The Integrative Taxonomic Approach Reveals Host Specific Species in an Encyrtid Parasitoid Species Complex

Integrated taxonomy uses evidence from a number of different character types to delimit species and other natural groupings. While this approach has been advocated recently, and should be of particular utility in the case of diminutive insect parasitoids, there are relatively few examples of its application in these taxa. Here, we use an integrated framework to delimit independent lineages in Encyrtus sasakii (Hymenoptera: Chalcidoidea: Encyrtidae), a parasitoid morphospecies previously considered a host generalist. Sequence variation at the DNA barcode (cytochrome c oxidase I, COI) and nuclear 28S rDNA loci were compared to morphometric recordings and mating compatibility tests, among samples of this species complex collected from its four scale insect hosts, covering a broad geographic range of northern and central China. Our results reveal that Encyrtus sasakii comprises three lineages that, while sharing a similar morphology, are highly divergent at the molecular level. At the barcode locus, the median K2P molecular distance between individuals from three primary populations was found to be 11.3%, well outside the divergence usually observed between Chalcidoidea conspecifics (0.5%). Corroborative evidence that the genetic lineages represent independent species was found from mating tests, where compatibility was observed only within populations, and morphometric analysis, which found that despite apparent morphological homogeneity, populations clustered according to forewing shape. The independent lineages defined by the integrated analysis correspond to the three scale insect hosts, suggesting the presence of host specific cryptic species. The finding of hidden host specificity in this species complex demonstrates the critical role that DNA barcoding will increasingly play in revealing hidden biodiversity in taxa that present difficulties for traditional taxonomic approaches

Cryptic Species? Patterns of Maternal and Paternal Gene Flow in Eight Neotropical Bats

Levels of sequence divergence at mitochondrial loci are frequently used in phylogeographic analysis and species delimitation though single marker systems cannot assess bi-parental gene flow. In this investigation I compare the phylogeographic patterns revealed through the maternally inherited mitochondrial COI region and the paternally inherited 7th intron region of the Dby gene on the Y-chromosome in eight common Neotropical bat species. These species are diverse and include members of two families from the feeding guilds of sanguivores, nectarivores, frugivores, carnivores and insectivores. In each case, the currently recognized taxon is comprised of distinct, substantially divergent intraspecific mitochondrial lineages suggesting cryptic species complexes. In Chrotopterus auritus, and Saccopteryx bilineata I observed congruent patterns of divergence in both genetic regions suggesting a cessation of gene flow between intraspecific groups. This evidence supports the existence of cryptic species complexes which meet the criteria of the genetic species concept. In Glossophaga soricina two intraspecific groups with largely sympatric South American ranges show evidence for incomplete lineage sorting or frequent hybridization while a third group with a Central American distribution appears to diverge congruently at both loci suggesting speciation. Within Desmodus rotundus and Trachops cirrhosus the paternally inherited region was monomorphic and thus does not support or refute the potential for cryptic speciation. In Uroderma bilobatum, Micronycteris megalotis and Platyrrhinus helleri the gene regions show conflicting patterns of divergence and I cannot exclude ongoing gene flow between intraspecific groups. This analysis provides a comprehensive comparison across taxa and employs both maternally and paternally inherited gene regions to validate patterns of gene flow. I present evidence for previously unrecognized species meeting the criteria of the genetic species concept but demonstrate that estimates of mitochondrial diversity alone do not accurately represent gene flow in these species and that contact/hybrid zones must be explored to evaluate reproductive isolation

Cryptic diversity, systematic and historical biogeography of the genus Manerebia Staudinger, 1897 (Satyrinae: Pronophilina) in the neotropics

