Metodologias de mapeamento de QTLs modernas empregam abordagem multivariada e se beneficiam da matriz de covariâncias fenotípicas para melhorar as estimativas de localização e efeitos de QTLs. No entanto, a correlação fenotípica pode ser em parte atribuída às relações de causalidade entre os fenótipos e mesmo as abordagens de mapeamento de QTLs multivariadas atuais têm desconsiderado tais relacionamentos. Dentre as metodologias científicas desenvolvidas para o estudo da causalidade em dados observacionais, destacam-se os modelos de equações estruturais e os modelos gráficos. Neste trabalho, foi estudado um conjunto de caracteres fenotípicos relacionados à morfologia de raízes, absorção de fósforo e acúmulo de biomassa em uma população composta de 145 linhagens endogâmicas recombinantes (RILs) do programa de melhoramento de milho da EMBRAPA Milho e Sorgo. O mapeamento de QTLs para os caracteres fenotípicos foi realizado utilizando mapeamento de múltiplos intervalos univariado (MIM) e multivariado (MT-MIM). A análise MIM revelou QTLs afetando diâmetro de raízes, área de superfície de raízes finas, peso seco da parte aérea e concentração de fósforo na parte aérea e nas raízes. A análise MT-MIM revelou 12 QTLs, com diferentes padrões de pleiotropia, com efeitos marginais para as sete variáveis analisadas. Um modelo de relacionamento causal entre os caracteres fenotípicos foi desenvolvido utilizando conhecimento prévio e modelagem de equações estruturais. O modelo de equações estruturais apresentou fluxo unidirecional de causalidade entre as variáveis, com as variáveis de morfologia de raízes exercendo efeito sobre as variáveis de acúmulo de biomassa, que por sua vez, têm efeito sobre as variáveis de absorção de fósforo. A aplicação do algoritmo PC para a descoberta de causalidade automatizada baseada nos padrões de independências condicionais não foi capaz de orientar todas as relações de causalidade descobertas, porém revelou um relacionamento mais complexo que o modelo de equações estruturais, com potenciais ciclos de retroalimentação causais. O emprego de algoritmos de descoberta de causalidade baseados em informações de QTLs, chamados QDG e QPSO, permitiu a orientação de todos os relacionamentos de causalidade encontrados pelo algoritmo PC e confirmou a existência de dois ciclos vizinhos de relacionamento causais entre as variáveis estudadas. Como regra geral, os QTLs pleiotrópicos detectados pela metodologia MT-MIM apresentaram efeitos sobre caracteres fenotípicos alinhados causalmente nos modelos propostos pelos algoritmos PC e QDG, sugerindo que alguns dos QTLs detectados são na realidade efeitos indiretos de QTLs situados em posição mais elevada no modelo causal. O emprego da abordagem MT-MIM aliada à análise de causalidade permitiu melhor compreensão da arquitetura genética dos caracteres de morfologia de raiz, acumulação de biomassa e aquisição de fósforo em milho. / Modern QTL mapping approaches are multivariate and take advantage of the phenotypic covariance matrix to improve estimates of QTL positions and effects. However, phenotypic correlation can also be assigned to the causal relationship among phenotypes, and even modern multivariate QTL analysis does not take these relationships into account. Structural equation models and graphical models are the main methodologies to study causality from observational data. We studied a set of phenotypes related to root morphology, biomass accumulation and phosphorus acquisition in maize. These phenotypes were measured in a maize population from the EMBRAPA breeding program composed of 145 recombinant inbred lines (RILs) derived from the crossing of two divergent lines for phosphorus acquisition efficiency. QTL mapping for the traits was performed using univariate (MIM) and multivariate (MT-MIM) multiple interval mapping. MIM analysis revealed QTL affecting root diameter, fine root surface area, shoot dry weight and root dry weight. MT-MIM analysis revealed 12 QTL with different pleiotropy patterns and QTL with marginal effects affecting all seven studied characters. A causal model for phenotype characters was developed using a priori knowledge and structural equation model techniques. The structural equation model presented an unidirectional causal flow among the variables, with root morphological traits exerting causal effects over biomass traits, which in turn cause phosphorus acquisition traits. Using PC algorithm for an automatic search of causal models based on conditional independence was not able to orient all discovered causal relationships among traits but revealed a more intricated relationship than the structural equation model, with potential causal feedback loops among the traits. Employing causal search algorithms based on QTL information (named QDG and QPSO) allowed the orientation of all causal relationships detected by PC algorithm and it has also confirmed the presence of two neighbor causal cycles among the studied traits. As a general rule, pleiotropic QTL detected by MT-MIM approach exerted effects over traits according to the causal model discovered by PC and QDG algorithms, suggesting that some of the QTL detected effects were indirect effects of QTL located upstream at the proposed causal model. Employing MT-MIM approach and causal analysis has allowed a better comprehension of genetic architecture underlying root morphology, biomass accumulation and phosphorus acquisition traits in maize.

