D’après un rapport de l’OMS, la tuberculose reste en 2015 l’une des 10 premières causes de décès à l’échelle mondiale. De ce fait, en matière de santé, éradiquer la maladie à l’horizon 2030 est un des objectifs majeurs fixés par les Nations Unies. La bactérie responsable de cette infection, Mycobacterium tuberculosis, est un pathogène obligatoire dont l’origine et l’évolution sont intrinsèquement liées à celles de son hôte principal, Homo sapiens. En effet, les souches actuelles de tuberculose présentent, tout comme l’homme, une forte structure phylogénétique, trace de leur origine géographique. Les pays pauvres et en développement sont les plus touchés par l’épidémie globale, favorisée par des systèmes de santé défaillants et une haute prévalence du VIH. Les pays occidentaux ne sont pas épargnés, menacés par l’émergence de souches de plus en plus résistantes aux antibiotiques provenant en grande partie de l’ex URSS. Au cours de cette thèse, j’analyse l’histoire évolutive, la propagation et l’acquisition de résistances aux antibiotiques de plusieurs épidémies de tuberculose en me basant sur des données génétiques et génomiques. Dans un premier temps je m’intéresse aux effets d’une campagne nationale de traitements en Asie Centrale sur le développement de souches multi-résistantes et met également en lumière le rôle clef de certaines mutations dans le succès des clones présentés. Ainsi cette campagne a été partiellement mise en échec par la présence de souches pré-résistantes, grâce à la survenue de mutations avant même la mise en place des traitements antibiotiques. Par la suite je me suis focalisé sur un clade particulier de souches multi-résistantes, le clone Russe W148. Je présente sa dispersion géographique et temporelle à travers l’Eurasie et démontre l’importance des mutations compensatoires dans son succès épidémique. De plus, la tuberculose ne touche pas seulement les hommes mais infecte également plusieurs autres mammifères. Afin d’appréhender les contraintes adaptatives accompagnants ces changements d’hôtes, j’ai effectué divers tests de sélection dans le but d’identifier les gènes impliqués. Pour finir, nous avons développé un indice souche spécifique, permettant de mesurer le succès épidémique de celles-ci à un niveau individuel. Dans le cadre d’études épidémiologiques, cette mesure peut être croisée avec des informations sur le patient, la souche ou même socio-économiques. / According to a 2015 WHO report, tuberculosis remains one of the top 10 causes of death worldwide. Despite considerable efforts by the United Nations to eradicate the disease by 2030, a global TB epidemic still persists. Its causative agent, the bacterium Mycobacterium tuberculosis, an obligate pathogen, has been plaguing humanity since it originated, and has coevolved with its main host, Homo sapiens, over thousands of years. Contemporary tuberculosis strains exhibit a structured phylogeographic pattern, carrying the genetic print of their geographic origin. The Koch bacillus infects and kills in large numbers, in poor and developing countries, where fragile health care systems, combined with high HIV prevalence, facilitate epidemic spread. In western countries, the major current threats are the multiplication and propagation of antibiotic resistant strains (MDR/XDR) coming predominantly from former Soviet republics. In this thesis, I unravel the evolutionary history, propagation, and acquisition of drug resistance-conferring mutations in different settings, by implementing multiple genetic and genomic data sets. First, focusing on Central Asia, using whole genome sequencing and Bayesian statistics, I assess the effects of a treatment campaign on the development of MDR strains and highlight key mutations in successful strains. More importantly, the success of DOTs campaigns was compromised by the genetic make-up of these outbreak clades (pre-treatment low frequency resistance SNPs). Special attention was also given to a particular outbreak of MDR strains, i.e. the Russian W148 clone. I present its westward spatial and temporal propagation at a continental scale during the last century, and underline the key contribution of compensatory mutations in its epidemic success. However, tuberculosis does not only infect humans, but also has experienced successive mammalian host jumps. To decipher the adaptive constraints accompanying such secondary events, a systemic gene screen with selection signature-detecting algorithms was implemented to identify putative targets during diversifying selection. Finally, novel mathematical tools and indices that reflect the epidemicity of a strain were developed, jumping from a population-driven approach to a strain specific one, with broader epidemiological applications. This allows us to correlate strain fitness with patient, lineage, and socio-economic information.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2018-04-09T14:42:24Z No. of bitstreams: 2 Dissertação - Lucas Donizetti Vieira - 2018.pdf: 2934046 bytes, checksum: 50730e1b6711826aafed66d22a56852f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-04-09T14:44:18Z (GMT) No. of bitstreams: 2 Dissertação - Lucas Donizetti Vieira - 2018.pdf: 2934046 bytes, checksum: 50730e1b6711826aafed66d22a56852f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-04-09T14:44:18Z (GMT). No. of bitstreams: 2 Dissertação - Lucas Donizetti Vieira - 2018.pdf: 2934046 bytes, checksum: 50730e1b6711826aafed66d22a56852f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-03-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Population genomics studies in Neotropical species are scarce, however, they are of great importance for the understanding of the neutral and adaptive variations, in order to develop strategies of management and conservation for the species. In this context our objective was to verify the role of natural selection in the distribution of populations diversity in Handroanthus serratifolius and Tabebuia aurea. Here, we study population genomics based in sequence capture using 24,751 pre-developed probes. For H serratifolius were sampled c. 64,000 SNPs in 144 individuals from 16 populations. For T aurea were sampled c. 18,000 SNPs in 48 individuals from 6 populations. Genetic differentiation was high (FST = 0.194, p <0.001) in H. serratifolius and intermediate (FST = 0.144, p <0.001) in T. aurea. The inbreeding coefficient was low in both species, however, in three populations of H. serratifolius it was intermediate or high. The genetic structure of populations presented two genetic groups for the populations sampled in the two species. H. serratifolius with high genetic differentiation and T. aurea with admixture in all populations. We did not find SNPs outliers in the distribution of FST values, due to the wide distribution of values found for the populations of the two species. In the search for correlation between environmental variables and changes in allelic frequency H. serratifolius presented only three SNPs, inside six genes and T. aurea presented 16 SNPs, inside 25 genes. The functional annotation of these genes showed relationship with molecular processes, transposable elements and one in T. aurea related to resistance to diseases. In conclusion, our data show a selection signal in H. serratifolius for only three loci with local adaptation signal. In T. aurea our data show a greater number of locos (16 locos) with local adaptation