Ectomycorrhizal fungal communities are dominated by mammalian dispersed truffle-like taxa in north-east Australian woodlands

Mycorrhizal fungi are very diverse, including those that produce truffle-like fruiting bodies. Truffle-like fungi are hypogeous and sequestrate (produced below-ground, with an enclosed hymenophore) and rely on animal consumption, mainly by mammals, for spore dispersal. This dependence links mycophagous mammals to mycorrhizal diversity and, assuming truffle-like fungi are important components of mycorrhizal communities, to plant nutrient cycling and ecosystem health. These links are largely untested as currently little is known about mycorrhizal fungal community structure and its dependence on mycophagous mammals. We quantified the mycorrhizal fungal community in the north-east Australian woodland, including the portion interacting with ten species of mycophagous mammals. The study area is core habitat of an endangered fungal specialist marsupial, Bettongia tropica, and as such provides baseline data on mycorrhizal fungi-mammal interactions in an area with no known mammal declines. We examined the mycorrhizal fungi in root and soil samples via high-throughput sequencing and compared the observed taxa to those dispersed by mycophagous mammals at the same locations. We found that the dominant root-associating ectomycorrhizal fungal taxa (> 90% sequence abundance) included the truffle-like taxa Mesophellia, Hysterangium and Chondrogaster. These same taxa were also present in mycophagous mammalian diets, with Mesophellia often dominating. Altogether, 88% of truffle-like taxa from root samples were shared with the fungal specialist diet and 52% with diets from generalist mammals. Our data suggest that changes in mammal communities, particularly the loss of fungal specialists, could, over time, induce reductions to truffle-like fungal diversity, causing ectomycorrhizal fungal communities to shift with unknown impacts on plant and ecosystem health

Diatom metabarcoding and microscopic analyses from sediment samples at Lake Nam Co, Tibet: The effect of sample-size and bioinformatics on the identified communities

Diatoms (Bacillariophyceae) are characterized by silicified cell walls that favor their long-term preservation in sediments, therefore widely used as bioindicators of present and past water conditions. Alongside with traditional morphological analyses, metabarcoding has become a valuable tool to study the community structures of various organisms, including diatoms. Here, we test whether the quantity of sediment sample used for DNA extraction affects the results obtained from high-throughput sequencing (metabarcoding) of the diatom rbcL region by isolating DNA from 10 g and 0.5 g (wet weight) of lake surface sediment samples. Because bioinformatics processing of metabarcoding data may affect the outcome, we also tested the consistency of the results from three different pipelines: 1) ESVs (exact sequence variants) pipeline; 2) clustering sequences at 95% sequence identity to form OTUs (operational taxonomic units; 95% OTUs); and 3) 97% OTUs pipeline. Additionally, the agreement between metabarcoding data and morphological inventories of corresponding samples were compared. Our results demonstrate highly uniform patterns between the diatom rbcL amplicons from 10 g and 0.5 g of sedimentary DNA (sedDNA) extracts (HTS 10 and HTS 0.5, respectively). Furthermore, after the careful curation of the sequencing data, metabarcoding results were highly consistent among the data sets produced by different bioinformatics pipelines. Comparing results from metabarcoding and microscopy, we identified some taxonomic mismatches: morphological analyses identified 59 diatom genera, whereas metabarcoding 49 to 54 genera. These mismatches are related to incompleteness of the sequence databases, but also to inconsistencies in diatom taxonomy in general and potential dissolution effects of diatom valves caused by high alkalinity of the investigated lake waters. Nevertheless, multivariate community analysis revealed consistent results between data sets identified by microscopy and metabarcoding – water depth and conductivity as the most significant variables in driving diatom communities in Lake Nam Co – further confirming that metabarcoding is a viable method for identifying diatom-environment relationships

Tree diversity and species identity effects on soil fungi, protists and animals are context dependent

