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Tedersoo, L.

Publications and source records attributed to Tedersoo, L..

15 recordsLinked to original sources

ClassifyITS: An R Package for assigning taxonomy to fungal ITS sequences using taxon-specific cutoff values

1. Fungi are key drivers of decomposition and nutrient cycling across the globe, yet accurate classification of environmental fungal internal transcribed spacer (ITS) sequences remains challenging. These persistent challenges reflect the variable evolutionary properties of ITS, limited representation of fungal diversity in reference databases, and the application of classifiers originally developed for more conserved prokaryotic markers. 2. Here, we present ClassifyITS, an R package that performs alignment-based taxonomic classification of full length fungal ITS sequences or individual ITS subregions (ITS1 or ITS2) using taxon-specific sequence identity thresholds. In addition to taxonomic assignments, ClassifyITS generates summary statistics and diagnostic visualizations to support interpretation and quality control. 3. Using a deep subsurface fungal ITS dataset containing many poorly characterized taxa, ClassifyITS outperformed the common classifiers SINTAX and DADA2, with higher agreement to expert curated assignments and lower rates of over classifying and under classifying sequences to taxonomic ranks. Across all classifiers and approaches, taxonomic accuracy increased strongly with sequence similarity to the reference database, emphasizing the importance of continued expansion and curation of fungal sequence databases. 4. By providing an accessible and reproducible R based workflow that improves taxonomic classification, ClassifyITS supports more accurate biodiversity monitoring and enhances downstream functional interpretation of fungal communities.

bioinformatics↗

Full-length COI barcodes improve eDNA metabarcoding data denoising relative to mini-barcodes

Animal COI (mitochondrial cytochrome oxidase I) metabarcoding of environmental DNA (eDNA) is increasingly used to assess biodiversity in complex substrates such as soil. However, due to read-length constraints of second-generation sequencing platforms, mini-barcodes have been used instead of the full barcode region. Long-read sequencing technologies now enable the recovery of full-length barcode sequences, and are more commonly applied for studying microbes, but their use for metabarcoding the full-length standard COI barcoding region in animals remains limited. In this study, we compared three COI amplicon sets - 313 bp, 660 bp, and 1,256 bp - amplified from soil eDNA samples and sequenced using Illumina and PacBio platforms to evaluate their overall concurrence, the effectiveness of identifying nuclear mitochondrial DNA segments (NUMTs) and chimeras, as well as their respective taxonomic resolution. The long-read datasets exhibited a higher identification rate of NUMTs and true chimeras, suggesting that longer sequences improve the detection of noise in COI metabarcoding data, thereby reducing the occurrence of spurious taxa. Taxonomy assignment confidence was similar between the 313 bp and 660 bp datasets, whereas extending the amplicon beyond the standard COI barcode region (1,256 bp) reduced confidence, likely because longer reads extend into regions poorly represented in barcode reference databases. Despite substantially lower sequencing depth in the 660 bp dataset, per-sample OTU richness did not differ significantly from that recovered with the Illumina 313 bp amplicon set. Similarly, the relationships between samples were strongly correlated across the detected OTU communities, indicating consistent ecological interpretations between short and long amplicons. We conclude that the standard [~]658 bp COI barcode is an optimal marker for soil animal metabarcoding from eDNA, balancing target recovery, artifact detection, taxonomic assignment and ecological interpretability. As COI eDNA metabarcoding becomes increasingly used in biodiversity assessment and is increasingly adopted in large-scale monitoring initiatives, this study provides methodological guidance for improving the robustness of soil animal community biomonitoring.

ecology↗

Benchmarking full-length ITS metabarcoding across Illumina 2x500, PacBio, and Oxford Nanopore sequencing using mock and soil communities

