Dissertations / Theses on the topic 'Molecular species delimitation'

The plant family Gesneriaceae is represented in Sri Lanka by six genera: Aeschynanthus, Epithema, Championia, Henckelia, Rhynchoglossum and Rhynchotechum, with 13 species (plus one subspecies/variety) of which ten are endemic including the monotypic genus Championia, according to the last revision in 1981. They are exclusively distributed in undisturbed habitats, and some have high ornamental value. The species are morphologically diverse, but face a problem of taxonomic delineation, which is further complicated by the presence of putative hybrids. Sri Lanka and Indian Peninsula, represent the Deccan plate of the ancient Gondwanan supercontinent. The presence of a relict flora may indicate the significance of the geological history of the Deccan plate for the evolution of angiosperms. The high degree of endemism here, along with their affinities to the global angiosperm flora paints a complex picture, but its biogeographic history is still unclear. The pantropical family Gesneriaceae distributed in Sri Lanka and South India is therefore an appropriate study group in this context. Besides, the family itself has a complex but largely unresolved biogeographical history especially concerning the origin and diversification of Old World Gesneriaceae. Modern approaches for the taxonomic studies were applied, integrating morphological and molecular data. Multiple samples were collected for each species across their geographical distribution. Nuclear ITS and chloroplast trnL-F sequences for the taxa from Sri Lanka were used to generate regional genus phylogenies of all six genera, using maximum parsimony. The rate of evolution of the nuclear ITS region versus chloroplast trnL-F was varied greatly across the six genera studied. Molecular delimitations were mostly congruent with the classical taxonomy. Over 65 taxonomic characters were studied in detail to recognize synapomorphies for clades and taxa. A complete taxonomic revision of Gesneriaceae in Sri Lanka, including lectotypification, was conducted based on both, the molecular and morphological data. This resulted in the recognition of 14 species in the six genera, including one newly described species H. wijesundarae Ranasinghe and Mich. Möller. Henckelia communis and H. angusta were not supported molecularly as two separate entities but are recognized as two species because of consistent morphological differences between them. Henckelia humboldtiana is proposed to represent a species complex due to its highly variable and inconsistent molecular and morphological diversity and overlap with H. incana and H. floccosa; more research is needed here. National conservation assessments were conducted, and all 14 species were recognized as threatened. Biogeographic affinities of Sri Lankan Gesneriaceae were elucidated, generating a dated phylogeny using an existing matrix of four plastid gene regions; trnL-F, matK, rps16 and ndhF, amended by sequences generated in this study. The final combined matrix included 175 taxa including newly generated sequences for the 13 Sri Lankan taxa. Phylogenetic trees were generated using parsimony, maximum likelihood and Bayesian inference. Molecular dating was carried out using BEAST and ancestral area reconstruction using BioGeoBears. These analyses indicated that the six genera of Gesneriaceae arrived in Sri Lanka separately and sometimes different time periods. One lineage dated back to the early diversification of the subfamily Didymocarpoideae (generally regarded as the Old World Gesneriaceae), which occurred around the KT boundary, before the Deccan plate was connected to Asia.

Les écosystèmes des eaux souterraines sont de plus en plus reconnus pour leur faune endémique, phylogénétiquement ancienne et écologiquement spécialisée. Avec plus de 425 espèces décrites, les amphipodes niphargidés constituent la famille des eaux souterraines la plus riche en espèces au monde et un système modèle intéressant pour la biologie de l'évolution. Cependant, les scientifiques doivent faire face à des données incomplètes et biaisées en raison de trois déficits majeurs: le déficit Linnéen pour la taxonomie, le déficit Darwinien pour la phylogénie, et le déficit Wallacien pour la biogéographie. La présente thèse vise à évaluer l'importance de ces déficits chez les niphargidés, ouvrant ainsi la voie pour y remédier. Le premier chapitre est une évaluation des effets de la découverte d'espèces cryptiques (une des causes du déficit Linnéen) sur notre compréhension des modèles de distribution à grande échelle de la diversité des niphargidés. Contrairement à ce que l'on attendait, les espèces cryptiques putatives sont réparties de manière homogène le long des gradients environnementaux, et leur découverte ne modifie donc pas notre compréhension des modèles de distribution. Le deuxième chapitre analyse l'importance de l'application des techniques moléculaires à la taxonomie des niphargidés. En étudiant le genre Microniphargus, la morphologie seule s'est avérée peu informative en raison de la pédomorphose et de l'homoplasie. L'utilisation de marqueurs ADN a permis d'attribuer le genre à une famille différente (Pseudoniphargidae), venant éclaircir les relations phylogénétiques au sein des Niphargidae (et contribuant ainsi à remédier au déficit Darwinien). Le troisième chapitre traite du rôle des régressions et transgressions marines sur la distribution des niphargidés en utilisant la biogéographie moléculaire et une modélisation biogéographique innovante (afin de remédier au déficit Wallacien). Les résultats soutiennent l'idée que la dispersion a joué un rôle essentiel dans la biogéographie historique des niphargidés, en montrant que leurs voies de dispersion sont corrélées à des événements paléogéographiques anciens. Enfin, le quatrième chapitre traite de la taxonomie, de la phylogénie et de la biogéographie d'un clade de niphargidés distribué dans la région des Alpes et des Carpates, et illustre un cas de discordance mitonucléaire dans la délimitation d'espèces vivant dans des zones affectées par les glaciations quaternaires. Une histoire complexe de divergence de lignées évolutives et de contacts secondaires pendant les fluctuations climatiques du Pléistocène explique la plus grande variabilité de l'ADN mitochondrial par rapport aux marqueurs nucléaires. Dans une telle situation, la description formelle d'espèces cryptiques basée sur le seul barcodage de l'ADN mitochondrial, comme dans certains articles récents sur les niphargidés, n'est pas recommandée. Cette thèse ouvre plusieurs perspectives pour des recherches futures basées sur la taxonomie intégrative et la modélisation biogéographique, permettant aux niphargidés très diversifiés de jouer un rôle majeur dans la surveillance des écosystèmes des eaux souterraines.
