Knoblich Marie, Gursinsky Torsten, Gago-Zachert Selma, Weinholdt Claus, Grau Jan, Behrens Sven-Erik
Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Charles Tanford Protein Centre, Kurt-Mothes-Str. 3A, 06120 Halle (Saale), Germany.
Institute of Computer Science, Martin Luther University Halle-Wittenberg, Von-Seckendorff-Platz 1, 06120 Halle (Saale), Germany.
Nucleic Acids Res. 2025 Feb 27;53(5). doi: 10.1093/nar/gkaf136.
RNA-mediated crop protection increasingly becomes a viable alternative to agrochemicals that threaten biodiversity and human health. Pathogen-derived double-stranded RNAs (dsRNAs) are processed into small interfering RNAs (siRNAs), which can then induce silencing of target RNAs, e.g. viral genomes. However, with currently used dsRNAs, which largely consist of undefined regions of the target RNAs, silencing is often ineffective: processing in the plant generates siRNA pools that contain only a few functionally effective siRNAs (esiRNAs). Using an in vitro screen that reliably identifies esiRNAs from siRNA pools, we identified esiRNAs against Cucumber mosaic virus (CMV), a devastating plant pathogen. Topical application of esiRNAs to plants resulted in highly effective protection against massive CMV infection. However, optimal protection was achieved with newly designed multivalent 'effective dsRNAs' (edsRNAs), which contain the sequences of several esiRNAs and are preferentially processed into these esiRNAs. The esiRNA components can attack one or more target RNAs at different sites, be active in different silencing complexes, and provide cross-protection against different viral variants-important properties for combating rapidly mutating pathogens such as CMV. esiRNAs and edsRNAs have thus been established as a new class of 'RNA actives' that significantly increase the efficacy and specificity of RNA-mediated plant protection.
RNA介导的作物保护日益成为一种可行的替代农用化学品的方法，农用化学品会威胁生物多样性和人类健康。病原体来源的双链RNA（dsRNA）被加工成小干扰RNA（siRNA），然后可以诱导靶RNA（如病毒基因组）的沉默。然而，对于目前使用的dsRNA，其大部分由靶RNA的未定义区域组成，沉默往往无效：在植物中加工会产生仅包含少数功能有效siRNA（esiRNA）的siRNA库。通过一种能从siRNA库中可靠鉴定esiRNA的体外筛选方法，我们鉴定出了针对黄瓜花叶病毒（CMV）的esiRNA，CMV是一种具有毁灭性的植物病原体。将esiRNA局部施用于植物可有效抵御大规模的CMV感染。然而，新设计的多价“有效dsRNA”（edsRNA）能实现最佳保护，edsRNA包含几种esiRNA的序列，并优先加工成这些esiRNA。esiRNA组分可以在不同位点攻击一个或多个靶RNA，在不同的沉默复合体中发挥作用，并提供针对不同病毒变体的交叉保护——这是对抗像CMV这样快速变异病原体的重要特性。因此，esiRNA和edsRNA已被确立为一类新的“RNA活性物质”，可显著提高RNA介导的植物保护的功效和特异性。
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