signal. / Estudos de genômica de populações em espécies Neotropicais são escassos, no entanto, apresentam grande importância para o entendimento das variações neutras e adaptativas, a fim de desenvolver estratégias de manejo e conservação das espécies. Nesse contexto nosso objetivo foi verificar o papel da seleção natural na distribuição da diversidade de populações de Handroanthus serratifolius e Tabebuia aurea. Aqui, nós estudamos a genômica de populações baseado em sequence capture utilizando 24.751 sondas previamente desenvolvidas. Para H serratifolius foram amostrados c. 64.000 SNPs em 144 indivíduos de 16 populações. Para T aurea foram amostrados c. 18.000 SNPs em 48 indivíduos de 6 populações. A diferenciação genética foi alta (FST = 0,194, p < 0,001) em H. serratifolius e intermediária (FST = 0,144, p < 0,001) em T. aurea. O coeficiente de endogamia foi baixo nas duas espécies, no entanto, em três populações de H. serratifolius foi intermediária ou alta. A estrutura genética das populações apresentou dois grupos genéticos para as populações amostradas nas duas espécies. H. serratifolius com alta diferenciação genética e T. aurea com mistura em todas as populações. Nós não encontramos SNPs outliers na distribuição de valores de FST, devido a ampla distribuição de valores encontrada para as populações das duas espécies. Na busca por correlação entre variáveis ambientais e mudanças na frequência alélica H. serratifolius apresentou apenas três SNPs, presentes em seis genes e T. aurea apresentou 16 SNPs, presentes em 25 genes. A anotação funcional desses genes mostrou relação com processos moleculares, elementos transponíveis e um em T. aurea ligado a resistência a doenças. Concluindo, nossos dados apresentam sinal de seleção em H. serratifolius para apenas três locos com sinal de adaptação local. Em T. aurea nossos dados mostram maior número de locos (16 locos) com sinal de adaptação local.

L’igname (Dioscorea spp) est un aliment de base de plus de 100 millions de personnes en Afrique. L’objectif de cette thèse était d'étudier la diversité génomique de l'igname, comprendre les bases génétiques de sa domestication, et d'étudier son adaptation à différentes zones climatiques. L’étude du processus de domestication de l’igname a été menée par une approche de génomique comparée entre l’espèce cultivée D. rotundata et deux espèces sauvages apparentées D. praehensilis et D. abyssinica, en utilisant des données de séquençage NGS génomique. Nous avons mis en évidence des sélections fortes de gènes de la voie de biosynthèse de l’amidon. Des gènes impliqués dans la morphologie des tubercules ou l’aptitude au phototropisme, ainsi que des gènes du complexe NADH deshydrogenase ont également été identifiés comme sélectionnés durant la domestication. Ce même complexe NADH-DH a également été identifié lors de la recherche de gènes associés à la distribution d’une collection d’ignames selon la variabilité climatique. Nous avons aussi créé la première banque de novo d’éléments transposables (ET) de l’igname. L’étude que nous avons menée sur les éléments répétés (ER) du génome de l’igname nous a permis d’identifier une forte corrélation entre la variabilité des abondances relatives d’un grand nombre d’ERs et la variabilité climatique. Enfin, nous avons pu proposer une hypothèse quant à l’origine de l’igname cultivée D. rotundata. La domestication de l'igname dériverait de l'espèce inféodée au milieu forestier, D. praehensilis. Ces résultats remettent en cause l’hypothèse d’une origine stricte en zone de savane pour les espèces cultivées et l’agriculture en Afrique de l'Ouest. / Yam (Dioscorea spp) is a major staple for more than 100 million people in Africa. The main objectives of the present PhD project were to study yam genomic diversity, its domestication, and to characterize the genomic determinism of its adaptation to different climatic zones. We investigated the genetic basis of yam domestication in a comparative genomic approach between the cultivated species D. rotundata and two wild close relatives D. praehensilis and D. abyssinica, by exploiting NGS sequencing data. We demonstrated that genes from the starch biosynthesis were selected during yam domestication. Genes related to tuber morphology or phototropism ability, as well as genes of the NADH dehydrogenase complex were also under selection. The same NADH-DH complex was also identified when assessing adaptation to climate variability. We also created the first de novo database of yam transposable elements (TEs). The study we performed on these repeat elements (REs) highlighted a strong correlation between the variability in relative abundances of numerous REs and climatic variability. Finally, we were able to propose an hypothesis on the origin of the cultivated yam D. rotundata. Our hypothesis identifies the origin of yam in the forest areas, with the species D. praehensilis as the putative progenitor. Our results question the generally admitted hypothesis of savannah origins for crops and agriculture in Africa.

This dissertation examines the politics of representation in biotechnosciences. Through web representations, I examine three emerging endeavors that propose to create large-scale human population genomic databases to study complex, common diseases and conditions. These projects were initiated in different nations (US, UK, and Iceland), created under different institutional configurations, and are at various stages of development. The websites, which are media technologies do not simply reflect and promote these endeavors. Rather, they help shape these database projects in which the science is uncertain and the technologies not yet built. Thus, they are constitutive technologies that affect the construction of these database projects. More needs to be done to explore how to interpret the 'virtual' realm and how it relates to the 'real' world and specific situations. By bringing hypertextuality into the analysis, I explore how knowledges, practices, and subjectivities are created. By adapting the methods of a number of science and technology (STS) authors, I develop a more dynamic lens in which to investigate web representations and 'emerging' biomedical projects. My concern however, is not only in what represents what, but how representations are constructed. The power of the latter derives from its invisibility. In re-conceptualizing representation and new media technologies, I show that these sites are techno-social spaces for creating knowledge, specific ways of seeing, and practicing biomedicine today. The narrowing time/space between generating data, releasing information, and incorporating publics into their endeavors raises crucial issues as to how biomedicine is represented and how broader audiences are engaged. In the dominant discourses, these projects are all situated within biomedical, (post)genomic, and information revolutions. Here, they hang on the technological object, the database, with the ability to contain what we are coming to understand as life/genetic/bio information. Through the moves of both treating these databases as part of a complex system and investigating them through a lens of representation, I begin to include potential participants and broader audiences into the analysis. Informatic bodies, populations, and subjects are co-created at, by, and through these sites as the developing database projects and information are (re)presented. / Ph. D.

