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<p>Cucumber mosaic virus (CMV) is an important plant virus that is classified within the <i>Bromoviridae</i> family, and has a broad host range. Among the multifunctional CMV proteins, <i>2b</i> plays a crucial role, which is essential for CMV virulence. Topical application of double-stranded RNAs (dsRNAs) to plants induces gene silencing, offering a strategy for developing biopesticides against plant pathogens. This study aimed to explore the potential induction of resistance against CMV through the application of exogenous small RNAs produced in tobacco plants. To achieve this goal, a gene construct containing a partial sequence of CMV-<i>2b</i> was transiently expressed in tobacco leaves using <i>Agrobacterium tumefaciens</i>, to generate <i>2b</i> small RNAs. Subsequently, total RNA was extracted from transiently transformed leaves, and after confirming <i>2b</i> small RNA expression through stem-loop PCR, the extracted total RNA was exogenously applied to infected tomato and bean plants. To enhance the stability of naked RNAs, the total RNA was loaded onto clay nanoparticles (LDH-RNA). The CMV titer was then assessed using quantitative PCR at 4 and 12 days post RNA and nanoparticle applications. Quantitative PCR analysis revealed a significant reduction in the CMV titer in tomato and common bean plants four days after RNA application, although the difference was not significant at 12 days post application. Evaluation of the CMV titer - at days 4 and 12 revealed a reduction of virus titer in bean plants when RNA loaded on clay nanoparticles. Thus, this research underscores the efficacy of exogenously applied small RNAs produced in plants for CMV control, highlighting the potential for enhanced RNA stability through loading onto clay nanoparticles.</p>

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Exogenous application of CMV-2b-derived interfering RNA (2biRNA) expressed in tobacco plants against cucumber mosaic virus

  • 期刊论文

  • 发表日期：

    2026年5月22日

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  • Fatemeh Gholami Zarneh,
  • Sajad Moradi,
  • Abdolbaset Azizi

摘要

Cucumber mosaic virus (CMV) is an important plant virus that is classified within the Bromoviridae family, and has a broad host range. Among the multifunctional CMV proteins, 2b plays a crucial role, which is essential for CMV virulence. Topical application of double-stranded RNAs (dsRNAs) to plants induces gene silencing, offering a strategy for developing biopesticides against plant pathogens. This study aimed to explore the potential induction of resistance against CMV through the application of exogenous small RNAs produced in tobacco plants. To achieve this goal, a gene construct containing a partial sequence of CMV-2b was transiently expressed in tobacco leaves using Agrobacterium tumefaciens, to generate 2b small RNAs. Subsequently, total RNA was extracted from transiently transformed leaves, and after confirming 2b small RNA expression through stem-loop PCR, the extracted total RNA was exogenously applied to infected tomato and bean plants. To enhance the stability of naked RNAs, the total RNA was loaded onto clay nanoparticles (LDH-RNA). The CMV titer was then assessed using quantitative PCR at 4 and 12 days post RNA and nanoparticle applications. Quantitative PCR analysis revealed a significant reduction in the CMV titer in tomato and common bean plants four days after RNA application, although the difference was not significant at 12 days post application. Evaluation of the CMV titer - at days 4 and 12 revealed a reduction of virus titer in bean plants when RNA loaded on clay nanoparticles. Thus, this research underscores the efficacy of exogenously applied small RNAs produced in plants for CMV control, highlighting the potential for enhanced RNA stability through loading onto clay nanoparticles.