Orientador: Alexandre Magno Sebbenn / Resumo: Cariniana estrellensis e Cariniana legalis são uma das maiores árvores dos biomas florestais da Amazônia e Mata Atlântica, sendo atualmente vulneráveis à extinção devido ao intenso desmatamento desses biomas. Estratégias para conservação in situ e ex situ são urgentes e estudos de diversidade genética e fluxo de genes são chaves e para esses propósitos. Assim, investigamos a diversidade genética, a estrutura genética espacial (SGS) e o fluxo histórico de genes em populações fragmentadas de ambas as espécies, utilizando marcadores de microssatélites. Todas as árvores encontradas nas populações foram mapeadas, medidas para o diâmetro na altura do peito (DAP) e amostrado o cambio de casca. O índice de fixação (F), em alguns casos, foi significativamente maior em árvores com menor DAP, indicando que as árvores menores apresentam um maior parentesco do que as maiores. Foi detectada SGS significativa para populações de ambas as espécies (60-350 m), indicando um padrão de dispersão de genes de isolamento pela distância (IBD). Para ambas as espécies, foi observada alta imigração de semente (38,5-61,5%) e pólen (80,1-100%), mostrando que as populações não são isoladas geneticamente. Não foi detectada autofecundação, mas o cruzamento entre árvores relacionadas foi detectado nas espécies (8,9-12,5%), sugerindo uma seleção mais forte contra árvores de autofecundação do que se originou do cruzamento entre árvores relacionadas. A distância de dispersão de pólen e sementes em C. estrellensi... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Cariniana estrellensis and Cariniana legalis, two of the largest trees in the Amazon and Atlantic Forest biomes, are currently vulnerable to extinction due to the intense deforestation of these biomes. Strategies for in and ex situ conservation are urgent, and studies of genetic diversity and gene flow are key aspects needed to develop these strategeis. Thus, we investigate the genetic diversity, spatial genetic structure (SGS), and historical gene flow in fragmented populations of both species, using microsatellite markers. All trees found in the study populations were mapped, measured for diameter at breast height (DBH), and sampled for bark cambium. Our results show that in some cases, fixation index (F) was significantly higher in trees with lower DBH, indicating that smaller trees have higher levels of inbreeding than larger ones. Significant SGS was detected in populations of both species (60-350 m), indicating a gene dispersal pattern of isolation by distance (IBD). For both species, we found high seed (38.5-61.5%) and pollen (80.1-100%) immigration demonstrating that populations are not genetically isolated. No self-fertilization was detected, but we did find evidence of mating among related trees (8.9-12.5%), suggesting stronger selection against selfed individuals than those originated from mating among relatives. Pollen and seed dispersal distance for C. estrellensis reached longer distances (> 3 km) than for C. legalis (maximum of 385 m). However, pollen and seeds... (Complete abstract click electronic access below) / Doutor

Submitted by FRANCINE BEATRIZ DE SOUZA null (francinnysouza@yahoo.com.br) on 2018-03-25T19:33:25Z No. of bitstreams: 1 Francine__Tese_VersãoFinal_2018.pdf: 1284636 bytes, checksum: 8130187eea228723fe2e75938a47a468 (MD5) / Approved for entry into archive by Cristina Alexandra de Godoy null (cristina@adm.feis.unesp.br) on 2018-03-26T12:25:02Z (GMT) No. of bitstreams: 1 souza_fb_dr_ilha.pdf: 1284636 bytes, checksum: 8130187eea228723fe2e75938a47a468 (MD5) / Made available in DSpace on 2018-03-26T12:25:03Z (GMT). No. of bitstreams: 1 souza_fb_dr_ilha.pdf: 1284636 bytes, checksum: 8130187eea228723fe2e75938a47a468 (MD5) Previous issue date: 2018-02-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Cariniana estrellensis e Cariniana legalis são uma das maiores árvores dos biomas florestais da Amazônia e Mata Atlântica, sendo atualmente vulneráveis à extinção devido ao intenso desmatamento desses biomas. Estratégias para conservação in situ e ex situ são urgentes e estudos de diversidade genética e fluxo de genes são chaves e para esses propósitos. Assim, investigamos a diversidade genética, a estrutura genética espacial (SGS) e o fluxo histórico de genes em populações fragmentadas de ambas as espécies, utilizando