Seleção balanceadora é um processo evolutivo que engloba diversos mecanismos: vantagem do heterozigoto, seleção dependente de frequência, pressões seletivas que variam ao longo do tempo ou do espaço, e alguns casos de pleiotropia. O estudo desses mecanismos em si foi e ainda é um tópico de grande interesse para os biólogos evolutivos, e moldou o estudo da evolução ao longo do último século. Antes de a teoria neutra ter sido proposta, acreditava-se que a seleção balanceadora fosse comum. A descoberta de que muita da diversidade genética observada podia ser explicada por evolução neutra motivou, portanto, uma melhor compreensão da seleção balanceadora como um regime seletivo capaz de manter variantes vantajosas nas populações. O estudo da seleção balanceadora, em seus primórdios, foi restrito a organismos que podiam ser manipulados em laboratório. Com o advento de métodos que permitiam quantificar a variabilidade genética - tais como a eletroforese de proteínas, sequenciamento em pequena escala e re-sequenciamento genômico de milhares de indivíduos -, a variabilidade genética humana passou a ser ativamente estudada e interpretada. Diversos estudos buscaram por assinaturas de seleção natural - i.e., padrões de variação genômica deixadas por tais regimes seletivos - e avaliaram seu significado comparando-as com o que seria esperado sob um cenário estritamente neutro. A maior parte desses esforços foram concentrados no estudo da seleção positiva, tida como o principal mecanismo responsável pela evolução adaptativa. Poucos estudos buscaram assinaturas de seleção balanceadora no genoma humano. Isso se deve em parte à escassez de métodos com alto poder para detectar tais assinaturas. Adicionalmente, estudos prévios não analisaram dados em escala genômica, ou se concentraram principalmente nas regiões codificadoras de proteínas. Aqui, nós descrevemos um método simples e com alto poder para detectar assinaturas de seleção balanceadora. Em humanos, esse método supera outros comumente usados para a detecção de tais assinaturas e, em teoria, poderia ser usado para detectá-las em outras espécies, desde que seu poder seja avaliado caso-a-caso através de simulações neutras. Nosso método (\"Non-Central Deviation\", NCD) é apresentado em duas versões: NCD2, que requer informação acerca dos polimorfismos da espécie analisada e das substituições entre essa espécie e um grupo externo, e NCD1, que requer apenas informação acerca dos polimorfismos da espécie analisada. Embora em humanos NCD2 supere NCD1, este último pode ser utilizado para espécies para as quais não haja informação de um grupo externo. Quando aplicamos NCD2 a dados humanos, usando chimpanzé como grupo ex- terno, encontramos mais de 200 genes codificadores de proteínas com forte assinatura de seleção balanceadora, dos quais apenas 1/3 tinha evidência prévia de seleção balanceadora. Encontramos também um enriquecimento para diversas categorias de ontologia gênica, das quais cerca da metade é relacionada à imunidade. Verificamos que dentre os genes com evidências de seleção balanceadora há um excesso de casos de expressão preferencial em tecidos tais como \"adrenal\" e \"pulmão\", e também um excesso de genes com expressão mono-alélica. No geral, vimos que as regiões selecionadas no genoma humano incluem tanto sítios codificadores quanto regulatórios. Não encontramos um excesso de assinaturas de seleção balanceadora em regiões regulatórias, ao contrário do que reportaram outros estudos. Finalmente, encontramos um excesso de polimorfismos não-sinônimos em relação aos sinônimos nos genes selecionados. Tendo documentado a ocorrência de seleção balanceadora no genoma humano e identificado genes que foram potencialmente alvos deste regime seletivo, nós investi- gamos as consequências evolutivas desse processo. Nós partimos da hipótese que a seleção balanceadora sobre um sítio reduz a eficiência com a qual a seleção purificadora elimina variantes deletérias em sítios vizinhos. Esse processo é uma consequência do quanto a seleção sobre um loco afeta, através de ligação genética, as frequências de sítios não-neutros adjacentes. Testamos essa hipótese examinando se os genes sob seleção balanceadora apresentam um excesso de variantes deletérias em relação a expectativas derivadas a partir do restante do genoma. Usando três diferentes métricas para determinadas se e/ou o quão deletéria é uma dada variante, identificamos um excesso de variantes deletérias dentro dos genes sob seleção balanceadora, e mostramos que tal padrão não pode ser atribuído a efeitos confundidores. Esse achado mostra que, juntamente com os benefícios associados à variação adaptativa, a seleção balanceadora aumenta o fardo de mutações deletérias no genoma humano. De forma geral, nossos achados sugerem que a seleção balanceadora provavelmente mantém variantes genéticas envolvidas em uma miríade de processos biológicos além da imunidade e que ela foi mais comum no genoma humano do que se acreditava anteriormente, afetando entre 1-8% dos genes codificadores de proteínas, bem como diversas regiões não-codificadoras. Adicionalmente, a seleção balanceadora parece ser importante para a evolução humana não apenas por seu efeito sobre a aptidão, mas também por ter sido uma importante força capaz de moldar a diversidade genética observada atualmente em humanos e a susceptibilidade a doenças / Balancing selection is an evolutionary process that encompasses several mechanisms: heterozygote advantage, negative frequency dependent selection, selective pressure that fluctuates in time or in space, and some cases of pleiotropy. The study of these mechanisms .per se has been and still is a topic of great interest for evolutionary biologists, and has shaped the study of evolution throughout the last century. Before the proposition of the neutral theory of molecular evolution, it was believed that balancing selection was pervasive. The realization that much of the observed genetic diversity could be explained by neutral evolution thus motivated a better understanding of balancing selection as a selective regime capable of maintaining adaptive variants in populations. The study of balancing selection, in its early stages, was restricted to organisms that could be manipulated in the laboratory. With the advent of methods that allowed quantification of genetic variation - such as protein electrophoresis, small scale sequencing and genome-wide re-sequencing of thousands of individuals - human variation started to be actively studied and interpreted. Several studies have looked for signatures of natural selection - i.e., patterns of genomic variation that selective regimes leave in the genome - and evaluated their significance by comparing them to what would be expected under a strictly neutral scenario. Most of these efforts focused on the study of positive selection, thought of as the prime mechanism responsible for adaptive evolution. Only a few studies looked for signatures of balancing selection in the human genome. This