Work

A Rapid Genomic DNA Extraction Method for Direct Quantitative Polymerase Chain Reaction-Based On-Site Detection of Soil-Borne Fungal Disease

Year: 2026

Type: article

Abstract: Soil-borne phytopathogenic fungi cause root rot, wilt, and damping-off in crops, leading to major yield losses worldwide. Because symptoms appear only after underground infection progresses, early detection is crucial. Here, a rapid 20-min genomic DNA extraction method was developed for eight pathogens-Alternaria tenuissima, Botryosphaeria dothidea, Fusarium oxysporum, Glomerella cingulata, Phytophthora cactorum, Rosellinia necatrix, Sclerotium rolfsii, and Sclerotinia sclerotiorum. The protocol uses a cetyltrimethylammonium bromide-based buffer, steel and glass beads, brief heating (95°C, 1 min), vortexing, and sequential purification with Q-Sepharose and magnetic beads. All pathogens were detected within 30 quantitative polymerase chain reaction cycles, while soil-only controls exceeded 30 Cq. Sclerotial DNA of S. rolfsii (Cq ≈ 25) was also detected, confirming applicability for overwintering inocula. This simple and low-cost protocol enables rapid, reliable detection of multiple soil-borne fungi directly from soil, providing a practical tool for on-site disease diagnosis and management.

Source: The Plant Pathology Journal

DOI: 10.5423/ppj.nt.10.2025.0163

Authors: Gudam Kwon, Dain Hong, Sen Lian, Jisuk Yu, Kook-Hyung Kim

Institutions: Seoul National University, Qingdao Agricultural University

Language: English

Topic: Plant Pathogens and Fungal Diseases

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Zero hunger

Open Access status: gold

Estimated APC paid: $850