Fungal Beta Diversity Research Articles

Effect of LSU and ITS genetic markers and reference databases on analyses of fungal communities

The effect of genetic markers and reference databases on analyses of fungal communities were estimated using fungal large subunit (LSU) and internal transcribed spacer (ITS) amplicon datasets in consecutive years of rhizosphere samples from three candidate biofuel crops, corn (Zea mays), switchgrass (Panicum virgatum), and miscanthus (Miscanthus × giganteus). These two marker genes were selected to contrast possible differences in biological conclusions. In addition, two ITS schemes based on two ITS reference databases were used to assess differences due to reference database composition. A taxonomy-supervised method was invoked using the Ribosomal Database Project naive Bayesian classifier that accesses all three databases. The UNITE classification scheme had the highest number of classified taxa in the raw classification result; however, it also had the highest proportion of unknown taxa (sequences that were classified to “unclassified,” “unidentified,” incertae sedis or, in the case of Warcup, to matches containing two unique names). After removal of these unknown taxa, LSU had the highest classification rate followed by Warcup and UNITE. As expected, the communities resolved using the two ITS databases, based on the same sequences, were relatively more similar than those from the lower-coverage LSU classification scheme. The choice of marker gene or even the same reads with different classification databases revealed different community patterns due to database coverage, e.g., the relative abundance of the most abundant groups changed or were only detected in one or two of the classification schemes, such as for Mortierella, Fusarium, and Phoma. No marked difference in fungal beta-diversity was identified among the three methods. Differentiation between the three biofuel crops and between the drought and normal rainfall years was apparent, regardless of method. Though classification rates, taxonomic conflicts, and coverage differences within the high-abundance fungal groups varied according to classification scheme, there was no overall impact on beta diversity among the three methods.

Macro and Microelements Drive Diversity and Composition of Prokaryotic and Fungal Communities in Hypersaline Sediments and Saline-Alkaline Soils.

Understanding the effects of environmental factors on microbial communities is critical for microbial ecology, but it remains challenging. In this study, we examined the diversity (alpha diversity) and community compositions (beta diversity) of prokaryotes and fungi in hypersaline sediments and salinized soils from northern China. Environmental variables were highly correlated, but they differed significantly between the sediments and saline soils. The compositions of prokaryotic and fungal communities in the hypersaline sediments were different from those in adjacent saline–alkaline soils, indicating a habitat-specific microbial distribution pattern. The macroelements (S, P, K, Mg, and Fe) and Ca were, respectively, correlated closely with the alpha diversity of prokaryotes and fungi, while the macronutrients (e.g., Na, S, P, and Ca) were correlated with the prokaryotic and fungal beta-diversity (P ≤ 0.05). And, the nine microelements (e.g., Al, Ba, Co, Hg, and Mn) and micronutrients (Ba, Cd, and Sr) individually shaped the alpha diversity of prokaryotes and fungi, while the six microelements (e.g., As, Ba, Cr, and Ge) and only the trace elements (Cr and Cu), respectively, influenced the beta diversity of prokaryotes and fungi (P < 0.05). Variation-partitioning analysis (VPA) showed that environmental variables jointly explained 55.49% and 32.27% of the total variation for the prokaryotic and fungal communities, respectively. Together, our findings demonstrate that the diversity and community composition of the prokaryotes and fungi were driven by different macro and microelements in saline habitats, and that geochemical elements could more widely regulate the diversity and community composition of prokaryotes than these of fungi.

Fungal community reveals less dispersal limitation and potentially more connected network than that of bacteria in bamboo forest soils.

