Development of high-resolution melting markers for assessment of the hybridization between Mauremys sinensis and Mauremys reevesii

Two freshwater turtle species of the genus Mauremys are known to occur in South Korea: M. sinensis and M. reevesii. Mauremys sinensis is an invasive alien species, whereas M. reevesii is a legally protected and native species to South Korea. They occur sympatrically and their hybrids have recently been found both in captivity and in the wild, posing a serious threat to the conservation of M. reevesii. To facilitate rapid identification of hybrids, we have developed a genotyping marker for high resolution melting (HRM) analysis that targets the intron I region. Three distinct HRM curves, representing M. reevesii, M. sinensis, and their hybrids, were obtained. These results were identical to those observed for R35, with the hybrid possessing both alleles. This demonstrates that HRM markers can effectively distinguish Mauremys species from their hybrids. The present study provides a valuable tool for rapid and accurate identification of hybrids for monitoring.