Multi-targeting of viral RNAs with synthetic trans-acting small interfering RNAs enhances plant antiviral resistance
Multi-targeting of viral RNAs with synthetic trans-acting small interfering RNAs enhances plant antiviral resistance
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DOI：
10.1111/tpj.14466
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发表时间：
2019-09-16
期刊：
影响因子：
7.2
通讯作者：
Daros, Jose-Antonio
中科院分区：
文献类型：
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作者：
Carbonell, Alberto;Lison, Purificacion;Daros, Jose-Antonio
基于RNA干扰（RNAi）的工具用于多种生物体中，以通过互补小RNA对靶RNA的序列特异性降解来诱导抗病毒抗性。在植物中，高度特异性的基于RNAi的抗病毒工具包括人工microRNA（amiRNA）和合成的反式作用小干扰RNA（syn-tasiRNA）。syn-tasiRNA已经成为一种有前景的抗病毒工具，其允许通过从单一前体同时表达几种syn-tasiRNA来多靶向病毒RNA。在这里，我们在番茄植物中比较了表达单个amiRNA的amiRNA构建体和表达四种不同syn-tasiRNA的syn-tasiRNA构建体对番茄斑萎病毒（TSWV）的影响，TSWV是影响全球番茄作物的经济上重要的病原体。大多数syn-tasiRNA系对TSWV具有抗性，而大多数amiRNA系是易感的，并且积累了在amiRNA靶位点具有突变的病毒后代。只有具有较高amiRNA积累的两个amiRNA系是抗性的，而syn-tasiRNA系中的抗性并不排除具有高syn-tasiRNA积累的系。总的来说，这些结果表明，syn-tasiRNA诱导增强的抗病毒抗性，因为每个单独的syn-tasiRNA的组合沉默效应，这最小化了病毒同时突变所有不同靶位点以完全逃避每个syn-tasiRNA的可能性。
RNA interference (RNAi)-based tools are used in multiple organisms to induce antiviral resistance through the sequence-specific degradation of target RNAs by complementary small RNAs. In plants, highly specific antiviral RNAi-based tools include artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs). syn-tasiRNAs have emerged as a promising antiviral tool allowing for the multi-targeting of viral RNAs through the simultaneous expression of several syn-tasiRNAs from a single precursor. Here, we compared in tomato plants the effects of an amiRNA construct expressing a single amiRNA and a syn-tasiRNA construct expressing four different syn-tasiRNAs against Tomato spotted wilt virus (TSWV), an economically important pathogen affecting tomato crops worldwide. Most of the syn-tasiRNA lines were resistant to TSWV, whereas the majority of the amiRNA lines were susceptible and accumulated viral progenies with mutations in the amiRNA target site. Only the two amiRNA lines with higher amiRNA accumulation were resistant, whereas resistance in syn-tasiRNA lines was not exclusive of lines with high syn-tasiRNA accumulation. Collectively, these results suggest that syn-tasiRNAs induce enhanced antiviral resistance because of the combined silencing effect of each individual syn-tasiRNA, which minimizes the possibility that the virus simultaneously mutates all different target sites to fully escape each syn-tasiRNA.