Exogenous dsRNA-mediated RNA interference suppresses potato virus Y (PVY) infection in tobacco by targeting and silencing the CP gene

doi:10.14088/j.cnki.issn0439-8114.2026.02.015

Abstract

Abstract: To evaluate the inhibitory effect of exogenous double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) on potato virus Y (PVY) infection in tobacco, the coat protein (CP) gene of PVY was cloned. Subsequently, a dsRNA fragment targeting this gene was designed based on sequence analysis. A 474-bp dsRNA targeting the PVY CP gene was synthesized in vitro and applied together with PVY onto tobacco plants. The relative mRNA level of the CP gene in the inoculated leaves was determined by quantitative real-time PCR (qRT-PCR), while viral symptom development was observed over time. Both the disease incidence and disease index were recorded to comprehensively evaluate the control efficacy of the dsRNA treatment against PVY. The results showed that exogenous application of PVY CP gene targeting dsRNA significantly suppressed the expression of the PVY CP gene in the inoculated leaves and effectively alleviated virus-induced symptoms, resulting in a control efficacy of 50.58% in pot trials.

Key words: potato virus Y (PVY), double-stranded RNA (dsRNA), RNA interference (RNAi), coat protein (CP), biocontrol

CLC Number:

XU Ru-bing, LUO Zhao-peng, LI Yan-yan, LIU Jun-xia, YANG Jun, QIN Yao. Exogenous dsRNA-mediated RNA interference suppresses potato virus Y (PVY) infection in tobacco by targeting and silencing the CP gene[J]. HUBEI AGRICULTURAL SCIENCES, 2026, 65(2): 101-106.

References

[1] 刘勇昌,耿丽,高强,等.基于叶片光谱分析的烟草马铃薯Y病毒病严重度诊断[J].烟草科技,2021,54(7):23-28.
[2] GUO Y S,WU W,ZHANG X L,et al.Triplex immunostrip assay for rapid diagnosis of tobacco mosaic virus, tobacco vein banding mosaic virus, and potato virus Y[J].Plant disease,2022, 106(12):3033-3039.
[3] TORRANCE L,TALIANKSY M E.Potato virus Y emergence and evolution from the Andes of south America to become a major destructive pathogen of potato and other solanaceous crops worldwide[J].Viruses,2020,12(12):1430.
[4] 蒋君梅,杜巧丽,陈美晴,等.马铃薯Y病毒衣壳蛋白的表达、纯化与结晶条件筛选[J].植物保护学报,2022,49(2):508-514.
[5] IVANOV K I,MÄKINEN K. Coat proteins, host factors and plant viral replication[J].Current opinion in virology,2012,2(6):712-718.
[6] SAMARSKAYA V O,SPECHENKOVA N,ILINA I,et al.A non-canonical pathway induced by externally applied virus-specific dsRNA in potato plants[J].International journal of molecular sciences,2023,24(21):15769.
[7] 刘静,陈雪梅,宋云枝,等.马铃薯Y病毒HC-Pro、CI、NIb和CP基因介导的病毒抗性比较研究[J].植物病理学报,2010,40(1):57-65.
[8] 刘天波,蔡海林,滕凯,等.病毒诱导的基因沉默防控烟草马铃薯Y病毒病研究[J].中国烟草学报,2020,26(5):82-89.
[9] 张文庆,王桂荣.RNA干扰——从基因功能到生物农药[M].北京:科学出版社,2021.
[10] AKBAR S,WEI Y,ZHANG M Q.RNA interference: Promising approach to combat plant viruses[J].International journal of molecular sciences,2022,23(10):5312.
[11] USLU V V,WASSENEGGER M.Critical view on RNA silencing-mediated virus resistance using exogenously applied RNA[J].Current opinion in virology,2020,42:18-24.
[12] NECIRA K,MAKKI M,SANZ-GARCÍA E,et al. Topical application of Escherichia coli-Encapsulated dsRNA induces resistance in Nicotiana benthamiana to potato viruses and involves RDR6 and combined activities of DCL2 and DCL4[J].Plants,2021,10(4):644.
[13] WEN Z Y,HU R J,PI Q L,et al.DEAD-box RNA helicase RH20 positively regulates RNAi-based antiviral immunity in plants by associating with SGS3/RDR6 bodies[J].Plant biotechnology journal,2024,22(12):3295-3311.
[14] KONAKALLA N C,KALDIS A,BERBATI M,et al.Exogenous application of double-stranded RNA molecules from TMV p126 and CP genes confers resistance against TMV in tobacco[J].Planta,2016,244(4):961-969.
[15] NIEHL A,SOININEN M,PORANEN M M,et al.Synthetic biology approach for plant protection using dsRNA[J].Plant biotechnology journal,2018,16(9):1679-1687.
[16] CHENG X Q,ZHOU Q,XIAO J D,et al.Nanoparticle LDH enhances RNAi efficiency of dsRNA in piercing-sucking pests by promoting dsRNA stability and transport in plants[J]. Journal of nanobiotechnology,2024,22(1):544.
[17] HORN T,BOUTROS M.E-RNAi: A web application for the multi-species design of RNAi reagents—2010 update[J].Nucleic acids research,2010,38(Web Server issue):W332-W339.
[18] KALDIS A,BERBATI M,MELITA O,et al.Exogenously applied dsRNA molecules deriving from the Zucchini yellow mosaic virus (ZYMV) genome move systemically and protect cucurbits against ZYMV[J].Molecular plant pathology,2018,19(4):883-895.
[19] AHN S J,DONAHUE K,KOH Y,et al.Microbial-based double-stranded RNA production to develop cost-effective RNA interference application for insect pest management[J]. International journal of insect science,2019,11:1179543319840323.
[20] MA Z Z,ZHOU H,WEI Y L,et al.A novel plasmid-Escherichia coli system produces large batch dsRNAs for insect gene silencing[J].Pest management science,2020,76(7):2505-2512.
[21] 关梅,晁子健,闫硕,等.RNA农药的研究现状和发展前景[J].现代农药,2023,22(2):11-18.