Species- and sex-specific multiple pcr amplifications of partial cytochrome b gene and Zfx/Zfy introns from invasive and non-invasive samples of Korean ungulates

Abstract

There are five representative ungulates (e.g., goral, Nemorhaedus caudatus, goat, Capra hircuä roe deer, Capreolus pygargus, water deer, Hydropotes inermiä, musk deer, Moschus moschiferus) in the wild of South Korea. Their fecal morphologies are similar to each other between species or sexes, and therefore it is very difficult to identify the species or sex. To distinguish the species and sex of the elusive animals, we developed species- and sex-specific multiple PCR amplifications using pairs of primer sets. Partial cytochrome b gene and Zfx/Zfy introns were targeted for the PCR amplifications. For species identification using a multiple primer set (BJGL-F1 / GT-Fl / RDF1 / WD-F1/MD-F1 and BJGL-R / GT-R / RD-R / WD-R / MD-R), all (n = 21/21) of the invasive samples were correctly identified. About 71% (n = 15 / 21) of the non-invasive samples were successfully identified. The multiple primer set could determine each species which showed a unique sized PCR fragment. For sex identification using a multiple primer set (BJSexX-F/Y-F and BJSex-Rl), about 90% (n = 19/2l) of invasive samples were correctly determined. About 62% (n=13/2l) of the non-invasive samples were successfully identified. Using the multiple primer set, both X- and Y-chromosome linked bands for males (n = 21) and only X-chromosome linked band for females (n = 11) could be detected. The results would be applicable to the species and sex identification of the Korean and the Far-East Asian wild ungulates.