is partially due to the paucity of powerful methods to detect its signatures. Moreover, previous studies either did not analyze data on genomic scale or focused primarily on protein-coding regions. Here, we describe a powerful and simple method to detect signatures of balancing selection. In humans, it outperforms other methods commonly used to detect such signatures and could in theory be used for other species, provided that its power is evaluated for each species through neutral simulations. Our method (\"Non-Central Deviation\", NCD) has two versions: NCD2, which requires polymorphism information on the ingroup species, as well as divergence information between the ingroup and an outgroup species, and NCD1, which only requires the ingroup information. Although NCD2 is more powerful for humans, NCD1 can be used for species that lack information from an outgroup. When applying NCD2 to human data, using chimpanzee as the outgroup, we found more than 200 protein-coding regions with strong signatures of balancing selection, only 1/3 of which had prior evidence for balancing selection. There was also an enrichment for several gene ontology categories, approximately half of which are related to immunity. We also found that among genes with evidence for balancing selection there was an excess of cases of preferential expression in specific tissues, such as \"adrenal\" and \"lung\", and an excess of genes with mono-allelic expression. Overall, we found that selected regions of the genome include both coding and regulatory sites. We failed to find a marked excess of balancing selection in regulatory regions, as reported in previous studies. Finally, we found an excess of nonsynonymous versus synonymous polymorphisms within the selected genes. Having documented the occurrence of balancing selection in the human genome and identified genes which were potential targets of this selective regime, we next investigated evolutionary consequences of this process. We hypothesized that balancing selection acting on a site reduces the efficiency with which purifying selection purges deleterious variants at nearby sites. This process is a consequence of how the dynamics of selection at one locus, mediated by linkage, can interfere with the frequencies of adjacent non-neutral sites. We tested this hypothesis by examining if the genes under balancing selection show an excess of deleterious variants with respect to expectations derived from the remainder of the genome. Using three different metrics to determine deleteriousness, we identified a significant excess of deleterious variants within balanced genes, and we show that this pattern cannot be attributed to confounding factors. This finding shows that together with the benefits associated with adaptive variation, balancing selection is increasing the burden of deleterious mutations in the human genome. Overall, our findings suggest that balancing selection likely maintains variation in a myriad of biological processes other than immunity and that it has been more common in the human genome than previously thought, affecting between 1-8% of human protein-coding genes, as well as a number of non-protein coding regions. Moreover, balancing selection appears to be important to human evolution not only because of its influence on fitness, but also because it has been an important force shaping current human genetic diversity and susceptibility to disease

A mandioca foi domesticada no sudoeste da bacia amazônica, e presentemente é o cultivo alimentício amazônico mais importante no mundo. Após a domesticação inicial pressões seletivas divergentes deram origem aos grupos de variedades de mandiocas mansas e bravas. A distribuição atual destes grupos é um tanto diferente ao longo da Amazônia, o que pode ser reflexo de padrões de dispersão distintos de variedades mansas e bravas ao longo da história da domesticação do cultivo. O objetivo deste estudo foi avaliar a diversidade e estrutura genética, genômica e a filogeografia de mandiocas cultivadas por agricultores tradicionais ao longo dos principais rios da bacia amazônica brasileira. Análises filogenéticas de linhagens matrilineares foi realizada com base no polimorfismo de quatro marcadores microssatélites cloroplastidiais (cpSSR). A diversidade e estrutura genética foram avaliadas com 14 marcadores microssatélites nucleares (ncSSR), enquanto que a abordagem genômica foi realizada com base em 5.871 polimorfismos de nucleotídeos únicos (SNPs). Foi observada considerável diferenciação [FST = 0,78 (cpSSR), 0,28 (ncSSR), e 0,37 (SNPs)] entre as variedades cultivadas e Manihot esculenta ssp. flabellifolia, o parente silvestre da mandioca. Não foram detectadas associações de haplótipos cpSSR com os grupos de variedades mansas e bravas ou com os rios. Apesar da ausência de padrões filogeográficos, as análises de agrupamento e estrutura genética com base nos três tipos de marcadores avaliados sugeriram que as variedades mansas e bravas são igualmente relacionadas à população silvestre. O segundo padrão mais importante de estruturação genética foi observado entre variedades mansas e bravas [FST = 0,08 (ncSSR) e 0,10 (SNPs)], no entanto houve um considerável grau de mistura entre variedades de ambos os grupos. Estes resultados, juntamente com a elevada variação genética observada dentro de variedades mansas e bravas são resultantes do manejo realizado por agricultores tradicionais ao longo da região amazônica. A ausência de padrões filogeográficos entre rios e regiões foi observada também com marcadores ncSSR e SNPs. Entretanto, quando a estrutura genética e genômica foi avaliada dentro das variedades mansas e bravas, alguns padrões contarstantes e tendências de estruturação entre rios foram observados. A ausência de padrões claros de estrutura genética e genômica entre diferentes rios não permitiu inferências sobre prováveis rotas de dispersão do cultivo a partir do seu centro de origem no sudoeste da Amazônia. No entanto, os padrões contrastantes de diferenciação genética e genômica dentro de variedades mansas e bravas podem estar associados a histórias de dispersão distintas para estes grupos de variedades de mandioca. Entre os locos genômicos, 658 SNPs estão possivelmente sob seleção positiva quando se considera a divergência entre variedades de mandioca cultivada e o parente silvestre. Destes, 202 SNPs podem estar especificamente associados com a seleção divergente entre variedades mansas e bravas. Estes locos podem estar em genes importantes para a domesticação inicial e seleção para características importantes do cultivo, e podem ser o ponto de partida para o melhor entendimento das bases genômicas da domesticação e diversificação da mandioca. / Manioc, or cassava, was domesticated in southwestern Amazonia, and is currently the most important staple crop in the world that originated there. After its initial domestication divergent selective pressures gave rise to the groups of sweet and bitter varieties. The current distribution of