ilustraciones, graficasLas mariposas Pronophilina Reuter, es una de las subtribus de la tribu Satyrini, son reconocidas como uno de los grupos de mariposas más diversificados en ambientes montañosos y presentan altos niveles de endemismo. Sin embargo, la determinación taxonómica precisa de las especies en muchos géneros de Pronophilina se ha visto afectada por la diversidad críptica y taxones taxonómicamente confusos como es el caso del género Manerebia Staudinger. Este género es un grupo de mariposas andinas, que se distribuye desde el norte de Argentina hasta Venezuela, y presenta una alta diversidad críptica y una variación fenotípica alta (polimorfismos). Se han descrito varias especies nuevas durante las últimas décadas, y otras aún esperan ser descritas. No obstante, la ubicación de Manerebia dentro de la subtribu Pronophilina debe considerarse provisional porque no hay un análisis filogenético y su monofilia aún no se ha evaluado. Además, aún se desconocen las relaciones filogenéticas de las especies dentro del género. Aunque, el género es de interés desde las perspectivas ecológica, evolutiva, biogeográfica y de conservación, pero como base necesaria para tales estudios se necesita un conocimiento sólido que ayude a comprender e inferir los patrones filogenéticos y biogeográficos sobre la historia evolutiva del género Manerebia en el Neotrópico. Por lo tanto, evaluamos la monofilia del género Manerebia, determinamos su posición taxonómica y las relaciones filogenéticas dentro de la tribu Satyrini, y proporcionamos una mejor comprensión de las relaciones a nivel de subtribu dentro de Satyrini. Encontramos a Manerebia como un grupo monofilético en Pronophilina y aclaramos sus relaciones filogenéticas. Descubrimos que el uso de un muestreo taxonómico más grande puede ayudar a mejorar los problemas al usar genes individuales y permite construir relaciones sistemáticas más sólidas. Con base en nuestros análisis, encontramos 48 especies distintas de nuestras 24 especies nominales muestreadas, de las cuales 14 son especies nuevas. Por lo tanto, de acuerdo con nuestra propuesta sistemática, el género Manerebia comprendería 58 especies nominales, pero por el momento algunas permanecen sin describir. Los análisis filogenéticos, junto con los métodos de delimitación de especies y los caracteres morfológicos, permitieron evaluar la alta diversidad críptica dentro del género. Además, nuestro análisis destaca la importancia de emplear el marco de taxonomía integradora para la detección de diversidad críptica en regiones como el Neotrópico. Generamos la primera hipótesis filogenética para el género Manerebia basada en datos de secuencias mitocondriales (COI) y utilizando herramientas filogenéticas. Se proponen nueve clados para el género Manerebia a lo largo de los Andes Central y del Norte, siendo el Norte de los Andes la zona con mayor riqueza para el género. Nuestros análisis nos permitieron aclarar algunas de las relaciones filogenéticas dentro del género a nivel de especie. Finalmente, nuestro estudio exploró la historia biogeográfica del género Manerebia estimando tiempos y tasas de diversificación de sus linajes y empleando un análisis biogeográfico parareconstruir su historia evolutiva. Nuestros resultados nos permitieron inferir que el tiempo de divergencia de Manerebia fue entre el Mioceno tardío y el Plioceno, y la mayoría de los linajes existentes ya habían aparecido en el Pleistoceno. El género tuvo un estallido temprano general en el límite del Mioceno tardío / Plioceno temprano seguido de una desaceleración debido a una disminución en la especiación a lo largo del Pleistoceno, y este patrón se refleja para todos los clados en Manerebia. Los eventos de dispersión fue posiblemente el proceso biogeográficos más común dentro del género, y nuestros resultados nos permiten confirmar el papel de la geomorfología andina en la evolución de la biodiversidad Neotropical. (Texto tomado de la fuente)The Pronophilina Reuter butterflies, one of the subtribes of the tribe Satyrini, are recognized as one of the most diversified groups of butterflies in mountain environments and present high levels of endemism. However, the accurate taxonomic determination of species in many genera of Pronophilina has been affected by the cryptic diversity and taxonomically confusing taxa as is the case of the genus Manerebia Staudinger. This genus is an Andean butterflies group, which is distributed from northern Argentina to Venezuela, and it presents a high cryptic diversity and a phenotypic variation (polymorphisms). Several new species have been described during the last few decades, and others still await description. Nevertheless, the placement of Manerebia within the subtribe Pronophilina is to be considered tentative because there isn't a phylogenetic analysis, and its monophyly is not evaluated yet. In addition, the species phylogenetic relationships within the genus are unknown yet. However, the genus is of interest from ecological, evolutionary, biogeographic, and conservation perspectives, but as a necessary base for such studies a robust knowledge is needed to help to understand and infer the phylogenetic and biogeographic patterns about the genus Manerebia evolutionary history in the Neotropic. Hence, we evaluated the monophyly of the genus Manerebia, determined its taxonomic position and phylogenetic relationships within the tribe Satyrini, and provided a better understanding of the at the subtribe level relationships within the Satyrini. We found Manerebia as a monophyletic group into Pronophilina and clarified its phylogenetic relationships. We found that using larger taxonomic sampling may help to improve the problems when using individual genes and it allows to build systematic relationships more robust. Based on our analyses we found 48 distinct species from our sampled 24 nominal species, where 14 are new species. Therefore, according to our systematic proposal, the genus Manerebia would comprise 58 nominal species, but for the moment some remain undescribed. The phylogenetic analyses, together with the species delimitation methods and the morphological characters, allowed us to evaluate the high cryptic diversity within the genus. In addition, our analysis highlights the importance of employe the integrative taxonomy framework for the detection of cryptic diversity in regions such as the Neotropics. We generated the first phylogenetic hypothesis for the genus Manerebia based on sequence data from mitochondrial (COI) using phylogenetic tools. Nine clades are proposed for the Manerebia along the Central and Northern Andes being the Northern Andes the zone with the most richness. Our analyses permitted us to clarify some of the phylogenetic relationships within the genus to species-level. Finally, our study explored the biogeographical history of the genus Manerebia estimating times and rates of diversification for its lineages and employing a biogeographical analysis in order to reconstruct its evolutionary history. Our results allowed us to infer that the divergence time of Manerebia was between the late Miocene and Pliocene, and most extant lineages had already appeared in the Pleistocene. The genus had an overall early burst in the late Miocene / early Pliocene boundary followed by deceleration due to a decrease in speciation along to Pleistocene, and this pattern is reflected for all clades in Manerebia. Dispersal events are possibly the most common process within the genus, and our results confirm the role of the Andean geomorphological inthe evolution of Neotropical biodiversity.This thesis was supported by an Internal Research Grant of the Institute of Zoology and Biomedical Research of the Jagiellonian University, BW/IZ/ADD/2005, and by NCN grant Harmonia-10 2018/30/M/NZ8/00293 “Evolutionary biogeography and diversification of the predominantly Andean butterfly subtribe Pronophilina (Nymphalidae, Satyrinae) based on phylogenetic data generated using modern molecular methods”. Molecular analysis was partly carried out in the laboratory of the Nature Education Centre, Jagiellonian University, Kraków, Poland (CEPUJ). Finally, PROM Programme International Scholarship Exchange of Ph.D. Students and Academics. Polish National Agency for Academic Exchange-NAWA. Jagiellonian University, Krakow, Poland. Agreement number: PPI/PRO/2018/1/00001/U/001-2019DoctoradoDoctor en Ciencias - BiologíaBiogeografi