Adult hippocampal neurogenesis is regulated at various levels and by various factors. Genetic influence is an important key determinant of adult neurogenesis and exerts its effects at all levels. In vivo studies have suggested that adult hippocampal neurogenesis is highly variable and heritable among different laboratory strains of mice. To dissect the genetic effect from other contributing factors, it is necessary to study adult neurogenesis under highly controlled environment conditions. We extracted adult hippocampal precursor cells (AHPCs) from 20 strains of the BXD set of recombinant inbred mice, cultured them and studied the effect of genetic background on neurogenesis. The BXD panel consists of mouse lines derived from an intercross between inbred parentals C57BL/6J and DBA/2J. Both of the parentals are fully sequenced and all the strains are well characterized in terms of genotypic and phenotypic characteristics. This allows us to use advanced genetic techniques to identify novel genomic loci and gene-gene interactions important in adult neurogenesis. Comparison of the AHPCs from 20 BXD strains, with respect to cell proliferation and neuronal and astrocytic differentiation in vitro, revealed a large variation for these traits across the strains. Proliferation, as measured by BrdU incorporation, showed over two- fold differences between the extremes. Similar differences were observed for neurogenic (4-fold) and astrogenic differentiation (2-fold). These three traits all showed strong heritability values indicating that the differences were mainly attributed to the genetic component. QTL mapping, with these phenotypic data, revealed that there was no major contribution from single loci controlling these traits. Instead, we found many loci with smaller effects associated with these traits. Gene expression profiling using RNA samples from proliferating cultures of the 20 BXD mice strains yielded two cis eQTL candidates that directly regulated proliferation, LRP6 and Chchd8. LRP6 is well known as a co-receptor of Wnt signaling, but the function of Chchd8 is not known. Further experimentation, using over expression and gene silencing demonstrated that LRP6 negatively regulates AHPCs proliferation. Thus, from this study using a system genetics approach, we were able to identify, LRP6 as a novel regulator of adult hippocampal neurogenesis.

No description available.

Parmi les contraintes abiotiques la salinité est considérée comme un problème majeur, qui affecte le fonctionnement des plantes, en particulier leur croissance et leur rendement. Afin d’étudier le contrôle génétique de la tolérance à la salinité chez Medicago truncatula, plante modèle de la famille des légumineuses, deux expérimentations ont été réalisées. La première expérimentation visait à étudier l’effet de la contrainte saline sur différents paramètres morpho-physiologiques pour un panel de génotypes de M. truncatula afin de déterminer les traits de phénotypage pour la tolérance à la salinité. Les génotypes A17, TN1.11, DZA315.16, A20, TN1.12 et F83005.5 ont été sélectionnés parmi des lignées originaires de différents pays méditerranéens, qui ont été déjà séquencées (http://www1.montpellierinra.fr/BRC-MTR/mauguio/mauguio.php). Les génotypes ont été étudiés sous 6 traitements salins (0, 30, 60, 90,120 et 150 mM NaCl) dans un essai factoriel sous forme de blocs complets aléatoires en trois répétitions. L’analyse de la variance montre des différences significatives entre les niveaux de salinité et une interaction entre les génotypes et les traitements salins concernant la plupart des caractères étudiés. Le génotype « DZA315.16 » présente les valeurs les plus importantes concernant les effets principaux pour les caractères morphologiques alors que « TN1.11 » présente les valeurs les plus faibles. La projection verticale de la surface foliaire de la plante (Leaf Area=LA), significativement corrélée à la biomasse des plantes, apparaît comme un trait d’intérêt pour le phénotypage de la tolérance à la salinité. La concentration saline la mieux adaptée pour démontrer les différences parmi les lignes étudiées se situe entre 90 et 120 mM NaCl. Le génotype « TN1.11 » contrairement à « DZA315.16 » et à « Jemalong-A17 » présente un maintien de la surface foliaire de la plante en réponse à la salinité. Pour la deuxième expérimentation, une population de cent lignées recombinantes (Recombinant Inbred Lines=RILs) produite par le croisement entre « TN1.11 » et « Jemalong-A17 » a été retenue pour l’analyse du contrôle génétique de la tolérance à la salinité. Les RILs ont été développés par la méthode de descendant mono graines (Single Seed descent= SSD) jusqu’ à la génération F6 à l’INP-ENSAT, France. Le plan d’experimentation est « Spli plots » , sous forme de blocs randomisés avec trois répétitions et deux conditions : traitement salin (100 mM NaCl) et témoin (eau). L’expérience a été menée pour déterminer la variabilité génétique et pour identifier les QTLs contrôlant les caractères morphologiques et physiologiques chez la population des lignées recombinantes (RILs). L’analyse de la variance a montré une large variation génétique et une ségrégation transgressive pour les caractères étudiés. La différence entre la moyenne des RILs et la moyenne de leurs parents n’est pas significative concernant tous les caractères étudiés dans les deux conditions, ce qui montre que les RILs utilisées dans notre expérimentation sont représentatives de toutes les lignées recombinantes possibles du croisement « TN1.11 x Jemalong-A17 ». 