A central aim of this microbial ecology research was to investigate the mechanisms shaping the assembly of soil microbial communities. Despite the importance of bacterial and fungal mediation of carbon cycling in forest ecosystems, knowledge concerning their distribution patterns and underlying mechanisms remains insufficient. Here, soils were sampled from six bamboo forests across the main planting area of Moso bamboo in southern China. The bacterial and fungal diversities were assessed by sequencing 16S rRNA and ITS gene amplicons, respectively, with an Illumina MiSeq. Based on structural equationmodelling, dispersal limitation had strongest impact on bacterial beta diversity, while the mean annual precipitation had a smaller impact by directly or indirectly mediating the soil organic carbon density. However, only the mean annual temperature and precipitation played direct roles in fungal beta diversity. Moreover, the co-occurrence network analyses revealed a possibly much higher network connectivity in the fungal network than in the bacteria. With less dispersal limitation, stronger environmental selection and a potentially more connected network, the fungal community had more important roles in the soil carbon metabolisms in bamboo forests. Fungal beta diversity and the clustering coefficient explained approximately 14.4% and 6.1% of the variation in the carbon metabolic profiles among sites, respectively, but that of bacteria only explained approximately 1.7% and 1.8%, respectively. This study explored soil microbial spatial patterns along with the underlying mechanisms of dispersal limitation, selection and connectivity of ecological networks, thus providing novel insights into the study of the distinct functional traits of different microbial taxa.

Plant diversity is coupled with beta not alpha diversity of soil fungal communities following N enrichment in a semi-arid grassland

Soil fungi and plants are tightly linked in pathogenic, commensal, and mutualistic ways. These interactions play a critical role in terrestrial ecosystem decomposition, nutrient cycling, and maintenance of plant productivity and diversity. A comprehensive understanding whether this fundamental plant-fungi relationship persists in ecosystems with increased N input is, however, still lacking. In this study, we investigated the relationships between plant and soil fungal diversity in a 6-year multi-level nitrogen addition experiment in a semi-arid grassland in northern China, using Illumina Miseq sequencing of the ITS1 barcode region for fungal identification. We hypothesized that N-induced changes in plant communities would be positively associated with those in soil fungal communities, and that the corresponding changes in both communities can be explained by the direct functional associations between plant and soil fungal communities and their shared environmental drivers. Our results showed that N-induced changes in plant alpha diversity, i.e., Shannon diversity, showed no significant relationship with those in soil fungal alpha diversity. The lack in significance of the relationship was primarily due to their contrasting correlates with N-induced soil physicochemical variables (i.e., soil available P (AP), inorganic N (NH4+_N and NO3−_N) and organic C content), and weakened plant-fungi functional associations. In contrast to the lack of relationship between plant and soil alpha diversity, we did find a significant positive relationship between plant and fungal beta diversity (compositional dissimilarity between plots) under N enrichment. Our results reveal that the same soil physicochemical variables, including soil AP, inorganic N, moisture, pH, and associated extractable cations, correlated with compositional changes in plant and fungi following N enrichment. The strong coupling of plant and fungal beta diversity could largely be driven by their consistent responses to these shared edaphic factors. As such, our results suggest that information on N-induced changes in plant diversity can be used to predict beta, but not alpha diversity of soil fungal communities in semi-arid grassland ecosystems.

Distinct seasonality of chytrid-dominated benthic fungal communities in the neritic oceans (Bohai Sea and North Yellow Sea)

Abstract Benthic fungal diversities in Bohai Sea and North Yellow Sea were investigated using pyrosequencing of 18S rDNA. Overall, Chytridiomycota dominated, followed by Basidiomycota, Ascomycota and Cryptomycota, in terms of alpha diversities and relative abundance. The beta diversity of benthic fungi showed a significant seasonality but no regional differences, accounted for by contrasting relative abundances of Chytridiomycota and Basidiomycota. Significantly seasonal changes in Chytridiomycota and Basidiomycota assemblage structure were also observed, but not for Ascomycota and Cryptomycota. Environmental filtering was more important than water depth and geographic distance in shaping the distribution of benthic fungi in the neritic oceans. The overall fungal beta diversity co-varied with concentration of chlorophyll a, pH, and salinity, distance from land, and water depth. The assemblage structure of benthic Chytridiomycota, Basidiomycota, Ascomycota and Cryptomycota co-varied with different sets of environmental parameters, suggesting their niche differentiations in the coastal sediments.

