Impact of sheep urine deposition and plant species on ammonia-oxidizing bacteria in upland grassland soil

The effects of different concentrations of synthetic sheep urine and plant species on ammonia-oxidizing bacterial (AOB) communities in an upland grassland soil were investigated using a microcosm approach. Plant species characteristic of unimproved and improved agricultural pastures (Agrostis capillaris and Lolium perenne, respectively) were planted in soil microcosms, and different levels of synthetic sheep urine were applied, with harvests 10 and 50 days following urine application. Shifts in the community structure of the AOB were investigated using terminal restriction fragment length polymorphism of amoA amplicons. Species richness and diversity were significantly altered by synthetic sheep urine addition and time depending on plant species type. Principal coordinate analysis revealed that AOB community structure was largely dependent on interactions between sheep urine deposition, plant species, and time after urine application, while significant changes in AOB structure were also revealed by similarity percentage analysis. The results of this study suggested that high levels of sheep urine, combined with floristic changes that are characteristic of agricultural intensification, can contribute to temporal and spatial changes in the structure of key bacterial communities in upland grassland soil. Changes in AOB community structure could potentially affect important soil processes, such as nitrification, with subsequent implications for nutrient cycling in agricultural systems.     

Soil Bacterial and Fungal Community Structure Across a Range of Unimproved and Semi-Improved Upland Grasslands

Changes in soil microbial community structure due to improvement are often attributed to concurrent shifts in floristic community composition. The bacterial and fungal communities of unimproved and semi-improved (as determined by floristic classification) grassland soils were studied at five upland sites on similar geological substrata using both broad-scale (microbial activity and fungal biomass) and molecular [terminal restriction fragment length polymorphism (TRFLP), automated ribosomal intergenic spacer analysis (ARISA)] approaches. It was hypothesized that microbial community structure would be similar in soils from the same grassland type, and that grassland vegetation classifications could thus be used as predictors of microbial community structure. Microbial community measurements varied widely according to both site and grassland type, and trends in the effect of grassland improvement differed between sites. These results were consistent with those from similar studies, and indicated that floristic community composition was not a stable predictor of microbial community structure across sites. This may indicate a lack of correlation between grassland plant composition and soil microbial community structure, or that differences in soil chemistry between sites had larger impacts on soil microbial populations than plant-related effects.     

Seasonal and management influences on bacterial community structure in an upland grassland soil

Floristically diverse Nardo-Galion upland grasslands are common in Ireland and the UK and are valuable in agricultural, environmental and ecological terms. Under improvement (inputs of lime, fertiliser and re-seeding), they convert to mesotrophic grassland containing very few plant species. The effects of upland grassland improvement and seasonality on soil microbial communities were investigated at an upland site. Samples were taken at five times in one year in order to observe seasonal trends, and bacterial community structure was monitored using automated ribosomal intergenic spacer analysis (ARISA), a DNA-fingerprinting approach. Differences in soil chemistry and bacterial community structure between unimproved and improved grassland soils were noted. Season was also found to cause mild fluctuations in bacterial community structure, with soil samples from colder months (October and December) more correlated with change in ribotype profiles than samples from warmer months. However, for the majority of seasons clear differences in bacterial community structures from unimproved and improved soils could be seen, indicating seasonal influences did not obscure effects associated with improvement.     

An Evaluation of 18S rDNA Approaches for the Study of Fungal Diversity in Grassland Soils

Fungal community structure and diversity in two types of agricultural grassland soil were investigated by amplified 18S ribosomal DNA restriction analysis (ARDRA) and 18S ribosomal DNA sequence analysis. These two grassland sites represent a species-rich old hay meadow and an agriculturally improved site with low floristic diversity. Two primer sets were used in combination to amplify approximately 550 bp of rDNA from three major fungal groups, the zygomycetes, basidiomycetes, and ascomycetes, and clone libraries were created for each site. 18S ARDRA was used to analyze 170 rDNA clones, and three diversity indices were calculated. A small-scale culturing analysis was also carried out and the most common isolates analyzed using ARDRA and sequence analysis. The soil fungal community revealed by the rDNA approaches was significantly different from that produced by this limited culture-based analysis. Twenty-eight soil-derived clones were sequenced, and many represented fungal taxa rarely reported in culture-based studies. The PCR-based techniques detected differences in diversity between the two fungal communities and changes in patterns of dominance that paralleled higher plant diversity. The results suggest that 18S rDNA-based approaches are a useful tool for initial screening of fungal communities, and that they represent a more comprehensive picture of the community than plate culturing.     

