Agroinfiltration Transient Assay to Evaluate the Potential of ihpRNA Constructs for Lily Mottle Virus Resistance

In order to obtain a resistant strain of Lily mottle virus (LMoV), three RNA interference (RNAi) vectors with intron hairpin RNA structure (ihpRNA) were constructed and transformed into Agrobacterium GV 3101 strain.The gene hotspots of LMoV coated protein (CP) and cylindrical inclusion (CI) were selected, and the positive and negative fragments of the target gene were inserted into both sides of the Pdk intron fragment of plant expression vector pRNAi-LIC by one-step, zero-background ligation-independent cloning (OZ-LIC) method.Antiviral analysis was performed using Agrobacterium-mediated transientexpression assay.RT-PCR and indirect ELISA results showed that the hairpin RNA from the reverse repeats of CI-517 bp and CI-295 bp could interfere the virus effectively, while the CP-400 bp hairpin's impact was not significant.The work illustrates that the genetically modified genome could LMoV.