Aphid-borne Viral Spread Is Enhanced by Virus-induced Accumulation of Plant Reactive Oxygen Species

Most known plant viruses are spread from plant to plant by insect vectors. There is strong evidence that nonpersistently transmitted viruses manipulate the release of plant volatiles to attract insect vectors, thereby promoting virus spread. The mechanisms whereby aphid settling and feeding is altered on plants infected with these viruses, however, are unclear. Here we employed loss-of-function mutations in cucumber mosaic virus (CMV) and one of its host plants, tobacco (Nicotiana tabacum), to elucidate such mechanisms. We show that, relative to a CMV Delta 2b strain with a deletion of the viral suppressor of RNAi 2b protein in CMV, plants infected with wild-type CMV produce higher concentrations of the reactive oxygen species (ROS) H2O2 in plant tissues. Aphids on wild-type CMV-infected plants engage in shorter probes, less phloem feeding, and exhibit other changes, as detected by electrical penetration graphing technology, relative to CMV Delta 2b-infected plants. Therefore, the frequency of virus acquisition and the virus load per aphid were greater on CMV-infected plants than on CMV Delta 2b-infected plants. Aphids also moved away from initial feeding sites more frequently on wild-type CMV infected versus CMV Delta 2b-infected plants. The role of H2O2 in eliciting these effects on aphids was corroborated using healthy plants infused with H2O2. Finally, H2O2 levels were not elevated, and aphid behavior was unchanged, on CMV-infected RbohD-silenced tobacco plants, which are deficient in the induction of ROS production. These results suggest that CMV uses its viral suppressor of RNAi protein to increase plant ROS levels, thereby enhancing its acquisition and transmission by vector insects.