Abstract
Transgenic plants expressing artificial microRNAs (amiRNAs) have been shown to confer specific resistance to corresponding viruses. Here, we generated Nicotiana benthamiana transgenic lines containing Oryza sativa miR528 as backbone, expressing amiRNAs targeting RNA-dependent RNA polymerase (RdRp) gene of Cymbidium mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV). The amiRNA transgenic lines could express amiR-CymMV and confer high percentage resistance to CymMV, while lack of detectable level of amiR-ORSV expression in amiR-ORSV transgenic N. benthamiana plants led to weak resistance to ORSV infection. In this project, we provide the first report of CymMV-resistant transgenic N. benthamiana plants based on amiRNA strategy. We believe that this amiRNA approach can be extended to generate CymMV-resistant transgenic orchids.
Highlights
RNA-mediated virus resistant transgenic plants have been successfully generated in several plant species with different RNA silencing strategies such as sense[7,8,9], antisense[7,10,11], double stranded[12,13] and hairpin[14] RNA
From the reported transgenic orchids, a few groups of researchers focused on the Cymbidium mosaic virus (CymMV)-resistant transgenic orchids by antisense approach to target the virus coat protein gene[27,28,29]
We designed plant expression vectors pG0229, driven by the double Cauliflower mosaic virus (CaMV) 35S promoter, targeting the region of the gene coding for the RNA-dependent RNA polymerase (RdRp) of CymMV or Odontoglossum ringspot virus (ORSV)
Summary
RNA-mediated virus resistant transgenic plants have been successfully generated in several plant species with different RNA silencing strategies such as sense[7,8,9], antisense[7,10,11], double stranded[12,13] and hairpin[14] RNA. RNA silencing, as a result of the formation of double-stranded RNA (dsRNA), leads to sequence-specific gene silencing. From the reported transgenic orchids, a few groups of researchers focused on the CymMV-resistant transgenic orchids by antisense approach to target the virus coat protein gene[27,28,29]. We designed plant expression vectors pG0229, driven by the double Cauliflower mosaic virus (CaMV) 35S promoter, targeting the region of the gene coding for the RNA-dependent RNA polymerase (RdRp) of CymMV or ORSV. ORSV-specific amiRNA transgenic N. benthamiana plants were weakly resistant to ORSV infection
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