21 QTLs ont été détectés dans la condition témoin et 19 QTLs ont été identifiés sous contrainte saline (100 mM NaCl). Le pourcentage de la variance phénotypique expliqué par les QTLs varie entre 4.60% et 23.01%. Certains de ces QTLs sont spécifiques à la condition saline, ce qui démontre l’existence du contrôle génétique de la tolérance à la salinité chez M. truncatula ; tandis que les autres ne sont pas spécifiques et contrôlent un même caractère dans les deux conditions. Des QTLs superposés concernant différents caractères ont été aussi observés. Les résultats fournissent des informations importantes en vue de futures analyses fonctionnelles de la tolérance à la salinité chez M.truncatula et pour d’autres espèces voisines. / Among abiotic stresses salinity is considered as a serious problem affecting plant functions especially growth and yield. In order to study the genetic control of salt stress in the model legume Medicago truncatula, two experiments were performed. The first experiment was conducted to study the effect of salt stress on some morpho-physiological parameters in M. truncatula genotypes and to determine the eventual use of some traits as tolerance criteria. Genotypes including A17, TN1.11, DZA315.16, A20, TN1.12 and F83005.5 are selected through a sequenced lines collection (http://www1.montpellierinra.fr/BRC-MTR/mauguio/mauguio.php) which are originated from different Mediterranean countries. Genotypes were studied under 6 salinity treatments (0, 30, 60, 90,120 and 150 mM NaCl) in a factorial design based on randomized complete blocks with three replications. Analysis of variance show significant differences among genotypes, salinity levels and interaction between genotypes and salt treatments for most of studied traits. “DZA315.16” genotype presents the highest main effect values for morphological traits whereas”TN1.11” has low values. Vertically projected leaf area (LA); show the highest variability through all studied salt concentrations. The best concentration to find differences between parental lines is 90 to 120 mM Nacl. A segregating population of recombinant inbred lines (100 RILs) of M.truncatula derived from a cross between TN1.11 and Jemalong-A17 was used for the second experiment. RILs were developed by single-seed descent until F6 generation at the INP-ENSAT, France. The experiment was undertaken to determine the genetic variability and to identify QTLs controlling several traits related to plant growth and physiology, in the population of recombinant inbred lines (RILs). Analyses of variance showed a large genetic variation and transgressive segregation for the traits studied. The difference between the mean of RILs and the mean of their parents was not significant for all of the traits in both conditions, showing that the RILs used in our experiment are representative of the possible recombinant lines from the cross TN1.11 x A17. A total of 21 QTLs were detected under control and 19 QTLs were identified under 100mM salt stress conditions. The percentage of total phenotypic variance explained by the QTLs ranged from 4.60% to 23.01%. Some of the QTLs were specific for one condition, demonstrating that the genetic control of a traits differed under control and salt stress conditions. Some others are non-specific and control a trait in both conditions. Overlapping QTLs for different traits were also observed. The results provide important information for further functional analysis of salt tolerance in M. truncatula

Aujourd'hui, alors que le nombre de marqueurs génétiques disponibles augmente rapidement, de nouvelles populations doivent être créées pour exploiter au mieux cette quantité d'informations dans le but de mieux comprendre l'architecture génétique de caractères complexes. Les populations de type MAGIC ont été créées pour rassembler les avantages des populations bi-parentales et des panels d'associations, la bonne puissance de détection et une localisation précise. L'objectif de cette thèse était d'étudier l'intérêt de la population MAGIC INRA pour l'analyse de l'architecture génétique de la précocité de floraison. Cette population a été créée à partir de 60 parents brassés durant 12 générations de panmixie grâce à l'introduction d'un gène de stérilité mâle (ms1b). Cette étude a été réalisée sur 56 parents toujours disponibles en banque de graines et 380 lignées dérivées de la population après les 12 générations de recombinaison. Cette population a été génotypée avec la puce 9K iSelect, représentant environ 5 000 SNPs localisés sur tout le génome, additionnée de 14 marqueurs localisés dans des gènes