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<p>Begomoviruses pose a major threat to mungbean and other economically important crops worldwide, causing severe yield losses. Among them, begomovirus vignaradiataindiaense is a major causative agent of yellow mosaic disease (YMD) in mungbean. Topical application of dsRNA has emerged as an innovative, non-transgenic approach for plant virus control by triggering RNA interference (RNAi)-mediated degradation of homologous viral RNAs. In this study, we explored the potential of dsRNA to confer resistance against begomovirus vignaradiataindiaense in YMD-susceptible mungbean plant. We designed hairpin RNA (hpRNA) constructs targeting key viral open reading frames (ORFs)—TR-1 (AC4/AC1), TR-2 (AC2/AC3/AC1), and a stacked TR-1 + 2 (AC4/AC1 + AC2/AC3/AC1)— and evaluated their efficacy through transient expression assays. Notably, only the hpTR-1 + 2 construct, which simultaneously targets multiple essential viral genes, conferred complete (100%) resistance and effectively restricted systemic virus movement. Building on this finding, we assessed the efficacy of in vivo produced hpRNA (hpTR-1 + 2) applied as a topical spray. The ability of hpRNA to trigger RNAi was confirmed by detecting 21 to 24 nt small interfering RNAs (siRNAs) in both locally treated and newly emerging leaves up to 12 days after application. Mungbean plants sprayed with hpTR-1 + 2 either on the day of infection or two days prior exhibited complete resistance to YMD. Furthermore, hpRNA treatment on plants already infected (two or four days post-infection) significantly reduced disease severity, demonstrating the theraeputic potential. Overall, our results demonstrate that the hpTR-1 + 2 multi-targeting strategy effectively controls YMD in mungbean and position topical dsRNA application as a sustainable, non-transgenic approach for plant disease management.</p>

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Hairpin-RNA spray confers resistance to mungbean yellow mosaic India virus in mungbean

  • 期刊论文

  • 发表日期：

    2025年9月18日

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  • Kiran Vilas Dhobale,
  • Lingaraj Sahoo

摘要

Begomoviruses pose a major threat to mungbean and other economically important crops worldwide, causing severe yield losses. Among them, begomovirus vignaradiataindiaense is a major causative agent of yellow mosaic disease (YMD) in mungbean. Topical application of dsRNA has emerged as an innovative, non-transgenic approach for plant virus control by triggering RNA interference (RNAi)-mediated degradation of homologous viral RNAs. In this study, we explored the potential of dsRNA to confer resistance against begomovirus vignaradiataindiaense in YMD-susceptible mungbean plant. We designed hairpin RNA (hpRNA) constructs targeting key viral open reading frames (ORFs)—TR-1 (AC4/AC1), TR-2 (AC2/AC3/AC1), and a stacked TR-1 + 2 (AC4/AC1 + AC2/AC3/AC1)— and evaluated their efficacy through transient expression assays. Notably, only the hpTR-1 + 2 construct, which simultaneously targets multiple essential viral genes, conferred complete (100%) resistance and effectively restricted systemic virus movement. Building on this finding, we assessed the efficacy of in vivo produced hpRNA (hpTR-1 + 2) applied as a topical spray. The ability of hpRNA to trigger RNAi was confirmed by detecting 21 to 24 nt small interfering RNAs (siRNAs) in both locally treated and newly emerging leaves up to 12 days after application. Mungbean plants sprayed with hpTR-1 + 2 either on the day of infection or two days prior exhibited complete resistance to YMD. Furthermore, hpRNA treatment on plants already infected (two or four days post-infection) significantly reduced disease severity, demonstrating the theraeputic potential. Overall, our results demonstrate that the hpTR-1 + 2 multi-targeting strategy effectively controls YMD in mungbean and position topical dsRNA application as a sustainable, non-transgenic approach for plant disease management.