Infection pattern of male-killing viruses alters phenotypes in the tea tortrix moth Homona magnanima

Male-killing is a microbe-induced reproductive manipulation in invertebrates whereby male hosts are eliminated during development. In the tea tortrix moth Homona magnanima, Osugoroshi viruses 1‒3 (OGVs), belonging to Partitiviridae induce male-killing. The infection patterns of OGVs are diverse; however, how the influence of these patterns of host phenotypes remains largely unknown. Using field-collected larvae, we established a OGV1 and OGV3 double-infection line, in addition to a triple-infection line, and examined the dsRNA segments, purified viral proteins, OGV density, and host phenotypes. PCR analysis demonstrated that the triple-infection line lost one dsRNA segment, whereas the double-infection line lost eight segments, including one RNA-dependent RNA polymerase (RdRp) gene. LC-MS analysis revealed three potential structural proteins in the OGVs. Males died at the larval stage in the triple-infection line and at the embryo-larval stage in the double-infection line of OGV1 and OGV3; the RNA load of female parents did not contribute to the developmental stage at which males died. These findings indicate that the pattern of viral infection, rather than viral RNA load transmitted from female parent, controls the stage of development at which male-killing occurs. Furthermore, the duration of the larval stage of the double-infection line was found to be significantly longer than that of the triple-infection line. The shorter duration of the larval stage of the triple-infection line could be advantageous over the double-infection line in maximizing transmission efficiency.