Doctorat en Sciences
info:eu-repo/semantics/nonPublished

Accurate species delimitation has critical implications for ecological and conservation studies; and for understanding factors driving diversification. However, a growing body of evidence indicates that morphology-based species circumspection in lichenized ascomycetes often fails to accurately represent the number of fungal species. The use of molecular data in lichen systematics provides an important alternative to traditional morphological characters for identifying natural groups and assessing evolutionary histories in challenging lichen taxa. In this work, I examined two common lichen-forming genera in western North America, Rhizoplaca and Xanthoparmelia, as models for investigating character evolution, species delimitation in morphologically and chemically diverse species, and identification of lineages in the early stages of divergence. Phylogenetic hypotheses were reconstructed to assess character evolution using sequence data from four nuclear ribosomal markers and fragments from two nuclear loci. I applied a multifaceted approach to delimit species in Rhizoplaca and Xanthoparmelia by assembling multiple lines of evidence using DNA sequence data, and genealogical and population genetic analyses. I have found that traditionally circumscribed species are not supported by molecular data. For example, in Rhizoplaca previously unrecognized lineages were identified within what has thus far been considered a single species. In contrast, morphologically and chemically distinct species within Xanthoparmelia were not supported by molecular data. Distinct medullary chemistries, growth forms, and the production of vegetative diaspores appear to have evolved independently multiple times in Xanthoparmelia. This work clearly indicates that morphological and chemical characters do not always accurately reflect lichen species diversity within even the best known and studied genera. My study of the Rhizoplaca melanophthalma species complex demonstrates that the genus Rhizoplaca, as presently circumscribed, is more diverse in western North American than previously thought. I present these analyses as a working example of species delimitation in morphologically cryptic lichenized fungi. In Xanthoparmelia diagnostic morphological and chemical characters have evolved in a highly homoplasious manner. In contrast to other studies documenting previously undiscovered fungal lineages masked within lichen species circumscribed by traditional morphological and chemical characters, my work suggests that species diversity has been overestimated in the lichen genus Xanthoparmelia.

China harbors 10% of the world's salamander species. Studying their evolutionary history provides critical insights into the evolution of the fauna of the Far East. The stout newts (Pachytriton, also known as paddle-tailed newts) are a genus of aquatic montane salamanders that are widely distributed in southeastern China. Despite their longstanding popularity among the global pet trade, little is known of their biology beyond external morphology. My thesis presents the first systematic study to elucidate phylogenetic relationships, character evolution, biogeographic patterns, species delimitation, and speciation mechanisms in this genus.

Molecular taxonomy is the science that classifies and identifies organism basing on molecular information. Its application to species-rich groups of organisms, as insects, could improve and accelerate taxa identification and delimitation. The efficiency and the limits of the methods for molecular taxonomy have to be properly evaluated for achieving unbiased results with this taxonomic approach. The main aims of this thesis are: i) to develop COI barcode libraries for Euro-Mediterranean Chrysomelidae identification and test DNA-barcoding efficiency in the identification of the species of the family; ii) to estimate group-specific nucleotide distance thresholds for the molecular identification of Chrysomelidae taxa and compare the error related to their use with the error related to the use of a general threshold value estimated for the whole family; iii) to apply molecular species delimitation methods to other insects taxa with the aim of resolving their taxonomic status in an integrative taxonomy framework; vi) to test the influence of factors intrinsic to the analysed data on molecular species delimitation efficiency. The results obtained confirmed the high efficiency of DNA-Barcoding as tool for Chrysomelidae molecular identification (94-99% efficiency) and the usefulness of molecular delimitation methods in integrative taxonomy for resolving taxonomic debated issues. The use of groups specific thresholds resulted to be related to a significantly lower identification error than the use of a general threshold for the whole Chrysomelidae, suggesting how the use of fixed thresholds for taxa identification is unwise. Finally, some factors, i.e. the species delimitation methods used, the geographic intraspecific distance among individuals collection localities and the difficulties in morphological identification of species, resulted to affect the efficiency of molecular species delimitation.

