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Abstract <p>Plant viruses significantly decrease the yield of cultivated plants; therefore, developing new methods of combating viral diseases and studying the effectiveness of these methods is relevant. The expression of the heterologous extracellular RNase gene of <i>Zinnia elegans</i> (<i>ZRNase&#xa0;II</i>) increases the resistance of transgenic plants to viruses. The authors have proposed a model system for visualizing the protective effect of the <i>ZRNase&#xa0;II</i> gene and studying the spread of GFP-marked viral particles in <i>Nicotiana benthamiana</i> plants. The&#xa0;presence of the target <i>ZRNase&#xa0;II</i> gene was confirmed in the plants obtained after <i>Agrobacterium</i>-mediated transformation. Four lines were selected and adapted to ex&#xa0;vitro conditions. Total RNase activity in all transgenic lines was four to seven times higher than in wild-type plants. The systemic spread of the Potato virus X-based PVX-GFP construct in these plants was studied. To this end, agroinfiltration was performed using the pICH27566 and pICH6692 vectors. Following agroinfiltration, transient expression of GFP occurred and its systemic spread throughout the plant was observed due to transport proteins within the virus-like particles. Expression of the heterologous RNase gene (<i>ZRNase&#xa0;II</i>) delayed the spread of viral particles by 5–30 days and decreased the accumulation of GFP-marked viral particles in all transgenic lines compared to the control. To assess the rapid systemic response of plants to agroinfiltration and virus expression, the content of chlorophyll and anthocyanins in <i>N.&#xa0;benthamiana</i> leaves was studied outside the zone of visual spread of GFP-marked viral particles. Differences were found between control and transgenic plants in anthocyanin content in response to infection. Expression of the <i>ZRNase&#xa0;II</i> gene in plants delayed the systemic spread of virus-like particles and changed the response to biotic stress at the plant secondary metabolism level.</p>

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Study of the Systemic Spreading of GFP-Labeled Virus-Like Particles in Transgenic Nicotiana benthamiana Plants Transformed with the Extracellular RNase II Gene

  • 期刊论文

  • 发表日期：

    2026年6月15日

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  • O. O. Ovcharenko,
  • A. O. Potrokhov,
  • O. M. Yaroshko,
  • S. V. Litvinov,
  • P. D. Maystrov,
  • V. A. Rudas,
  • M. V. Kuchuk

摘要

Abstract

Plant viruses significantly decrease the yield of cultivated plants; therefore, developing new methods of combating viral diseases and studying the effectiveness of these methods is relevant. The expression of the heterologous extracellular RNase gene of Zinnia elegans (ZRNase II) increases the resistance of transgenic plants to viruses. The authors have proposed a model system for visualizing the protective effect of the ZRNase II gene and studying the spread of GFP-marked viral particles in Nicotiana benthamiana plants. The presence of the target ZRNase II gene was confirmed in the plants obtained after Agrobacterium-mediated transformation. Four lines were selected and adapted to ex vitro conditions. Total RNase activity in all transgenic lines was four to seven times higher than in wild-type plants. The systemic spread of the Potato virus X-based PVX-GFP construct in these plants was studied. To this end, agroinfiltration was performed using the pICH27566 and pICH6692 vectors. Following agroinfiltration, transient expression of GFP occurred and its systemic spread throughout the plant was observed due to transport proteins within the virus-like particles. Expression of the heterologous RNase gene (ZRNase II) delayed the spread of viral particles by 5–30 days and decreased the accumulation of GFP-marked viral particles in all transgenic lines compared to the control. To assess the rapid systemic response of plants to agroinfiltration and virus expression, the content of chlorophyll and anthocyanins in N. benthamiana leaves was studied outside the zone of visual spread of GFP-marked viral particles. Differences were found between control and transgenic plants in anthocyanin content in response to infection. Expression of the ZRNase II gene in plants delayed the systemic spread of virus-like particles and changed the response to biotic stress at the plant secondary metabolism level.