Metabarcoding is a powerful tool for biodiversity comparisons, where standard-size DNA barcodes (>500 bases) offer better taxonomic resolution than shorter ones. Still, the choice of sequencing platforms and bioinformatics pipelines may strongly affect inferred diversity due to various technical biases. We assessed the relative performance of Illumina MiSeq i100 (2x500 paired-end), PacBio Revio and Oxford Nanopore MinION sequencing and bioinformatics pipelines, using full-length ITS amplicon sequencing datasets from a 103-species mock community and 45 composite soil samples. Despite numerous low-quality reads, PacBio yielded the lowest overall error rate and highest number of taxa. Illumina revealed the highest proportion of chimeric and index-switched reads, along with a strong bias towards shorter amplicons. MinION data analysed using PRONAME and Minovar - a bioinformatics pipeline presented here - had the largest proportion of low-quality data, and rare taxa were lost during data filtering and read polishing steps. Although Minovar enabled amplicon sequence variant (ASV) level precision for common taxa, we recommend clustering ASVs into OTUs. For PacBio, standard filtering approaches outperformed the ASV approach because they retained rare taxa. For Illumina, a stringent ASV approach or removal of rare OTUs would limit artefacts. Across all platforms, excess PCR cycles promoted chimeric and low-quality reads and lost quantitativity in biodiversity assessments. With moderate differences in effect sizes, all analytical approaches supported the conclusion that sampling design determines how we see soil biodiversity responses to land use. For biodiversity surveys based on the full-length ITS metabarcoding, we recommend using PacBio sequencing with standard, non-ASV pipelines.

bioinformatics↗

Global population structures and demographic history of Suillus luteus, a pine co-introduced ectomycorrhizal fungus associated with exotic forestry and invasion.

Human colonization since the 19th century has resulted in the global spread of pines across the Southern Hemisphere, well beyond their original northern boreal distribution. Such introductions moved not only the pines but also expanded the distribution of their symbiotic partners. Although the introduction of pines is documented through historical records, little is known about the introduction history of their ectomycorrhizal fungi, which are critical symbionts for the survival and invasion of pines. Using Suillus luteus as an example, population genomic analyses of 208 individuals across both native and introduced ranges showed that all introductions originated from Europe, likely mediated by human activities along with pine introductions. With the exception of North America, introduced populations were genetically differentiated from the Europe population, with varying magnitudes of population expansion in different introduced regions, often linked to forestry practices. Genetic variation within the native European population followed isolation by distance, but not in the introduced range, highlighting the disparity in the spatial genetic patterns of native versus exotic habitats. This study provides insight into the population genetics of a globally introduced ectomycorrhizal fungus whose introduction process is likely applicable to other pine-co-introduced ectomycorrhizal fungi.

evolutionary biology↗

Splitting and filling the gaps: a reorganization of Corymbiglomeraceae and new taxa from trans-Pacific tropical regions

Diversisporales comprises species with worldwide distribution that produce glomoid, otosporoid, or tricisporoid spores. The recent reorganization of the order by Oehl et al. (2016) recognizes two families, Diversisporaceae and Corymbiglomeraceae, comprising one and five genera, respectively. Several Glomeromycotan specimens collected in northern and southeastern Mexico and in French Polynesian atolls were characterized using both morphological and molecular analyses. Phylogenetic inference revealed that they represent new members in the Diversisporales, supporting the reorganization of the genus Redeckera into three independent lineages: Albocarpum gen. nov., with A. arenaceum sp. nov., A. leptohyphum sp. nov., and A. fulvum comb. nov., Pulvinocarpum pulvinatum gen. et comb. nov., and Redeckera, which retains five species, including R. varelae sp. nov. In addition, we described Melanocarpum mexicanum gen. et sp. nov. and Diversispora papillosa sp. nov. A broader phylogeny, based on eDNA sequences and representative of Diversisporales species, including the newly described taxa, further supported the split of Redeckera and suggested three additional clades likely corresponding to a new family and two new genera, awaiting the discovery of representative morphospecies to be formally described. Using eDNA sequences metadata, the occurrences of the newly described taxa were mapped, allowing to recognize distribution patterns, mostly in the pantropical zone, distinguish widespread and rare species, and suggest possible endemisms. Finally, the coexistence of species forming large sporocarps (A. fulvum and A. leptohyphum) alongside species forming spores in loose aggregates (A. arenaceum), prompted us to propose a possible sporulation dimorphism in Albocarpum, an argument previously raised to explain the nested placement of Corymbiglomus and Paracorymbiglomus within the Redeckera clade.