Plant species richness and the presence of certain influential species (sampling effect) drive the stability and functionality of ecosystems as well as primary production and biomass of consumers. However, little is known about these floristic effects on richness and community composition of soil biota in forest habitats owing to methodological constraints. We developed a DNA metabarcoding approach to identify the major eukaryote groups directly from soil with roughly species-level resolution. Using this method, we examined the effects of tree diversity and individual tree species on soil microbial biomass and taxonomic richness of soil biota in two experimental study systems in Finland and Estonia and accounted for edaphic variables and spatial autocorrelation. Our analyses revealed that the effects of tree diversity and individual species on soil biota are largely context dependent. Multiple regression and structural equation modelling suggested that biomass, soil pH, nutrients and tree species directly affect richness of different taxonomic groups. The community composition of most soil organisms was strongly correlated due to similar response to environmental predictors rather than causal relationships. On a local scale, soil resources and tree species have stronger effect on diversity of soil biota than tree species richness per se

Global patterns in endemicity and vulnerability of soil fungi

Fungi are highly diverse organisms, which provide multiple ecosystem services. However, compared with charismatic animals and plants, the distribution patterns and conservation needs of fungi have been little explored. Here, we examined endemicity patterns, global change vulnerability and conservation priority areas for functional groups of soil fungi based on six global surveys using a high-resolution, long-read metabarcoding approach. We found that the endemicity of all fungi and most functional groups peaks in tropical habitats, including Amazonia, Yucatan, West-Central Africa, Sri Lanka, and New Caledonia, with a negligible island effect compared with plants and animals. We also found that fungi are predominantly vulnerable to drought, heat and land-cover change, particularly in dry tropical regions with high human population density. Fungal conservation areas of highest priority include herbaceous wetlands, tropical forests, and woodlands. We stress that more attention should be focused on the conservation of fungi, especially root symbiotic arbuscular mycorrhizal and ectomycorrhizal fungi in tropical regions as well as unicellular early-diverging groups and macrofungi in general. Given the low overlap between the endemicity of fungi and macroorganisms, but high conservation needs in both groups, detailed analyses on distribution and conservation requirements are warranted for other microorganisms and soil organisms

Global patterns in endemicity and vulnerability of soil fungi

Fungi are highly diverse organisms, which provide multiple ecosystem services. However, compared with charismatic animals and plants, the distribution patterns and conservation needs of fungi have been little explored. Here, we examined endemicity patterns, global change vulnerability and conservation priority areas for functional groups of soil fungi based on six global surveys using a high-resolution, long-read metabarcoding approach. We found that the endemicity of all fungi and most functional groups peaks in tropical habitats, including Amazonia, Yucatan, West-Central Africa, Sri Lanka, and New Caledonia, with a negligible island effect compared with plants and animals. We also found that fungi are predominantly vulnerable to drought, heat and land-cover change, particularly in dry tropical regions with high human population density. Fungal conservation areas of highest priority include herbaceous wetlands, tropical forests, and woodlands. We stress that more attention should be focused on the conservation of fungi, especially root symbiotic arbuscular mycorrhizal and ectomycorrhizal fungi in tropical regions as well as unicellular early-diverging groups and macrofungi in general. Given the low overlap between the endemicity of fungi and macroorganisms, but high conservation needs in both groups, detailed analyses on distribution and conservation requirements are warranted for other microorganisms and soil organisms

Microbiomes from feces vs. gut in aquatic vertebrates: distinct community compositions between substrates and preservation methods