marcadores de microssatélites. Todas as árvores encontradas nas populações foram mapeadas, medidas para o diâmetro na altura do peito (DAP) e amostrado o cambio de casca. O índice de fixação (F), em alguns casos, foi significativamente maior em árvores com menor DAP, indicando que as árvores menores apresentam um maior parentesco do que as maiores. Foi detectada SGS significativa para populações de ambas as espécies (60-350 m), indicando um padrão de dispersão de genes de isolamento pela distância (IBD). Para ambas as espécies, foi observada alta imigração de semente (38,5-61,5%) e pólen (80,1-100%), mostrando que as populações não são isoladas geneticamente. Não foi detectada autofecundação, mas o cruzamento entre árvores relacionadas foi detectado nas espécies (8,9-12,5%), sugerindo uma seleção mais forte contra árvores de autofecundação do que se originou do cruzamento entre árvores relacionadas. A distância de dispersão de pólen e sementes em C. estrellensis atingiu longa distância (> 3 km) do que em C. legalis (máximo de 385 m). No entanto, o pólen e as sementes em C. estrellensis e o pólen em C. legalis foram dispersos em um padrão de IBD. Os resultados sugerem que as populações estudadas são adequadas para conservação in situ e ex situ. / Cariniana estrellensis and Cariniana legalis, two of the largest trees in the Amazon and Atlantic Forest biomes, are currently vulnerable to extinction due to the intense deforestation of these biomes. Strategies for in and ex situ conservation are urgent, and studies of genetic diversity and gene flow are key aspects needed to develop these strategeis. Thus, we investigate the genetic diversity, spatial genetic structure (SGS), and historical gene flow in fragmented populations of both species, using microsatellite markers. All trees found in the study populations were mapped, measured for diameter at breast height (DBH), and sampled for bark cambium. Our results show that in some cases, fixation index (F) was significantly higher in trees with lower DBH, indicating that smaller trees have higher levels of inbreeding than larger ones. Significant SGS was detected in populations of both species (60-350 m), indicating a gene dispersal pattern of isolation by distance (IBD). For both species, we found high seed (38.5-61.5%) and pollen (80.1-100%) immigration demonstrating that populations are not genetically isolated. No self-fertilization was detected, but we did find evidence of mating among related trees (8.9-12.5%), suggesting stronger selection against selfed individuals than those originated from mating among relatives. Pollen and seed dispersal distance for C. estrellensis reached longer distances (> 3 km) than for C. legalis (maximum of 385 m). However, pollen and seeds of C. estrellensis and pollen of C. legalis were dispersed in an IBD pattern. The results suggest that the studied populations are suitable for in and ex situ conservation.

The Atlantic cod (Gadus morhua L.) fishery is of great economic importance to the Faroese economy. There are two separately managed cod stocks around the Faroe Islands, the Faroe Plateau and the Faroe Bank cod. Both have experienced dramatic decreases in size and informed management decisions are vital for both stock viability and exploitation. The stocks are geographically isolated by an 800 m deep channel and water temperatures are on average 1 – 2 ºC higher on the Faroe Bank than on the Faroe Plateau. There are clear phenotypic differences between the stocks; in particular, the markedly higher growth rate for the Faroe Bank cod has caught public and scientific attention. There is continuing debate regarding the relative importance of genetics and environmental contributions to the contrasting phenotypes. Analyses of reproductive parameters (field data and experimental captive spawnings) as well as analyses of microsatellite and single nucleotide polymorphism (SNP) markers were undertaken to better resolve the issue. Field data as well as data from experimental captive spawnings provided evidence of reproductive differences between Faroe Plateau and Faroe Bank cod. Peak spawning occurred earlier on the Faroe Plateau than on the Faroe Bank and this difference in timing of spawning was maintained in captivity. In particular, differences in sizes of eggs (average diameters of 1.40 and 1.30 mm for Faroe Plateau and Faroe Bank cod