College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China.
State Key Laboratory of Agro-Biotechnology and Key Laboratory of Pest Monitoring and Green Management-MOA, China Agricultural University, Beijing 100193, China.
Viruses. 2018 Nov 23;10(12):664. doi: 10.3390/v10120664.
RNA silencing is a conserved surveillance mechanism against invading viruses in plants, which involves the production of virus-derived small interfering RNAs (vsiRNAs) that play essential roles in the silencing of viral RNAs and/or specific host transcripts. However, how vsiRNAs function to target viral and/or host transcripts is poorly studied, especially in maize ( L.). In this study, a degradome library constructed from (SCMV)-inoculated maize plants was analyzed to identify the cleavage sites in viral and host transcripts mainly produced by vsiRNAs. The results showed that 42 maize transcripts were possibly cleaved by vsiRNAs, among which several were involved in chloroplast functions and in biotic and abiotic stresses. In addition, more than 3000 cleavage sites possibly produced by vsiRNAs were identified in positive-strand RNAs of SCMV, while there were only four cleavage sites in the negative-strand RNAs. To determine the roles of vsiRNAs in targeting viral RNAs, six vsiRNAs were expressed in maize protoplast based on artificial microRNAs (amiRNAs), of which four could efficiently inhibit the accumulations of SCMV RNAs. These results provide new insights into the genetic manipulation of maize with resistance against virus infection by using amiRNA as a more predictable and useful approach.
RNA 沉默是植物中一种针对入侵病毒的保守监测机制,涉及产生病毒衍生的小干扰 RNA (vsiRNA),它们在病毒 RNA 和/或特定宿主转录本的沉默中发挥重要作用。然而,vsiRNA 如何靶向病毒和/或宿主转录本的功能研究甚少,尤其是在玉米(Zea mays L.)中。在本研究中,分析了由(SCMV)接种的玉米植物构建的降解组文库,以鉴定主要由 vsiRNA 产生的病毒和宿主转录本中的切割位点。结果表明,42 种玉米转录本可能被 vsiRNA 切割,其中一些涉及叶绿体功能以及生物和非生物胁迫。此外,在 SCMV 的正链 RNA 中鉴定出了 3000 多个可能由 vsiRNA 产生的切割位点,而在负链 RNA 中只有四个切割位点。为了确定 vsiRNA 在靶向病毒 RNA 中的作用,基于人工 microRNA (amiRNA) 在玉米原生质体中表达了 6 个 vsiRNA,其中 4 个可以有效地抑制 SCMV RNA 的积累。这些结果为利用 amiRNA 作为一种更可预测和有用的方法对玉米进行抗病毒感染的遗传操作提供了新的见解。