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Related Experiment Video

Updated: May 21, 2026

Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants

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Methods for DNA barcoding of fungi.

Ursula Eberhardt¹

¹CBS-KNAW Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands. u.eberhardt@cbs.knaw.nl

Methods in Molecular Biology (Clifton, N.J.)

|June 12, 2012

Summary

This study details DNA barcoding methods for fungi, focusing on macro-fungi and aged specimens. It outlines nuclear ribosomal gene sequencing (ITS and LSU D1/D2) for fungal identification.

Area of Science:

Mycology
Molecular Biology
Genetics

Background:

Accurate fungal identification is crucial for ecological and taxonomic studies.
DNA barcoding offers a molecular approach for species identification.
Standardized fungal DNA barcoding loci are still under development.

Purpose of the Study:

To describe current DNA barcoding methodologies applicable to fungi.
To provide guidance on collecting and processing fungal samples, including aged herbarium material.
To outline laboratory protocols for medium-throughput DNA barcoding using a 96-well format.

Main Methods:

Focus on collection procedures for macro-fungi.
Laboratory methods adapted for medium-throughput DNA barcoding (96-well format).

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Amplification and sequencing of nuclear ribosomal internal transcribed spacer (ITS) regions.

Amplification and sequencing of the large ribosomal subunit (LSU) D1/D2 region.

Main Results:

The described methods are suitable for identifying fungi from various environments.
Nuclear ribosomal genes ITS and LSU D1/D2 are presented as widely used markers.
Protocols are detailed for non-robotic, medium-throughput processing.

Conclusions:

The chapter provides practical guidance for fungal DNA barcoding.
ITS and LSU D1/D2 are effective markers for fungal identification in the absence of a single standard.
These methods can be applied to both fresh and aged fungal specimens.