candidats. Ce jeu de données moléculaires a été complété par des données fines de phénotypage de la précocité de floraison. Suite aux 12 générations de panmixie, le DL de cette population a été très réduit, à longue comme à moyenne distance (<10cM). Ce faible DL nous a amené à développer un algorithme basé uniquement sur le DL qui ordonne les marqueurs de manière à avoir un DL décroissant monotone avec la distance. L'algorithme ordonne globalement de la même manière que la carte génétique les marqueurs à longue distance mais à courte distance le DL est moins lié à la distance génétique. La différence réside sur l'équilibre entre les effets de la recombinaison et de la dérive génétique sur le DL. L'intérêt de la population MAGIC INRA pour détecter des QTLs a ensuite été étudié avec deux approches : une approche évolutionniste et une approche de génétique d'association. La première approche détecte les loci soumis à sélection par comparaison des fréquences alléliques de la population initiale (G0) et de la population évoluée (G12) grâce à une nouvelle méthode. La population initiale est composée des parents pondérés par une contribution estimée avec une nouvelle méthode bayésienne. 26 régions génomiques soumises à sélection ont été détectées. Une analyse de génétique d'association avec les marqueurs détectés sous sélection a montré que respectivement cinq et trois zones étaient associées à la précocité avec un semis d'automne et au caractère printemps/hiver. Une analyse phénotypique a effectivement mis en évidence la précocification de la date de floraison et une augmentation de la proportion de plantes de type printemps. Une analyse de génétique d'association a ensuite été réalisée sur les lignées SSD sur 12 caractères x environnements i.e. la date d'épiaison et le temps de remplissage du grain mesurés dans six environnements. Les tests d'association ont aussi été réalisés avec des variables synthétisant l'information présente dans plusieurs traits phénotypiques soit avec une ACP, soit avec un modèle écophysiologique. Au total, toutes ces analyses ont détecté six QTLs dont trois correspondants à des gènes majeurs. Parmi ces six QTLs, deux sont spécifiques des caractères mesurés avec un semis d'automne et deux avec ceux mesurés avec un semis de printemps. / Nowadays, with the dramatically increase of available molecular markers, there is a deep need for new populations allowing to exploit all of this information to better understand the genetic architecture of complex traits. MAGIC populations as they are built to bring together bi-parental populations and association panel advantages, provide such powerful detection and fine mapping capacities. The aim of these PhD was to study the MAGIC INRA population usefulness for the study of genetic architecture of earliness. This population is derived from 12 cycles of random crosses between 60 founders, turning wheat from selfing to outcrossing thanks to the use of a nuclear male sterility gene (ms1b, Probus donor). This population is composed of 56 parents still available and 380 SSD lines. Parents and SSD lines were genotyped using the 9K iSelect SNPs array, providing around 5 000 SNPs on the whole genome, as well as 14 addition markers located in candidate genes. They were also finely phenotyped for earliness traits. With the 12 panmictic generations, the population LD decreased strongly, especially at long and medium distance (<10cM). This allowed us to develop an algorithm mapping markers on the sole pairwise LD information, ordering markers in a way to have the LD decreasing along the distance. When considering long distances, overall the results were consistent with the order found on genetic maps while at short distance LD was poorly linked to genetic distance. These differences between long and short distances were linked to the balance between recombination and drift effects on LD. The usefulness of the MAGIC INRA population for QTL detection was analyzed with two approaches: an evolutionary approach and an association genetics approach. The first one detects loci under selection by identifying high shift in allelic frequency with a new method. The initial population was composed of founders weighted by a contribution estimated with a new Bayesian method. 26 genomic areas under selection were detected. An association genetics analysis with the markers detected as under selection showed respectively five and three genomic regions associated with earliness and growth habit. Actually the G12 population was found phenotypically earlier than the G0 and with more spring individuals. A broader association genetics analysis was performed on G12 population, studying 12 traits x environments i.e. heading date and grain filling time, both observed in six environmental conditions. Two additional integrated traits from either PCA or ecophysiological model were also analyzed. In all, these different analyses detected six QTLs, three of them corresponding to candidate genes. Among these six QTLs, two were specific to autumn sowing and two specific to spring sowing.

