradation of PAHs in the treatment of both produced waters.

Impacts of methamidophos, copper, and their combinations on bacterial community structure and function in black soil

The potential ecotoxicologial risks of methamidophos, copper, and their combinations on microbial community of black soil ecosystem in the Northeast China were assessed in species richness and structures by using 16S rDNA-PCR-DGGE analysis approach, and functional characteristics at community levels by using BIOLOGGN system analysis method as well as two conventional methods(DHA and SIR). All results of DGGE banding fingerprint patterns(amplified by bacterial specific 16S rDNA V3 high variable region universal primer) indicated that the species richness of bacterial community in tested soil was significantly decreased to different extents by using different concentrations of single methamidophos, copper, especially some of their combinations had worse effects than their corresponding single factors. In addition,the structures of soil bacterial community had been disturbed under all stresses applied in this study because of the enrichment of some species and the disappearance of other species from the bacterial community. The effects of the single factors with lower concentrations on the communiy structure were weaker than those with higher concentrations. Moreover, the bacterial community structures under the combined stresses of methamidophos and copper were significantly different from those of control and their corresponding single factors. The change of DHA and carbon source substrate utilizing fingerprint patterns based on BIOLOGGNsystem were two relatively sensitive directors corresponding to the stress presented in this study. Between methamodophos and copper, there happened the significant joint-toxic actions when they were used in combination on DHA and carbon source substrate utilizing fingerprint patterns of soil bacterial communities. The DHA of soil under the combined stresses was lower than that of the control and that under the single factors, and the BIOLOGGN substrate utilizing patterns of soil treated by combinations were distinctively differentiated from the control and their corresponding single factors. From all of above, the methamidophos, copper, especially their combinations had the clearly potential ecotoxicological risks to influence the natural soil microbial ecological system by changing the structure, richness, and the functional characteristics of microbial community.

Functional profiles of soil microbial communities in the alpine and temperate grasslands of China

Grassland ecosystems in cold regions are typical of short growing seasons and limited primary productivity,rendering soil microorganisms as major ecosystem engineers in governing biogeochemical cycling.Climate warming and extensive livestock grazing have dramatically influenced soil microbial diversity and function in grassland worldwide,but it remains elusive how functional microbial communities exist and respond to global changes.Here,we present a review to highlight similarities and differences in soil functional microbial communities between alpine grasslands in the Qinghai-Tibet Plateau and temperate grasslands in the Inner Mongolian Plateau,both of which are major plateaus in China,but differ substantially in geography.We show that many specialized functional groups thrive under harsh conditions,exhibiting a high functional diversity.Their community compositions mirror the heterogeneity and complexity of grassland soils.Moreover,functional microbial responses to environmental changes have been extremely variable,with few consistent patterns across both plateaus.Because we identify a lack of technical standardization that prevents in-depth comparative studies for functional microbial communities,we conclude the review by outlining several research gaps that need to be filled in future studies.

Lowered water table causes species substitution while nitrogen amendment causes species loss in alpine wetland microbial communities

Alpine wetlands are hotspots of carbon(C)storage and methane emission,and they could be key contributors to global warming.In recent years,rapid warming has lowered the water table in alpine wetlands on the Tibetan Plateau,concurrent with intensified nitrogen(N)deposition via anthropogenic activities.We carried out a field experiment to investigate the ecological impacts of these two factors on soil bacterial and functional communities,which are essential drivers of greenhouse gas emissions.Nitrogen amendment alone decreased the phylogenetic alpha-diversity of bacterial communities which could be offset by lowered water table.In contrast,microbial functional alpha-diversity,revealed by a high-throughput microarray,remained unchanged.Both bacterial and functional beta-diversity responded to lowered water table,but only bacterial community responded to N amendment.The alpha-Proteobacteria,beta-Proteobacteria,and Bacteroidetes were the major responsive bacterial lineages,and C degradation,methanogenesis,alkaline shock,and phosphorus oxidation were the major responsive functional processes.Partitioning analysis revealed that N amendment changed bacterial community structure mainly via species loss processes but did not affect bacterial functional communities,with soil pH and ammonium as the key factors influencing changes in bacterial community structure.Conversely,lowered water table altered bacterial and functional communities through species substitution processes linked to soil pH and soil moisture.According to our results,the response mechanisms of microbial communities to lowered water table and N amendment are fundamentally different in alpine wetlands.

