Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions

Susanna Keriö; Eeva Terhonen; Jared M LeBoldus

doi:10.21769/BioProtoc.3634

Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions

Bio Protoc. 2020 Jun 5;10(11):e3634. doi: 10.21769/BioProtoc.3634.

Authors

Susanna Keriö^{1

2}, Eeva Terhonen³, Jared M LeBoldus^{1

4}

Affiliations

¹ Department of Botany and Plant Pathology, Oregon State University, 2082 Cordley Hall, Corvallis OR, 97331, USA.
² Department of Forestry and Horticulture, Connecticut Agricultural Experiment Station, 123 Huntington Street, New Haven CT, 06511, USA.
³ Department of Forest Botany and Tree Physiology, Büsgen Institute, Büsgenweg 2, 37077 Göttingen, Germany.
⁴ Department of Forest Engineering, Resources and Management, Oregon State University, 210 Snell Hall, Corvallis OR, 97331, USA.

Abstract

We present a safe and low-cost method suitable for DNA extraction from mycelium and tree tissue samples. After sample preparation, the extraction takes about 60 min. Method performance was tested by extracting DNA from various tree tissue samples and from mycelium grown on solid and liquid media. DNA was extracted from juvenile and mature host material (Picea abies, Populus trichocarpa, Pseudotsuga menziesii) infected with different pathogens (Heterobasidion annosum, Heterobasidion parviporum, Leptographium wagenerii, Sphaerulina musiva). Additionally, DNA was extracted from pure cultures of the pathogens and several endophytic fungi. PCR success rate was 100% for young poplar material and fungal samples, and 48-72% for conifer and mature broadleaved plant samples. We recommend using 10-50 mg of fresh sample for the best results. The method offers a safe and low-cost DNA extraction alternative to study tree-fungus interactions, and is a potential resource for teaching purposes.

Keywords: DNA extraction; Forest pathology; Fungal DNA; Low-cost; Non-toxic; Plant DNA; Plant-microbe interactions.