Infective second-stage juveniles of the obligate plant-parasitic root-knot and cyst nematodes invade plant roots to induce specialized feeding structures. Here, we present data on the distribution of plasmodesmata in cell walls of syncytia and giant cells induced by cyst and root-knot nematodes. An Arabidopsis and a tobacco line were used, containing viral movement proteins fused to green fluorescent protein as a localization marker for plasmodesmata. Plasmodesmata were detected in walls between giant cells but also in walls toward neighboring cells. In syncytia, plasmodesmata were mainly detected at later stages. In young syncytia, few plasmodesmata were observed and a specific temporal callose deposition along plasmodesmata indicated impaired symplasmic exchange. In order to study the relevance of callose deposition for successful cyst nematode development in Arabidopsis, two mutant lines inhibited in callose synthesis and degradation, respectively, were used in nematode infection assays. Histological analyses showed that syncytia were smaller when callose degradation was reduced, indicating a significant importance of this process to cyst nematode development.