Comparison of the Genetic Diversity of the Captive and Wild Populations of the Tsushima Leopard Cat Using a GRAS-Di Analysis

SIMPLE SUMMARY: The Tsushima leopard cat (Prionailurus bengalensis euptilurus) is exclusively found on the Tsushima Island in Japan; however, its population has been declining in recent years and is in danger of extinction. Hence, effort is underway for both in situ and ex situ conservation. Genetic management is also important in the management of captive populations, and various studies are being conducted regarding the same. In our previous study, we developed a reference genome of the Tsushima leopard cat and used GRAS-Di analysis, a genome-wide analysis, to genetically evaluate the wild populations. In this study, we attempted to improve the Tsushima leopard cat reference genome and compared the captive and wild populations by GRAS-di analysis. The results showed that the captive population had almost the same genetic diversity as the wild population and managed to remain in good condition. ABSTRACT: The Tsushima leopard cat (Prionailurus bengalensis euptilurus) (TLC) is a regional population of the Amur leopard cat (P. bengalensis euptilurus) that lives only on the Tsushima Island in Japan and is threatened with extinction. Because the TLC population is small, genetic management is important. In this study, we obtained the draft genome of the TLC and identified single-nucleotide polymorphism (SNP) markers using a genotyping by random amplicon sequencing-direct (GRAS-Di) analysis. We genotyped 31 captive individuals and 50 wild individuals, of which 48 were from a previous study. The identified SNPs were used to clarify the genetic diversity and genetic structure of the wild and captive populations of the TLC. The size of the genome was estimated to be about 2.42 Gb. The number of SNP markers developed was 139, and although PID and probability of exclusion obtained using these SNP markers were not as high as those reported in the studies of other wild species, these SNP markers could be used to identify individuals and parentage. Moreover, the genetic diversity indices of the captive population were similar to those of the wild population. These SNP markers will be useful for understanding the ecology of the TLC and planning conservation strategies.