evolutionary biology↗

Sampling design and sample processing affect soil biodiversity assessments

Biodiversity surveys require an appropriate sampling design for optimal performance and comparability across space and time and across studies. Based on PacBio and Illumina amplicon sequencing of animals, bacteria and fungi, we assessed and compared various soil sampling designs from widely used continental and global metabarcoding-based biodiversity projects. Sampling designs revealed up to 27-fold, 6-fold and 15-fold differences in biodiversity estimates for animals, bacteria and fungi, respectively. Taxonomic coverage depended mostly on the number of subsamples but not sampling area within the 347-1790 m2, 428-1924 m2, 327-1790 m2 plot size range for animals, bacteria and fungi, respectively. Additional sampling of subsoil did not add significantly to diversity estimates of animals and fungi, although there was a slight positive effect on bacteria. Both soil pooling (compositing subsamples before DNA extraction) and DNA pooling (combining DNA extracts prior to PCR) reduced differences between sampling designs by decreasing diversity estimates in designs with many subsamples while increasing them in designs with fewer subsamples. However, pooling did not eliminate the influence of sampling factors (soil depth, sampling area and sample size). DNA pooling outperformed soil pooling in inventorying animals and fungi but not bacteria. Pooling had no effect on recovering rare biological species or sequencing artefacts. Soil pooling saved from 79.5% to 98.3% of labour and analytical costs compared with no pooling, depending on the number of pooled subsamples. The remarkable impact of sampling design on community diversity and composition should be considered during data collection and meta-analyses compiling data from different sampling designs.

microbiology↗

Variation of carbon, nitrogen and phosphorus content in fungi reflects their ecology and phylogeny

Fungi are an integral part of the nitrogen and phosphorus cycling in trophic networks, as they participate in biomass decomposition and facilitate plant nutrition through root symbioses. Nutrient content varies considerably between the main fungal habitats, such as soil, plant litter or decomposing dead wood, but there are also large differences within habitats. While some soils are heavily loaded with N, others are limited by N or P. One way in which nutrient availability can be reflected in fungi is their content in biomass. In this study, we determined the C, N, and P content (in dry mass) of sporocarps of 214 fungal species to inspect how phylogeny and membership in ecological guilds (soil saprotrophs, wood saprotrophs, and ectomycorrhizal fungi) affect the nutrient content of fungal biomass. The C content of sporocarps (415 {+/-} 25 mg g-1) showed little variation (324-494 mg g-1), while the range of N (46 {+/-} 20 mg g-1) and P (5.5 {+/-} 3.0 mg g-1) contents was within one order of magnitude (8-103 mg g-1 and 1.0-18.9 mg g-1, respectively). Importantly, the N and P contents were significantly higher in the biomass of soil saprotrophic fungi compared to wood saprotrophic and ectomycorrhizal fungi. While the average C/N ratio in fungal biomass was 11.2, values exceeding 40 were recorded for some fungi living on dead wood, typically characterized by low N content. The N and P content of fungal mycelium also showed a significant phylogenetic signal, with differences in nutrient content being relatively low within species and genera of fungi. A strong correlation was found between N and P content in fungal biomass, while the correlation of N content and the N-containing fungal cell wall biopolymer - chitin showed only weak significance. The content of macronutrients in fungal biomass is influenced by the fungal life style and nutrient availability and is also limited by phylogeny.

microbiology↗

Soil health is linked to primary productivity across Europe

A healthy soil is at the core of sustainable management and policy, but its importance for plant productivity across environmental gradients and land-use types remains poorly understood. To address this gap, we conducted a pan-European field study including 588 sites from 27 countries to investigate the link between soil health and primary productivity across three major land-use types: woodlands, grasslands, and croplands. We found that mean soil health (a composite index based on soil properties, biodiversity, and plant disease control) in woodlands was 31.4% higher than in grasslands, and 76.1% higher than in croplands. Soil health was positively linked to cropland and grassland productivity at the continental scale. Woodland productivity was best explained by climate. Among microbial diversity indicators, we observed a positive association between the richness of Acidobacteria, Firmicutes, and Proteobacteria and primary productivity. Among microbial functional groups, we found that nitrogen-fixing bacteria and mycorrhizal fungi positively related to primary productivity in croplands and grasslands, while plant pathogens showed a negative relationship. Together, our results point to the importance of soil biodiversity and soil health for maintaining primary productivity across contrasting land-use types.

ecology↗

What mycologists should talk about when they are talking about the International Code of Nomenclature for algae, fungi, and plants