Orientador: Antonia Cecilia Zacagnini Amaral
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Espécies são unidades fundamentais da biologia e sua identificação é essencial para a pesquisa nos mais diversos campos. Esta tarefa, no entanto, é dificultada por limites interespecíficos naturalmente mal definidos, especialmente em ambientes marinhos, onde complexos de espécies crípticas são comuns. Assim, a delimitação de espécies tem recebido grande atenção nos últimos anos e técnicas moleculares têm se mostrado de grande importância para a questão. Corbula (Bivalvia: Corbulidae) é um gênero frequente e ecologicamente importante em comunidades bentônicas marinhas de sublitoral. A taxonomia do grupo é bastante confusa, em parte devido à plasticidade fenotípica das conchas que dificulta o estabelecimento de limites morfológicos entre as espécies. O presente estudo teve como objetivo estudar, com base em sequências de dois marcadores moleculares (COI e 16S), os limites entre seis espécies de Corbula morfologicamente identificadas da costa sudeste e sul do Brasil, de forma a testar a delimitação morfológica. Como se trata de espécies predominantemente de sublitoral, o material analisado encontrava-se preservado em álcool, havendo sido fixado em formol. Dessa forma, fez-se necessário o desenvolvimento de protocolos específicos de extração e amplificação. Uma combinação de extração orgânica com adsorção em sílica mostrou-se o melhor método de extração de DNA total. Para as reações de amplificação, a utilização de nested PCR produziu resultados superiores à PCR direta. As análises de delimitação utilizaram quatro métodos diferentes, dois baseados em árvores (GMYC e Brownie) e dois não (regra das 4x e ABGD). Os resultados divergiram entre métodos e marcadores, mas a combinação das diferentes linhas de evidência permitiu corroborar a delimitação morfológica de três espécies (Corbula caribaea, Corbula tryoni e Corbula lyoni). Os indivíduos identificados como Corbula patagonica dividiram-se em duas espécies distintas. O único indivíduo de Corbula aequivalvis foi considerado distinto das outras espécies e um indivíduo atribuído a Corbula sp1 não pôde ser distinguido de C. caribaea
Abstract: Species are fundamental unities in many biological studies and, being so, their identification is essential for researches in many different fields. This task, however, is complicated by badly defined interspecies boundaries, especially in the sea, where cryptic species are quite common. Species delimitation has been receiving much attention, and molecular techniques have been proved of great value to the matter. Corbula (Bivalvia, Corbulidae) is frequent and ecologically important genus in benthic marine communities. Nevertheless, its taxonomy is confusing, in part due to a plastic shell, which makes it difficult to establish species boundaries. This study aimed to analyze the COI and 16S sequences of six morphologically identified Corbula species occurring off the South-Southeastern Brazilian coast. Being a mainly sublittoral genus, most of the analyzed material had been previously sampled, fixed in formalin and preserved in alcohol. Hence, initially specific protocols for DNA extraction and PCR were developed. Better results were obtained with an extraction protocol combining organic extraction and silica adsorption. The nested PCR yielded more product than the direct PCR. Delimitation analyses were conducted with four different methods: two tree based (GMYC and Brownie) and two non-tree based (4x rule and ABGD). Different methods and markers produced different delimitations, but the combined evidence supports the morphological delimitation of three species: Corbula caribaea, Corbula tryoni and Corbula lyoni. Individuals assigned to Corbula patagonica were separated into two molecular species. Only one individual of Corbula aequivalvis was analyzed and it was distinguished from other species. One individual assigned to Corbula sp1 could not be distinguish from C. caribaea
Doutorado
Ecologia
Doutora em Ecologia

Understanding the processes that generate diversity is key to interpreting the patterns we see in the present. New developments in modelling these processes have promised unprecedented prospects for unravelling the evolutionary past. However, the empirical behaviour of these models in many of their practical applications is not well understood. This thesis investigates the influence of diversification models in a variety of contexts. First, I consider the Generalised Mixed Yule-Coalescent (GMYC) method for molecular species delimitation. This method identifies transition points between species- and population-level diversification processes on a time-resolved evolutionary tree. I show that this method is sensitive to the choice of mitochondrial marker used, and that the best marker can vary widely across study groups. Next, I investigate the influence of diversification models used to place prior distributions on time-resolved trees in molecular dating. Specifically, I look at the influence of the tree prior in analysing data sets with multiple individuals per species. These data sets can arise by accident where species boundaries are not well understood, and violate the assumptions of both population- and species-level tree priors. I use simulation to show that molecular date estimates can be seriously affected by the choice of tree prior in some circumstances, but are remarkably robust in general. Finally, I extend the analysis of tree prior sensitivity to new methods for dating the origins of human language families. I show that these methods are also robust to the choice of tree prior, and that speciation priors are preferred for language data sets regardless of taxonomic scale. My work will contribute to an improved understanding of the role of diversification models in empirical studies and will increase confidence in these methods across multiple realms of enquiry.

Orientadores: Karina Lucas da Silva Brandão, André Victor Lucci Freitas
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: O gênero Hermeuptychia Forster (Nymphalidae, Satyrinae, Euptychiina) está amplamente distribuído no continente Americano, desde a Argentina até o sul dos Estados Unidos. O gênero foi anteriormente considerado um complexo de espécies, e atualmente são reconhecidas oito espécies . Todas as espécies possuem um padrão alar muito parecido, o que compromete a identificação taxonômica correta. Em adição, a posição filogenética do gênero dentro da subtribo Euptychiina permanece incerta. Para o presente estudo foram obtidos espécimens de 45 localidades de cinco países, com maior ênfase em uma amostragem no Brasil. Três marcadores moleculares, dois do DNA mitocondrial (cox1 5' e nad6) e um do DNA nuclear (RpS5) foram utilizados para gerar hipóteses filogenéticas (Máxima Parcimônia e Inferência Bayesiana), para delimitar espécies, e para gerar estimativas de tempo de divergência e distribuição ancestral. Adicionalmente, o desempenho da região anterior da cox1 como 'barcode - código de barras' para delimitar as espécies de Hermeuptychia foi testado. Além disso, análise morfológica da genitália masculina foi empregada para a delimitação e identificação de espécies. Os indivíduos amostrados agruparam-se em dez clados nas análises moleculares, correspondendo a sete espécies reconhecidas mais H. gisella, que havia sido anteriormente sinonimizada com H. cucullina. A análise morfológica dos indivíduos