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首页> 外文期刊>International Journal of Molecular Sciences >Hairpin RNA Targeting Multiple Viral Genes Confers Strong Resistance to Rice Black-Streaked Dwarf Virus
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Hairpin RNA Targeting Multiple Viral Genes Confers Strong Resistance to Rice Black-Streaked Dwarf Virus

机译：针对多个病毒基因的发夹RNA赋予对水稻黑纹矮化病毒的强大抵抗力

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Rice black-streaked dwarf virus (RBSDV) belongs to the genus Fijivirus in the family of Reoviridae and causes severe yield loss in rice-producing areas in Asia. RNA silencing, as a natural defence mechanism against plant viruses, has been successfully exploited for engineering virus resistance in plants, including rice. In this study, we generated transgenic rice lines harbouring a hairpin RNA (hpRNA) construct targeting four RBSDV genes, S1 , S2 , S6 and S10 , encoding the RNA-dependent RNA polymerase, the putative core protein, the RNA silencing suppressor and the outer capsid protein, respectively. Both field nursery and artificial inoculation assays of three generations of the transgenic lines showed that they had strong resistance to RBSDV infection. The RBSDV resistance in the segregating transgenic populations correlated perfectly with the presence of the hpRNA transgene. Furthermore, the hpRNA transgene was expressed in the highly resistant transgenic lines, giving rise to abundant levels of 21–24 nt small interfering RNA (siRNA). By small RNA deep sequencing, the RBSDV-resistant transgenic lines detected siRNAs from all four viral gene sequences in the hpRNA transgene, indicating that the whole chimeric fusion sequence can be efficiently processed by Dicer into siRNAs. Taken together, our results suggest that long hpRNA targeting multiple viral genes can be used to generate stable and durable virus resistance in rice, as well as other plant species.
机译：水稻黑条矮缩病毒（RBSDV）属于呼肠孤病毒科的斐济病毒属，在亚洲水稻产区造成严重的产量损失。 RNA沉默作为一种针对植物病毒的天然防御机制，已经成功地用于工程改造包括水稻在内的植物的抗病毒性。在这项研究中，我们产生了具有针对四个RBSDV基因S1，S2，S6和S10的发夹RNA（hpRNA）构建体的转基因水稻品系，该基因编码RNA依赖性RNA聚合酶，推定的核心蛋白，RNA沉默抑制剂和外部衣壳蛋白。三代转基因品系的田间苗圃和人工接种试验均显示它们对RBSDV感染具有较强的抵抗力。分离的转基因种群中的RBSDV抗性与hpRNA转基因的存在完全相关。此外，hpRNA转基因在高度抗性的转基因品系中表达，从而产生了丰富的21-24 nt小干扰RNA（siRNA）。通过小RNA深度测序，抗RBSDV的转基因品系从hpRNA转基因中的所有四个病毒基因序列中检测到siRNA，这表明Dicer可将整个嵌合融合序列有效地加工成siRNA。综上所述，我们的结果表明，靶向多种病毒基因的长hpRNA可用于在水稻以及其他植物物种中产生稳定而持久的病毒抗性。

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