rolfsii、S. delphinii和S. rolfsii两个种内类群分化的高分辨率熔化分析。

IF 2 4区 生物学 Q2 AGRONOMY
Mycobiology Pub Date : 2025-09-26 eCollection Date: 2025-01-01 DOI:10.1080/12298093.2025.2561285
Dong Jae Lee, Young-Joon Choi
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引用次数: 0

摘要

菌核菌(=Agroathelia) rolfsii和S. delphinii是土传真菌病原体负责南方枯萎病和衣领腐病在世界各地的许多作物。准确检测和鉴定这些相关病原体对病害管理和抗性植物品种的选择至关重要。本研究通过参考基因组序列分析,确定了一个多态性简单序列重复(SSR)位点,并建立了基于定量PCR (qPCR)的高分辨率融化(HRM)方法来区分S. delphinii, S. rolfsii以及S. rolfsii的两个种内类群(G1和G2)。在没有探针的封闭管系统中使用单个引物对,HRM分析通过熔化温度分布来区分目标群体,并检测到少至1pg的基因组DNA。在番茄病株的纯培养、人工接种土壤和根际土壤上验证了该方法的有效性。该qPCR- hrm方法为S. rolfsii G1、S. rolfsii G2和S. delphinii的检测和区分提供了一种快速、经济、强大的诊断工具,补充了传统的PCR和基于探针的qPCR诊断。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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.

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来源期刊
Mycobiology
Mycobiology AGRONOMYMYCOLOGY-MYCOLOGY
CiteScore
3.90
自引率
5.30%
发文量
41
审稿时长
22 weeks
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.
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