Differential responses of soil microbial biomass, diversity, and compositions to altitudinal gradients depend on plant and soil characteristics

Alt'itudinal gradients strongly affect plant biodiversity, but the effects on microbial patterns remain unclear, especially in the large scale. We therefore designed an altitudinal gradient experiment that covered three climate zones to monitor soil microbial community dynamics and to compare those with plant and soil characteristics. Illumina sequencing of the 16S rRNA gene and ITS gene was used to analyze soil microbial (bacterial and fungal) diversity and composition, and fumigation–extraction was used to determine microbial biomass; the plant community metrics (i.e., percent cover, Shannon-Wiener, grass biomass, and carbon/nitrogen in leaf and biomass) and soil properties (i.e., soil moisture, soil temperature, bulk density, organic carbon, total nitrogen, and available nitrogen) were determined. The results showed that carbon/nitrogen in microbial biomass was higher at medium altitude and was positively related to carbon and nitrogen in both soil and grass biomass along the altitudinal gradients. Soil bacterial alpha diversity was significantly higher at medium altitude but fungal alpha diversity did not affected by altitudinal gradients; the effect of altitudinal gradients on bacterial beta diversity was larger than that on fungal beta diversity, although both groups were significantly affected by altitudinal gradients. Moreover, Alpha-proteobacteria, Beta-proteobacteria, and Gemmatimonadetes were significantly more abundant in higher altitude than in lower altitude, both Acidobacteria and Actinobacteria significantly declined with increasing altitude; other bacterial taxa such as Chloroflexi, Nitrospirae, Gamma-proteobacteria, and Delta-proteobacteria were significantly higher at medium altitudes. For fungal taxa, Basidiomycota and Ascomycota were the dominant phyla and responded insignificantly to the altitudinal gradients. The responses of microbial alpha diversity were mostly associated with plant Shannon index, organic carbon, and total nitrogen, whereas microbial beta diversity and composition mainly depended on soil moisture and temperature. Overall, these results suggest that soil bacteria rather than fungi can reflect changes in plant and soil characteristics along altitudinal gradients.

Effects of ecological restoration on soil microbial diversity in a temperate grassy woodland

Soil microbial communities are often overlooked in the context of ecological restoration. Given their central role in a broad range of ecosystem processes, however, understanding their response to restoration activities is critical to predicting restoration trajectories. In this study, we quantified the response of soil bacterial and fungal communities to restoration treatments, variation in microhabitat elements and vegetation in a critically endangered Australian box-gum grassy woodland ecosystem. Restoration treatments included the addition of coarse woody debris (CWD) and reduced grazing pressure. Four years after applying restoration treatments, we found no significant effect of CWD addition on soil microbial diversity, while reduced grazing significantly affected composition of the fungal, but not the bacterial, communities. Both bacterial and fungal communities responded to microhabitat element (open ground vs. old logs and trees), overlying vegetation and soil edaphic properties, and strong aboveground-belowground linkages were observed. Plant alpha diversity was positively correlated to soil bacterial, but not fungal, alpha diversity and plant community composition was a good predictor of both soil bacterial and fungal beta diversity. Co-occurrence network analysis identified numerous complex, non-linear associations between soil bacteria, fungi, edaphic properties and overlying plants. Soil microbes affected by restoration treatments included fungal saprotrophs and Actinobacteria, likely involved in litter breakdown, as well as bacteria likely involved in soil N cycling. Although the directions of the observed plant-microbe relationships remain unclear, we demonstrated the possibility of inducing changes to soil microbial communities to enhance restoration outcomes in box-gum grassy woodland ecosystems.

Distinctive Tropical Forest Variants Have Unique Soil Microbial Communities, But Not Always Low Microbial Diversity.

