Virus-derived small interfering RNAs affects the accumulations of viral and host transcripts in maize

【作者】 夏子豪； Zhao Zhenxing； Jiao Zhiyuan； Zhou Tao； Wu Yuanhua； 范在丰；

【Author】 Xia Zihao;Zhao Zhenxing;Jiao Zhiyuan;Zhou Tao;Wu Yuanhua;Fan Zaifeng;College of Plant Protection,Shenyang Agricultural University;Department of Plant Pathology,China Agricultural University;

【机构】 College of Plant Protection,Shenyang Agricultural University； Department of Plant Pathology,China Agricultural University；

【摘要】 RNA silencing is a conserved surveillance mechanism against invading viruses,which involves the production of virus-derived small interfering RNAs(vsiRNAs) that play essential roles in silencing of viral RNAs and specific host transcripts.However,how vsiRNAs function to target viral and host transcripts is poorly studied,especially in maize(Zea mays L.).In this study,a degradome library constructed from Sugarcane mosaic virus(SCMV)-inoculated maize plants was sequenced and analysed to identify the cleavage sites in viral and host transcripts mainly produced by vsiRNAs.The results showed that forty-two maize transcripts were possibly cleaved by vsiRNAs.In addition,more than three thousand possible cleavage sites existed in positive-strand RNAs of SCMV.To determine the roles of vsiRNAs in targeting viral RNAs,artificial microRNAs(amiRNAs) using a rice miRNA(osa-miR528a)precursor backbone were designed and expressed in maize protoplast.Six vsiRNAs were selected to target different sequence elements of SCMV positive-strand RNAs,of which four could efficiently inhibit the accumulation of SCMV RNAs.All of six vsiRNAs could be detected by Northern blotting in both SCMV-infected maize plants and plasmid-transfected maize protoplast.These results provide new insights into maize crop engineering against virus infection by making amiRNA a more predictable and useful technology.更多还原

【Abstract】 RNA silencing is a conserved surveillance mechanism against invading viruses,which involves the production of virus-derived small interfering RNAs(vsiRNAs) that play essential roles in silencing of viral RNAs and specific host transcripts.However,how vsiRNAs function to target viral and host transcripts is poorly studied,especially in maize(Zea mays L.).In this study,a degradome library constructed from Sugarcane mosaic virus(SCMV)-inoculated maize plants was sequenced and analysed to identify the cleavage sites in viral and host transcripts mainly produced by vsiRNAs.The results showed that forty-two maize transcripts were possibly cleaved by vsiRNAs.In addition,more than three thousand possible cleavage sites existed in positive-strand RNAs of SCMV.To determine the roles of vsiRNAs in targeting viral RNAs,artificial microRNAs(amiRNAs) using a rice miRNA(osa-miR528a)precursor backbone were designed and expressed in maize protoplast.Six vsiRNAs were selected to target different sequence elements of SCMV positive-strand RNAs,of which four could efficiently inhibit the accumulation of SCMV RNAs.All of six vsiRNAs could be detected by Northern blotting in both SCMV-infected maize plants and plasmid-transfected maize protoplast.These results provide new insights into maize crop engineering against virus infection by making amiRNA a more predictable and useful technology.更多还原