Methodology for Extracting High-Molecular-Weight DNA from Field Collections of Macrofungi

Area of Science:

• Mycology
• Genomics
• Molecular Biology

Background:

• Many macrofungi cannot be cultured, necessitating environmental DNA (eDNA) isolation for genomic studies.
• Collecting samples in remote locations often precludes refrigeration, requiring alternative preservation methods.
• Fungal secondary metabolites can interfere with DNA purification, demanding optimized extraction protocols.

Purpose of the Study:

• To develop and describe a reliable DNA extraction protocol for environmental macrofungi samples.
• To enable high-quality genome assembly from non-refrigerated fungal specimens.
• To provide a practical method for researchers working with challenging fungal samples.

Main Methods:

• Samples collected in isopropanol at ambient temperature.
• Chromatin isolation strategy employed.
• Sequential removal of unwanted molecular components for DNA purification.

Main Results:

• Successful DNA isolation from diverse Australasian ectomycorrhizal and saprotrophic macrofungi.
• Protocol adaptable to species-specific metabolite profiles.
• Yielded DNA suitable for long-read sequencing and genome assembly.

Conclusions:

• The described protocol is effective for extracting high-quality DNA from environmental macrofungi.
• This method overcomes limitations of culturing and sample preservation.
• Facilitates advancements in fungal genomics and biodiversity research.