these groups is somewhat different across Amazonia, which may be due to distinct dispersal patterns of sweet and bitter varieties during the crop\'s domestication history. The aim of the present study was to evaluate genetic diversity and structure, genomics, and phylogeography of manioc cultivated by traditional farmers along the major rivers of Brazilian Amazonia. Phylogenetic analyses among matrilineages were performed based on the polymorphism of four chloroplastidial microsatellite markers (cpSSR). The evaluation of genetic diversity and structure were performed with 14 nuclear microsatellite markers (ncSSR), and a genomics approach was performed based on 5,871 single nucleotide polymorphism markers (SNPs). Considerable differentiation [FST = 0.78 (cpSSR), 0.28 (ncSSR), and 0.37 (SNPs)] was observed between cultivated varieties and Manihot esculenta ssp. flabellifolia, manioc\'s wild relative. No associations of cpSSR haplotypes with the groups of sweet and bitter varieties, nor with rivers were detected. Despite the lack of phylogeographic patterns, the analyses of genetic structure and relationships suggested that sweet and bitter varieties are equally related to wild populations. The second most important pattern of genetic structuring was observed between sweet and bitter varieties [FST = 0.08 (ncSSR) and 0.10 (SNPs)], although there was considerable overlap between groups. These results, combined with the high levels of genetic variability observed within sweet and bitter varieties, are due to the traditional management practices of smallholder farmers across Amazonia. The lack of phylogeographical patterns among rivers and regions were also observed with ncSSR and SNP markers. However, when the genetic and genomic structures were separately evaluated within sweet and bitter varieties, some contrasting patterns and tendencies of genetic structuring among the rivers was observed. The absence of clear patterns of genetic and genomic structure among different rivers did not permit inferences on probable routes of dispersal of the crop from its center of origin in southwestern Amazonia. Nevertheless, the contrasting patterns of genetic and genomic differentiation within sweet and bitter varieties may be associated with distinct dispersal histories for these groups of manioc varieties. Among the genomic loci, 658 SNPs are possibly under positive selection when considering the divergence between cultivated varieties of manioc and the wild relative. Of these, 202 SNPs may be specifically associated with divergent selection between sweet and bitter varieties. These loci may be located in genes important for initial domestication and selection for important characteristics of the crop, and may be a starting point for better comprehension of the genomic bases of manioc domestication and diversification.

Les progrès technologiques récents nous permettent d'accéder à la variation des génomes complets ce qui nous ouvre la porte d'une meilleure compréhension de leur diversification via des approches de génomique des populations et de génomique du paysage. Ce travail de thèse se base sur l'analyse des données de génomes complets (WGS) pour caractériser la diversité génétique des petits ruminants (chèvre et moutons) et rechercher les bases génétiques d'adaptations locales.Dans un premier temps, ce travail appréhende un aspect méthodologique et examine la précision et le biais de différentes approches d'échantillonnage des génomes pour caractériser la variabilité génétique, en les comparant aux données WGS. Nous mettons en évidence un fort biais des approches classiques (i.e. puces à ADN, capture de l'exome) ainsi que des séquençages de génomes à faibles taux de couverture (1X et 2X), et nous suggérons des alternatives basées sur un échantillonnage aléatoire de marqueurs dont la densité est variable selon les objectifs d'étude (évaluation de la diversité neutre, déséquilibre de liaison, signatures de sélection). Le jeu de données produit a permis d'évaluer l'état des ressources génétiques de différentes populations domestiques (races locales marocaines, iraniennes, races industrielles) et sauvages (aegagre, mouflon asiatique). Nous relevons une très forte diversité génétique dans les populations indigènes et sauvages qui constituent des réservoirs d'allèles et peuvent jouer un rôle important pour préserver le potentiel adaptatif des petits ruminants domestiques dans un contexte de changement climatique. L'étude plus approfondie des populations de chèvres du Maroc montre une forte diversité génétique faiblement structurée géographiquement, et met en évidence des portions de génome présentant des signaux de sélection. Leur étude montre l'existence de mécanismes adaptatifs potentiellement différents selon les populations (e.g. transpiration/halètement dans l'adaptation probable à la chaleur).Enfin, nous explorons les bases génétiques de l'adaptation locale à l'environnement chez les moutons et chèvres via une approche de génomique de paysage. En scannant les génomes de 160 moutons et 161 chèvres représentant la diversité éco-climatique du Maroc, nous identifions de nombreux variants et gènes candidats qui permettent d'identifier les voies physiologiques potentiellement sous-jacentes à l'adaptation locale. En particulier, il apparait que les mécanismes respiratoires et les processus cardiaques joueraient un rôle clé dans l'adaptation à l'altitude. Les résultats suggèrent que les chèvres et moutons ont probablement développé différents mécanismes adaptatifs pour répondre aux mêmes variations environnementales. Cependant, nous identifions plusieurs cas probables de voies adaptatives communes à plusieurs espèces. Par ailleurs, nous avons caractérisé les patrons de variations du niveau de différenciation de régions chromosomiques sous sélection en fonction de l'altitude. Cela nous permet de visualiser la diversité des réponses adaptatives selon les gènes (par exemple, sélection de variants à faible et/ou haute altitude). Ainsi, ce travail pose les bases de la compréhension de certains mécanismes d'adaptation locale. / Recent technological developments allow an unprecedented access to the whole genome variation and would increase our knowledge on genome diversification using population and landscape genomics. This work is based on the analysis of Whole Genome Sequence data (WGS) with the purpose of characterising genetic diversity in small ruminants (sheep and goats) and exploring genetic bases of local adaptation.First, we addressed a methodological aspect by investigating the accuracy and possible bias in the widely used genotyping approaches to characterize genetic variation in comparison with WGS data. We highlighted strong bias in conventional approaches (SNP chips and exome capture) and also in low-coverage whole genome re-sequencing (1X and 2X), and we suggested effective solutions based on sampling panels of random markers over the genome depending the purpose of the study (assessing neutral diversity, linkage