Expression of artificial microRNAs in tomato confers efficient and stable virus resistance in a cell-autonomous manner |
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Authors: | Xiaohui Zhang Hanxia Li Junhong Zhang Chanjuan Zhang Pengjuan Gong Khurram Ziaf Fangming Xiao Zhibiao Ye |
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Institution: | (1) National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China;(2) Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan, China;(3) Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, Idaho 83844-3052, USA; |
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Abstract: | Expression of artificial microRNAs (amiRNAs) in plants can target and degrade the invading viral RNA, consequently conferring
virus resistance. Two amiRNAs, targeting the coding sequence shared by the 2a and 2b genes and the highly conserved 3′ untranslated
region (UTR) of Cucumber mosaic virus (CMV), respectively, were generated and introduced into the susceptible tomato. The transgenic tomato plants expressing amiRNAs
displayed effective resistance to CMV infection and CMV mixed with non-targeted viruses, including tobacco mosaic virus and tomato yellow leaf curl virus. A series of grafting assays indicate scions originated from the transgenic tomato plant maintain stable resistance to CMV
infection after grafted onto a CMV-infected rootstock. However, the grafting assay also suggests that the amiRNA-mediated
resistance acts in a cell-autonomous manner and the amiRNA signal cannot be transmitted over long distances through the vascular
system. Moreover, transgenic plants expressing amiRNA targeting the 2a and 2b viral genes displayed slightly more effective
to repress CMV RNA accumulation than transgenic plants expressing amiRNA targeting the 3′ UTR of viral genome did. Our work
provides new evidence of the use of amiRNAs as an effective approach to engineer viral resistance in the tomato and possibly
in other crops. |
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