Flight Dispersal in Supratidal Rockpool Beetles.

Flight dispersal is ecologically relevant for the survival of supratidal rockpool insects. Dispersal has important consequences for colonisation, gene flow, and evolutionary divergence. Here, we compared the flight dispersal capacity of two congeneric beetle species (Ochthebius quadricollis and Ochthebius lejolisii) that exclusively inhabit these temporary, fragmented, and extreme habitats. We estimated flight capacity and inferred dispersal in both species using different approaches: experimental flying assays, examination of wing morphology, and comparison of microsatellite markers between species. Our findings revealed that both species exhibited similar flight behaviour, with 60 to 80% of the individuals flying under water heating conditions. Notably, females of both species had larger body sizes and wing areas, along with lower wing loading, than males in O. quadricollis. These morphological traits are related to higher dispersal capacity and more energetically efficient flight, which could indicate a female-biassed dispersal pattern. The wing shapes of both species are characterised by relatively larger and narrower wings in relation to other species of the genus, suggesting high flight capacity at short distances. Molecular data revealed in both species low genetic divergences between neighbouring populations, non-significant differences between species, and no isolation by distance effect at the study scale (<100 km). These results point to passive dispersal assisted by wind.

Novel Microsatellite Loci, Cross-Species Validation of Multiplex Assays, and By-Catch Mitochondrial Genomes on Ochthebius Beetles from Supratidal Rockpools.

Here we focus on designing, for the first time, microsatellite markers for evolutionary and ecological research on aquatic beetles from the genus Ochthebius (Coleoptera, Hydraenidae). Some of these non-model species, with high cryptic diversity, exclusively inhabit supratidal rockpools, extreme and highly dynamic habitats with important anthropogenic threats. We analysed 15 individuals of four species (O. lejolisii, O. subinteger, O. celatus, and O. quadricollis) across 10 localities from the Mediterranean coasts of Spain and Malta. Using next-generation sequencing technology, two libraries were constructed to interpret the species of the two subgenera present consistently (Ochthebius s. str., O. quadricollis; and Cobalius, the rest of the species). Finally, 20 markers (10 for each subgenus) were obtained and successfully tested by cross-validation in the four species under study. As a by-catch, we could retrieve the complete mitochondrial genomes of O. lejolisii, O. quadricollis, and O. subinteger. Interestingly, the mitochondrial genome of O. quadricollis exhibited high genetic variability compared to already published data. The novel SSR panels and mitochondrial genomes for Ochthebius will be valuable in future research on species identification, diversity, genetic structure, and population connectivity in highly dynamic and threatened habitats such as supratidal coastal rockpools.