Insect endosymbionts influence many important metabolic and developmental processes of their host. It has been speculated that they may also help to manipulate and suppress plant defenses to the benefit of herbivores. Recently, endosymbionts of the root herbivore Diabrotica virgifera virgifera have been reported to suppress the induction of defensive transcripts in maize roots, which may explain the finding of another study that once attacked plants become more susceptible to subsequent D. v. virgifera attack. To test this hypothesis, we cured D. v. virgifera from its major endosymbiont Wolbachia and tested whether endosymbiont-free individuals elicit different defense responses in maize roots. The presence of Wolbachia did not alter the induction of defense marker genes and resistance in a susceptible maize hybrid and a resistant line. Furthermore, attacked maize plants emitted the same amount of (E)-β-caryophyllene, a volatile signal that serves as foraging cue for both entomopathogenic nematodes and D. v. virgifera. Finally, the effectiveness of the entomopathogenic nematode Heterorhabditis bacteriophora to infest D. v. virgifera was not changed by curing the larvae from their endosymbionts. These results show that the defense mechanisms of maize are not affected by Wolbachia. Consequently, D. v. virgifera does not seem to derive any plant-defense mediated benefits from its major endosymbiont.