syn-tasiRnas targeting the coat protein of potato virus Y confer antiviral resistance in Nicotiana benthamiana

• Published in: Plant signaling & behavior 2024-12, Vol.19 (1), p.2358270-2358270
• Main Authors: Zhao, Xingyue, Gao, Qian, Wang, Haijuan, Yue, Jianying, An, Derong, Li, Bin, Yan, Fangfang, Carmen, Simon-Mateo, Zhao, Yuanzheng, Zhou, Hongyou, Zhao, Mingmin
• Format: Article
• Language: English
• Subjects: Capsid Proteins - genetics; Capsid Proteins - metabolism; Disease Resistance - genetics; Nicotiana - genetics; Nicotiana - immunology; Nicotiana - virology; Plant Diseases - genetics; Plant Diseases - immunology; Plant Diseases - virology; Plants, Genetically Modified - virology; Potato; potato virus Y; Potyvirus - physiology; Research Paper; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; synthetic sirnas; trans-acting small interfering RNA
• ISSN: 1559-2316; 1559-2324; 1559-2324
• DOI: 10.1080/15592324.2024.2358270

Trans-acting small interfering RNAs (tasiRNAs) are 21-nt phased (phased siRNAs) resulting from successive DCL-catalyzed processing from the end of a double-stranded RNA substrate originating from the RDR of an AGO-catalyzed cleaved RNA at a micro RNA target site. Plant tasiRNAs have been synthesized to produce synthetic tasiRNAs (syn-tasiRNAs) targeting viral RNAs that confer viral resistance. In this study, we engineered syn-tasiRNAs to target potato virus Y (PVY) infection by replacing five native siRNAs of TAS1c with 210-bp fragments from the coat protein (CP) region of the PVY genome. The results showed that the transient expression of syn-tasiR-CPpvy2 in Nicotiana benthamiana (N. benthamiana) plants conferred antiviral resistance, supported by the absence of PVY infection symptoms and viral accumulation. This indicated that syn-tasiR-CPpvy2 successfully targeted and silenced the PVY CP gene, effectively inhibiting viral infection. syn-tasiR-CPpvy1 displayed attenuated symptoms and decreased viral accumulation in these plants However, severe symptoms of PVY infection and a similar amount of viral accumulation as the control were observed in plants expressing syn-tasiR-CPpvy3. syn-tasiR-CPpvy/pvx, which targets both PVY and potato virus X (PVX), was engineered using a single precursor. After the transient expression of syn-tasiR-CPpvy/pvx3 and syn-tasiR-CPpvy/pvx5 in N. benthamiana, the plants were resistant to both PVY and PVX. These results suggested that engineered syn-tasiRNAs could not only specifically induce antiviral resistance against one target virus but could also be designed for multi-targeted silencing of different viruses, thereby preventing complex virus infection in plants.