There has been little study of whether different variants of tropical rainforest have distinct soil microbial communities and levels of diversity. We compared bacterial and fungal community composition and diversity between primary mixed dipterocarp, secondary mixed dipterocarp, white sand heath, inland heath, and peat swamp forests in Brunei Darussalam, Northwest Borneo by analyzing Illumina Miseq sequence data of 16S rRNA gene and ITS1 region. We hypothesized that white sand heath, inland heath and peat swamp forests would show lower microbial diversity and relatively distinct microbial communities (compared to MDF primary and secondary forests) due to their distinctive environments. We found that soil properties together with bacterial and fungal communities varied significantly between forest types. Alpha and beta-diversity of bacteria was highest in secondary dipterocarp and white sand heath forests. Also, bacterial alpha diversity was strongly structured by pH, adding another instance of this widespread pattern in nature. The alpha diversity of fungi was equally high in all forest types except peat swamp forest, although fungal beta-diversity was highest in primary and secondary mixed dipterocarp forests. The relative abundance of ectomycorrhizal (EcM) fungi varied significantly between forest types, with highest relative abundance observed in MDF primary forest. Overall, our results suggest that the soil bacterial and fungal communities in these forest types are to a certain extent predictable and structured by soil properties, but that diversity is not determined by how distinctive the conditions are. This contrasts with the diversity patterns seen in rainforest trees, where distinctive soil conditions have consistently lower tree diversity.

The role of stochasticity differs in the assembly of soil- and root-associated fungal communities

Spatial patterns in community structure are known to exist at a range of spatial scales. By understanding the relative importance of spatial and environmental variation in shaping community structure, it is possible to gain insight into the complexity of the underlying mechanisms driving community assembly. Here, we investigated how fungal community assembly is impacted by species sorting across a strong environmental filter in the same geographic space. Using a spatially explicit sampling design, root and soil associated fungal communities were sampled from eight study sites over a 500 km transect along the south-east coast of Australia. The relative importance of spatial and environmental factors in shaping fungal beta diversity was determined across a range of scales, along with the importance of deterministic and stochastic processes in shaping fungal beta diversity changes for each community type. Fungal community composition patterns were found to be generally similar in both root and soil associated communities, across the scales included in the sampling design (40 m–500 km). However, the mechanisms responsible for driving community assembly differed in the root and soil associated communities. Niche-based processes were found to be influential in shaping soil fungal community assembly, while the patterns in root-associated community structure were consistent with neutral assembly. This may be due to factors such as niche breadth, environmental heterogeneity, dispersal, and immigration differing in importance among the two environments. Our results indicate that both niche and neutral processes can be active in fungal community assembly and provide insight into the conditions under which each class of processes dominate.

454 Pyrosequencing Analysis of Fungal Assemblages from Geographically Distant, Disparate Soils Reveals Spatial Patterning and a Core Mycobiome

Identifying a soil core microbiome is crucial to appreciate the established microbial consortium, which is not usually subjected to change and, hence, possibly resistant/resilient to disturbances and a varying soil context. Fungi are a major part of soil biodiversity, yet the mechanisms driving their large-scale ecological ranges and distribution are poorly understood. The degree of fungal community overlap among 16 soil samples from distinct ecosystems and distant geographic localities (truffle grounds, a Mediterranean agro-silvo-pastoral system, serpentine substrates and a contaminated industrial area) was assessed by examining the distribution of fungal ITS1 and ITS2 sequences in a dataset of 454 libraries. ITS1 and ITS2 sequences were assigned to 1,660 and 1,393 Operational Taxonomic Units (OTUs; as defined by 97% sequence similarity), respectively. Fungal beta-diversity was found to be spatially autocorrelated. At the level of individual OTUs, eight ITS1 and seven ITS2 OTUs were found in all soil sample groups. These ubiquitous taxa comprised generalist fungi with oligotrophic and chitinolytic abilities, suggesting that a stable core of fungi across the complex soil fungal assemblages is either endowed with the capacity of sustained development in the nutrient-poor soil conditions or with the ability to exploit organic resources (such as chitin) universally distributed in soils.

