| Summary: | Grapevine leafroll disease is one of the most important virus diseases of grapevines and occurs in every major grape-growing region of the world. The vector-transmission mechanisms of the causative agent, <i>Grapevine leafroll-associated virus 3</i> (GLRaV-3), remain poorly understood. We show that the vine mealybug, <i>Planococcus ficus</i>, feeds through a membrane feeding system on GLRaV-3 viral purifications from both <i>V. vinifera</i> and <i>N. benthamiana</i> and transmits the virus to test plants from plants from both species. Building on this strategy, we used an immunofluorescence approach to localize virions to two retention sites in <i>P. ficus</i> mouthparts. Assays testing molecules capable of blocking virus transmission demonstrated that GLRaV-3-transmission by <i>P. ficus</i> could be disrupted. Our results indicate that our membrane feeding system and transmission-blocking assays are a valid approach and can be used to screen other candidate blocking molecules.
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