Phylogenetic study of extirpated Korean leopard using mitochondrial DNA from an old skin specimen in South Korea

Abstract

The leopard, Panthera pardus, is a threatened species in its range throughout the world. Although, historically, the Korean Peninsula had a high population density of leopards, they were extirpated from South Korea by 1970, leaving almost no genetic specimens. Traditionally, Korean leopards are classified as Panthera pardus orientalis; however, their classification is based only on locality and morphology. Therefore, there is a need for genetic studies to identify the phylogenetic status of Korean leopards at the subspecies level. Presently, no extant wild specimen is available from South Korea; therefore, we extracted genetic material from the old skin of a leopard captured in Jirisan, South Korea in the 1930s and conducted the first phylogenetic study of the South Korean leopard. A total of 726 bp of mitochondrial DNA, including segments of the NADH5 and control region, were amplified by PCR. A phylogenetic analysis of the fragment, along with sequences of nine leopard subspecies from GenBank revealed that the extinct South Korean leopard belonged to the Asian leopard group and in the same clade as the Amur leopard (Panthera pardus orientalis). Thus, the leopard that inhabited South Korea in the past was of the same subspecies as the Amur leopard population currently inhabiting the transboundary region of Russia, China, and North Korea. These results emphasize the importance of conserving the endangered wild Amur leopard population (estimated to be about 60–80 individuals) in Russia and China, for future restoration of leopards in the Korean Peninsula.

Efficient and cost-effective non-invasive population monitoring as a method to assess the genetic diversity of the last remaining population of Amur leopard (Panthera pardus orientalis) in the Russia Far East

“…Two haplotypes differed at one variable site with A/T transversion at position 15596 ( Table 3 ) in the part of cytochrome b [ 44 ] and some samples showed both A and T peaks, suggesting the heteroplasmy as mentioned by Sugimoto et al [ 37 ] (Type A, n = 6; Type T, n = 14; heteroplasmy, n = 3). We did not report Amur leopard specific ORI 1, ORI 2 and KOR 1 haplotypes described by Uphyrkina et al [ 38 ] and Hyun et al [ 12 ]. Consistent with the results of previous studies, the haplotype diversity of the population was very low.…”

“…The Amur leopard ( Panthera pardus orientalis Schlegel, 1857), the northernmost leopard subspecies, is critically endangered [ 11 , 12 ] and has severely low genetic diversity [ 13 ]. Historically, it was distributed over a wide range of East Asia including the Korean Peninsula, northeastern China, and the southern part of the Russian Far East [ 14 , 15 ].…”

“…Historically, it was distributed over a wide range of East Asia including the Korean Peninsula, northeastern China, and the southern part of the Russian Far East [ 14 , 15 ]. A rapid population decline was observed in the 20 th century mainly due to excessive hunting, declining of prey species and marginalization of habitat resources, and only a small population currently exists in the mountain-forest region near the borders of Russia, China, and North Korea [ 12 , 13 , 16 , 17 ]. According to snow tracking surveys conducted in Russia from the 1990s to early 2000s, the number of leopards has been retained at 20–40 individuals in the southwestern part of Primorsky Krai [ 18 – 20 ].…”

Whole genome survey of big cats (Genus: Panthera) identifies novel microsatellites of utility in conservation genetic study

“…Incremental adoption of genetic tools and techniques for wildlife conservation and management have been observed globally in the past 25 years mainly due to the development of the robust protocols for DNA extraction and PCR (polymerase chain reaction) 13 – 16 . DNA tools are now increasingly employed for establishing species-level identity 17 , 18 , resolving taxonomic ambiguities 6 , 19 , 20 , wildlife conflict mitigation 21 , 22 , and more recently, establishing the source of origin 23 – 25 . Microsatellites or short tandem repeats (STR) are neutral, co-dominantly inherited, widely distributed, hypervariable, short repetitive nuclear DNA units that have been regarded as the best candidate to develop a genetic signature of the individual (DNA fingerprint), population, and subspecies 16 , 26 – 29 .…”

Whole genome survey of big cats (Genus:Panthera) identifies novel microsatellites of utility in conservation genetic study

“…Incremental adoption of genetic tools and techniques for wildlife conservation and management have been observed globally in the past 25 years mainly due to the development of the robust protocols for DNA extraction and PCR (Polymerase chain reaction) [29][30][31][32] . DNA tools are now increasingly employed for establishing species-level identity 33,34 , resolving taxonomic ambiguities 6,35,36 , wildlife conflict mitigation 37,38 , and more recently, establishing the source of origin [39][40][41] . Microsatellites or short tandem repeats (STR) are neutral, codominantly inherited, widely distributed, hypervariable, short repetitive nuclear DNA units that have been regarded as the best candidate to develop a genetic signature of the individual (DNA fingerprint), population, and subspecies.…”