The spatial scaling of saprotrophic fungal beta diversity in decomposing leaves

Assembly of fungal communities remains poorly understood in part because of the daunting range of spatial scales that may be involved in this process. Here, we use individual leaves as a natural sampling unit, comprising spatially distinct habitat and/or resource patches with unique histories and suites of resources. Spatial patterns in fungal beta diversity were tested for consistency with the metacommunity paradigms of species sorting and neutral dynamics. Thirty senesced leaves were collected from the forest floor (O horizon) in replicate upland forest, riparian forest and vernal pool habitats. We quantified spatial distance between leaves, and fungal community composition was assayed by terminal restriction fragment length polymorphism. Significant distance-decay relationships were detected at all but one upland site. This is the first study where changes in fungal community composition were quantified across discrete adjacent habitat patches, providing evidence that fungal distance decay is operational at a scale of centimetres. Although leaves of differing lignin contents were sampled from each site, leaf type was not consistently important in explaining variation in fungal community composition. However, depth of a leaf within the forest floor significantly influenced community composition at five of six sites. Environmental heterogeneity associated with depth could include moisture gradients, relative influence of soil or spore colonization, and impact of forest floor biotic community (i.e. collembola and earthworms). Because the influence of spatial distance and depth on fungal community composition could not be disentangled, both species-sorting and neutral processes may be embedded within the distance-decay relationships that we found.

Contrasting Diversity Patterns of Crenarchaeal, Bacterial and Fungal Soil Communities in an Alpine Landscape

BackgroundThe advent of molecular techniques in microbial ecology has aroused interest in gaining an understanding about the spatial distribution of regional pools of soil microbes and the main drivers responsible of these spatial patterns. Here, we assessed the distribution of crenarcheal, bacterial and fungal communities in an alpine landscape displaying high turnover in plant species over short distances. Our aim is to determine the relative contribution of plant species composition, environmental conditions, and geographic isolation on microbial community distribution.Methodology/Principal FindingsEleven types of habitats that best represent the landscape heterogeneity were investigated. Crenarchaeal, bacterial and fungal communities were described by means of Single Strand Conformation Polymorphism. Relationships between microbial beta diversity patterns were examined by using Bray-Curtis dissimilarities and Principal Coordinate Analyses. Distance-based redundancy analyses and variation partitioning were used to estimate the relative contributions of different drivers on microbial beta diversity. Microbial communities tended to be habitat-specific and did not display significant spatial autocorrelation. Microbial beta diversity correlated with soil pH. Fungal beta-diversity was mainly related to soil organic matter. Though the effect of plant species composition was significant for all microbial groups, it was much stronger for Fungi. In contrast, geographic distances did not have any effect on microbial beta diversity.Conclusions/SignificanceMicrobial communities exhibit non-random spatial patterns of diversity in alpine landscapes. Crenarcheal, bacterial and fungal community turnover is high and associated with plant species composition through different set of soil variables, but is not caused by geographical isolation.

Assessment of soil fungal diversity in different alpine tundra habitats by means of pyrosequencing

Studying fungal diversity is vital if we want to shed light on terrestrial ecosystem functioning. However, there is still poor understanding of fungal diversity and variation given that Fungi are highly diversified and that most of fungal species remain uncultured. In this study we explored diversity with 454 FLX sequencing technology by using the Internal Transcribed Spacer 1 (ITS1) as the fungal barcode marker in order to evaluate the effect of 11 environmental conditions on alpine soil fungal diversity, as well as the consistency of those results by taking into account rare or unidentified Molecular Operational Taxonomic Units (MOTUs). In total we obtained 205131 ITS1 reads corresponding to an estimated fungal gamma diversity of between 5100 and 12 000 MOTUs at a 98% similarity threshold when considering respectively only identified fungal and all MOTUs. Fungal beta-diversity patterns were significantly explained by the environmental conditions, and were very consistent for abundant/rare and fungal/unidentified MOTUs confirming the ecological significance of rare/unidentified MOTUs, and therefore the existence of a fungal rare biosphere. This study shows that a beta-diversity estimation based on pyrosequencing is robust enough to support ecological studies. Additionally, our results suggest that rare MOTUs harbour ecological information. Thus the fungal rare biosphere may be important for ecosystem dynamics and resilience.