Water Content Differences Have Stronger Effects than Plant Functional Groups on Soil Bacteria in a Steppe Ecosystem

Many investigations across natural and artificial plant diversity gradients have reported that both soil physicochemical factors and plant community composition affect soil microbial communities. To test the effect of plant diversity loss on soil bacterial communities, we conducted a five-year plant functional group removal experiment in a steppe ecosystem in Inner Mongolia (China). We found that the number and composition type of plant functional groups had no effect on bacterial diversity and community composition, or on the relative abundance of major taxa. In contrast, bacterial community patterns were significantly structured by soil water content differences among plots. Our results support researches that suggest that water availability is the key factor structuring soil bacterial communities in this semi-arid ecosystem.     

Links between plant and rhizoplane bacterial communities in grassland soils, characterized using molecular techniques

Molecular analysis of grassland rhizosphere soil has demonstrated complex and diverse bacterial communities, with resultant difficulties in detecting links between plant and bacterial communities. These studies have, however, analyzed "bulk" rhizosphere soil, rather than rhizoplane communities, which interact most closely with plants through utilization of root exudates. The aim of this study was to test the hypothesis that plant species was a major driver for bacterial rhizoplane community composition on individual plant roots. DNA extracted from individual roots was used to determine plant identity, by analysis of the plastid tRNA leucine (trnL) UAA gene intron, and plant-related bacterial communities. Bacterial communities were characterized by analysis of PCR-amplified 16S rRNA genes using two fingerprinting methods: terminal restriction fragment length polymorphisms (T-RFLP) and denaturing gradient gel electrophoresis (DGGE). Links between plant and bacterial rhizoplane communities could not be detected by visual examination of T-RFLP patterns or DGGE banding profiles. Statistical analysis of fingerprint patterns did not reveal a relationship between bacterial community composition and plant species but did demonstrate an influence of plant community composition. The data also indicated that topography and other, uncharacterized, environmental factors are important in driving bacterial community composition in grassland soils. T-RFLP had greater potential resolving power than DGGE, but findings from the two methods were not significantly different.     

Relationship between assemblages of mycorrhizal fungi and bacteria on grass roots

Soils support an enormous microbial diversity, but the ecological drivers of this diversity are poorly understood. Interactions between the roots of individual grass species and the arbuscular mycorrhizal (AM) fungi and bacteria in their rhizoplane were studied in a grazed, unimproved upland pasture. Individual root fragments were isolated from soil cores, DNA extracted and used to identify plant species and assess rhizoplane bacterial and AM fungal assemblages, by amplifying part of the small-subunit ribosomal RNA gene, followed by terminal restriction fragment length polymorphism analysis. For the first time we showed that AM fungal and bacterial assemblages are related in situ and that this relationship occurred at the community level. Principal coordinate analyses of the data show that the AM fungi were a major factor determining the bacterial assemblage on grass roots. We also report a strong influence of the composition of the plant community on AM fungal assemblage. The bacterial assemblage was also influenced by soil pH and was spatially structured, whereas AM fungi were influenced neither by the bacteria nor by soil pH. Our study shows that linkages between plant roots and their microbial communities exist in a complex web of interactions that act at individual and at community levels, with AM fungi influencing the bacterial assemblage, but not the other way round.     