Soja (Glycine max (L.) Merrill) é uma das mais importantes oleaginosas no mundo e sua relevância pode ser também mensurada pela extensão da sua produção no Brasil, onde representa quase a metade do total da área plantada com grãos. A importância da soja está também associada à multiplicidade de utilização do grão, como por exemplo, na alimentação humana e animal, na indústria química e na geração de energia como biodiesel. A produtividade da soja tem sido frequentemente impactada devido ao ataque de pragas e doenças. Dentre as espécies de fitonematoides, o nematoide-das-lesões-radiculares (Pratylenchus brachyurus Godfrey), tem gerado significativas perdas econômicas aos produtores, variando entre 30 a 50%, dependendo da infestação da cultura. Este trabalho teve como objetivo identificar QTLs (Quantitative Trait Loci), a partir de um conjunto de fenótipos associados à resistência ao nematoide-das-lesões-radiculares utilizando a abordagem de mapeamento multivariado. Uma população de 174 indivíduos F2, obtidos a partir do cruzamento entre duas linhagens de soja FTPG06A e FTPG12X (com baixo fator de reprodução do nematoide), foi utilizada para a obtenção dos valores genéticos preditos (BLUPs - Best Linear Unbiased Predictions) de cinco características estudadas e também para a genotipagem com o SoySNP6k Bead Chip. As características avaliadas foram: fator de reprodução (FR), peso fresco (PA) e comprimento (CA) da parte aérea, peso fresco (PR) e comprimento (CR) da raiz. Um total de 1.240 marcadores SNP (Single Nucleotide Polymorphism) foram mapeados nos 20 grupos de ligação (GLs) da soja. O comprimento total do mapa foi de 3.084,46 centiMorgans (cM), com intervalo médio de 2,54 cM entre marcadores adjacentes. A identificação de QTLs (Quantitative Trait Loci) para os caracteres fenotípicos foi realizada utilizando-se o mapeamento de intervalos múltiplos univariado (MIM) e multivariado (MT-MIM), com estimativa dos efeitos principais dos QTLs e análises de epistasia envolvendo pares de QTLs. Na abordagem MIM foram identificados três QTLs associados à variável CR, nos GLs (Grupos de Ligação) A2 e E (2 QTLs), explicando um total de 34,22% da variação fenotípica dessa variável para a população em estudo. Na abordagem MT-MIM, foram selecionados dois conjuntos de variáveis, de acordo com a correlação entre as mesmas. Três regiões genômicas foram reveladas, sendo estatisticamente significativas para as variáveis CR, FR e CA. A comparação dos seis QTLs identificados no presente estudo com o banco de dados de QTLs do Soybase forneceu evidências de que cinco QTLs, ainda não foram descritos na literatura. Uma busca por sequências candidatas em duas regiões de interesse, associadas à variável FR, foi realizada, com base na plataforma de dados genômicos PlantGDB. Várias sequências candidatas indicam relação com mecanismos importantes na resposta das plantas a estresses bióticos. Dessa forma, os resultados obtidos no presente estudo forneceram informações para auxiliar na melhor compreensão da arquitetura genética dos caracteres quantitativos analisados. / Soybean (Glycine max (L.) Merrill) is one of the most important oil crop worldwide and its relevance can also be measured by the extension of soybean grain production in Brazil, where it represents almost half of the total planting area with grains crops. Importance of soybean is also related to a multiplicity of usage of the grain, for example, in human consumption, animal feed, chemical industry and for energy generation as biofuel. Soybean yield has been frequently reduced by occurrence of pest and diseases. Among phytonematodes species, root lesion nematode (Pratylenchus brachyurus Godfrey) has caused significant economic losses to farmers, ranging from 30 to 50%, depending on crop infestation. This research aimed to identify QTLs (Quantitative Trait Loci), from a set of phenotypes associated with resistance to root lesion nematode, using the multivariate multiple interval mapping. A mapping population of 174 F2 plants derived from a by-parental cross between two soybean breeding lines FTPG06A and FTPG12X (with low nematode reproduction factor), was used for prediction of genetic values (BLUPs - Best Linear Unbiased Predictions) for five traits studied and also for genotyping with SoySNP6k Bead Chip. Traits evaluated were reproduction factor (FR), shoot weight (PA), shoot length (CA), root weight (PR) and root length (CR). A total of 1,240 SNP (Single Nucleotide Polymorphism) markers were mapped into 20 soybean linkage groups (LG). A total map length was 3,084.46 centiMorgans (cM) with an average of 2.54 cM between flanking markers. QTL mapping for those traits was performed using univariate (MIM) and multivariate (MT-MIM) multiple-interval mapping, with main QTL effects estimates and epistasis analysis between QTL pairs. MIM analysis identified three QTLs associated to CR trait at LG A2 and LG E (2 QTL), explaining 34,22% of phenotypic variation estimated for this mapping population. For MT-MIM analysis, two sets of traits were selected, according to the correlation among them. Three genomic regions statistically significant for CR, FR and CA traits were identified. Comparison between six identified QTL and QTL database at Soybase provided evidence that five QTL have not been published yet. Search for candidate sequences located in two regions associated with FR trait were further performed on the PlantGDB genomic data platform. Several candidate sequences indicate relationships with important plant response mechanisms to biotic stresses. Thus, results obtained in the present study provided information to improve knowledge of the genetic architecture of the analyzed quantitative traits.

