Kwon Gudam, Kim Kook-Hyung
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Korea.
Plant Genomics and Breeding Institute, Seoul National University, Seoul 08826, Korea.
Plant Pathol J. 2025 Aug;41(4):532-538. doi: 10.5423/PPJ.NT.05.2025.0062. Epub 2025 Aug 1.
Soil-borne pathogenic fungi cause substantial economic losses worldwide by infecting the underground parts of plants. In fruit trees, infections are especially damaging, as they often result in the death of the entire plant. Therefore, early detection is essential for effective disease management caused by soil-borne pathogens. In this study, we designed and validated real-time PCR primers targeting eight soil-borne and apple tree-associated phytopathogenic fungi. Each primer set successfully detected 20 ng of target genomic DNA (gDNA) within 25 cycles, while the same amount of non-target gDNA mixture was detected only after 35 cycles of amplification. Moreover, target DNA amplification remained unaffected in the presence of mixed non-target gDNA background, confirming the high specificity of the primers. Sensitivity test showed that 1 fg of plasmid DNA, corresponding to about 290 copies, was detectable around 30 cycles with all primer sets. These primers support accurate pathogen detection and early diagnosis in various environmental samples.
土壤传播的致病真菌通过感染植物的地下部分在全球范围内造成重大经济损失。在果树上，感染的危害尤其大，因为它们常常导致整株植物死亡。因此，早期检测对于有效管理由土壤传播病原体引起的病害至关重要。在本研究中，我们设计并验证了针对八种与土壤和苹果树相关的植物病原真菌的实时PCR引物。每个引物组均在25个循环内成功检测到20 ng的目标基因组DNA（gDNA），而相同量的非目标gDNA混合物仅在35个循环的扩增后才被检测到。此外，在存在混合非目标gDNA背景的情况下，目标DNA扩增不受影响，证实了引物的高特异性。灵敏度测试表明，所有引物组在约30个循环时均可检测到1 fg的质粒DNA，相当于约290个拷贝。这些引物有助于在各种环境样品中进行准确的病原体检测和早期诊断。