eggs, respectively) and indirect evidence of greater volumes spawned by the Faroe Bank females suggested stock differences with respect to egg size – egg number trade-off. It was hypothesised that the strategy adopted by cod on the Faroe Bank, with a higher number of smaller eggs, evolved in response to a more hostile environment (bare seabed and higher exposure to predators) experienced by early life stages in this area. Experimental captive spawnings with Faroe Bank cod showed a large interfamily skew in survival rates of cod eggs and fry. Egg size was identified as a useful indicator of survival rates in the egg stage, but egg survival rates could not be used to predict viability in later developmental stages, thus highlighting the importance of employing some sort of genetic monitoring of cod fry to ensure sufficient family representation in the progeny. While no tank effect was evident concerning fry survival, a significant tank effect was identified concerning body sizes of fry. Microsatellite data were analysed using large sample sizes of Faroe Plateau and Faroe Bank cod with the Faroe Plateau divided into two locations, Faroe Plateau North-East and Faroe Plateau West (cod from each of the two were known to belong to separate spawning grounds). Two Norwegian coastal cod samples were included as outlier populations. While no genetic differentiation was detected between the two Faroe Plateau locations, these analyses revealed a detectable, albeit relatively modest, degree of genetic differentiation between cod from the Faroe Plateau and the Faroe Bank (FST = 0.0014 and 0.0018; DJost_EST = 0.0027 and 0.0048; P < 0.0001 and P < 0.001 for the Faroe Plateau North-East – Faroe Bank and the Faroe Plateau West – Faroe Bank comparisons). These values were several times smaller than those between Faroese and Norwegian coastal cod (pairwise FST and DJost_EST values in the range of 0.0061 – 0.0137 and 0.0158 – 0.0386, respectively). Despite recent reductions in census population sizes for Faroe Plateau and, particularly, Faroe Bank cod, genetic diversity estimates were comparable to the ones observed for Norwegian coastal cod and there was no evidence of significant genetic bottlenecks. Lastly, data for one of the markers (Gmo132) indicated genotype-dependent vertical distribution of cod (as investigated for Faroe Plateau North-East cod). Contrary to some previously published studies, analysis of SNPs of two candidate genes for adaptive divergence, the hemoglobin gene Hb-ß1 and the transferrin gene Tf1, failed to detect differentiation between samples of Faroe Plateau and Faroe Bank cod analysed in this thesis. Of 3533 novel SNPs simultaneously discovered and genotyped by restriction-site associated DNA (RAD) sequencing, 58 showed evidence of genetic differentiation between Faroe Plateau North-East and Faroe Bank cod (P < 0.05). No single locus was fixed for different alleles between Faroe Plateau and Faroe Bank cod. A set of eight informative SNPs (FST values between Faroe Plateau and Faroe Bank samples > 0.25; P < 0.0005) were selected for validation in larger samples, that included cod from both Faroe Plateau areas and the Faroe Bank as well as Norwegian coastal and White Sea cod. Six out of the eight loci amplified successfully with a PCR-based method and there was 100 % concordance between genotypes of individuals screened by both techniques. Due to ascertainment bias, the SNPs should only be applied with caution in a broader geographical context. Nonetheless, these SNPs did confirm the genetic substructure suggested for Faroese cod by microsatellite analyses. While no genetic differentiation was evident between the two Faroe Plateau locations, significant genetic differentiation was evident between Faroe Plateau and Faroe Bank cod at five of the SNPs (FST values in the range of 0.0383 – 0.1914). This panel of five SNPs could confidently be used to trace groups of Faroe Plateau and Faroe Bank cod to their population of origin. In conclusion, multiple lines of evidence demonstrate that Faroe Plateau and Faroe Bank cod are truly two genetically distinct populations. While the findings contribute to a broader understanding of the biology and the genetics of Faroe Plateau and Faroe Bank cod, the novel SNPs developed may provide a valuable resource for potential future demands of i.e. genetic stock identification methods.

