Immunofluorescence localization and ultrastructure of Stewart's wilt disease bacterium Pantoea stewartii in maize leaves and in its flea beetle vector Chaetocnema pulicaria (Coleoptera: Chrysomelidae)

Abstract

Pantoea stewartii is the causal agent of Stewart's wilt of sweet corn, the most serious bacterial disease of sweet corn and maize in the North-Central and Eastern USA. P. stewartii is transmitted mainly by the corn flea beetle Chaetocnema pulicaria (Coleoptera: Chrysomelidae) and this bacterium is assumed to overwinter in its vector beetle. Using immunofluorescence confocal and transmission electron microscopy (TEM) we localized P. stewartii in its host plant and insect vector. In infected maize, P. stewartii was found mainly in xylem vessels of major and minor veins as well as in intercellular spaces of infected leaves. In the vector beetle, P. stewartii was found by immunofluorescence in the foregut, midgut and hindgut up to 12 days post-acquisition (post-feeding on infected plants for 2 days). TEM of thin sections in the beetle's gut four and eight days post-acquisition revealed bacterial cells similar to those of P. stewartii in the gut lumen, close to or associated with the gut microvilli, as well as inside epithelial cells of the midgut. This suggests a stronger biological relationship between this bacterium and its vector, and that the corn flea beetle may carry P. stewartii intracellularly as well as extracellularly within the gut, which has implications in the persistence and overwintering of this bacterium in the vector and on the epidemiology of Stewart's wilt disease of corn.