Grapevine leafroll-associated virus 3 (GLRaV-3) is associated with the economically damaging grapevine leafroll disease, and is transmitted in a semi-persistent manner by several mealybug species. We performed the first controlled field study of vector-mediated inoculations with GLRaV-3 in a commercial vineyard with previously asymptomatic vines, and monitored the vines during four growing seasons. We then compared the outcome of vector-mediated inoculations in the field study to an analogous laboratory study. In the vineyard, about half of all inoculated plants became infected with GLRaV-3, fewer than in the controlled laboratory inoculations. Mealybugs had lower settling and feeding success in the field than in the laboratory inoculations. Our study suggests that laboratory studies may overestimate transmission efficiency. All successfully inoculated vineyard plants first became symptomatic one growing season after inoculations, and berry quality declined within one year after inoculations. Vineyard plants were effective virus sources one year after inoculations. Our findings generally indicate that newly symptomatic vines in commercial vineyards probably became infected during the previous growing season, and a decline in berry quality can be expected during the same year in which symptoms appear.