Resistance to CymMV and ORSV in artificial microRNA transgenic Nicotiana benthamiana plants

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.

Dual resistance of transgenic plants against Cymbidium mosaic virus and Odontoglossum ringspot virus

“…This phenomena may explain why we could generate more resistant lines to CymMV but few to ORSV or both in our study. Previously, different strategies for designing transgenes have been attempted, including using the sense or antisense CymMV CP gene 14,21 and amiRNA targeting RdRp genes of CymMV and ORSV 17 , but most transgenic studies have successfully conferred resistance to only CymMV. To date, no transgenic lines with resistance to ORSV or efficient dual resistance to both viruses have been reported.…”

“…(2018) using Oryza sativa miR528 as a backbone, expressed artificial miRNA (amiRNA) targeting replicase genes of CymMV and ORSV. AmiRNA transgenic N. benthamiana lines conferred high resistance to CymMV infection but weak resistance to ORSV infection 17 . No transgenic plants completely resistant to both CymMV and ORSV have been reported.…”

An Algorithmic framework for genome-wide identification of Sugarcane (Saccharum officinarumL.)-encoded microRNA targets against SCBV

“…We have designed amiRNA-based gene construct of SCBV which contains modified miRNA/miRNA* sequence in duplex of precursor (sof-MIR159) with artificial sequence to create mature amiRNA as shown in (Figure 10). The amiRNA-based silencing technology is successfully validated in many crop plants to control emerging plant viruses [18,19,21]. In summary, our computational work for the silencing of the SCBV genome could offer new approach for the remodel of existing strategy as antiviral agents.…”

“…Artificial microRNA (amiRNA)-mediated silencing of invading viruses in plants was first time reported by Niu [18]. This amiRNA-based silencing strategy was applied effectively in many plants to combat plant viruses: cotton leaf curl Kokhran virus (CLCuKoV-Bu) [19], Cucumber mosaic virus (CMV) [20], CymMV and ORSV [21]. Serological method, ELISA, immunosorbent electron microscopy (ISEM) [22], IC-PCR [23], RT-PCR [24], rolling circle amplification (RCA) [25] are the most widely used reliable detection techniques for SCBV.…”

Peer Review #1 of "A novel computational approach to the silencing of Sugarcane Bacilliform Guadeloupe A Virus determines potential host-derived MicroRNAs in sugarcane (Saccharum officinarum L.) (v0.1)"

“…Several studies have been successful in generating resistance against plant viruses using amiRNA-technology in crop plants such as tomato (Chen et al 2019;Zhang et al 2011) and cotton (Akmal et al 2017;Ali et al 2013). Most recently, amiRNA-mediated gene silencing has been demonstrated against Potexvirus and Tobamovirus successfully (Petchthai et al 2018). miRNA: mRNA target identification, validation and interactions are the basis for discerning the important functions of miRNAs in the greater perspective of regulatory network leading to different biotic developments.…”