Evidence map›Paper›PMID 41876377›Full record

ArticleMolecular ecology resources2026

Limitations of Common Molecular Markers in Fungal Biodiversity Analysis and the Benefits of Their Synergistic Use.

Vasilii Shapkin, Tomáš Zelenka, Tomáš Větrovský, Martin Kostovčík, Ivana Eichlerová, Petr Kohout, Lucia Žifčáková, Jan Borovička, Michal Tomšovský, Slavomír Adamčík and 2 more

Abstract readEvaluation Study
In one paragraph

Article in Molecular ecology resources, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Morphology and Multigene Phylogeny Reveal Six New Species ofJournal of fungi (Basel, Switzerland) · 2026
    Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

12 authors.

Vasilii ShapkinInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Tomáš ZelenkaInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Tomáš VětrovskýInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Martin KostovčíkInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Ivana EichlerováInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Petr KohoutInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Lucia ŽifčákováInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.
Jan BorovičkaInstitute of Geology of the Czech Academy of Sciences, Prague, Czech Republic.
Michal TomšovskýDepartment of Forest Protection and Wildlife Management, Faculty of Forestry and Wood Technology, Mendel University in Brno, Brno, Czech Republic.ORCID https://orcid.org/0000-0002-9505-6175
Slavomír AdamčíkLaboratory of Molecular Ecology and Mycology, Institute of Botany, Plant Science and Biodiversity Centre, Slovak Academy of Sciences, Bratislava, Slovakia.
Petr BaldrianInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.ORCID https://orcid.org/0000-0002-8983-2721
Miroslav KolaříkInstitute of Microbiology of the Czech Academy of Sciences, Praha, Czech Republic.ORCID https://orcid.org/0000-0003-4016-0335

Funding

Agentúra na Podporu Výskumu a Vývoja APVV-24-0473Akademie Věd České RepublikyCharles University Grant Agency 360015Grantová Agentura České Republiky 21-17749SGrantová Agentura České Republiky 25-17553SMinistry of Education, Youth and Sports of the Czech Republic CZ.02.01.01/00/22_008/0004597Ministry of Education, Youth and Sports of the Czech Republic CZ.02.01.01/00/22_08/0004635
6 · The paper itself

Abstract

High-throughput sequencing of the Internal Transcribed Spacer (ITS) regions is the primary method for estimating fungal diversity from environmental DNA. However, reliance solely on ITS markers is complicated by its high variability in sequence length and the presence of multiple variants within a single genome, which can bias diversity estimates. This study compares the performance of the ITS1 and ITS2 molecular markers with five alternative markers targeting protein-coding genes using a defined mock community of 413 fungal species from 32 orders. We show that none of the markers provided a reliable estimate of fungal diversity and two failed to produce sufficient data. While the ITS2 marker showed the highest proportion of individual species detections at 75.3%, a multi-marker approach significantly improved this result. Combining the top three markers (ITS1, ITS2 and elongation factor 1-alpha gene) increased this number to 92.2%. This approach also provided a crucial corrective for relative sequence abundance (RSA) estimates, as combining markers helps to offset the individual biases of each of them resulting in a more balanced picture of the community composition. Although the ITS region remains a cornerstone in fungal metabarcoding studies, we recommend combining both ITS1 and ITS2 markers with at least one alternative DNA marker to achieve more accurate species richness and RSA estimates. The broader application of these alternative markers will depend on the continued development of comprehensive, curated reference databases.

Indexed as

BiodiversityFungiDNA, FungalDNA, Ribosomal SpacerGenetic MarkersPeptide Elongation Factor 1Sequence Analysis, DNADNA, FungalDNA, Ribosomal SpacerGenetic MarkersPeptide Elongation Factor 1alternative markersfungihigh throughput sequencingmock communityrelative sequence abundance

Identifiers

PMID41876377
PMCPMC13013093

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.