The effects of grape-tobacco intercropping patterns on populations of grape phylloxera, Daktulosphaira vitifoliae Fitch, as well as on the growth and development of the infested vines were evaluated in the field and the impact of an aqueous tobacco root extract on grape phylloxera was evaluated using a laboratory bioassay. The aqueous tobacco root extract exhibited biological activity against this pest. The egg mortality, nymph mortality, development period, life span and female fecundity were significantly affected. In the field trial, grape phylloxera populations were clearly lower as compared to the monoculture pattern. However, the rates of newly developed roots and newly infested grape roots were significantly higher and lower, in intercropping patterns than in the vine monoculture, respectively. The grape phylloxera population number on the grape roots decreased each year, and the vine trees gradually renewed upon continuous intercropping with tobacco over three years. These results confirmed that intercropping grapes with tobacco can effectively control grape phylloxera in an infested vineyard. The results also indicated that additional crops could be intercropped with grapes and are effective against grape phylloxera, which should be explored as an integrated approach for controlling the pest.