Genetics and extreme confinement of three overlooked butterfly species in Romania call for immediate conservation actions

A good knowledge of species distributions and their genetic structure is essential for numerous types of research such as population genetics, phylogeography, or conservation genetics. We document the presence of extremely local populations of three butterfly species (Iolana iolas, Satyrus ferula and Melanargia larissa) in the Romanian fauna. Satyrus ferula and M. larissa are reported for the first time in the country, while I. iolas is rediscovered following presumed extinction. Based on mitochondrial DNA (cytochrome c oxidase subunit 1—COI sequences), we assessed the genetic structure of these populations and placed them into a broader context through comparisons with other populations from across the range of these species. Each of the three species had a single haplotype in Romania, suggesting low female effective population size possibly under genetic erosion. Two of the populations (S. ferula and M. larissa) are genetically unique, displaying endemic haplotypes in south-western Romania. The Romanian populations of the three species likely remained unnoticed due to their extremely limited extent of occurrence. Their restricted range, close to the northern limits of distribution in the Balkans, their apparent low female effective population size, the presence of endemic haplotypes, and habitat vulnerability (especially for I. iolas) highlight the need for monitoring and conservation measures for the safeguarding of these populations. IMPLICATIONS FOR INSECT CONSERVATION: We discover and study the genetics of previously unknown populations of three butterfly species (Iolana iolas, Satyrus ferula and Melanargia larissa) in Romania, which deserve conservation measures. This study exemplifies how biodiversity exploration and genetic information can contribute to informed decisions regarding the management and protection of threatened populations.