possibilitou o estabelecimento de caracteres diagnósticos para todas as espécies de Hermeuptychia - incluindo H. cucullina, que não está presente nas análises moleculares - e concordou com os agrupamentos obtidos através das análises moleculares. As relações filogenéticas entre as espécies de Hermeuptychia permanecem incertas, possivelmente devido a um padrão de evolução rápida, descrito anteriormente para outros Satyrini. Entretanto, dois grupos de espécies-irmãs podem ser identificados, H. pimpla + H. harmonia, e H. gisella + H. fallax, ambos sustentados por ocorrência simpátrica. Em adição, H. gisella e H. fallax parecem apresentar um isolamento reprodutivo incompleto, com formação de híbridos. Algumas espécies de Hermeuptychia estão distribuídas amplamente, como H. atalanta, H. hermes e H. gisella, sendo que H. atalanta é a espécie mais comum encontrada no Brasil. H. fallax é uma espécie restrita a Mata Atlântica; H. pimpla e H. harmonia são espécies restritas à região andina, encontradas em altitudes moderadas; H. maimoune pode ser encontrada na região andina e no sul da Amazônia brasileira, correspondendo a duas espécies crípticas; H. cucullina foi encontrada no centro-oeste brasileiro e na região andina, e é a espécie mais rara de Hermeuptychia; e H. sosybius pode ser encontrada do sul dos Estados Unidos até a região norte da Colômbia. O gênero diversificou-se de seu grupo-irmão a cerca de 8,2 milhões de anos (mya), a diversificação das espécies ocorreu entre 3,5 e 1,4 mya, e a distribuição ancestral estimada é a cordilheira dos Andes. Apenas com a região 'barcode' e análise de distância usando Neighbor-Joining, foi possível separar as espécies de Hermeuptychia com uma taxa de 2% de erro. O limite entre as distâncias intra e interespecíficas estimado fica em torno de 2% de divergência genética
Abstract: The Hermeuptychia genus Forster (Nymphalidae: Satyrinae: Euptychiina) is widely distributed in the American continent, from Argentina to South United States. Previously considered a species complex, the genus presents eight valid species taxa at the moment. Wing pattern is very similar in all Hermeuptychia species resulting in difficult and prone to error taxonomic identification. Additionally its position within the subtribe Euptichiina remains uncertain. Samples from 45 locations in five countries, with major emphasis in Brazilian territory sampling, were obtained for the present study. Three molecular markers, two from mitochondrial DNA (cox1 5' and nad6) and one from nuclear DNA (RpS5), were used to generate phylogenetic hypothesis (Maximum Parsimony and Bayesian Inference), to delimit species, and to estimate divergence time and ancestral distributions. The 'barcode' region performance (cox1 5') was tested for Hermeuptychia species. Male genitalia morphology was also used to identify and delimitate species. Sampled individuals are grouped in ten molecular clusters, corresponding to seven valid species and H. gisella, previously synonymized to H. cucullina. Morphological analysis of individuals revealed morphological diagnose traits to identify all Hermeuptychia species, including H. cucullina, which is not present in the molecular analysis, and was congruent with molecular analysis. Phylogenetic relationships among Hermeuptychia species remain unresolved due to a possible rapid evolutionary pattern common to Satyrini. However, two pairs of sister species could be identified: H. pimpla + H. harmonia and H. gisella + H. fallax, both sympatric. Additionally, H. gisella + H. fallax present incomplete reproductive isolation, with hybrids. Some Hermeuptychia are widely distributed as H. atalanta, H. hermes, and H. gisella, and H. atalanta is the most common species found in Brazil. H. fallax is restricted to Atlantic Forest; H. pimpla and H. harmonia are restricted to Andes region, at moderately high altitudes; H. maimoune is found in the Andine and in the Amazonian regions, corresponding to two cryptic species; H. cucullina is the rarest Hermeuptychia and was found in Central Brazil and in Andes; and H. sosybius is the only Hermeuptychia found in North America, been present from South United States to north Colombia. The genus diverged from its sister group around 8.2 my, species diversification occurred between 3.5 and 1.4 my and the ancestral estimate distribution is Andine region. Only the 'barcode' region was able to identify each Hermeuptychia species, with an 2% error rate and the molecular threshold for intra and interspecific distance was around 2% of genetic divergence
Mestrado
Ecologia
Mestre em Ecologia

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Financiadora de Estudos e Projetos
The family Threskiornithidae includes 13 genera and 32 species, but the relationships among genera, species or subspecies have been little studied. This family is traditionally divided into two subfamilies: Plataleinae and Threskiornithinae. One of the more interesting taxonomical questions within this group is the case of the species Eudocimus ruber and Eudocimus albus. They are usually considered as separate species, but they show similar behavior and there are also records of hybridization in nature. This study aims to reconstruct the phylogenetic relationships within the family Threskiornithidae as well as assess the level of genetic differentiation between Eudocimus albus and E. rubber, using mitochondrial and nuclear gene sequences. DNA was extracted from blood and tissue samples from 13 species of Threskiornithidae (seven genera) and two outgroups. For the Eudocimus study were extracted 10 individuals of each species. We sequenced the 16S rRNA and the intron 7 of β- Fibrinogen for all species. For Eudocimus Cytochrome B, Cytochrome Oxidase I, intron 11 of Glyceraldehyde-3-Phosphate Desidrogenase, intron 4 of the Myelin Proteolipid Protein and intron 2 of Myoglobin were also sequenced. Sequences for other five species of the family were obtained from GenBank. Phylogenetic trees were constructed using Neighbor-Joining, Maximum Parsimony, Maximum Likelihood, Bayesian inference and Bayesian inference of species tree. Networks and genetic distances were determined for Eudocimus haplotypes. Several approaches for species delimitation using multilocus data were applied. All analyses strongly supported the family Threskiornithidae (18 species) as a monophyletic group. However, the current classification of two subfamilies was not supported by our data: Plataleinae formed a monophyletic group, but nested within Threskiornithinae (a paraphyletic group). Tests of monophyly rejected the hypothesis of monophyly of Threskiornithinae. The family Threskiornithidae also showed a division into two groups: one with only genera endemic to the American continent (Theristicus and Eudocimus) and another with the remaining species. Within the latter clade, species of genus Plegadis are observed in a basal position, while subfamily Plataleinae was grouped with the remaining species. This pattern of species distribution suggests an initial Gondwana division and subsequent colonization by species from the Old to the New World. The divergence within the family was estimated at 35-40 million years, which is before the separation between America and Antarctica. Mitochondrial genetic analysis showed Eudocimus species as two different lineages. Multilocus analysis based on nuclear genes revealed a strong signal of speciation despite the polyphyly found in three of the four markers.