Plant diversity predicts beta but not alpha diversity of soil microbes across grasslands worldwide

Aboveground–belowground interactions exert critical controls on the composition and function of terrestrial ecosystems, yet the fundamental relationships between plant diversity and soil microbial diversity remain elusive. Theory predicts predominantly positive associations but tests within single sites have shown variable relationships, and associations between plant and microbial diversity across broad spatial scales remain largely unexplored. We compared the diversity of plant, bacterial, archaeal and fungal communities in one hundred and forty‐five 1 m² plots across 25 temperate grassland sites from four continents. Across sites, the plant alpha diversity patterns were poorly related to those observed for any soil microbial group. However, plant beta diversity (compositional dissimilarity between sites) was significantly correlated with the beta diversity of bacterial and fungal communities, even after controlling for environmental factors. Thus, across a global range of temperate grasslands, plant diversity can predict patterns in the composition of soil microbial communities, but not patterns in alpha diversity.     

Links between Plant and Rhizoplane Bacterial Communities in Grassland Soils, Characterized Using Molecular Techniques

Molecular analysis of grassland rhizosphere soil has demonstrated complex and diverse bacterial communities, with resultant difficulties in detecting links between plant and bacterial communities. These studies have, however, analyzed “bulk” rhizosphere soil, rather than rhizoplane communities, which interact most closely with plants through utilization of root exudates. The aim of this study was to test the hypothesis that plant species was a major driver for bacterial rhizoplane community composition on individual plant roots. DNA extracted from individual roots was used to determine plant identity, by analysis of the plastid tRNA leucine (trnL) UAA gene intron, and plant-related bacterial communities. Bacterial communities were characterized by analysis of PCR-amplified 16S rRNA genes using two fingerprinting methods: terminal restriction fragment length polymorphisms (T-RFLP) and denaturing gradient gel electrophoresis (DGGE). Links between plant and bacterial rhizoplane communities could not be detected by visual examination of T-RFLP patterns or DGGE banding profiles. Statistical analysis of fingerprint patterns did not reveal a relationship between bacterial community composition and plant species but did demonstrate an influence of plant community composition. The data also indicated that topography and other, uncharacterized, environmental factors are important in driving bacterial community composition in grassland soils. T-RFLP had greater potential resolving power than DGGE, but findings from the two methods were not significantly different.     

Bacterial succession during 500?years of soil development under agricultural use

Soil bacterial succession under intensive anthropogenic disturbances is not well known. Using terminal restriction fragment length polymorphisms and 454 pyrosequencing of 16S rRNA genes, this study investigated how soil bacterial diversity and community structure changed under two agricultural land uses (paddy rice and upland cropping) in relation to soil development along a 500-year chronosequence created by intermittent reclamation of estuarine salt marshes. Multivariate analysis revealed orderly changes in soil physicochemical properties and bacterial community structure with time, confirming the occurrence of soil development and bacterial succession. Patterns of soil development and bacterial succession resembled each other, with recent land uses affecting their trajectories but not the overall direction. Succession of bacterial community structure was mainly associated with changes in ??-Proteobacteria and Verrucomicrobia. Two stages of bacterial succession were observed, a dramatic-succession stage during the first several decades when bacterial diversity increased evidently and bacterial community structure changed rapidly, and a long gradual-succession stage that lasted for centuries. Canonical correspondence analysis identified soil Na⁺, potentially mineralizable nitrogen, total phosphorous, and crystallinity of iron oxyhydrates as potential environmental drivers of bacterial succession. To conclude, orderly succession of soil bacterial communities occurred along with the long-term development of agroecosystems, which in turn was associated with soil physicochemical changes over time.     

Avian Composition Co‐varies with Floristic Composition and Soil Nutrient Concentration in Amazonian Upland Forests