Effect of Trichoderma viride biofertilizer on ammonia volatilization from an alkaline soil in Northern China

Ammonia（NH3） volatilization is one of the primary pathways of nitrogen（N） loss from soils after chemical fertilizer is applied, especially from the alkaline soils in Northern China, which results in lower efficiency for chemical fertilizers. Therefore, we conducted an incubation experiment using an alkaline soil from Tianjin（p H 8.37–8.43） to evaluate the suppression effect of Trichoderma viride（T. viride） biofertilizer on NH3 volatilization, and compared the differences in microbial community structure among all samples. The results showed that viable T. viride biofertilizer（T） decreased NH3 volatilization by 42.21% compared with conventional fertilizer（（CK）, urea）, while nonviable T. viride biofertilizer（TS） decreased NH3 volatilization by 32.42%. NH3 volatilization was significantly higher in CK and sweet potato starch wastewater（SPSW） treatments during the peak period. T. viride biofertilizer also improved the transfer of ammonium from soil to sweet sorghum. Plant dry weights increased 91.23% and 61.08% for T and TS, respectively, compared to CK. Moreover, T. viride biofertilizer enhanced nitrification by increasing the abundance of ammonium-oxidizing archaea（AOA） and ammonium-oxidizing bacteria（AOB）. The results of high-throughput sequencing indicated that the microbial community structure and composition were significantly changed by the application of T. viride biofertilizer. This study demonstrated the immense potential of T. viride biofertilizer in reducing NH3 volatilization from alkaline soil and simultaneously improving the utilization of fertilizer N by sweet sorghum.

Point process models, the dimensions of biodiversity and the importance of small-scale biotic interactions

Aims Recent mechanistic explanations for community assembly focus on the debates surrounding niche-based deterministic and dispersalbased stochastic models.This body of work has emphasized the importance of both habitat filtering and dispersal limitation,and many of these works have utilized the assumption of species spatial independence to simplify the complexity of the spatial modeling in natural communities when given dispersal limitation and/or habitat filtering.One potential drawback of this simplification is that it does not consider species interactions and how they may influence the spatial distribution of species,phylogenetic and functional diversity.Here,we assess the validity of the assumption of species spatial independence using data from a subtropical forest plot in southeastern China.Methods We use the four most commonly employed spatial statistical models—the homogeneous Poisson process representing pure random effect,the heterogeneous Poisson process for the effect of habitat heterogeneity,the homogenous Thomas process for sole dispersal limitation and the heterogeneous Thomas process for joint effect of habitat heterogeneity and dispersal limitation—to investigate the contribution of different mechanisms in shaping the species,phylogenetic and functional structures of communities.Important Findings Our evidence from species,phylogenetic and functional diversity demonstrates that the habitat filtering and/or dispersal-based models perform well and the assumption of species spatial independence is relatively valid at larger scales(50×50 m).Conversely,at local scales(10×10 and 20×20 m),the models often fail to predict the species,phylogenetic and functional diversity,suggesting that the assumption of species spatial independence is invalid and that biotic interactions are increasingly important at these spatial scales.

How the development of barren land into orchards affects soil ecosystem in Tibet,China

Fruit production is an important strategy for alleviating poverty on the Tibetan Plateau and leads to the conversion of natural barren land into orchards.This study aimed to understand how the conversion of barren land to peach(Prunus persica)orchards affects soil nutrients,heavy metals,and fungal communities in the 0–40 cm profile(at 20 cm intervals)in an experiment including three treatments,barren land(BL),peach orchards planting for 4 years(Y4),and peach orchards planting for 10 years(Y10).Results of the experiment showed that compared with BL,Y4 reduced the availability of some macronutrients(N and K)and micronutrients(Fe and Mn)due to the exclusive application of chemical fertilizer at the seedling stage.Conversely,Y10,which included six years of green cultivation management,using a combination of sheep manure and chemical fertilizer as well as alfalfa(Medicago sativa Linn)intercropping,effectively improved soil macronutrients,but did not enhance the availability of Fe and Mn.Although the investigated heavy metals(As,Hg,Pb,Cr,and Cd)in both the Y4 and Y10 soils were found to pose a low risk to food safety and soil environment,Hg,Cr,and As tended to accumulate in the subsoil(20–40 cm).Furthermore,the variations in the fungal community composition and functional groups were mainly driven by the interaction effects of macronutrients,micronutrients,and heavy metals,but their independent contribution to specific key functional groups cannot be overlooked.For example,Y4 and Y10 decreased the relative abundance of soil saprotrophic and lichenized fungi,mainly due to the loss of micronutrients(Fe and Mn).However,as a result of macronutrient input and dung saprotrophic fungi enrichment,orchard soils promoted the growth of pathogens that play critical roles in fungal co-occurrence networks.These findings indicate that supplementation with N or K fertilizer or manure at the seedling stage and fertilizers rich in Fe and Mn throughout the growth period would be beneficial to the balance of soil nutrients and provide insights into linking the variations in soil nutrients and heavy metals to the function of the fungal community during the conversion of barren land to orchards in alpine soil ecosystems.The risks posed by heavy metal accumulation and fungal pathogen enrichment should be actively prevented.