disequilibrium, selection signatures). The various datasets produced allowed assessing genetic resources in various domestic (Moroccan and Iranian indigenous breeds and industrials) and wild populations (bezoars and Asiatic mouflons). We identified a very high diversity in indigenous and wild populations. They constitute a reservoir of alleles allowing them to play a possible key role in the preservation of these species in the context of global changes. The deep study of Moroccan goats showed a high diversity that is weakly structured in geography and populations, and highlighted numerous genomic regions showing signatures of selection. These regions identified different putative adaptive mechanisms according to the population (e.g. panting/sweating to adapt to warm/desert environment).Then, we explored genetic bases of local adaptation to the environment in sheep and goats using a landscape genomics framework. We scanned genomes of 160 sheep and 161 goats representing the eco-climatic Moroccan-wide diversity. We identified numerous candidate variants and genes, which allowed for identifying physiological pathways possibly underlying local adaptation. Especially, it seems that respiration and cardiac process have key roles in the adaptation to altitude. Our results suggest dissimilar adaptive mechanisms for the same environment in sheep and goats. However, we highlighted several cases of common metabolic pathways in different species. Moreover, we characterized some patterns for the variation of genetic differentiation in some candidate genomic regions over environmental gradients. This allowed us to visualise different adaptive reaction depending genes. This work points the way towards a better understanding of some mechanisms underlying local adaptation.

L'adaptation locale est un phénomène micro-évolutif qui peut survenir lorsque des populations d'une même espèce sont exposées à des conditions environnementales différentes.Si cet environnement exerce une pression sous forme de sélection naturelle, qu'il existe un potentiel adaptatif au sein des populations et que le flux de gènes est suffisamment modéré, les populations vont alors tendre vers un optimum adaptatif local.Dans cette thèse, je m'intéresse aux moyens méthodologiques de l'étude de l'adaptation locale d'une part, et à l'étude de ce phénomène le long d'un gradient d'altitude chez la plante alpine Arabis alpina d'autre part.Dans la première partie méthodologique, je montre que les méthodes de scan génomique pour détecter les marqueurs génétiques sous sélection peuvent souffrir de forts taux de faux positifs lorsqu'exposées à des jeux de données complexes, mais réalistes.Je présente ensuite une méthode statistique de détection de marqueurs génétiques sous sélection qui, contrairement aux méthodes existantes, utilisent à la fois la notion de différentiation génétique (ou Fst) et une information environnementale.Cette méthode a été développée de manière à limiter son taux de faux positifs de manière générale.J'offre enfin une perspective concernant les liens entre une expérience ancienne en biologie évolutive (l'expérience de jardin commun) et les nouveaux développements moléculaires et statistiques modernes.Dans la seconde partie empirique, je présente une analyse de la démographie d'A. alpina dans six populations naturelles. Outre qu'elle révèle des caractéristiques biologiques intéressantes sur cette espèce (faible espérance de vie, reproduction et survie très différentielle...), cette analyse montre que la croissance diminue et la survie augmente chez cette espèce avec la baisse de la température moyenne (donc avec l'altitude).Puisque ces analyses ne permettent pas d'exclure des hypothèses de dérive et de plasticité phénotype, je présente une analyse en jardin commun sur A. alpina qui permet de lisser les problèmes de plasticité phénotypique et qui, combinée à des analyses moléculaires, permettent d'exclure l'hypothèse de dérive.Les résultats montrent qu'il existe un syndrome phénotypique adaptatif lié à la température moyenne qui tend à des plantes plus petites, plus compactes, qui croissent et se reproduisent moins, dans les milieux froids.À l'aide des données moléculaires et de méthodes de scan génomique, je présente une liste de 40 locus qui peuvent être impliqués dans ce processus.Pour finir, je discute l'ensemble de ces résultats empiriques dans un contexte plus général d'écologie alpine. Je résume ensuite les principaux obstacles méthodologiques à l'étude de l'adaptation locale et je fourni quelques perspectives méthodologiques. / Local adaptation is a micro-evolutionary phenomenon, which arises when populations of the same species are exposed to contrasted environmental conditions.If this environment exert some natural selection pressure, if an adaptive potential exists among the populations and if the gene flow is sufficiently mild, populations are expected to tend toward a local adaptive optimum.In this thesis, I study the methodological means of the study of local adaptation on the one hand, and I investigate this phenomenon along an elevation gradient in the alpine plant Arabis alpina on the other hand.In the first, methodological part, I show that the genome scan methods to detect selection using genetic markers might suffer strong false positive rates when confronted to complex but realistic datasets.I then introduce a statistical method to detect markers under selection, which, contrary to existing methods, make use of both the concept of genetic differentiation (or Fst) and environmental information.This method has been developed in order to reduce its global false positive rate.Finally, I present some perspectives regarding the relationships between the relatively old ``common garden'' experiment and the new developments in molecular biology and statistics.In the second, empirical part, I introduce an analysis of the demographic characteristics of A. alpina in six natural populations. Besides providing interesting biological information on this species (low life expectancy, strongly contrasted reproduction and survival...), these analyses show that growth increase and survival decrease with the decrease of average temperature (hence with altitude).Since these analyses do not allow us to rule out hypotheses such as drift and phenotypic plasticity, I show the results of a common garden experiment which enable us to smooth phenotypic plasticity and, when combined with molecular data, enable us to rule out the hypothesis of drift.The results show the existence of an adaptive phenotypic syndrome, in which plants are smaller, are more compact, grow slower and reproduce less in cold temperature environments.Using the molecular data, I draw a list of 40 locus which might be involved in this adaptive process.In the end, I discuss these empirical findings as a whole to place them in a more general context of alpine ecology. I sum up the main methodological challenges when studying local adaptation and offer some methodological perspectives.

