Genetic diversity and population structure of <i>Cybister rugosus</i> based on the mitochondrial COI gene and microsatellite markers

The aquatic beetle Cybister rugosus has experienced ongoing habitat loss, a decline in population numbers and confirmed extinctions among insular populations in Japan. It has recently been classified on the Red List as endangered by the Japanese Ministry of Environment and has been designated a specified class II nationally rare species of wild fauna and flora. To design effective conservation strategies for this species, it is essential to compile data on its genetic variation to ascertain its genetic diversity and population structure. Previous studies found low levels of genetic variation in the COI gene among Japanese insular populations and failed to detect a fine population structure. Thus, we developed ten novel microsatellite markers for C. rugosus, using whole-genome shotgun sequencing. The degree of polymorphism for these markers was characterized using summary statistics describing the genetic variation in 49 individuals from populations in Cambodia and on the islands of the Ryukyu Archipelago. Microsatellite data indicated differentiation among the insular populations in Japan, which could not be clearly shown in the COI data, and some insular populations showed low levels of genetic diversity. The newly developed microsatellite markers will contribute to future ecological and evolutionary studies on this species and to conservation research.