Spatial heterogeneity in the plant species composition of tropical forests is expected to influence animal species abundance and composition because vegetation constitutes the primary habitat feature for forest animals. Floristic variation is tied to variation in soils, so edaphic properties should ultimately influence animal species composition as well. The study of covariation in floristic and faunistic turnover has been hindered by the difficulty of completing coordinated surveys in hyperdiverse tropical communities, but this can be overcome with the use of a few plant taxa that function as surrogates for general floristic turnover. We used avian and plant transect surveys and soil sampling in a western Amazonian upland (terra firme) forest landscape to test whether spatial variation in bird community composition is associated with floristic turnover and corresponding edaphic gradients. Partial Mantel tests and Non‐metric Multidimensional Scaling showed floristic distinctiveness between two forest types closely associated with differences in soil cation concentrations, and differences in both floristic composition and cation concentrations were further linked to compositional differences in avian species, independent of geographic distances among sites. Ten percent of bird species included in Indicator Species Analyses showed significant associations with one of the two forest types. The upland forest types that we sampled, each corresponding to a different geological formation, are intermediate relative to edaphically extreme environments in the region. Models of avian diversification should take into account this environmental heterogeneity, as should conservation planning approaches that aim to represent faunal diversity. Abstract in Spanish is available in the online version of this article.     

The bacterial biogeography of British soils

Despite recognition of the importance of soil bacteria to terrestrial ecosystem functioning there is little consensus on the factors regulating belowground biodiversity. Here we present a multi-scale spatial assessment of soil bacterial community profiles across Great Britain (> 1000 soil cores), and show the first landscape scale map of bacterial distributions across a nation. Bacterial diversity and community dissimilarities, assessed using terminal restriction fragment length polymorphism, were most strongly related to soil pH providing a large-scale confirmation of the role of pH in structuring bacterial taxa. However, while α diversity was positively related to pH, the converse was true for β diversity (between sample variance in α diversity). β diversity was found to be greatest in acidic soils, corresponding with greater environmental heterogeneity. Analyses of clone libraries revealed the pH effects were predominantly manifest at the level of broad bacterial taxonomic groups, with acidic soils being dominated by few taxa (notably the group 1 Acidobacteria and Alphaproteobacteria). We also noted significant correlations between bacterial communities and most other measured environmental variables (soil chemistry, aboveground features and climatic variables), together with significant spatial correlations at close distances. In particular, bacterial and plant communities were closely related signifying no strong evidence that soil bacteria are driven by different ecological processes to those governing higher organisms. We conclude that broad scale surveys are useful in identifying distinct soil biomes comprising reproducible communities of dominant taxa. Together these results provide a baseline ecological framework with which to pursue future research on both soil microbial function, and more explicit biome based assessments of the local ecological drivers of bacterial biodiversity.     

Comparison of Rhizosphere Bacterial Communities in Arabidopsis thaliana Mutants for Systemic Acquired Resistance

Systemic acquired resistance (SAR) is an inducible systemic plant defense against a broad spectrum of plant pathogens, with the potential to secrete antimicrobial compounds into the soil. However, its impact on rhizosphere bacteria is not known. In this study, we examined fingerprints of bacterial communities in the rhizosphere of the model plant Arabidopsis thaliana to determine the effect of SAR on bacterial community structure and diversity. We compared Arabidopsis mutants that are constitutive and non-inducible for SAR and verified SAR activation by measuring pathogenesis-related protein activity via a β-glucoronidase (GUS) reporter construct driven by the β-1-3 glucanase promoter. We used terminal restriction fragment length polymorphism (T-RFLP) analysis of MspI- and HaeIII-digested 16S rDNA to estimate bacterial rhizosphere community diversity, with Lactobacillus sp. added as internal controls. T-RFLP analysis showed a clear rhizosphere effect on community structure, and diversity analysis of both rhizosphere and bulk soil operational taxonomic units (as defined by terminal restriction fragments) using richness, Shannon–Weiner, and Simpson’s diversity indices and evenness confirmed that the presence of Arabidopsis roots significantly altered bacterial communities. This effect of altered soil microbial community structure by plants was also seen upon multivariate cluster analysis of the terminal restriction fragments. We also found visible differences in the rhizosphere community fingerprints of different Arabidopsis SAR mutants; however, there was no clear decrease of rhizosphere diversity because of constitutive SAR expression. Our study suggests that SAR can alter rhizosphere bacterial communities, opening the door to further understanding and application of inducible plant defense as a driving force in structuring soil bacterial assemblages.     

