This thesis comprises studies of the phylogenetic relationships in the flowering plant clade Potentilleae in Rosaceae. The relationships were elucidated by using DNA sequence data from the nuclear genome as well as from the plastid genome. In particular, the focus of the studies was the investigation of allopolyploidy, i.e. speciation as a result of hybridization and subsequent chromosome doubling. A phylogenetic method was used for identifying allopolyploidy through comparison of trees resulting from the analyses of different DNA sequences. Five sub-clades were investigated. First, both the sister clades that together contain all of Potentilleae: Fragariinae and Potentilla. Secondly, three subclades of Fragariinae, namely Alchemilla in wide sense, Sibbaldia and relatives, and Fragaria. The aim was to unravel the phylogenetic relationships, including instances of allopolyploidy. Classification issues were discussed in relation to the phylogenetic results. The split between Potentilla (=Potentillinae) and Fragariinae received better support than in previous studies. The phylogeny of Fragariinae was found to be consistent with classifying ten genera: Alchemilla in wide sense (incl. Aphanes and Lachemilla), Comarum, Sibbaldia, Sibbaldianthe, Sibbaldiopsis, Chamaerhodos, Drymocallis, Dasiphora, Potaninia, Fragaria, and also including a few orphan Potentilla species. The segregated genera Ivesia, Horkelia, Horkeliella and Duchesnea were found to be nested within Potentilla, corroborating earlier studies, while the segregated genus Argentina (P. anserina and close relatives) showed an ambiguous position. Plastid and nuclear (ribosomal) phylogenies were compared and incongruences were detected as potential instances of allopolyploid speciation. Five strongly supported incongruences were detected in Fragariinae and four of them were considered to be potentially caused by allopolyploidy. In addition, five supported incongruences were found in Potentilla. Alchemilla in the wide sense was found to contain four major clades, African Alchemilla, Eurasian Alchemilla, Lachemilla and Aphanes. Both Lachemilla and Aphanes were nested within Alchemilla and it was suggested that the name Alchemilla should be used in the wide sense, i.e. including both the genera Lachemilla and Aphanes. The genus Sibbaldia as commonly classified was shown to be polyphyletic in five different places in Potentilleae. Three Sibbaldia clades ended up in Fragariinae and two in Potentilla. A phylogeny of Fragaria, based on a nuclear low/single copy DNA region was estimated. The gene copy phylogeny was used to construct a reticulate tree hypothesizing allopolyploid speciation events. The evolution of Fragaria was shown to have been shaped by polyploidy. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 4: Manuscript. Paper 5: Manuscript.