A família Threskiornithidae inclui 13 gêneros e 32 espécies e suas relações interespecíficas, assim como as designações dos gêneros, espécies ou subespécies foram pouco estudadas. A família tem sido dividida em duas subfamílias: Plataleinae e Threskiornithinae. O caso de Eudocimus ruber e Eudocimus albus é uma das questões interessantes da taxonomia do grupo, pois têm sido consideradas como espécies, mas mostram similaridades no comportamento e há registros de hibridização na natureza. Os objetivos do presente estudo foram reconstruir as relações filogenéticas dentro da família Threskiornithidae e avaliar o nível de diferenciação genética entre Eudocimus albus e E. ruber, baseando-se nos dados de sequências de genes mitocondriais e nucleares. DNA foi extraído de amostras de sangue e de tecidos de 13 espécies de Threskiornithidae, representantes de sete gêneros da família e de dois grupos externos. Para o estudo do caso de Eudocimus foram analisadas amostras de 10 indivíduos de cada espécie. Foram sequenciados os genes 16S rRNA e o íntron 7 do β-fibrinogênio para todas as espécies. Nos indivíduos de Eudocimus foram sequenciados também os genes Citocromo B, Citocromo Oxidase I, o íntron 11 da Gliceraldeído-3-Fosfato Desidrogenase, o íntron 4 da Proteína Proteolipídica da Mielina e o íntron 2 da Mioglobina. Sequências para outras cinco espécies da família foram obtidas do GenBank. Árvores filogenéticas foram construídas pelos métodos de inferência de Neighbor-Joining, Máxima Parcimônia, Máxima Verossimilhança, Análise Bayesiana e Estimativa Bayesiana de árvore de espécies. Redes de haplótipos e distâncias genéticas foram determinadas para as sequências de Eudocimus. Diversas abordagens de delimitação de espécies usando informação multilocus foram realizadas. A família Threskiornithidae com 18 espécies se apresentou como grupo monofilético fortemente sustentado em todas as análises. A classificação atual das duas subfamílias não foi corroborada: Plataleinae se apresentou como grupo monofilético, mas agrupada dentro dos Threskiornithinae, sendo este último um grupo parafilético. Os testes de monofilia rejeitaram a hipótese de Threskiornithinae ser um grupo monofilético. A família Threskiornithidae pode ser dividida em dois grupos: o primeiro agrupando somente gêneros endêmicos do continente americano (Theristicus e Eudocimus) e o outro com as demais espécies. Dentro deste ultimo clado, observa-se em posição basal as espécies do gênero Plegadis e a subfamília Plataleinae agrupada com o restante das espécies. Este padrão de distribuição de espécies concorda com uma divisão inicial Gondwânica e uma posterior colonização por espécies do velho ao novo mundo. A divergência da família foi estimada em 35 40 milhões de anos, data anterior à separação do continente Americano da Antártica. As análises genéticas mitocondriais mostraram as espécies de Eudocimus como duas linhagens diferentes. Nas análises multilocus baseadas nos genes nucleares foi possível recuperar um forte sinal de especiação apesar da polifilia encontrada em três dos quatro marcadores.

A delimitação de espécies é um aspecto de fundamental importância dentro da Biologia Evolutiva, bem como para a conservação da biodiversidade. No entanto, delimitar espécies com base em morfologia é extremamente complicado, especialmente em grupos que tiveram uma radiação recente e que apresentam pouca descontinuidade morfológica entre os táxons, como a tribo Andropogoneae (Poaceae). A presente tese utiliza sequências de DNA, como genes nucleares de cópia-única (low-copy nuclear loci) e sequenciamento completo do plastoma, para resolver a circunscrição de complexos de espécies em Andropogoneae, particularmente dos gêneros Saccharum e Eriochrysis, bem como para investigar o posicionamento filogenético dos mesmos em relação aos demais gêneros da tribo. Aspectos filogenéticos, taxonômicos e nomenclaturais foram investigados. Do ponto de vista nomenclatural, foi possível esclarecer que Andropogoneae Dumortier é o nome correto para a tribo, e não Sacchareae Martinov, como recentemente sugerido por diversos autores. As análises filogenéticas realizadas corroboram a hipótese de uma diversificação inicial rápida em Andropogoneae e a não monofilia da subtribo Saccharinae. A origem alopoliploide do gênero Saccharum foi demonstrada a partir de evidências de genes nucleares. Saccharum s.l. é polifilético e Tripidium deve ser reconhecido como um gênero distinto. As análises filogenéticas também foram capazes de resolver a circunscrição interna de Saccharum s.l., confirmando a ocorrência de três espécies nativas na América do Sul: S. angustifolium, S. asperum e S. villosum. A ocorrência de híbridos naturais entre S. villosum e S. angustifolium foi documentada. As análises filogenéticas de Eriochrysis confirmaram a monofilia do gênero e resolveram a circunscrição de suas espécies: E. villosa é um táxon distinto de E. cayennensis, bem como E. laxa é uma espécie distinta de E. warmingiana. Híbridos naturais entre E. laxa e E. villosa também foram documentados. Eriochrysis villosa é citada pela primeira vez para o Uruguai e E. laxa para o estado do Rio Grande do Sul. A presente tese demonstrou a eficiência dos genes nucleares de cópia única na delimitação de espécies e gêneros da tribo Andropogoneae, mesmo na presença de poliploidia, evolução reticulada e radiação recente. O sequenciamento completo do plastoma também se mostrou uma ferramenta extremamente promissora para inferências filogenéticas em Andropogoneae.