A mandioca foi domesticada no sudoeste da bacia amazônica, e presentemente é o cultivo alimentício amazônico mais importante no mundo. Após a domesticação inicial pressões seletivas divergentes deram origem aos grupos de variedades de mandiocas mansas e bravas. A distribuição atual destes grupos é um tanto diferente ao longo da Amazônia, o que pode ser reflexo de padrões de dispersão distintos de variedades mansas e bravas ao longo da história da domesticação do cultivo. O objetivo deste estudo foi avaliar a diversidade e estrutura genética, genômica e a filogeografia de mandiocas cultivadas por agricultores tradicionais ao longo dos principais rios da bacia amazônica brasileira. Análises filogenéticas de linhagens matrilineares foi realizada com base no polimorfismo de quatro marcadores microssatélites cloroplastidiais (cpSSR). A diversidade e estrutura genética foram avaliadas com 14 marcadores microssatélites nucleares (ncSSR), enquanto que a abordagem genômica foi realizada com base em 5.871 polimorfismos de nucleotídeos únicos (SNPs). Foi observada considerável diferenciação [FST = 0,78 (cpSSR), 0,28 (ncSSR), e 0,37 (SNPs)] entre as variedades cultivadas e Manihot esculenta ssp. flabellifolia, o parente silvestre da mandioca. Não foram detectadas associações de haplótipos cpSSR com os grupos de variedades mansas e bravas ou com os rios. Apesar da ausência de padrões filogeográficos, as análises de agrupamento e estrutura genética com base nos três tipos de marcadores avaliados sugeriram que as variedades mansas e bravas são igualmente relacionadas à população silvestre. O segundo padrão mais importante de estruturação genética foi observado entre variedades mansas e bravas [FST = 0,08 (ncSSR) e 0,10 (SNPs)], no entanto houve um considerável grau de mistura entre variedades de ambos os grupos. Estes resultados, juntamente com a elevada variação genética observada dentro de variedades mansas e bravas são resultantes do manejo realizado por agricultores tradicionais ao longo da região amazônica. A ausência de padrões filogeográficos entre rios e regiões foi observada também com marcadores ncSSR e SNPs. Entretanto, quando a estrutura genética e genômica foi avaliada dentro das variedades mansas e bravas, alguns padrões contarstantes e tendências de estruturação entre rios foram observados. A ausência de padrões claros de estrutura genética e genômica entre diferentes rios não permitiu inferências sobre prováveis rotas de dispersão do cultivo a partir do seu centro de origem no sudoeste da Amazônia. No entanto, os padrões contrastantes de diferenciação genética e genômica dentro de variedades mansas e bravas podem estar associados a histórias de dispersão distintas para estes grupos de variedades de mandioca. Entre os locos genômicos, 658 SNPs estão possivelmente sob seleção positiva quando se considera a divergência entre variedades de mandioca cultivada e o parente silvestre. Destes, 202 SNPs podem estar especificamente associados com a seleção divergente entre variedades mansas e bravas. Estes locos podem estar em genes importantes para a domesticação inicial e seleção para características importantes do cultivo, e podem ser o ponto de partida para o melhor entendimento das bases genômicas da domesticação e diversificação da mandioca. / Manioc, or cassava, was domesticated in southwestern Amazonia, and is currently the most important staple crop in the world that originated there. After its initial domestication divergent selective pressures gave rise to the groups of sweet and bitter varieties. The current distribution of these groups is somewhat different across Amazonia, which may be due to distinct dispersal patterns of sweet and bitter varieties during the crop\'s domestication history. The aim of the present study was to evaluate genetic diversity and structure, genomics, and phylogeography of manioc cultivated by traditional farmers along the major rivers of Brazilian Amazonia. Phylogenetic analyses among matrilineages were performed based on the polymorphism of four chloroplastidial microsatellite markers (cpSSR). The evaluation of genetic diversity and structure were performed with 14 nuclear microsatellite markers (ncSSR), and a genomics approach was performed based on 5,871 single nucleotide polymorphism markers (SNPs). Considerable differentiation [FST = 0.78 (cpSSR), 0.28 (ncSSR), and 0.37 (SNPs)] was observed between cultivated varieties and Manihot esculenta ssp. flabellifolia, manioc\'s wild relative. No associations of cpSSR haplotypes with the groups of sweet and bitter varieties, nor with rivers were detected. Despite the lack of phylogeographic patterns, the analyses of genetic structure and relationships suggested that sweet and bitter varieties are equally related to wild populations. The second most important pattern of genetic structuring was observed between sweet and bitter varieties [FST = 0.08 (ncSSR) and 0.10 (SNPs)], although there was considerable overlap between groups. These results, combined with the high levels of genetic variability observed within sweet and bitter varieties, are due to the traditional management practices of smallholder farmers across Amazonia. The lack of phylogeographical patterns among rivers and regions were also observed with ncSSR and SNP markers. However, when the genetic and genomic structures were separately evaluated within sweet and bitter varieties, some contrasting patterns and tendencies of genetic structuring among the rivers was observed. The absence of clear patterns of genetic and genomic structure among different rivers did not permit inferences on probable routes of dispersal of the crop from its center of origin in southwestern Amazonia. Nevertheless, the contrasting patterns of genetic and genomic differentiation within sweet and bitter varieties may be associated with distinct dispersal histories for these groups of manioc varieties. Among the genomic loci, 658 SNPs are possibly under positive selection when considering the divergence between cultivated varieties of manioc and the wild relative. Of these, 202 SNPs may be specifically associated with divergent selection between sweet and bitter varieties. These loci may be located in genes important for initial domestication and selection for important characteristics of the crop, and may be a starting point for better comprehension of the genomic bases of manioc domestication and diversification.