A soja é a cultura agrícola brasileira que mais se expandiu nas últimas três décadas. Atualmente é uma commodity comercializada em larga escala na forma de grão, farelo e óleo. Por ser uma importante fonte de proteína, possui papel fundamental na indústria alimentícia, tanto humana quanto animal. O monocultivo em extensas áreas tem ocasionado o aumento da vulnerabilidade da cultura a patógenos e insetos-praga, com consequências relevantes na produção dos grãos. Dentre tais insetos-praga, os percevejos fitófagos representam um dos grupos de maior relevância para a cultura, pois podem comprometer o rendimento, a qualidade e a sanidade dos grãos. As perdas no rendimento da cultura devido ao ataque por percevejos são superiores a 30%, e o comprometimento no valor germinativo das sementes pode ser superior a 50%. Este trabalho teve como objetivo identificar QTLs (Quantitative Trait Loci), a partir de um conjunto de fenótipos associados à resistência aos percevejos e a caracteres agronômicos, utilizando a abordagem de mapeamento multivariado. Uma população de 228 indivíduos F2, obtida a partir do cruzamento entre as cultivares IAC-100 e CD-215, foi utilizada para a obtenção dos dados genotípicos. Os caracteres agronômicos avaliados na geração F2:3 foram: número de dias para o florescimento (NDF), altura da planta no florescimento (APF), número de dias para a maturidade (NDM), altura da planta na maturidade (APM), acamamento (AC), valor agronômico (VA) e produtividade de grãos (PG). Oito caracteres associados à resistência aos percevejos foram avaliados: período de granação (PEG), retenção foliar (RF), número de vagens por planta (NVP), índice percentual de dano nas vagens (IPDV), número de sementes (NS), peso de cem sementes (PCS), peso de sementes boas (PSB) e peso de sementes manchadas (PSM). O mapa genético obtido, representando os 20 grupos de ligação (GLs) da soja, foi constituído por 417 marcadores SNP (Single Nucleotide Polymorphism), 61 SSR (Simple Sequence Repeat), 30 AFLP (Amplified Fragment Length Polymorphism) e 8 marcadores TRAP (Target Region Amplification Polymorphism). A cobertura do genoma da soja foi de 2.814,82 centiMorgans (cM), com um intervalo médio de 5,46 cM entre marcadores adjacentes. A identificação de QTLs (Quantitative Trait Loci) para os caracteres fenotípicos foi realizada utilizando-se o mapeamento de intervalos múltiplos univariado (MIM) e multivariado (MT-MIM), com estimativa dos efeitos principais dos QTLs. A abordagem MIM revelou um total de 60 QTLs, distribuídos por 13 GLs da soja, sendo 29 QTLs associados a caracteres de resistência aos percevejos e 31 QTLs relacionados a caracteres agronômicos. A percentagem da variação fenotípica explicada pelos QTLs (R2) variou de 14,27% para AC a 65,45% para NDM. Na abordagem MT-MIM, foram selecionados nove conjuntos de variáveis, de acordo com a correlação entre as mesmas. Foram reveladas 20 regiões genômicas distintas, com uma alta tendência de concentração de QTLs em posições similares. No geral, para a maioria das características, os valores marginais de R2 obtidos para os modelos MT-MIM foram superiores em relação aos modelos MIM, variando de 27,98% para APF a 65,30% para NDM. A abordagem MT-MIM permitiu a identificação de 13 novas posições genômicas, com efeito em pelo menos uma das variáveis analisadas, que não foram reveladas nos modelos MIM. Uma comparação com o banco de dados do Soybase forneceu evidências de que muitos QTLs, identificados nesta população em estudo, coincidem com QTLs descritos em outros backgrounds genéticos. No entanto, 56 QTLs identificados no presente estudo ainda não foram descritos na literatura. A maioria dos QTLs identificados explicam, individualmente, até 10% da variação fenotípica das características avaliadas. No entanto, QTLs presentes em oito novas regiões identificadas pela abordagem MT-MIM e oito novos QTLs identificados pela abordagem MIM contribuíram para explicar uma maior percentagem da variação dos fenótipos estudados. Esses QTLs devem ser melhor investigados considerando sua relevância para a seleção simultânea de características de interesse, permitindo uma maior eficiência de seleção e um maior ganho genético. Os resultados obtidos no presente estudo forneceram informações para auxiliar na melhor compreensão da arquitetura genética dos caracteres quantitativos analisados, bem como sobre a relação genética entre os mesmos. / Soybean is the Brazilian crop with the most expansion along the past three decades. Currently, it is a commodity commercialized in large scale as grain, bran and oil. Because it is an important source of protein, it plays a fundamental role in the food industry, both human and animal. Soybean monoculture in large areas has increased crop vulnerability to pathogens and insect pests, with significant consequences on grain production. Among such pest insects, stink bugs are considered a major pest of soybean crop, feeding directly on seeds, reducing yield and seed quality. Losses in crop yield due to stink bugs attack are greater than 30% and seed germination compromising can be greater than 50%. This study aimed to identify QTL (Quantitative Trait Loci), for stink bug resistance traits and agronomic traits using the multivariate multiple interval mapping. An F2 mapping population of 228 plants derived from