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No. of bitstreams: 0 Previous issue date: 2017-03-03 / Lippia alba (Verbenaceae), é uma espécie herbácea tropical com grande plasticidade fenotípica e genômica, amplamente utilizada na medicina popular. Recentemente, a espécie foi descrita como um novo complexo autopoliploide contendo cinco números cromossômicos (2x=30, 2x=30+8, 3x=45, 4x=60 e 6x=90), e esforços têm sido feitos a fim de entender sua origem e evolução. No presente trabalho, foram descritos perfis cariotípicos mais detalhados da espécie, por meio de mapeamento cromossômico utilizando sondas espécie-específicas e análises do comportamento meiótico e de viabilidade polínica. A partir do sequenciamento genômico de baixa cobertura (IIlumina MiSeq), foram desenvolvidos novos marcadores citogenéticos (denominados CL66 e CL98) os quais foram utilizados para o mapeamento cromossômico em acessos representando os cinco citótipos do complexo. Para a análise meiótica, seis estágios da divisão (metáfase I; anáfase I + telófase I; metáfase II; anáfase II + telófase II) foram quantificados, e aproximadamente, 100 células foram avaliadas para cada estágio. Os mesmos acessos foram avaliados quanto à viabilidade polínica (1.000 grãos de pólen foram quantificados para cada indivíduo). Os resultados da Hibridização Fluorescente in situ (FISH) revelaram que ambas as repetições satélite estão localizadas na porção terminal dos cromossomos. Em geral, a repetição CL98 mostrou um padrão uniforme nos diferentes acessos. Foram observados dois, três, quatro e seis cromossomos marcados em diploides, triploides, tetraploides e hexaploide, respectivamente, revelando que o número de cromossomos marcados variou proporcionalmente, de acordo com o nível de ploidia do acesso. Por outro lado, a repetição CL66 apresentou-se polimórfica. Variações foram observadas entre os acessos, principalmente, entre os indivíduos diploides. Com relação às análises meióticas, alto percentual de irregularidade foi observado nos citótipos poliploides. Entretanto, alguns acessos 2x também mostraram consideráveis erros durante a microsporogênese. Entre as irregularidades encontradas, destacam-se: pareamento cromossômico anormal; segregação cromossômica desigual; cromossomos perdidos; tríades e políades. Os resultados da viabilidade polínica corroboraram os dados da meiose. A partir do conjunto de dados obtidos foi possível concluir que 1) a metodologia para o desenvolvimento de marcadores cromossômicos específicos para L. alba mostrou-se eficiente; 2) as repetições satélite exibiram diferentes comportamentos (estável e dinâmico) no genoma de L. alba; 3) a ocorrência de microsporogênese irregular em diploides, associada à viabilidade polínica, sugerem que os acessos 2x sejam elementos importantes na formação do complexo poliploide e 4) a ampla variação cariotípica observada na espécie pode ser consequência de múltiplos e independentes eventos de duplicação genômica, aliado a rearranjos cromossômicos. Possivelmente, L. alba encontra-se em processo de estabilização do seu cariótipo tornando a espécie, um importante modelo para estudos de poliploides naturais nos trópicos. / Lippia alba (Verbenaceae) is a tropical aromatic shrub with extensive phenotypical and genomic plasticity widely used in traditional medicine. Recently, the species was described as a new natural autopolyploid complex with five distinct chromosome numbers (2x=30, 2x=30+8, 3x=45, 4x=60 and 6x=90). Strides have been done in order to understand the cytotypes origin and species evolution. In this study, a detailed karyotype of L. alba using Fluorescence in situ Hybridization (FISH) with species-specific probes was described. We also report the meiosis behavior and pollen viability in sixty accessions. Using massive parallel sequencing (IIlumina MiSeq platform) new cytogenetic landmarks (CL66 and CL98) were chosen for probing all cytotypes described for the species. For meiotic analysis, the percentage of abnormalities was quantified, evaluating around 100 cells in six stages (metaphase I; anaphase I + telophase I; metaphase II; anaphase II + telophase II). Around 1,000 pollen per accession were used to estimate pollen viability. FISH results revealed that both satDNA arrays are located preferentially on terminal sites of the chromosomes. In general, the CL98 repeat showed a uniform pattern in different accessions. We observed 2, 3, 4, and 6 marked chromosomes respectively in diploid, triploid, tetraploid and hexaploid accessions revealing that the number of depicted chromosomes varied proportionally according to the ploidy level. On the other hand, the CL66 repeat was polymorphic. Great variations were observed among the accessions mainly within the diploids. In general, the meiotic analysis revealed higher index of abnormalities in polyploid cytotypes. However, some 2x accessions also showed considerable irregularities during the microsporogenesis. Desynapsis, unequal segregation, lost chromosomes, triads and polyads were the most common irregularities observed. Pollen viability analysis corroborated the meiosis data. It was possible to conclude that 1) the development of specific landmarks for L. alba was efficient; 2) the karyotypic profiles of both satDNA revealed different behavior; 3) microsporogenesis analysis and pollen viability of 2x accessions suggest that diploids are the key point for the origin of the polyploid complex and 4) independent and multiples events of genome duplication associated to chromosome rearrangements may have generated great karyotypic variation in the species. L. alba karyotype is possibly under stabilization process making the species an important model to study natural polyploids in the tropics.

Ray-finned fishes (Actinopterygii) exhibit the greatest biodiversity among vertebrates. The vast majority of extant actinopterygian fish species belong to clade Teleostei - a lineage whose significant evolutionary success might have resulted from a teleost specific whole- genome duplication (TSGD) that occurred at the onset of this group, subsequent to its divergence from the rest of actinopterygian lineages. Despite the growing body of sequenced fish genomes and analyses of their transcriptomes, the largest contribution to understanding fish genomes comes from analyses of DNA content and from cytogenetics. Genomes of ray-finned fishes and especially those of Teleostei exhibit vast diversity and rapid dynamics of repetitive DNA sequences whose variability is reflected in a wide range of fish genome sizes and in the dynamics behind karyotype differentiation. Therefore, ray-finned fishes offer a unique opportunity to study genome variability as a driving force underlying morphological and ecological diversification, evolution and adaptation. Particularly, the mapping of repetitive DNA sequences by means of fluorescence in situ hybridization (FISH) has proven to be a very useful and informative approach during the last two decades and contributed greatly to our understanding of the fish genome...

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