RNAi-Mediated Resistance to Cucumber Mosaic Virus (CMV) in Genetically Engineered Tomato

Abstract

Cucumber mosaic virus is one of the most constraints to the production of tomato and other vegetable crops worldwide. Here, we generated an RNAi construct containing inverted repeat of 1138 bp fragment of a partial replicase gene of CMV-O and used it to produce transgenic tomato plants expressing CMV-specific dsRNA of the replicase gene. Inoculation of transgenic plants with CMV strain O discriminated three categories of plants: plants that showed complete resistance, which were free of symptoms; highly resistant plants, which had mild symptoms, but later recovered because new leaves that emerged were free of symptoms; and susceptible plants, which showed severe symptoms similar to wild-type plants. The completely resistant lines were selected and challenged with a closely related strain, CMV-Y. Interestingly, the transgenic plant lines either remained immune or showed high levels of resistance to the strain. No virus could be detected in uninoculated new leaves of the resistant lines after RT-PCR and Dot immunobinding assay (DIBA) analyses. We could show that the resistance is correlated with post-transcriptional gene silencing because of the production of transgenic specific siRNA.