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Giant viruses, like pandoraviruses and mimiviruses, have been discovered from diverse environments, and their broad global distribution has been established. Here, we report two new isolates of  Pandoravirus  spp. and one  Mimivirus  sp., named  Pandoravirus hades ,  Pandoravirus persephone , and  Mimivirus  sp. isolate styx, co-isolated from riverbank soil in Japan. We obtained nearly complete sequences of the family B DNA polymerase gene ( polB ) of  P. hades  and  P. persephone;  the former carried two known intein regions, while the latter had only one. Phylogenetic analysis revealed that the two new pandoravirus isolates are closely related to  Pandoravirus dulcis . Furthermore, random amplified polymorphic DNA analysis revealed that  P. hades  and  P. persephone  might harbor different genome structures. Based on phylogenetic analysis of the partial  polB  sequence,  Mimivirus  sp. isolate styx belongs to mimivirus lineage A. DNA staining suggested that the  Pandoravirus  spp. asynchronously replicates in amoeba cells while  Mimivirus  sp. replicates synchronously. We also observed that  P. persephone-  or  Mimivirus  sp. isolate styx-infected amoeba cytoplasm is extruded by the cells. To the best of our knowledge, we are the first to report the isolation of pandoraviruses in Asia. In addition, our results emphasize the importance of virus isolation from soil to reveal the ecology of giant viruses.

Co-Isolation and Characterization of Two Pandoraviruses and a Mimivirus from a Riverbank in Japan