a biparental cross between IAC-100 and CD-215 was used for genotyping. An F2:3 population was developed to evaluate eight stink bug resistant traits such as graining period, leaf retention, pod number per plant, percentage of pod damage, number of seeds, hundred seed weight, weight of healthy seeds and spotted seed weight. Other seven agronomic traits were evaluated such as number of days to flowering, plant height at flowering, number of days to maturity, plant height at maturity, lodging, agronomic value and grain yield. A total of 417 SNP (Single Nucleotide Polymorphism) markers, 61 SSR (Simple Sequence Repeat), 30 AFLP (Amplified Fragment Length Polymorphism) and 8 TRAP (Target Region Amplification Polymorphism) markers were mapped into 20 soybean linkage groups. The total map length was 2,814.82 cM with an average of 5.46 cM between markers. QTL mapping for those traits was performed using univariate (MIM) and multivariate (MT-MIM) multiple interval mapping, with main QTL effects estimates. MIM analysis identified a total of 60 QTL, through 13 soybean linkage groups, with 29 QTL related to stink bug resistant traits and 31 QTL related to agronomic traits. Phenotypic variation explained by QTL ranged from 14.27% for lodging to 65.45% for number of days to maturity. The traits were divided into nine groups for MT-MIM analysis considering their correlation coefficient. Twenty different genomic regions were identified showing a very high QTL clustering. For most of the traits phenotypic variation estimates for MT-MIM models were higher than MIM models, ranging from 27.98% to 65.30% for plant height at flowering and number of days to maturity, respectively. MT-MIM analysis showed 13 genomic regions controlling at least one of the evaluated traits which were not identified at MIM analysis. Comparison between identified QTL and QTL database at Soybase demonstrated that some QTL were similar to those described in different genetic background. However, 56 QTL detected in the present study were described for the first time in literature. Most of the QTL identified explain, individually, less than 10% of phenotypic variation. However, eight genomic regions identified with MT-MIM analysis and eight QTL identified with MIM analysis explain a great amount of phenotypic variation. These QTL should be investigated considering their importance for simultaneous selection for a high genetic gain. Results obtained in the present study provided information for a better understand of genetics architecture underlying quantitative traits studied and the genetic relation among them.

Com a disponibilidade de tecnologias de genotipagem robustas, fornecendo milhares de marcadores moleculares a um baixo custo por amostra, tornou-se mais acessível escanear o genoma todo. Logo, as abordagens de mapeamento associativo, baseado no cálculo do desequilíbrio de ligação, e a busca de regiões do genoma que apresentam sinais de seleção foram favorecidas. Essas ferramentas apresentam um grande potencial a ser explorado em programas de melhoramento de soja, auxiliando na obtenção de informações valiosas em relação à arquitetura genética dos caracteres de interesse agronômico e à dinâmica dos processos de seleção. Portanto, os objetivos deste estudo foram: (i) verificar os efeitos da relação genética entre genótipos no desequilíbrio de ligação; (ii) obter informações em relação a arquitetura genética para dez caracteres por meio da abordagem de mapeamento associativo; e (iii) identificar regiões sob seleção diferencial em uma população representada por cultivares brasileiras comparadas a de uma população de genótipos exóticos de diferentes origens geográficas. Para isto, 95 genótipos de soja foram genotipados utilizando a plataforma da Affymettrix (180 K Axiom&reg; Soybean Genotyping Array). O efeito da estrutura populacional sobre DL foi investigado utilizando como correções as matrizes resultantes do software STRUCTURE, DAPC, PCA e a matriz de parentesco. Para o mapeamento associativo, os genótipos foram fenotipados para dez caracteres. Foram realizadasvárias abordagens para detecção de sinais de seleção: estimação da diferenciação populacional nas regiões do genoma por meio do índice de fixção (FST) e cross-population composite likelihood ratio test (XP-CLR); identificação de regiões do genoma com redução de diversidade nucleotídica (&mu;); e a presença de blocos de haplótipos. Ao utilizar medidas de correções para estimação do DL no genoma da soja observamos a influência da estrutura da população e do parentesco nos padrões de DL. Pelo mapeamento associativo, foram identificados 181 marcadores associados para dez caracteres avaliados em soja. Além disso, foi verificada a complexa interação entre regiões envolvidas no controle de caracteres quantitativos (QTL) e ambientes. Regiões diferencialmente selecionadas foram identificadas entre a população de genótipos brasileiros e de materiais de origens diversas, demonstrando que essas passaram por processos de seleção divergente. / The availability of robust genotyping technologies providing thousands of markers with a low-cost per sample, whole-genome scans are becoming more accessible. Hence, associative mapping approaches, based on the calculation of linkage disequilibrium (LD), and the detection of signals of selection were