La biologie de la conservation est un domaine de recherche en pleine expansion en raison de la perte accélérée de la biodiversité à l’échelle mondiale. Pour mieux comprendre les processus et les menaces au maintien des populations de petite taille et les effets des facteurs anthropiques sur la biodiversité, la génétique est fréquemment utilisée en conservation. Des analyses génétiques peuvent, par exemple, nous informer sur les tendances à long terme, la diversité des populations et les stratégies de reproduction d’une espèce. La tortue des bois (Glyptemys insculpta) est une espèce endémique à l’Amérique du Nord qui est en danger d’extinction selon l’Union internationale pour la conservation de la nature. Dans le cadre de ma thèse, j’avais comme objectif de caractériser la diversité génétique de cette espèce menacée au Canada. À cet effet, j’ai analysé la génétique des populations de tortues des bois à plusieurs échelles spatiales et temporelles, afin de mieux cerner les processus ayant un impact sur la diversité des populations. Dans un premier temps, les relations de parentalité ont été reconstruites au sein d’une population de tortues des bois pour estimer la fréquence de paternité multiple et de paternité répétée. Les résultats de mes travaux suggèrent que l’emmagasinement de sperme chez la femelle et la reproduction multiple avec les mêmes partenaires pour plus d’une saison de reproduction pourraient expliquer ces phénomènes. Ces stratégies de reproduction pourraient dans ce cas être induites par la faible densité de la population à l’étude, ou encore par la fidélité au site d’hibernation où la majorité des évènements de copulation ont lieu. Par la suite, je me suis intéressée à la diversité génétique des populations de tortues de bois. J’ai voulu comprendre les effets de la configuration spatiale des éléments du paysage et les évènements de dispersion géographique sur la diversité des populations. À l’aide d’une approche de génétique du paysage, mes analyses montrent que la division des populations par bassins versants explique une large fraction de la diversité génétique interpopulations. Ces résultats confirment également que les bassins versants représentent des unités de gestion propices à la protection des populations de tortues des bois. Finalement, des analyses de réseaux ont été utilisées pour mieux cerner la dynamique de flux génique entre les populations de la rive nord et de la rive sud du fleuve Saint-Laurent. Plus spécifiquement, la rive nord se caractérise par un réseau robuste de populations isolées, alors que les populations de la rive sud présentent plutôt une structure de métapopulation. En utilisant les réseaux construits à partir de données génétiques, des scénarios hypothétiques furent comparés pour explorer la sélection de populations à l’aide du logiciel BRIDES. Les résultats de ces analyses ont permis de cibler l’importance de certaines populations de tortues des bois pour la connectivité du réseau. L’importance de ces populations n’aurait pu être prédite par les résultats de la diversité et de la différenciation génétique, les indices de centralité et les analyses d’élimination de nœuds. Grâce à la génétique, cette thèse apporte de nouvelles connaissances sur la tortue des bois, les stratégies de reproduction des différents sexes, le flux génique, la connectivité et l’influence du réseau hydrographique sur la diversité des populations. Ces résultats nous permettent d’avoir une meilleure compréhension des processus affectant la diversité génétique de cette espèce afin de mieux la protéger. Toutes les analyses réalisées pour cette thèse sont directement applicables à l’ensemble des autres espèces longévives avec des générations chevauchantes. / Conservation biology is a rapidly expanding field of research due to the accelerating loss of global biodiversity. To better understand the processes and threats to the persistence of small populations and the effects of anthropogenic factors on biodiversity, genetic approaches are frequently used in conservation. Genetic analyzes can, for example, inform us about long-term trends, population diversity and reproductive strategies of a species. The wood turtle (Glyptemys insculpta) is a species endemic to North America that is endangered according to the International Union for the Conservation of Nature. As part of my thesis, my objective was to characterize the genetic diversity of this threatened species in Canada. In order to better understand the impact of reproductive strategy and landscape structure on population diversity, I analyzed the genetics of wood turtle populations at several spatial and temporal scales. First, parentage relationships were reconstructed in a population of wood turtles to estimate the frequency of multiple and repeated paternity. The results of my work suggest that sperm storage in females and multiple reproduction with the same partners for more than one breeding season could explain these phenomena. These reproduction strategies could in this case be induced by the low density of the study population, or by fidelity to the overwintering site where the majority of copulation events take place. Subsequently, I assessed the genetic diversity of wood turtle populations. I wanted to understand the effects of the landscape configuration and geographic dispersion events on the diversity of populations. Using a landscape genetics approach, my analyzes show that the division of populations by watershed explains a large fraction of the genetic diversity between populations. These results also confirm that watersheds represent management units conducive to the protection of wood turtle populations. Finally, network analysis was used to better understand the gene flow dynamics among populations located on the north and south shores of the St. Lawrence River. More specifically, the north shore is characterized by a robust network of isolated populations, whereas the populations on the south shore present more of a metapopulation structure. Using population graphs, hypothetical scenarios were compared to explore the node selection process using the BRIDES algorithm. The results of these analyzes made it possible to point out specific populations of wood turtles, considering their importance for network connectivity. This could have not been predicted by using genetic diversity and distinctiveness estimates, node-based metrics, and node removal analysis for these populations. Thanks to genetics, this thesis brings new knowledge on the wood turtle, the reproductive strategies of both sexes, the gene flow, the connectivity and the influence of the hydrographic network on population diversity. These results allow us to have a better understanding of the processes affecting the genetic diversity of this species in order to better protect it. All analyses performed for this thesis are directly applicable to other long-lived species with overlapping generations.