Species delimitation is a vital issue concerning evolutionary biology and conservation of biodiversity. However, delimiting species based on morphology is a difficult task especially in plant groups with an evolutionary history involving rapid radiation and little morphological discontinuity between taxa, as the tribe Andropogoneae (Poaceae). The present thesis uses DNA sequences, such as low-copy nuclear genes and complete plastome sequencing, to resolve the taxonomic circumscriptions of species complexes in Andropogoneae, particularly from genera Saccharum and Eriochrysis, and to investigate their phylogenetic affinities to other genera of the tribe. Phylogenetic, taxonomic, and nomenclatural aspects were investigated. We clarified that Andropogoneae Dumortier is the correct name for the tribe, rather than Sacchareae Martinov, as recently suggested by several authors. The present phylogenetic analyses support the hypothesis of an initial rapid diversification in Andropogoneae and the non-monophyly of subtribe Saccharinae. The allopolyploid origin of Saccharum was demonstrated using evidence from nuclear genes. Saccharum s.l. is polyphyletic and Tripidium should be recognized as a distinct genus. The phylogenetic analyses were also able to define the circumscriptions of the species of Saccharum s.l., confirming the occurrence of three native species in South America: S. angustifolium, S. asperum and S. villosum. The occurrence of natural hybrids between S. villosum and S. angustifolium was documented. The phylogenetic analyses of Eriochrysis confirmed the monophyly of the genus and resolved the circumscriptions of its species: E. villosa is distinct from E. cayennensis, and E. laxa is distinct from E. warmingiana. Natural hybrids between E. laxa and E. villosa were also documented. Eriochrysis villosa is reported here for the first time for Uruguay and E. laxa for the State of Rio Grande do Sul. The present thesis has demonstrated the efficiency of low-copy nuclear genes in the delimitation of species and genera from tribe Andropogoneae, even in presence of polyploidy, reticulate evolution and recent radiation. The complete plastome sequencing is also a promising tool for phylogenetic inferences in Andropogoneae.

Genomes provide windows into the evolutionary histories of species. The recent accessibility of genome-scale data in non-model organisms and the proliferation of powerful statistical models are now providing unprecedented opportunities to uncover evolutionary relationships and to test hypotheses about the processes that generate and maintain biodiversity. This dissertation work reveals shallow-scale species boundaries and population genetic structure in two imperiled groups of salamanders and demonstrates that the number and information content of genomic regions used in species delimitation exert strong effects on the resulting inferences. Genome scans are employed to test hypotheses about the mechanisms of genetic sex determination in cryptobranchid salamanders, suggesting a conserved system of female heterogamety in this group. At much deeper scales, phylogenetic analyses of hundreds of protein-coding genes across all major amphibian lineages are employed to reveal the backbone topology and evolutionary timescales of the amphibian tree of life, suggesting a new set of hypotheses for relationships among extant amphibians. Yet, genomic data on their own are no panacea for the thorniest questions in evolutionary biology, and this work also demonstrates the power of a model testing framework to dissect support for different phylogenetic and population genetic hypotheses across different regions of the genome.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
The Cryptonanus genus, in Didelphidae family, is currently constituted by five recognized species restricted to the open vegetation areas of South America: C. agricolai, C. chacoensis, C. guahybae, C. ignitus e C. unduaviensis. Morphological analyses on specimens from Cerrado, Caatinga and Pantanal, indicate the need of taxonomic revision for this group, especially considering its wide range of phenotypic variation and geographic distribution. This study aims to contribute in Cryptonanus species delimitation by evaluating possible patterns of intra and interspecific genetic variation, by the analysis of the phylogenetic relationships from the genetic lineages and their geographic distribution. Combined analysis using mtDNA potentially informative sequences, from cytochrome b (cytb) and subunit I of cytochrome oxidase c (coI) genes, were performed for 51 Cryptonanus specimens, in addition to independent analysis of each marker, 64 specimens for cytb and 54 for coI, sampling a total of 42 localities in South America. Genetic distances were estimated based on a DNA barcode approach. Phylogenetic relationships were retrieved from the results of Maximum Parsimony, Maximum Likelihood and Bayesian Inference methods. The topologies recovered were congruent relative to the position of the genus Cryptonanus as a monophyletic cluster, subdivided in nine clades. Regarding species delimitation, two evolutionary hypotheses were tested: 1- the maintenance of four from five valid taxa (except C. ignitus, since this species was not included in the analyses), based on the characterization of intraspecific genetic variation ranged from 0 to 5,3% (cytb) and from 0 to 5,1% (coI), while interspecific variation ranging from 4,1 to 13,2% (cytb) and from 4,7 to 9,6% (coI), related with genetically structured populations in C. agricolai and C. chacoensis clusters; 2- extended biological diversity, related with the identification of five new species besides the five currently recognized. On this scenario, intraspecific genetic variation ranges from 0 to 2,4% (cytb) and from 0 to 3% (coI), while interespecific variation ranges from 2,1 to 13,2% for cytb and 1,6 to 9,6% for coI. The hypothesis 2 is argued to provide a better picture for species delimitation in Cryptonanus, based on the congruence between geographic structure, intra and interspecific genetic variation ranges and the recovery of reciprocal monophyletic genetic lineages in every phylogenetic analyses.
