Scholarlink

<p><i>Ustilaginoidea virens</i>, the causative agent of rice false smut, poses a significant threat to global rice production, reducing grain yield and quality. Our objective was to develop rapid and sensitive detection assays that enable timely detection and management. BLASTn analysis was conducted on the whole genome of <i>U. virens</i> Uv-8b against NCBI NR database in order to find sequences exclusive to <i>U. virens</i>. Only candidate genes that were 100% identical to the genomes of <i>U. virens</i> and had no homology to non-target species were retained. Finally, the <i>Uv-05919</i> gene was selected and used to develop conventional PCR, quantitative PCR, Recombinase Polymerase Amplification (RPA), and Loop-Mediated Isothermal Amplification (LAMP) assays. The specificity of the assays was evaluated using multiple <i>U. virens</i> isolates as well as non-<i>U. virens</i> pathogens. All the developed assays exhibited high specificity to <i>U. virens</i> with no cross-reactivity with non-target organisms. The sensitivity thresholds were 100 pg / 50 µL in RPA, 10 pg / 25 µL in LAMP, 12 plasmid copies/ 20 µL in qPCR, and 100 pg/ 25 µL in conv-PCR. All assays reliably detected <i>U. virens</i> in infected rice panicles but not in healthy controls, demonstrating their field applicability.</p> Graphical Abstract <p></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

A Multi-assay molecular toolkit for rapid and sensitive detection of Ustilaginoidea virens the causative agent of rice false smut using PCR, qPCR, RPA, and LAMP assays

  • 期刊论文

  • 发表日期：

    2026年3月18日

查看全文
  • Mohamad Ayham Shakouka,
  • Sapna Sharma,
  • Sanghmitra Aditya,
  • Dwijesh Chandra Mishra,
  • Deepak Singh Bisht,
  • Anirban Roy,
  • Mahender Singh Saharan,
  • Gopala Krishnan Subbaiyan,
  • Bishnu Maya Bashyal

摘要

Ustilaginoidea virens, the causative agent of rice false smut, poses a significant threat to global rice production, reducing grain yield and quality. Our objective was to develop rapid and sensitive detection assays that enable timely detection and management. BLASTn analysis was conducted on the whole genome of U. virens Uv-8b against NCBI NR database in order to find sequences exclusive to U. virens. Only candidate genes that were 100% identical to the genomes of U. virens and had no homology to non-target species were retained. Finally, the Uv-05919 gene was selected and used to develop conventional PCR, quantitative PCR, Recombinase Polymerase Amplification (RPA), and Loop-Mediated Isothermal Amplification (LAMP) assays. The specificity of the assays was evaluated using multiple U. virens isolates as well as non-U. virens pathogens. All the developed assays exhibited high specificity to U. virens with no cross-reactivity with non-target organisms. The sensitivity thresholds were 100 pg / 50 µL in RPA, 10 pg / 25 µL in LAMP, 12 plasmid copies/ 20 µL in qPCR, and 100 pg/ 25 µL in conv-PCR. All assays reliably detected U. virens in infected rice panicles but not in healthy controls, demonstrating their field applicability.

Graphical Abstract