Hotspot siRNA Confers Plant Resistance against Viral Infection.
Hotspot siRNA Confers Plant Resistance against Viral Infection.
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热点 siRNA 赋予植物抵抗病毒感染的能力
DOI：
10.3390/biology11050714
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发表时间：
2022-05-06
期刊：
影响因子：
4.2
通讯作者：
Hong, Yiguo
中科院分区：
文献类型：
--
作者：
Mohamed, Atef;Jin, Zhenhui;Osman, Toba;Shi, Nongnong;Tor, Mahmut;Jackson, Stephen;Hong, Yiguo
抗病毒RNAi的一个标志是病毒siRNA （vsiRNA）的产生。对vsirna的分析表明，病毒基因组的某些热点区域或转录的病毒rna更容易发生rnai介导的切割。然而，热点vsirna与宿主先天防御的生物学相关性仍有待阐明。在这里，我们证明了通过合成vsiRNA直接靶向热点使植物对病毒感染具有抗性。基于非洲木薯花叶病毒（ACMV）的vsiRNA谱，合成了热点和冷点vsiRNA。然而，只有双链热点vsiRNA保护植物免受ACMV感染，病毒DNA复制和病毒mRNA水平无法检测。我们进一步证明，热点vsirna介导的病毒抗性具有阈值效应，需要活性RDR6。这些数据表明，热点vsirna在抗病毒RNAi上具有功能意义，可能具有作为外源保护剂控制破坏性植物病毒病害的潜力。抗病毒RNA干扰（RNAi）的一个标志是产生病毒小干扰RNA （vsiRNA）。对vsiRNA的分析表明，病毒RNA基因组或转录的病毒RNA的某些区域（称为vsiRNA热点）更容易被rnai介导的vsiRNA生物发生裂解。然而，热点vsirna与宿主先天防御病原体的生物学相关性仍有待阐明。在这里，我们展示了通过合成的vsiRNA直接靶向热点赋予宿主对病毒感染的抗性。利用Northern blotting和RNAseq技术，我们获得了非洲木薯花叶病毒（ACMV）的vsirna谱。合成了热点和冷点vsiRNA对应的小rna的正义链和反义链。本烟与双链热点siRNA共接种可保护植物免受ACMV感染，在ACMV感染中，病毒DNA复制和病毒mRNA积累无法检测到。热点vsiRNA的正义链和反义链，以及冷点vsiRNA的双链和单链形式都没有病毒保护活性。我们进一步证明，热点vsirna介导的病毒抗性具有阈值效应，需要活性RDR6。这些数据表明，热点vsirna在抗病毒RNAi上具有一定的功能意义，可能具有作为控制植物破坏性病毒病害的外源保护剂的潜力。
A hallmark of antiviral RNAi is the production of viral siRNA (vsiRNA). Profiling of vsiRNAs indicates that certain hotspot regions of viral genome or transcribed viral RNAs are more prone to RNAi-mediated cleavage. However, the biological relevance of hotspot vsiRNAs to the host innate defence remains to be elucidated. Here, we show that direct targeting a hotspot by synthetic vsiRNA confers plant resistance to virus infection. Hotspot and coldspot vsiRNAs, based on vsiRNA profile of the African cassava mosaic virus (ACMV), were synthesised. However, only the double-stranded hotspot vsiRNA protected plants from ACMV infection with undetectable levels of viral DNA replication and viral mRNA. We further demonstrated that the hotspot vsiRNA-mediated virus resistance had a threshold effect and required an active RDR6. These data show that hotspot vsiRNAs bear a functional significance on antiviral RNAi, suggesting that they may have the potential as exogenous protection agents for controlling destructive plant viral diseases. A hallmark of antiviral RNA interference (RNAi) is the production of viral small interfering RNA (vsiRNA). Profiling of vsiRNAs indicates that certain regions of viral RNA genome or transcribed viral RNA, dubbed vsiRNA hotspots, are more prone to RNAi-mediated cleavage for vsiRNA biogenesis. However, the biological relevance of hotspot vsiRNAs to the host innate defence against pathogens remains to be elucidated. Here, we show that direct targeting a hotspot by a synthetic vsiRNA confers host resistance to virus infection. Using Northern blotting and RNAseq, we obtained a profile of vsiRNAs of the African cassava mosaic virus (ACMV), a single-stranded DNA virus. Sense and anti-sense strands of small RNAs corresponding to a hotspot and a coldspot vsiRNA were synthesised. Co-inoculation of Nicotiana benthamiana with the double-stranded hotspot siRNA protected plants from ACMV infection, where viral DNA replication and accumulation of viral mRNA were undetectable. The sense or anti-sense strand of this hotspot vsiRNA, and the coldspot vsiRNA in both double-stranded and single-stranded formats possessed no activity in viral protection. We further demonstrated that the hotspot vsiRNA-mediated virus resistance had a threshold effect and required an active RDR6. These data show that hotspot vsiRNAs bear a functional significance on antiviral RNAi, suggesting that they may have the potential as an exogenous protection agent for controlling destructive viral diseases in plants.
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