{"title":"rolfsii、S. delphinii和S. rolfsii两个种内类群分化的高分辨率熔化分析。","authors":"Dong Jae Lee, Young-Joon Choi","doi":"10.1080/12298093.2025.2561285","DOIUrl":null,"url":null,"abstract":"<p><p><i>Sclerotium</i> (=<i>Agroathelia</i>) <i>rolfsii</i> and <i>S. delphinii</i> are soilborne fungal pathogens responsible for southern blight and collar rot in numerous crops worldwide. Accurate detection and identification of these related pathogens are critical for disease management and the selection of resistant plant cultivars. In this study, we identified a polymorphic simple sequence repeat (SSR) locus through reference genome sequence analysis and developed a quantitative PCR (qPCR)-based high-resolution melting (HRM) assay to distinguish <i>S. delphinii</i>, <i>S. rolfsii</i>, and two intraspecific groups (G1 and G2) of <i>S. rolfsii</i>. Using a single primer pair in a closed-tube system without probes, HRM analysis differentiated the target groups by melting temperature profiles and detected as little as 1 pg of genomic DNA. The assay was validated on pure cultures, artificially inoculated soil, and rhizosphere soil from diseased tomato plants. This qPCR-HRM method provides a rapid, cost-effective, and robust diagnostic tool for the detection and discrimination of <i>S. rolfsi</i>i G1, <i>S. rolfsii</i> G2, and <i>S. delphinii</i>, complementing conventional PCR and probe-based qPCR diagnostics.</p>","PeriodicalId":18825,"journal":{"name":"Mycobiology","volume":"53 6","pages":"761-769"},"PeriodicalIF":2.0000,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12477771/pdf/","citationCount":"0","resultStr":"{\"title\":\"High-Resolution Melting Analysis for Differentiation of <i>Sclerotium</i> (=<i>Agroathelia</i>) <i>rolfsii</i>, <i>S. delphinii</i>, and Two Intraspecific Groups of <i>S. rolfsii</i>.\",\"authors\":\"Dong Jae Lee, Young-Joon Choi\",\"doi\":\"10.1080/12298093.2025.2561285\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><i>Sclerotium</i> (=<i>Agroathelia</i>) <i>rolfsii</i> and <i>S. delphinii</i> are soilborne fungal pathogens responsible for southern blight and collar rot in numerous crops worldwide. Accurate detection and identification of these related pathogens are critical for disease management and the selection of resistant plant cultivars. In this study, we identified a polymorphic simple sequence repeat (SSR) locus through reference genome sequence analysis and developed a quantitative PCR (qPCR)-based high-resolution melting (HRM) assay to distinguish <i>S. delphinii</i>, <i>S. rolfsii</i>, and two intraspecific groups (G1 and G2) of <i>S. rolfsii</i>. Using a single primer pair in a closed-tube system without probes, HRM analysis differentiated the target groups by melting temperature profiles and detected as little as 1 pg of genomic DNA. The assay was validated on pure cultures, artificially inoculated soil, and rhizosphere soil from diseased tomato plants. This qPCR-HRM method provides a rapid, cost-effective, and robust diagnostic tool for the detection and discrimination of <i>S. rolfsi</i>i G1, <i>S. rolfsii</i> G2, and <i>S. delphinii</i>, complementing conventional PCR and probe-based qPCR diagnostics.</p>\",\"PeriodicalId\":18825,\"journal\":{\"name\":\"Mycobiology\",\"volume\":\"53 6\",\"pages\":\"761-769\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-09-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12477771/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Mycobiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1080/12298093.2025.2561285\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q2\",\"JCRName\":\"AGRONOMY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mycobiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1080/12298093.2025.2561285","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"AGRONOMY","Score":null,"Total":0}
High-Resolution Melting Analysis for Differentiation of Sclerotium (=Agroathelia) rolfsii, S. delphinii, and Two Intraspecific Groups of S. rolfsii.
Sclerotium (=Agroathelia) rolfsii and S. delphinii are soilborne fungal pathogens responsible for southern blight and collar rot in numerous crops worldwide. Accurate detection and identification of these related pathogens are critical for disease management and the selection of resistant plant cultivars. In this study, we identified a polymorphic simple sequence repeat (SSR) locus through reference genome sequence analysis and developed a quantitative PCR (qPCR)-based high-resolution melting (HRM) assay to distinguish S. delphinii, S. rolfsii, and two intraspecific groups (G1 and G2) of S. rolfsii. Using a single primer pair in a closed-tube system without probes, HRM analysis differentiated the target groups by melting temperature profiles and detected as little as 1 pg of genomic DNA. The assay was validated on pure cultures, artificially inoculated soil, and rhizosphere soil from diseased tomato plants. This qPCR-HRM method provides a rapid, cost-effective, and robust diagnostic tool for the detection and discrimination of S. rolfsii G1, S. rolfsii G2, and S. delphinii, complementing conventional PCR and probe-based qPCR diagnostics.
期刊介绍:
Mycobiology is an international journal devoted to the publication of fundamental and applied investigations on all aspects of mycology and their traditional allies. It is published quarterly and is the official publication of the Korean Society of Mycology. Mycobiology publishes reports of basic research on fungi and fungus-like organisms, including yeasts, filamentous fungi, lichen fungi, oomycetes, moulds, and mushroom. Topics also include molecular and cellular biology, biochemistry, metabolism, developmental biology, environmental mycology, evolution, ecology, taxonomy and systematics, genetics/genomics, fungal pathogen and disease control, physiology, and industrial biotechnology using fungi.