Genomic differentiation and niche divergence in the Hetaerina americana (Odonata) cryptic species complex.

The evolution of reproductive barriers, that is, the speciation process, implies the limitation of gene flow between populations. Different patterns of genomic differentiation throughout the speciation continuum may provide insights into the causal evolutionary forces of species divergence. In this study, we analysed a cryptic species complex of the genus Hetaerina (Odonata). This complex includes H. americana and H. calverti; however, in H. americana two highly differentiated genetic groups have been previously detected, which, we hypothesize, may correspond to different species with low morphological variation. We obtained single nucleotide polymorphism (SNP) data for 90 individuals belonging to the different taxa in the complex and carried out differentiation tests to identify genetic isolation. The results from STRUCTURE and discriminant analysis of principal components (DAPC), based on almost 5000 SNPs, confirmed the presence of three highly differentiated taxa. Also, we found FST values above 0.5 in pairwise comparisons, which indicates a considerable degree of genetic isolation among the suggested species. We also found low climatic niche overlap among all taxa, suggesting that each group occurs at specific conditions of temperature, precipitation and elevation. We propose that H. americana comprises two cryptic species, which may be reproductively isolated by ecological barriers related to niche divergence, since the morphological variation is minimal and, therefore, mechanical barriers are probably less effective compared to other related species such as H. calverti.

Morphological variation and reproductive isolation in the Hetaerina americana species complex.

Incomplete premating barriers in closely related species may result in reproductive interference. This process has different fitness consequences and can lead to three scenarios: niche segregation, sexual exclusion, or reproductive character displacement. In morphologically cryptic species, isolation barriers can be difficult to recognize. Here, we analyzed the morphological, behavioral, and genetic differences between two sympatric cryptic species of the genus Hetaerina to determine the characters that contribute the most to reproductive isolation and the effect of the high rates of behavior interference between the species. We found complete genetic isolation and significant differences in the morphometry of caudal appendages and wing shape, as well as body size variation between species. In contrast, we did not find clear differences in the coloration of the wing spot and observed high rates of interspecific aggression. Our results suggest that divergence in the shape of the caudal appendages is the principal pre-mating barrier that prevents interspecific mating. Moreover, a scenario of character displacement on body size was found. Nevertheless, size could play an important role in both inter- and intrasexual interactions and, therefore, we cannot differentiate if it has resulted from reproductive or aggressive interference.

Hetaerina calverti (Odonata: Zygoptera: Calopterygidae) sp. nov., a new cryptic species of the American Rubyspot complex.

Hetaerina americana Fabricius, 1798 has a long and irresolute taxonomic history. Several synonyms have been suggested (H. californica Hagen in Selys-Longchamps, 1859, H. basalis Hagen in Selys-Longchamps, 1859, H. texana Walsh, 1863, H. scelerata Walsh, 1863, etc.), related to the variation in the size of the wing spots as well as to the morphology of the male cerci. However, Calvert (1901) suggested that H. americana represents one variable species. Nevertheless, Vega-Sánchez et al. (2019) through a genetic and morphological analysis presented evidence to propose that H. americana represents a species complex. In the present work, we describe a new species that belongs to this complex: H. calverti sp. nov. The morphological characteristics by which males and females of H. calverti differ from H. americana are highlighted. The most important character for the differentiation of males is the shape of the cerci and the size of the individuals (when the two species are in sympatry). In females, the main differences are in the shape of the intersternites and the medio-dorsal carina of the last segment of the abdomen. Some generalities about the biology of the species are presented, including geographical distribution patterns and genetic divergence data. [urn:lsid:zoobank.org:act:F5C329AE-7A00-4979-8A0D-A13D869E54B1].

Complex evolutionary history of the American Rubyspot damselfly, Hetaerina americana (Odonata): Evidence of cryptic speciation.

Analyzing the magnitude and distribution of genetic variation within and among populations allows for hypothesis testing about historical demographic size changes, secondary contacts, refugia, and speciation patterns. Species distribution and genetic structure are greatly influenced by the complex life cycle and behavior of odonates. Hetaerina americana has been widely used as a model system in behavioral studies, but its population genetic structure has not been analyzed, except for a single study that included only three populations but identified the presence of markedly differentiated genetic groups, suggesting the existence of cryptic species. Here, we tested this hypothesis by assessing throughout the distribution range of H. americana the patterns of genetic and morphological variation in the male caudal appendages, due to the great importance of these structures in mate recognition. As molecular markers we used sequences of the mitochondrial cytochrome oxidase I (COI) gene and the nuclear internal transcribed spacer (ITS) region, as well as six nuclear microsatellites. We found very high population genetic differentiation (ΦST > 0.51) in the three sets of markers but with strong mitonuclear discordance. A neutrality test suggested that the mitochondrial genome might be under purifying selection in association to climatic variables (temperature seasonality). The assignment of individuals to nuclear genetic groups showed little admixture and complete congruence with morphological differentiation in the male caudal appendages. Hence, the results suggest that H. americana represents at least two different cryptic species which are isolated reproductively.