Fungal metabarcoding of substrates such as soil, wood, and water are uncovering an unprecedented number of fungal species that do not seem to produce tangible morphological structures and that defy our best attempts at cultivation, thus falling outside of the ambit of the International Code of Nomenclature for algae, fungi, and plants. The present study uses the new, ninth release of the species hypotheses of the UNITE database to show that species discovery through environmental sequencing vastly outpaces traditional, Sanger sequencing-based efforts in a strongly increasing trend over the last five years. Our findings challenge the present stance of the mycological community - that "the code" works fine and that these complications will somehow sort themselves out given enough time and a following wind - and suggest that we should be discussing not whether to allow DNA-based descriptions (typifications) of species and by extension higher ranks of fungi, but what the precise requirements for such DNA-based typifications should be. We submit a tentative list of such criteria for further discussion. However, the present authors fear that no waves of change will be lapping the shores of mycology for the foreseeable future, leaving the overwhelming majority of extant fungi without formal names and thus scientific and environmental agency. It is not clear to us who benefits from that, but neither fungi nor mycology are likely to be on the winning side.

scientific communication and education↗

MycoPhylo experiment reveals how mycorrhiza types and phylogenetic relationships affect soil biodiversity and functioning

O_LINatural forests and abandoned agricultural lands are increasingly replaced by monospecific forest plantations that have poor capacity to support biodiversity and ecosystem services. Natural forests harbour plants belonging to different mycorrhiza types that differ in their microbiome and carbon and nutrient cycling properties. C_LIO_LIHere we describe the MycoPhylo field experiment that encompasses 116 woody plant species from three mycorrhiza types and 237 plots, with plant diversity and mycorrhiza type diversity ranging from one to four and one to three per plot, respectively. C_LIO_LIThe MycoPhylo experiment enables us to test hypotheses about the plant species, species diversity, mycorrhiza type, and mycorrhiza type diversity effects and their phylogenetic context on soil microbial diversity and functioning and soil processes. C_LIO_LIAlongside with other experiments in the TreeDivNet consortium, MycoPhylo will contribute to our understanding of the tree diversity effects on soil biodiversity and ecosystem functioning across biomes, especially from the mycorrhiza type and phylogenetic conservatism perspectives. C_LI

plant biology↗

Towards understanding diversity, endemicity and global change vulnerability of soil fungi

Fungi play pivotal roles in ecosystem functioning, but little is known about their global patterns of diversity, endemicity, vulnerability to global change drivers and conservation priority areas. We applied the high-resolution PacBio sequencing technique to identify fungi based on a long DNA marker that revealed a high proportion of hitherto unknown fungal taxa. We used a Global Soil Mycobiome consortium dataset to test relative performance of various sequencing depth standardization methods (calculation of residuals, exclusion of singletons, traditional and SRS rarefaction, use of Shannon index of diversity) to find optimal protocols for statistical analyses. Altogether, we used six global surveys to infer these patterns for soil-inhabiting fungi and their functional groups. We found that residuals of log-transformed richness (including singletons) against log-transformed sequencing depth yields significantly better model estimates compared with most other standardization methods. With respect to global patterns, fungal functional groups differed in the patterns of diversity, endemicity and vulnerability to main global change predictors. Unlike -diversity, endemicity and global-change vulnerability of fungi and most functional groups were greatest in the tropics. Fungi are vulnerable mostly to drought, heat, and land cover change. Fungal conservation areas of highest priority include wetlands and moist tropical ecosystems.

ecology↗

DNA barcoding of fungal specimens using long-read high-throughput sequencing

Molecular methods are increasingly used to identify species that lack conspicuous macro- or micromorphological characters. Taxonomic and ecological research teams barcode large numbers of collected voucher specimens annually. In this study we assessed the efficiency of long-read high throughput sequencing (HTS) as opposed to the traditionally used Sanger method for taxonomic identification of multiple vouchered fungal specimens, and providing reference information about intra-individual allele polymorphism. We developed a workflow based on a test-set of 423 fungal specimens (representing 205 species), PacBio HTS method, and ribosomal rRNA operon internal transcribed spacer (ITS) and 28S rRNA gene (LSU) markers. PacBio HTS had a higher success rate than Sanger sequencing at a comparable cost. Species identification based on PacBio reads was usually straightforward, because the dominant operational taxonomic unit (OTU) typically represented the targeted organism. Unlike the Sanger method, PacBio HTS enabled detecting widespread allele polymorphism within the ITS marker in the studied specimens. We conclude that multiplex DNA barcoding of the fungal ITS and LSU markers using a PacBio HTS is a useful tool for taxonomic identification of large amounts of collected voucher specimens at competitive price. Furthermore, PacBio HTS accurately recovers various alleles, which can provide crucial information for species delimitation and population-level studies.

