Genetic Diversity and Population Dynamics of Leptobrachium leishanense (Anura: Megophryidae) as Determined by Tetranucleotide Microsatellite Markers Developed from Its Genome

SIMPLE SUMMARY: More than 41 percent of amphibians evaluated by International Union for Conservation of Nature are threatened. It is vitally important to establish scientific and effective protection strategies for these organisms. Leishan Spiny Toad is endemic to China and it has a narrow distribution area. Long-term intentional human use and habitat destruction has caused the species to suffer. Here, we developed newly reliable and efficient molecular markers based on its genome to assess its genetic diversity and population history and provided support for conservation of this toad. Our results show that this toad still possesses high genetic diversity, but population decline may increase the possibility of inbreeding, which could work against persisting survival. Recovering the toad’s habitat and strengthening the publicity and education of wildlife protection can be helpful to its sustainability. ABSTRACT: Persisting declination of amphibians around the world has resulted in the public attaching importance to the conservation of their biodiversity. Genetic data can be greatly helpful in conservation planning and management, especially in species that are small in size and hard to observe. It is essential to perform genetic assessments for the conservation of Leptobrachium leishanense, an endangered toad and receiving secondary protection on the list of state-protected wildlife in China. However, current molecular markers with low reliability and efficiency hinder studies. Here, we sampled 120 adult toes from the population in the Leishan Mountain, 23 of which were used to develop tetranucleotide microsatellite markers based on one reference L. leishanense genome. After primer optimization, stability detection, and polymorphism detection, we obtained 12 satisfactory microsatellite loci. Then, we used these loci to evaluate the genetic diversity and population dynamics of the 120 individuals. Our results show that there is a low degree of inbreeding in the population, and it has a high genetic diversity. Recently, the population has not experienced population bottlenecks, and the estimated effective population size was 424.3. Accordingly, stabilizing genetic diversity will be key to population sustainability. Recovering its habitat and avoiding intentional human use will be useful for conservation of this species.