16S rRNA gene analyses of bacterial community structures in the soils of evergreen broad-leaved forests in south-west China

Bacterial community structure was studied in humus and mineral soils of evergreen broad-leaved forests in Ailaoshan and Xishuangbanna, representing subtropical and tropical ecosystems, respectively, in south-west China using sequence analysis and terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Clone sequences affiliated to Acidobacteria were retrieved as the predominant bacterial phylum in both forest soils, followed by those affiliated to members of the Proteobacteria, Planctomycete and Verrucomicrobia. Despite higher floristic richness at the Xishuangbanna forest than at the Ailaoshan forest, soil at Xishuangbanna harbored a distinctly high relative abundance of Acidobacteria-affiliated sequences (80% of the total clones), which led to a lower overall bacterial diversity than at Ailaoshan. Bacterial communities in humus and mineral soils of the two forests appeared to be well differentiated, based on 16S rRNA gene phylogeny, and correlations were found between the bacterial T-RFLP community patterns and the organic carbon and nutrient contents of the soil samples. The data reveal that Acidobacteria dominate soil bacterial communities in the evergreen broad-leaved forests studied here and suggest that bacterial diversity may be influenced by soil carbon and nutrient levels, but is not related to floristic richness along the climatic gradient from subtropical to tropical forests in south-west China.     

Floristic composition and spatial distribution assessment of montane mesophilous grasslands in the central Apennines,Italy: A multi-scale and diachronic approach

Abstract

Evaluation of past land use offers valuable information in seeking to understand the distribution patterns of plant communities, insofar as such activity may have altered soil features, causing anthropogenic soil patchiness and marked plant diversity both locally and regionally. This study sought to provide a general overview of the relationships between the different syntaxonomical units and the soil features in a sub-Mediterranean grassland landscape, starting from assessment of soil parameters. In addition, we wanted to verify whether soil features and hierarchical landscape assessments of grasslands under homogeneous grazing disturbance are sufficient to explain the floristic differentiation of the plant communities, and whether study of land use history helps explain the distribution patterns of these plant communities. A clear understanding of the relationships between environmental factors and the floristic composition of plant communities is a strong basis for future inquiries into how disturbance variations (grazing, mowing, manuring, etc.) and past land use variations have affected grassland structures. This study examined nine syntaxa that develop on semi-flat slopes or those with northern exposure.     

东祁连山退化高寒草地土壤细菌群落与土壤环境因子间的相互关系

为探究东祁连山不同退化高寒草地细菌群落分布特征与土壤环境因子间的相互关系,采用高通量测序技术对轻度、中度和重度退化草地的土壤细菌群落结构变化及其多样性进行分析,运用CANOCO 4.5软件对土壤细菌群落与土壤环境因子间关系进行冗余分析(RDA).结果表明:不同退化高寒草地土壤理化性质间均差异显著,高通量测序共得到257125条有效序列,180826条优质序列,4790个OTUs.细菌群落Chao1指数依次为轻度>中度>重度;Shannon指数依次为轻度>重度>中度.系统发育分析表明,各样地土壤细菌类群分属于33个门,其中放线菌门、变形菌门和厚壁菌门是3种不同退化草地土壤中的优势类群.对不同退化草地土壤细菌各门所占比例分析发现,放线菌门、酸杆菌门和变形菌门随着退化程度加剧先减少后增加,厚壁菌门反之.RDA分析结果显示,细菌优势类群与蔗糖酶、纤维素酶和磷酸酶呈极显著相关,与pH、电导率、速效氮、速效钾呈显著相关.说明东祁连山不同退化高寒草地土壤细菌群落间差异明显,土壤环境因子是影响土壤细菌群落分布的重要因素.     

The impact of zinc and lead concentrations and seasonal variation on bacterial and actinobacterial community structure in a metallophytic grassland soil

The diversity and structure of bacterial and actinobacteral diversity and communities were determined in a metallophytic grassland soil from an upland site in northern England. The community profiles were subjected to multivariate analyses using correspondence and cluster analyses. The total bacterial community diversities and structures were not significantly affected by Pb and Zn concentration in the soil. However, the community structure did show changes between winter and summer samples. Raup and Crick analysis indicated that deterministic selection lead to winter profiles exhibiting significant similarity. The actinobacterial community was also unaffected by Pb and Zn concentration. However, seasonal changes were apparent as diversity were significantly lower in winter compared to summer profiles. Moreover, the community structure showed evidence of changes of structure based on the seasonal samples with winter samples showing significant similarity to each other.     