An investigation of Onychophora (velvet worms) of the Illawarra region

Onychophorans are a fascinating phylum of terrestrial invertebrates that form a group with the Arthropoda and Tardigrada known as Panarthropoda (Campbell et al. 2011). Diversity among the Onychophora has been grossly underestimated as their conserved morphology results in the prevalence of cryptic species, whereby two or more species may be grouped under one species name if gross morphology alone has been used as the only means of identification. In recent years, more detailed morphological analyses and the application of molecular analyses has shed more light on diversity among the Onychophora. Within this phylum, two families have been recognised, the Peripatopsidae and Peripatidae, with a lack of any higher taxonomic ranks. The family Peripatopsidae are found in Australia, New Zealand, South Africa, and New Guinea and currently consists of many monotypic genera. One of these, Anoplokaros, with its single species Anoplokaros keerensis Reid, 1996, was originally suggested to comprise a likely cryptic species-complex (Reid 1996). This hypothesis was tested within this study, employing an integrative taxonomic approach, involving both molecular and morphological analyses. It was revealed that three putative novel species are present within the Illawarra region, that were previously believed to be populations of A. keerensis. Unique morphological characters were observed in most of the populations examined, although these were not necessarily congruent with the mtDNA COI sequences. Morphological descriptions were also prepared for additional specimens that were not included in the phylogenetic analyses that could be used as a baseline to facilitate future revisions of these putative A. keerensis populations

Cryptic diversity in the leptothecate genera Laodicea and Tiaropsis (Cnidaria: Hydrozoa)

