Autophagy mediates a direct synergistic interaction during cotransmission of two distinct arboviruses by insect vectors

Multiple viral infections in insect vectors with synergistic effects are common in nature,but the underlying mechanism remains elusive.Here,we find that rice gall dwarf reovirus(RGDV)facilitates the transmission of rice stripe mosaic rhabdovirus(RSMV)by co-infected leafhopper vectors.RSMV nucleoprotein(N)alone activates complete anti-viral autophagy,while RGDV nonstructural protein Pns11 alone induces pro-viral incomplete autophagy.In co-infected vectors,RSMVexploits Pns11-induced autophagosomes to assemble enveloped virions via N-Pns11-ATG5 interaction.Furthermore,RSMV could effectively propagate in Sf9 cells.Expression of Pns11 in Sf9 cells or leafhopper vectors causes the recruitment of N from the ER to Pns11-induced autophagosomes and inhibits N-induced complete autophagic flux,finally facilitating RSMV propagation.In summary,these results demonstrate a previously unappreciated role of autophagy in the regulation of the direct synergistic interaction during co-transmission of two distinct arboviruses by insect vectors and reveal the functional importance of virus-induced autophagosomes in rhabdovirus assembly.