Sample type and preservation methods are likely to influence microbiome analysis results. Relatively few studies have explored the differences between feces and gut as well as ethanol-stored and frozen samples. Here, we sampled the same individuals of three aquatic vertebrates from the Qinghai-Tibetan Plateau non-invasively for feces, and subsequently for hindgut through dissection. Our study species, two fishes (Gymnocypris cf. namensis and Triplophysa sp.) and one amphibian (tadpoles of Nanorana parkeri), were all collected at the same time and site. Gut and fecal samples were stored in ethanol, and additionally, part of the gut samples were frozen, but temporarily thawed during transport as it often happens under difficult field conditions. Our results showed that both substrate (gut content vs. feces) as well as preservation method can influence the analysis of intestinal microbiomes. Frozen gut samples strongly differed from ethanol-stored samples, and especially in Nanorana most frozen samples were dominated (in relative abundance) by a set of Proteobacteria OTUs that were completely absent from the ethanol-stored samples. This blooming of contaminant bacteria occurred after less than 12 h of thawing, thus caution should be taken when constancy of cold temperatures cannot be maintained in the field for sample preservation purposes. Among ethanol-stored samples, bacterial communities from feces differed from those recovered from guts, but in part recovered similar patterns, such as a higher bacterial richness in the more herbivorous Nanorana tadpoles. Although our results argue against combining gut and fecal samples in analyses of host-specific microbiome differences, they also confirm that non-invasive sampling of feces can provide useful information of gut microbiomes in aquatic vertebrates, which may be important especially when working with endangered species

Tree species identity and diversity drive fungal richness and community composition along an elevational gradient in a Mediterranean ecosystem

Ecological and taxonomic knowledge is important for conservation and utilization of biodiversity. Biodiversity and ecology of fungi in Mediterranean ecosystems is poorly understood. Here, we examined the diversity and spatial dis- tribution of fungi along an elevational gradient in a Mediterranean ecosystem, using DNA metabarcoding. This study provides novel information about diversity of all eco- logical and taxonomic groups of fungi along an elevational gradient in a Mediterranean ecosystem. Our analyses revealed that among all biotic and abiotic variables tested, host species identity is the main driver of the fungal richness and fungal community composition. Fungal richness was strongly asso- ciated with tree richness and peaked in Quercus-dominated habitats and Cistus-dominated habitats. The highest taxonom- ic richness of ectomycorrhizal fungi was observed under Quercus ilex, whereas the highest taxonomic richness of saprotrophs was found under Pinus. Our results suggest that the effect of plant diversity on fungal richness and community composition may override that of abiotic variables across environmental gradients

Acricotopus indet. morphotype incurvatus: description and genetics of a new Orthocladiinae (Diptera: Chironomidae) larval morphotype from the Tibetan Plateau

High mountain ranges such as the Tibetan Plateau with an average altitude above 4500 m are topographically complex formations. Elevational gradients, physiographic diversity and climatic heterogeneity have led to highly biodiverse ecosystems in these regions. Mountain ranges can be seen as cradles of evolution and harbour, due to their unique characteristics, a high number of highly adapted species. At the same time these areas are hard to access and therefore taxonomic information is limited. Here we describe a new Acricotopus (Diptera: Chironomidae: Orthocladiinae) larval morphotype occurring in lakes and ponds of differing salinity and water depths located on the Southern and Central Tibetan Plateau. The description is based on larvae and their genetics (ribosomal 18S, 28S and mitochondrial COI sequences) collected from a shallow pond in close proximity to the large saline lake Selin Co. Larvae of Acricotopus indet. morphotype incurvatus are characterized by a mentum with a cluster of lateral teeth, partially folded inwards, a mandible with a toothed lobe in addition to four inner teeth and a sclerotized plate positioned behind the mentum. Up to now, these morphological features have only been found in early instars of other Acricotopus species. The proposed morphotype name is inspired by the peculiar form of the mentu

The effects of geographic origin and genotype on fungal diversity of silver birch (Betula pendula)

Abstract Soil microbes, especially root symbiotic fungi, often have drastic effects on the successful growth and establishment of plants. While plant intraspecific genetic variation is known to affect many ecosystem processes and functions, the effect it has on root fungal communities has received less attention. To determine the effect plant origin and genotype have on root fungal communities, we used high-throughput amplicon sequencing of ITS-regions to detect fungi from the roots of 64 clonally propagated silver birch (Betula pendula) trees representing four different geographical origins and 16 genotypes, all grown together in a common garden. We found that fungal alpha and beta-diversity but not community composition differ by silver birch genotype. Some birch genotypes are potentially more plastic in terms of their fungal interactions, which could make them more robust against environmental changes and provide a competitive advantage especially in disturbed habitats

Temperature and pH define the realised niche space of arbuscular mycorrhizal fungi

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