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Updated: Dec 15, 2025

Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants
Published on: May 5, 2023
How to resolve cryptic species of polypores: an example in Fomes
Ursula Peintner1, Regina Kuhnert-Finkernagel1, Viana Wille1
1University Innsbruck, Institute of Microbiology, Technikerstr. 25, 6020 Innsbruck, Austria.
Abstract:
Species that cannot be easily distinguished based on morphology, but which form distinct phylogenetic lineages based on molecular markers, are often referred to as cryptic species. They have been proposed in a number of fungal genera, including the basidiomycete genus Fomes. The main aim of this work was to test new methods for species delimitation in cryptic lineages of polypores, and to define useful characters for species identification. A detailed examination of a number of different Fomes strains that had been collected and isolated from different habitats in Italy and Austria confirmed the presence of distinct lineages in the Fomes fomentarius clade. Our zero hypothesis was that the Mediterranean strains growing on Quercus represent a species which can be delimited based on morphological and physiological characters when they are evaluated in statistically relevant numbers. This hypothesis was tested based on phylogenetic analysis of the rDNA ITS region, morphological characters of basidiomes and pure cultures, growth rates and optimum growth temperature experiments, mycelial confrontation tests, enzyme activity tests and volatile organic compound (VOC) production. The Mediterranean lineage can unambiguously be delimited from F. fomentarius. A syntype of an obscure and previously synonymized name, Polyporus inzengae, represents the Mediterranean lineage that we recognize as Fomes inzengae, a distinct species. The rDNA ITS region is useful for delimitation of Fomes species. Moreover, also a variety of morphological characters including hymenophore pore size, basidiospore size, and diameter of skeletal hyphae are useful delimiting characters. The ecology is also very important, because the plant host appears to be a central factor driving speciation. Physiological characters turned also out to be species-specific, e.g. daily mycelial growth rates or the temperature range of pure cultures. The production of VOCs can be considered as a very promising tool for fast and reliable species delimitation in the future.
Insights
Cryptic fungal species, like those in the Fomes genus, can now be identified using new molecular and morphological methods. This research confirms a distinct Mediterranean lineage, Fomes inzengae, previously hidden within Fomes fomentarius.
Area of Science:
- Mycology
- Molecular Systematics
- Fungal Taxonomy
Background:
- Cryptic species, morphologically similar but genetically distinct, pose challenges in fungal taxonomy.
- The basidiomycete genus Fomes is known to harbor cryptic species, necessitating refined delimitation methods.
- Previous classifications may have overlooked distinct lineages within well-known Fomes species.
Purpose of the Study:
- To develop and test novel species delimitation techniques for cryptic polypore lineages.
- To identify reliable morphological and physiological characters for distinguishing Fomes species.
- To investigate the phylogenetic and ecological distinctiveness of Mediterranean Fomes strains.
Main Methods:
- Phylogenetic analysis of the ribosomal DNA Internal Transcribed Spacer (rDNA ITS) region.
- Detailed morphological examination of basidiomes and pure cultures.
- Physiological assessments including growth rates, temperature optima, enzyme activity, and volatile organic compound (VOC) production.
- Mycelial confrontation tests.
Main Results:
- Distinct phylogenetic lineages within the Fomes fomentarius clade were confirmed.
- The Mediterranean lineage, associated with Quercus hosts, was unambiguously delimited from Fomes fomentarius.
- Morphological characters (pore size, spore size, hyphal diameter), ecological factors (host plant), and physiological traits (growth rate, temperature range) proved valuable for species identification.
- The obscure name Polyporus inzengae was resurrected for the Mediterranean lineage, now recognized as Fomes inzengae.
Conclusions:
- The rDNA ITS region is a reliable marker for Fomes species delimitation.
- A combination of molecular, morphological, ecological, and physiological data is crucial for resolving cryptic species.
- Fomes inzengae is recognized as a distinct species, highlighting the importance of host-plant association in fungal speciation.
- Volatile organic compound analysis shows promise for rapid fungal species identification.
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