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Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants
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Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions.
Susanna Keriö1,2, Eeva Terhonen3, Jared M LeBoldus1,4
1Department of Botany and Plant Pathology, Oregon State University, 2082 Cordley Hall, Corvallis OR, 97331, USA.
Bio-Protocol
|March 4, 2021
Summary
This study introduces a safe, affordable DNA extraction method for tree and fungal samples, yielding results in about an hour. It
Area of Science:
- Forest pathology
- Mycology
- Plant science
Background:
- Accurate DNA extraction is crucial for studying tree-pathogen interactions.
- Existing methods can be costly or hazardous.
- A need exists for accessible DNA extraction protocols for ecological and educational purposes.
Purpose of the Study:
- To develop and validate a safe, low-cost DNA extraction method.
- To assess the method's efficacy across diverse tree tissues and fungal samples.
- To provide a reliable protocol for research and teaching in forest science.
Main Methods:
- Developed a novel DNA extraction protocol from mycelium and tree tissues.
- Tested the method on various tree species (Picea abies, Populus trichocarpa, Pseudotsuga menziesii) and fungal pathogens.
- Evaluated DNA yield and quality for downstream PCR applications.
Main Results:
- Successful DNA extraction achieved from multiple tree tissues and fungal cultures.
- PCR success rates varied: 100% for young poplar and fungal samples, 48-72% for conifers and mature broadleaves.
- Optimal sample input is 10-50 mg of fresh material.
Conclusions:
- The presented method is a safe, cost-effective alternative for DNA extraction.
- It is suitable for analyzing tree-fungus interactions and diverse fungal samples.
- The protocol is a valuable resource for scientific research and educational settings.