Depuis la découverte de Phaeodactylum tricornutum par Bohlin en 1897, sa classification au sein de l'arbre de la vie a été controversée. En utilisant des morphotypes ovales et fusiformes Lewin a décrit en 1958 plusieurs traits caractéristiques de cette espèce rappelant la structure des diatomées mettant ainsi fin à la controverse sur la classification de P. tricornutum au sein des Bacillariophycées. Pour se faire, trois morphotypes (ovale, fusiforme et triradié) de Phaeodactylum tricornutum ont été observés. Au cours d’une centaine d’années environ, de 1908 à 2000, 10 souches de Phaeodactylum tricornutum (appelées écotypes) ont été collectées et stockées soit de manière axénique ou en l’état avec leur populations naturelles de bactéries dans les centres des ressources génétiques pour algues, cryo-préservées quand cela est possible. Divers outils cellulaires et moléculaires ont été établis pour disséquer et comprendre la physiologie et l'évolution de P. tricornutum, et/ou les diatomées en général. Grâce à des décennies de recherche et les efforts déployés par de nombreux laboratoires que P. tricornutum est aujourd’hui considérée comme une espèce modèle des diatomées. Le sujet de ma thèse traite majoritairement de la composition génétique et épigénétique du génome de P. tricornutum ainsi que de la diversité morphologique et physiologique sousjacente au sein des populations naturelles prospectées à différents endroits du globe. Pour se faire, j’ai généré les profils chromatiniens en utilisant différentes marques des modifications post-traductionnelles des histones (chapitres 1 et 2) et a également comparé la variation naturelle dans la distribution de certaines marques clés entre deux populations d’écotypes (chapitre 4). Nous avons également généré une carte de la diversité génétique à l’échelle du génome chez 10 écotypes de P. tricornutum révélant ainsi la présence d'un complexe d'espèces dans le genre Phaeodactylum comme la conséquence d’une hybridation ancienne (chapitre 3). Sur la base de nombreux rapports antérieurs et des observations similaires au sein de P. tricornutum, nous proposons l’hybridation naturelle comme une base solide et une possibilité plausible pour expliquer la diversité des espèces chez lest diatomées. De plus, nous avons mis à jour les annotations fonctionnelles et structurelles du génome de P. tricornutum (Phatr3, chapitre 2) et mis au point un algorithme de logiciel convivial pour aller chercher les cibles CRISPR du système d’édition du génome CRISPR / cas9 chez 13 génomes de phytoplancton incluant P. tricornutum (chapitre 5). Pour accomplir tout cela, j'ai utilisé diverses méthodes à la pointe de l’état de l’art comme la spectrométrie de masse, l’immunoprécipitation de la chromatine suivie de séquençage à haut débit ainsi que les séquençages du génome entier, de l'ARN et des protocoles d'édition du génome CRISPR et plusieurs logiciels / pipelines de calcul. Ainsi, le travail de thèse fournit une plate-forme complète qui pourra être utilisée à l’avenir pour des études épigénétiques, de génétiques moléculaires et fonctionnelles chez les diatomées en utilisant comme espèce modèle Phaeodactylum tricornutum. Ce travail est pionnier et représente une valeur ajoutée importante dans le domaine de la recherche sur les diatomées en répondant à des questions nouvelles ouvrant ainsi de nouveaux horizons à la recherche en particulier en épigénétique qui joue un rôle important mais pas encore assez apprécié dans le succès écologique des diatomées dans les océans actuels. / Since the discovery of Phaeodactylum tricornutum by Bohlin in 1897, its classification within the tree of life has been controversial. It was in 1958 when Lewin, using oval and fusiform morphotypes, described multiple characteristic features of this species that resemble diatoms structure, the debate to whether classify P. tricornutum as a member of Bacillariophyceae was ended. To this point three morphotypes (oval, fusiform and triradiate) of Phaeodactylum tricornutum have been observed. Over the course of approximately 100 years, from 1908 till 2000, 10 strains of Phaeodactylum tricornutum (referred to asecotypes) have been collected and stored axenically as cryopreserved stocks at various stock centers. Various cellular and molecular tools have been established to dissect and understand the physiology and evolution of P. tricornutum, and/or diatoms in general. It is because of decades of research and efforts by many laboratories that now P. tricornutum is considered to be a model diatom species. My thesis majorly focuses in understanding the genetic and epigenetic makeup of P. tricornutum genome to decipher the underlying morphological and physiological diversity within different ecotype populations. To do so, I established the epigenetic landscape within P. tricornutum genome using various histone post-translational modification marks (chapter 1 and chapter 2) and also compared the natural variation in the distribution of some key histone PTMs between two ecotype populations (chapter 4). We also generated a genome-wide genetic diversity map across 10 ecotypes of P. tricornutum revealing the presence of a species-complex within the genus Phaeodactylum as aconsequence of ancient hybridization (Chapter 3). Based on the evidences from many previous reports and similar observations within P. tricornutum, we propose natural hybridization as a strong and potential foundation for explaining unprecedented species diversity within the diatom clade. Moreover, we updated the functional and structural annotations of P. tricornutum genome (Phatr3, chapter 2) and developed a user-friendly software algorithm to fetch CRISPR/Cas9 targets, which is a basis to perform knockout studies using CRISPR/Cas9 genome editing protocol, in 13 phytoplankton genomes including P. tricornutum (chapter 5). To accomplish all this, I used various state-of-the-art technologies like Mass-Spectrometry, ChIPsequencing, Whole genome sequencing, RNA sequencing, CRISPR genome editing protocols and several computational softwares/pipelines. In brief, the thesis work provides a comprehensive platform for future epigenetic, genetic and functional molecular studies in diatoms using Phaeodactylum tricornutum as a model. The work is an addon value to the current state of diatom research by answering questions that have never been asked before and opens a completely new horizon and demand of epigenetics research that underlie the ecological success of diatoms in modern-day ocean.