Postponed access: the file will be accessible after 2022-09-01Although they are an ecologically and economically important group of animals, hydrozoans are understudied because of their difficult identification due to their small size and fragility. Hydrozoans as a group show a great diversity in life strategies, and often include both a polyp and a medusa stage in their life cycle, which complicates their taxonomy because both stages may be needed for a correct identification. For the two leptothecate hydrozoan genera Laodicea (Family Laodiceidae) and Tiaropsis (Family Tiaropsidae), the polyp stage is very small and easy to overlook, while the medusa stage is conspicuous and relatively straightforward to identify, at least to genus level. Only one species of each of these genera is believed to occur in Norwegian waters, Laodicea undulata (Forbes &Goodsir, 1853) and Tiaropsis multicirrata (M. Sars, 1835), but preliminary data suggest that the diversity of these taxa in the region is higher than previously thought. In this study, I used DNA barcoding and different molecular species delimitation methods (based on mitochondrial markers 16S and COI and the nuclear marker ITS) in combination with a detailed morphological analysis of both the hydroid and medusa stages to assess the species diversity of the genera Laodicea and Tiaropsis in Norway. Based on molecular evidence, Laodicea undulatais shown to comprise two molecularly distinct Norwegian clades, which appear not to be sister species. Specimens morphologically identified as T. multicirrata split up into three distinct clades in Norway according to the evaluated molecular markers, indicating cryptic diversity in Tiaropsis for the studied region. For both Tiaropsis and Laodicea, the results suggest that the observed clades correspond to undescribed species, but further work is necessary to place them in a broader phylogenetic perspective and to identify any potential morphological characters that define them.Master's Thesis in BiologyBIO399MAMN-BIOMAMN-HAVS

Its the shape that matters! The diverse world of genitalia: A taxonomic and evolutionary exploration of the neotropical genus Eois Habner (Lepidoptera: Geometridae: Larentiinae)

Eois Hübner (Geometridae: Larentiinae) is a hyperdiverse genus of moths containing 267 valid species, but with estimates of 1,000 or more Neotropical species yet to be described. The paucity of contemporary descriptive and distributional data for these moths not only limits monitoring and conservation efforts of potentially vulnerable populations, but also hinders investigations into ecological and evolutionary factors underlying the diversity of Eois. To begin to remedy shortcomings in our knowledge of this moth group, I conducted research on three different aspects of Eois systematics and evolution.1) I conducted morphometric analysis of cryptic species at a site in the Ecuadoran Andes, evaluating relationships between genetic and genitalic variation of morphologically similar (i.e., cryptic) species in the Eois olivacea clade, and investigating the extent that elevation and host plant associations influence evolutionary patterns across Ecuadorian populations. Based on 170 individuals sampled from different elevations and host plants at a single site within the Ecuadorian Andes, population genetic analyses revealed that samples can be assigned to four distinct taxa, with genetic divergence among taxa associated with different host plants. Morphometric analysis indicated that the adult samples belong to three distinct taxa, and molecular dating analysis implied that these taxa form a monophyletic clade that began diverging approximately five million years ago. 2) I circumscribed and described 16 new species of the Eois olivacea clade based on traditional morphological techniques using specimens from various institutional collections (UNR, AMNH, BMNH, USNM and McGuire Center), employing a data matrix of 107 morphological characters. I defined the clade based on wing pattern and other morphological features, and then provided detailed diagnoses and descriptions of each new species, as well as re-examining the four previously described species in the clade. 3) I analyzed male vs. female features of the genitalia to further our understanding of sexual traits and evolution of genitalia in Eois. I considered different mechanisms of diversification of genitalia and differences among the sexes, including genetic drift, pleiotropy, female choice, cryptic female choice, male to male competition, sexual conflict, and the lock-and-key hypothesis. I also explored variation of female versus male traits. Using the morphological data matrix mentioned above, I developed a phylogeny for a sample of 99 species (94 Eois and five outgroup taxa), using maximum parsimony and maximum likelihood methods, and separate dendrograms based on male-only and female-only characters. An examination among trees revealed discordance between dendrograms based on male-only and female-only traits, suggesting at least partially independent evolution of traits between the sexes. The Neotropical moth genus Eois is a remarkable group, and includes dazzling species with complex ecologies and fascinating evolutionary patterns. Nevertheless, understanding that diversity has been challenging, and if estimates of the species richness are correct, the genus is among the most species-rich in all of Lepidoptera. The results presented here provide a starting point in undertaking the challenging endeavor of describing the hundreds of new species of Eois. The results also represent a foundation for investigating sexual trait evolution in the genus, largely because knowledge of functional morphology of Eois genitalia is limited, as is information on mating interactions, mating costs and benefits, the physical interaction of male and female genital structures, and rates of evolutionary divergence of these animals