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<p>Tomato mottle mosaic virus (ToMMV; <i>Tobamovirus maculatessellati</i>, family&#xa0;<i>Virgaviridae</i>), first reported in Mexico in 2009, is emerging as a global threat to solanaceous crops. International trade of ToMMV-contaminated seeds may be associated with its global spread; however, the epidemiological features of virus-contaminated seeds remain unclear, and no SYBR Green-based real-time RT-PCR (SYBR-RT-qPCR) assay to detect ToMMV in seeds has been available. Therefore, we developed a SYBR-RT-qPCR that detects ToMMV and is 10–100 times more sensitive than other available methods; up to 1.54 copies were detected without amplifying other tobamoviruses that commonly infect Solanaceae species. In virus-contaminated seeds obtained from two cultivars of ToMMV-infected tomato plants, a direct immunostaining assay revealed ToMMV in the seed coat and hairs of both cultivars. Although ToMMV was detected in all tested seeds, seed-to-seedling transmission of ToMMV was only 0.42% for tomato cv. Carol Passion and 0.93% for cv. Rejina. Using these seed lots harboring seed-to-seedling-transmissible ToMMV, we confirmed the efficacy of this SYBR-RT-qPCR method for detecting ToMMV in a bulk sample of 400 seeds that included one ToMMV-contaminated seed. Therefore, our detection method is effective for seed inspection. This is the first report of a practical ToMMV-specific seed inspection assay based on SYBR-RT-qPCR; moreover, crucial properties related to seed transmission of ToMMV have been revealed. The insights and testing methods reported here should help prevent the spread of ToMMV through virus-contaminated seeds.</p>

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Development of a new SYBR Green-based real-time PCR method for detecting tomato mottle mosaic virus and the assessment of seed transmission properties of the virus in tomato

  • 期刊论文

  • 发表日期：

    2025年11月10日

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  • Yuya Imamura,
  • Fumino Nito,
  • Yuji Fujiwara,
  • Takayuki Matsuura,
  • Hironobu Yanagisawa

摘要

Tomato mottle mosaic virus (ToMMV; Tobamovirus maculatessellati, family Virgaviridae), first reported in Mexico in 2009, is emerging as a global threat to solanaceous crops. International trade of ToMMV-contaminated seeds may be associated with its global spread; however, the epidemiological features of virus-contaminated seeds remain unclear, and no SYBR Green-based real-time RT-PCR (SYBR-RT-qPCR) assay to detect ToMMV in seeds has been available. Therefore, we developed a SYBR-RT-qPCR that detects ToMMV and is 10–100 times more sensitive than other available methods; up to 1.54 copies were detected without amplifying other tobamoviruses that commonly infect Solanaceae species. In virus-contaminated seeds obtained from two cultivars of ToMMV-infected tomato plants, a direct immunostaining assay revealed ToMMV in the seed coat and hairs of both cultivars. Although ToMMV was detected in all tested seeds, seed-to-seedling transmission of ToMMV was only 0.42% for tomato cv. Carol Passion and 0.93% for cv. Rejina. Using these seed lots harboring seed-to-seedling-transmissible ToMMV, we confirmed the efficacy of this SYBR-RT-qPCR method for detecting ToMMV in a bulk sample of 400 seeds that included one ToMMV-contaminated seed. Therefore, our detection method is effective for seed inspection. This is the first report of a practical ToMMV-specific seed inspection assay based on SYBR-RT-qPCR; moreover, crucial properties related to seed transmission of ToMMV have been revealed. The insights and testing methods reported here should help prevent the spread of ToMMV through virus-contaminated seeds.