molecular biology↗

Relative performance of Oxford Nanopore MinION vs. Pacific Biosciences Sequel third-generation sequencing platforms in identification of agricultural and forest pathogens

Culture-based molecular characterization methods have revolutionized detection of pathogens, yet these methods are either slow or imprecise. The second-generation sequencing tools have much improved precision and sensitivity of detection, but the analysis processes are costly and take several days. Of third-generation techniques, the portable Oxford Nanopore MinION device has received much attention because of its small size and possibility of rapid analysis at reasonable cost. Here, we compare the relative performance of two third-generation sequencing instruments, MinION and Pacific Biosciences Sequel in identification and diagnostics of pathogens from conifer needles and potato leaves and tubers. We demonstrate that Sequel is efficient in metabarcoding of complex samples, whereas MinION is not suited for this purpose due to the high error rate and multiple biases. However, we find that MinION can be utilized for rapid and accurate identification of dominant pathogenic organisms from plant tissues following both amplicon-based and metagenomics-based approaches. Using the PCR-free approach with shortened extraction and incubation times, we performed the entire MinION workflow from sample preparation through DNA extraction, sequencing, bioinformatics and interpretation in two and half hours. We advocate the use of MinION for rapid diagnostics of pathogens, but care needs to be taken to control or account for all potential technical biases.\n\nIMPORTANCEWe develop new and rapid protocols for MinION-based third-generation diagnostics of plant pathogens that greatly improves the speed and precision of diagnostics. Due to high error rate and technical biases in MinION, PacBio Sequel platform is more useful for amplicon-based metabarcoding from complex biological samples.

microbiology↗

Human-induced decrease of ectomycorrhizal vegetation led to loss in global soil carbon content

Vegetation impacts on ecosystem functioning are mediated by mycorrhiza, a plant-fungal association formed by most plant species. Ecosystems dominated by distinct mycorrhizal types differ strongly in their biogeochemistry. Quantitative analyses of mycorrhizal impacts on ecosystem functioning are hindered by the absence of information on mycorrhizal distribution. We present the first global high-resolution maps of vegetation biomass distribution among main types of mycorrhizal associations. Arbuscular, ecto-, ericoid and non-mycorrhizal vegetation store 241{+/-}15, 100{+/-}17, 7{+/-}1.8 and 29 {+/-} 5.5 GT carbon in aboveground biomass, respectively. Soil carbon stocks in both topsoil and subsoil are positively related to the biomass fraction of ectomycorrhizal plants in the community, though the strength of this relationship varies across biomes. We show that human-induced transformations of Earths ecosystems have reduced ectomycorrhizal vegetation, with potential knock-on effects on terrestrial carbon stocks. Our work provides a benchmark for spatially explicit global quantitative assessments of mycorrhizal impacts on ecosystem functioning and biogeochemical cycles.\n\nOne Sentence SummaryFirst maps of the global distribution of mycorrhizal plants reveal global losses of ectomycorrhizal vegetation, and quantitative links between mycorrhizal vegetation patterns and terrestrial carbon stocks.

ecology↗

Proposal for practical multi-kingdom classification of eukaryotes based on monophyly and comparable divergence time criteria

Much of the ecological, taxonomic and biodiversity research relies on understanding of phylogenetic relationships among organisms. There are multiple available classification systems that all suffer from differences in naming, incompleteness, presence of multiple non-monophyletic entities and poor correspondence of divergence times. These issues render taxonomic comparisons across the main groups of eukaryotes and all life in general difficult at best. By using the monophyly criterion, roughly comparable time of divergence and information from multiple phylogenetic reconstructions, I propose an alternative classification system for the domain Eukarya to improve hierarchical taxonomical comparability for animals, plants, fungi and multiple protist groups. Following this rationale, I propose 32 kingdoms of eukaryotes that are treated in 10 subdomains. These kingdoms are further separated into 43, 115, 140 and 353 taxa at the level of subkingdom, phylum, subphylum and class, respectively (http://dx.doi.org/10.15156/BIO/587483). Most of the names have been used previously or these were deduced from those of the type taxa to be able to unambiguously link genera to higher taxonomic levels. In the era of phylogenomics, understanding about the phylogenetic relationships among organisms is rapidly increasing. Classifications systems must keep pace with this race to serve the research community by consistent improvements in precision in terms of taxonomic resolution and maintaining monophyly of the ingredient taxa.

evolutionary biology↗