内蒙古草原典型植物对土壤微生物群落的影响

为了分析内蒙古草原不同植物物种对土壤微生物群落的影响, 采用实时荧光定量PCR (real-time PCR)以及末端限制性片段长度多态性分析(terminal restriction fragment length polymorphism, T-RFLP)等分子生物学技术, 测定了退化-恢复样地上几种典型植物的根际土壤和非根际土壤中细菌和真菌的数量及群落结构。结果表明, 不同植物物种对根际和非根际细菌及根际真菌数量均有显著影响。根际土壤中的细菌和真菌数量普遍高于非根际土壤, 尤其以真菌更为明显。对T-RFLP数据进行多响应置换过程(multi-response permutation procedures, MRPP)分析和主成分分析(principal component analysis, PCA), 结果表明, 大多数物种的根际细菌及真菌的群落结构与非根际有明显差异, 并且所有物种的真菌群落可以按根际和非根际明显分为两大类群。此外, 细菌和真菌群落结构在一定程度上存在按物种聚类的现象, 以细菌较为明显。这些结果揭示了不同植物对土壤微生物群落的影响特征, 对理解内蒙古草原地区退化及恢复过程中植被演替引起的土壤性质和功能的变化有一定的帮助。     

植茶年限对土壤微生物群落结构及多样性的影响

为探明植茶年限对土壤微生物群落结构及多样性的影响,以0、20、25、38和48年茶园土壤表层(0～20 cm)、亚表层(20～40 cm)土壤样品为研究对象,采用T-RFLP技术及qPCR方法对土壤细菌(B)、真菌(F)群落进行分析。结果表明: 植茶后土壤理化性质明显改变,随植茶年限的增加土壤有机碳、碱解氮及有效磷含量呈先升高后降低的趋势,表层土壤有机碳和全氮含量均显著高于亚表层土壤。不同植茶年限土壤细菌群落组分存在差异且多样性指数随植茶年限的增加呈下降趋势,而不同植茶年限土壤真菌群落组分差异不明显且多样性指数无显著差异。总体来看,土壤细菌群落对植茶年限的响应比真菌群落敏感。随植茶年限的增加,茶园土壤微生物群落有从F/B较低的“细菌型”向F/B较高的“真菌型”转变的趋势。     

Axial dynamics, stability, and interspecies similarity of bacterial community structure in the highly compartmentalized gut of soil-feeding termites (Cubitermes spp.)

The highly compartmentalized gut of soil-feeding termites is characterized by pronounced axial dynamics in physicochemical conditions and microbial processes. In a companion paper (D. Schmitt-Wagner, M. W. Friedrich, B. Wagner, and A. Brune, Appl. Environ. Microbiol. 69:6007-6017, 2003), we demonstrated that the variety of physicochemical conditions in the different gut compartments of Cubitermes spp. is reflected in the diversity of the respective intestinal microbial communities. Here, we used molecular fingerprints of 16S rRNA genes of the bacterial community, obtained by terminal restriction fragment length polymorphism (T-RFLP) analysis, to describe the axial dynamics of the bacterial community structure in the different gut sections. Comparison of the T-RFLP profiles with the predicted terminal restriction fragments of the clones in clone libraries of the gut segments in Cubitermes orthognathus confirmed that all hindgut sections harbored distinct bacterial communities. Morisita indices of community similarity, calculated by comparing the different patterns, revealed large differences between the bacterial communities of soil, gut, and nest material and also among the individual gut sections. By contrast, comparison of the homologous gut segments of different Cubitermes species indicated that the three termite species investigated possessed a similar, gut-specific microbiota that remained comparatively stable even during several months of maintenance in the laboratory.