O gênero Cryptonanus, pertencente à família Didelphidae, é composto por cinco espécies válidas, restritas às formações abertas da América do Sul: C. agricolai, C. chacoensis, C. guahybae, C. ignitus e C. unduaviensis. De acordo com análises morfológicas de espécimes do Cerrado, Caatinga e Pantanal, é necessária uma revisão taxonômica do grupo, considerando todo o espectro de variação morfológica encontrada e a amplitude da distribuição geográfica. Este estudo tem como objetivo auxiliar na delimitação das espécies de Cryptonanus a partir da avaliação de possíveis padrões de distribuição da variação genética intra e interespecífica, da análise das relações filogenéticas obtidas entre as diferentes linhagens genéticas e da distribuição geográfica destas linhagens. Devido ao seu potencial informativo em estudos de delimitação taxonômica, foram realizadas análises combinadas de sequências dos genes citocromo b (cytb, 781 pb) e subunidade I da citocromo oxidase c (coI, 743 pb) do DNA mitocondrial para um conjunto de 51 espécimes de Cryptonanus, além de análises em separado para cada marcador, 64 espécimes para o cytb e 54 para o coI, amostrando 42 localidades da America do Sul. As estimativas de distância genética foram realizadas em análises utilizando a estratégia de DNA barcode. As relações filogenéticas foram analisadas com base nas inferências resultantes dos métodos de Máxima Parcimônia, Máxima Verossimilhança e Inferência Bayesiana. As topologias obtidas apresentaram resultados congruentes em relação ao posicionamento do gênero Cryptonanus em um agrupamento monofilético, o qual foi subdividido em nove clados. Em relação à delimitação de espécies, foram investigadas duas hipóteses evolutivas: 1- a manutenção de quatro dos cinco táxons válidos (excluindo-se C. ignitus, que não integra este estudo), a partir da caracterização de intervalos de variação genética intraespecífica entre 0 a 5,3% (cytb) e entre 0 a 5,1% (coI) e de variação interespecífica entre 4,1 a 13,2% (cytb) e 4,7 a 9,6% (coI), com estruturação genética nas populações de C. agricolai e C. chacoensis; 2- ampliação da diversidade do gênero com a identificação de cinco novas espécies, além das cinco espécies atualmente válidas, caracterizado por valores de variação intraespecífica entre 0 a 2,4% (cytb) e entre 0 a 3% (coI) e valores de variação interespecífica nos intervalos de 2,1 a 13,2% para cytb e 1,6 a 9,6% para coI. A hipótese 2 é defendida como sendo a condição que melhor representa a delimitação das espécies de Cryptonanus com base na congruência entre a estruturação geográfica, os intervalos de variação genética intra e interespecíficos válidos para outras espécies e a recuperação de linhagens genéticas reciprocamente monofiléticas em todas as análises filogenéticas.

Emplectonema gracile (Johnston 1837) is a hoplonemertean of marine intertidal hard-bottom communities and is distributed throughout the Northern Hemisphere. Although possessing a planktonic larval stage in its life history, the range of such cosmopolitan marine invertebrate species is often explained by cryptic speciation and anthropogenic transport. The purpose of this study is to test for possible cryptic species using mtDNA markers (COI and 16S rDNA) and to investigate population structure in E. gracile over a portion of its geographic range using mtDNA markers and ddRADseq nuclear SNP data. The results of both phylogenetic- and tree-based species delimitation revealed that E. gracileis a morphotype containing cryptic species. Three North Atlantic and one Pacific coast population are inferred as one species (E. gracile sensu stricto) and two Pacific coast populations (Akkeshi, Japan and Charleston, Oregon) are inferred as another species (Emplectonemasp 1), strongly confirming an earlier study and extending the range of the latter species to the Pacific coast of Japan. Anthropogenic transport is suggested as the likely mode of transport for E. gracile.Both Fst, PCA and haplotype network analyses suggest a lack of differentiation between E. gracile populations separated by large geographic distances.In contrast corresponding analyses forEmplectonemasp. 1 indicate differentiation between the two populations sampled. Further research will be necessary to reveal if rare anthropogenic transport or natural dispersal (larval transport, rafting) between geographically adjacent yet to be delimitedE. gracile morphotype populations is responsible for its seemingly disjunct distribution.

