Molecular phylogeny and morphology of Mycosphaerella nawae, the causal agent of circular leaf spot on persimmon

In this study, the phylogeny and morphology of Mycosphaerella nawae (Dothideomycetes, Ascomycota) were examined using Korean and Japanese isolates, to establish the phylogenetic relationship between M. nawae and its allied species. Korean and Japanese isolates of M. nawae were collected from circular leaf spot-diseased leaves and were confirmed based on internal transcribed spacer (ITS) sequence data. Phylogenetic analysis was conducted using multiple genes, including the ITS region, 28S rDNA, ß-tubulin, translation elongation factor-1a, and actin genes. Our results revealed that M. nawae is closely related to members of the genus Phaeophleospora but are distant from the Ramularia spp. In addition, microscopic analysis revealed pseudothecia on the adaxial and abaxial surface of overwintered diseased leaves (ODL) and only on the abaxial surface of diseased leaves. Ascospores are oval to fusiform, one-septate, tapered at both ends, 1.7~3.1 × 8.1~14.1 μm, and were observed in ODL. Conidia are oval, guttulate, one-septate, 3.5~4.9 × 12.8~19.8 μm, and barely discernable on 30-day cultures. To our knowledge, this is the first report on the phylogeny of M. nawae, which is closely related to the genus Phaeophleospora, especially P. scytalidii.