Research article on microsatellite marker development and genetic diversity analysis of Japanese wrinkled frogs using Nanopore sequencing technology published

A collaborative research project on the development of microsatellite markers and genetic diversity of the Japanese wrinkled frog (Glandirana rugosa) with Professor Teruhiko Takahara and Ms. Fukatsu (graduate student) from the Faculty of Life and Environmental Science at Shimane University has been published in Ecological Research.

While we have previously conducted collaborative research on sex-determining genes in Japanese wrinkled frogs, which possess different sex chromosomes depending on their geographic region, genetic markers capable of population dynamics analysis and individual identification had not been reported until now. Professor Takahara and his team have been investigating the population dynamics of Japanese wrinkled frog regional populations, including those on the Oki Islands. To develop microsatellite markers as genetic tools for this purpose, they performed genome sequencing using a Nanopore sequencer and isolated microsatellite loci. Although Nanopore sequencers are cost-effective next-generation sequencers excellent for long-read sequencing, sequence accuracy has been a concern. However, sequencing data obtained with SQK-LSK110 reagents provided sufficient accuracy for microsatellite loci isolation and primer design, successfully developing markers for 14 loci. Furthermore, population analysis using these markers revealed that detailed genetic relationships among adjacent regional populations could be elucidated.

Fukatsu, Y., T. Igawa, K. Sasaki, S. Yamagishi, and T. Takahara. 2025. “ Genetic Diversity in the Wrinkled Frog, Glandirana rugosa, Evaluated Using Microsatellite Markers Identified by Nanopore Sequencing.” Ecological Research 40, no. 6: e70014. https://doi.org/10.1111/1440-1703.70014.