Vector insect cells infected with Rice dwarf virus had vesicular compartments containing viral particles located adjacent to the viroplasm when examined by transmission electron and confocal microscopy. Such compartments were often at the periphery of infected cells. Inhibitors of vesicular transport, brefeldin A and monensin, and an inhibitor of myosin motor activity, butanedione monoxime, abolished the formation of such vesicles and prevented the release of viral particles from infected cells without significant effects on virus multiplication. Furthermore, the actin-depolymerizing drug, cytochalasin D, inhibited the formation of actin filaments without significantly interfering with formation of vesicular compartments and the release of viruses from treated cells. These results together revealed intracellular vesicular compartments as a mode for viral transport in and release from insect vector cells infected with a plant-infecting reovirus.In vector cells grown in monolayers (VCMs), Rice dwarf virus (RDV), a phytoreovirus, multiplies and spreads from primarily infected cells to neighbouring cells (Wei et al., 2006a) in addition to spreading via mature, free viral particles. Infection via free viral particles was protected by the addition of neutralizing antibodies to the cell culture medium. As part of integrated studies on RDV proliferation in vector insects, we have focused our study on the accumulation of the virus in cells of the insect vector and on the subsequent release of the virus.In electron micrographs of thin sections of insect tissues infected with plant-pathogenic reoviruses, viral particles were sequestered in spherical vesicular compartments (Fukushi et al., 1962;Shikata & Maramorosch, 1965;Shikata, 1969;Vidano, 1970;Omura et al., 1985). However, the biological significance of these inclusions in RDV infection has not been clarified because, for the most part, the tissues examined were in the late stage of infection which made it difficult to gather details on the formation of these compartments. Similar vesicular compartments appear to play a role in the release of viral particles from cultured cells infected with animal viruses, such as severe acute respiratory syndrome coronavirus (Ng et al., 2003) and human immunodeficiency virus (HIV) (Nydegger et al., 2003).A method for the continuous culture of cells from the leafhopper vector Nephotettix cincticeps, consisting of VCMs, has allowed the study of infection with RDV because such infection results in asymptomatic but persistent infection (Peterson & Nuss, 1985;Kimura, 1986). VCMs have been used to reveal fundamental aspects of viral activity at the cellular level, which suggests details of the transmission, multiplication and cytopathology of RDV in vector insects (Wei et al., 2006a(Wei et al., , b, c, 2007(Wei et al., , 2008. In the present study with this system, we investigated the role of vesicular compartments in the transport of RDV and its release from infected VCMs by confocal and electron microscopy, and the use of drugs that i...