Scholarlink

<p><i>Verticillium longisporum</i> is a highly destructive soil-borne pathogen responsible for verticillium wilt, a significant disease impacting numerous brassicaceous crops in various regions. Accurate species-level identification of <i>V. longisporum</i> is crucial for developing effective management strategies against verticillium wilt. However, the high sequence identity between <i>V. longisporum</i> and <i>Verticillium dahliae</i>, particularly at commonly used molecular markers like the Internal Transcribed Spacer (ITS) and Actin (ACT) genes, presents a significant challenge for accurate species-level differentiation. Here, we conducted a comprehensive comparative genomics analysis for <i>Verticillium</i> species. This approach, combined with targeted PCR validation, led to the identification of a novel V. longisporum-specific genomic fragment, designated VLspe2. Leveraging the sequence of VLspe2, we developed a precise Enzyme Mediated Duplex Exponential Amplification (EmDEA) assay for the rapid detection of <i>V. longisporum</i>, integrated with a fast DNA extraction protocol. The developed EmDEA assay demonstrated a minimum detection limit of 5&#xa0;fg in a 20 µL reaction volume. This isothermal EmDEA assay offers accurate and rapid detection of <i>V. longisporum</i> at a low constant temperature (37–42&#xa0;°C), providing invaluable guidance for on-site detection in agricultural fields and ports, ultimately contributing to plant disease management and food safety.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Rapid detection of the plant pathogenic fungi Verticillium longisporum using species-specific fragment and enzyme mediated duplex exponential amplification (EmDEA) assay

  • 期刊论文

  • 发表日期：

    2026年4月1日

查看全文
  • Yang Chen,
  • Xiaofei Lu,
  • Zhenchuan Mao,
  • Yan Li,
  • Jianlong Zhao,
  • Yuhong Yang,
  • Guohua Chen,
  • Jian Ling

摘要

Verticillium longisporum is a highly destructive soil-borne pathogen responsible for verticillium wilt, a significant disease impacting numerous brassicaceous crops in various regions. Accurate species-level identification of V. longisporum is crucial for developing effective management strategies against verticillium wilt. However, the high sequence identity between V. longisporum and Verticillium dahliae, particularly at commonly used molecular markers like the Internal Transcribed Spacer (ITS) and Actin (ACT) genes, presents a significant challenge for accurate species-level differentiation. Here, we conducted a comprehensive comparative genomics analysis for Verticillium species. This approach, combined with targeted PCR validation, led to the identification of a novel V. longisporum-specific genomic fragment, designated VLspe2. Leveraging the sequence of VLspe2, we developed a precise Enzyme Mediated Duplex Exponential Amplification (EmDEA) assay for the rapid detection of V. longisporum, integrated with a fast DNA extraction protocol. The developed EmDEA assay demonstrated a minimum detection limit of 5 fg in a 20 µL reaction volume. This isothermal EmDEA assay offers accurate and rapid detection of V. longisporum at a low constant temperature (37–42 °C), providing invaluable guidance for on-site detection in agricultural fields and ports, ultimately contributing to plant disease management and food safety.