Os camarões do gênero Atya Leach, 1816 são os maiores camarões da família Atyidae, sendo que as 13 espécies reconhecidas estão distribuídas em rios e riachos das regiões tropicais e subtropicais da América (vertentes atlântica e pacífica) e oeste da África. O primeiro relato de uma Atya ocorreu no séc. XVII e, desde então, novas espécies foram descritas e descrições prévias revisadas, produzindo um histórico de instabilidade e reclassificações. Embora ao longo do séc. XX revisões taxonômicas tenham estabilizado a sistemática do gênero, a variabilidade morfológica e distribuição geográfica trans-ístmica da espécie A. innocous gerou questionamentos. Além disso, mais recentemente trabalhos de filogenia molecular da família Atyidae que incluíram representantes de Atya suscitaram questões em relação à sistemática do gênero (possível não monofilia) e de algumas espécies como A. gabonensis, A. margaritacea e A. scabra. Visto que o uso de marcadores moleculares nunca foi empregado para a delimitação das espécies de Atya e que seu uso de forma complementar à morfologia poderia aperfeiçoar a sistemática do gênero, o objetivo do presente estudo foi avaliar por meio de dados moleculares as hipóteses taxonômicas das espécies A. gabonensis, A. innocous, A. margaritacea e A. scabra. Sequências dos genes mitocondriais 16S e Citocromo Oxidase I e gene nuclear Histona 3 foram geradas por meio de protocolos de extração e sequenciamento de DNA a partir do tecido de espécimes obtidos em empréstimos/doações. Potenciais espécies evidenciadas pelas análises de similaridade nucleotídicas (distâncias genéticas), compartilhamento de caracteres em um contexto evolutivo (reconstruções filogenéticas), Automatic Barcode Gap Discovery, Poisson Tree Processes e Generalized Mixed Yule Coalescence foram confrontadas com as hipóteses taxonômicas específicas atuais. A avaliação sistemática com dados moleculares aqui realizada, adicionalmente às informações morfológicas existentes na literatura sustentaram A. gabonensis como uma espécie de distribuição anfi-atlântica, mas não corroborou a hipótese de A. innocous como uma espécie trans-ístmica. Assim, o uso do nome A. innocous para as populações do Mar do Caribe e A. tenella para aquelas restritas ao Pacífico é sugerido. A espécie A. margaritacea, distribuída ao longo da costa pacífica da América foi considerada uma espécie válida e distinta de A. scabra, amplamente distribuída na vertente atlântica da América do Sul, África e Mar do Caribe. Contudo, é discutida a possibilidade de uma espécie críptica restrita no Golfo do México existir. Adicionalmente, o conhecimento existente e pertinente para futuros estudos de sistemática e taxonomia sobre os camarões do gênero Atya foram sumarizados e são apresentados.
The genus Atya Leach, 1816 shrimps are the largest of the Atyidae family, and the 13 acknowledge species are geographically distributed in rivers and stream in the tropical and subtropical regions of America (Atlantic and Pacific drainages) and West Africa. The first registry of an Atya was in the XVII century and since then new species were described and previous description revised in an eventful taxonomic historic. Although throughout the XX century taxonomic revisions stabilized the genus systematics, the morphological variability and the trans-isthmic geographic distribution of A. innocous caused questioning. Moreover, molecular phylogenetic studies that included Atya representatives raised doubt on the genus systematics (possibly non-monophyletism) and some species A. gabonensis A. margaritacea and A. scabra hypothesis. As molecular markers have never been used concerning Atya species delimitation complementary to the morphology and it could improve the genus systematics, the goal of this study was to evaluate with molecular markers the taxonomic hypothesis of the species A. gabonensis, A. innocous, A. margaritacea e A. scabra. Sequences of the mitochondrial genes 16S and Cytochrome Oxidase I and nuclear gene Histone 3 were generated by means of DNA extraction and sequence protocols from specimens obtained in loans/donations. Putative species evidenced by the analysis of nucleotide similarity (genetic distances), character sharing (phylogenetic reconstitutions), Automatic Barcode Gap Discovery, Poisson Tree Processes and Generalized Mixed Yule Coalescence were compared to the prevailing taxonomic hypothesis. The systematic evaluation with the molecular data of this study, in addition with the morphological information in the literature sustain A. gabonensis as an amphi-atlantic distributed species, but do not corroborated A. innocous hypothesis as an trans-isthmian species. In this sense, the use of A. innocous stricto sensu for the Caribbeans Sea populations and A. tenella to that restricted to the pacific drainage of America is suggested. Atya margaritacea, distributed along the pacific drainage of America, is considered a valid species distinct from A. scabra, widespread distributed in the Atlantic drainage of America and Africa, besides Caribbean Sea. However, the possibility of a cryptic species in the Gulf of Mexico population is discussed. Aditionally, the relevant knowledge to future systematic and taxonomy studies about the shrimps of the genus Atya were summarized and are shown.

碩士
國立臺灣大學
漁業科學研究所
96
Terrestrial hermit crabs are represented by the Coenobitidae, which includes only two genera, Birgus and Coenobita. Coconut crabs (Birgus) are among the world’s largest terrestrial arthropods, having a crab-like morphology and a strongly calcified exoskeleton. The shell-carrying terrestrial hermit crabs (Coenobita), in contrast, have depended on gastropod shells for protection and as an aid in respiration. The asymmetrical pleons of terrestrial hermit crabs have been considered an evidence of marine hermit crab ancestry. Shell-carrying has constrained the morphological evolution of hermit crabs by requiring a decalcified asymmetrical pleon capable of coiling into gastropod shells. The target purpose of this work is to explore the species identification, phylogenetic relationships and evolutionary processes of terrestrial hermit crabs based on the morphological and molecular information. Comparative analysis of morphological characters and molecular sequences in a phylogenetic effort efficiently improves the sensitivity and accuracy of evolutionary inference, producing more precise results than single dataset analyses can provide. The partial sequences from the mtDNA COI and 16S rDNA genes of Coenobita species, nine of which are presented here for the first time (C. brevimanus, C. cavipes, C. clypeatus, C. hilgendorfi, C. perlatus, C. pseudorugosus, C. purpureus, C. rugosus, C. violascens), demonstrate how morphological patterns can illuminate the processes of evolutionary transition at the genus scale. Besides, there have been several taxonomic ambiguities among these Coenobita species. We reexamined and reidentified reliable morphological characters for species identification by referring to the patristic distances between each species. And, most important of all, we present evidences of molecular and morphological studies that the evolutionary route of terrestrial adaptation in the Coenobita phylogeny was from the inland to the shore, and not, as some carcinologists would have believed, gradually from the shore to the inland.