favored. These tools have a great potential to be explored in soybean breeding programs, helping to obtain valuable information about the genetic architecture of the characters of agronomic interest and of the dynamics of selection processes. Therefore, the objectives of this study were: (i) to verify the effects of the genetic relationship among genotypes on soybean linkage disequilibrium; (ii) to obtain information about the genetic architecture in ten soybean traits with an association mapping approach; and (iii) to identify regions under differential selection between a population represented by Brazilian cultivars compared to a population of exotic genotypes from different geographic origins. For this, 95 soybean genotypes were genotyped using an Affymetrix (180 K Axiom&reg; Soybean Genotyping Array) platform. The effect of the population structure on LD was investigated using as corrections the matrices resulting from the software STRUCTURE, DAPC, PCA and a kinship matrix. We phenotyped ten soybean traits in order to carry the association mapping. Several approaches were carried aiming to detect selection signals: estimation of population genetic differentiation in the genomic regions with fixation index (FST) and cross-population composite likelihood ratio test (XP-CLR); identification of genomic regions with reduced nucleotide diversity (&mu;); and the presence of haplotype blocks.Using the LD corrections, it was showed the influence of population structure and kinship on LD patterns in soybean. By the association mapping, we identified 181 markers associated with the ten traits evaluated in soybean. In addition, complex interactions between QTL and environments were verified. Differentially selected regions were identified between theBrazilian genotypes population and the population of materials from diverse origins, demonstrating that they underwent divergent selection processes.

La connaissance de la diversité génétique de R. solanacearum autant que la connaissance du déterminisme génétique de la résistance sont importantes pour établir une stratégie de lutte contre cette bactérie ubiquiste. Elles permettent de choisir et d'orienter des méthodes de lutte et le développement d'outils de contrôle de quarantaine adaptés. Dans un contexte scientifique de plus en plus tourné en génétique par l'approche plante modèle et dans lequel les Solanacées occupent la place de plante pionnière d'une part, et d'autre part par l'émergence de contournement des résistances liées à la grande diversité génotypique et phénotypique de R. solanacearum (au point où on parle ici de complexe d'espèce), l'objectif de cette thèse est de contribuer à établir les bases d'un programme d'amélioration des Solanacées contre cette phytobactériose majeure au Cameroun. En recourant à une approche moléculaire, notamment la méthode de caractérisation phylophasique qui décrit la diversité génétique des souches de R. solanacearum en quatre phylotypes et séquevars, nous avons révélé l'existence de trois phylotypes dans ce pays d'Afrique dont la diversité agro-climatique lui vaut le surnom d'" Afrique en miniature ". Ces phylotopes sont le I (Asiaticum), II (Americanum) et III (Africanum). Les souches du phylotype III du Cameroun se sont révélées génétiquement distinctes des souches africaines de référence originaires du Zimbabwe et de l'Océan Indien. Les souches isolées sur pomme de terre dans les hauts plateaux de l'Ouest du Cameroun se classent dans les deux phylotypes, à savoir le II (r3-bv2) et le III. Ces souches du phylotype II et III des hauts plateaux de l'Ouest infectent aussi bien la pomme de terre que la tomate, et pourraient constituer une menace économique pour ces cultures dans cette région. Dans cette étude, les souches du phylotype I émergent en zone forestière humide comme une contrainte majeure à la culture des Solanacées. Ce constat a orienté le choix d'une souche de ce phylotype (CMR143) dans les travaux de la cartographie génétique de la résistance chez le piment. Pour ce faire, deux géniteurs de résistance différents CM334 et PM687 ont été considérés. Ces études génétiques ont montré une forte héritabilité de la résistance au flétrissement bactérien chez le piment. La détection de QTL par analyse de variance (ANOVA), par cartographie d'intervalle, simple (SIM) et composite (CIM), ont révélé l'existence de 3 à 6 QTLs et d'interactions digéniques dans le contrôle de la résistance du piment au flétrissement bactérien. Ces résultats confirment le caractère oligogénique de la résistance du piment au flétrissement bactérien. En mettant ensemble ces QTLs et ces interactions digéniques, 47 à 64 % de la variance phénotypique sont expliqués selon le caractère de résistance considéré. Par ailleurs, il est montré que la résistance du piment au flétrissement bactérien est fortement associée à la limitation de l'envahissement de la tige par la bactérie. De plus, cette étude a mis en évidence le caractère phylotype-spécifique de la résistance, car seulement 33% de facteurs génétiques contribuent au contrôle du flétrissement bactérien lors de tests avec le phylotype 2. La localisation de ces derniers permet de conclure en validant partiellement l'hypothèse de synténie entre la tomate et le piment.

Made available in DSpace on 2015-03-26T13:42:33Z (GMT). No. of bitstreams: 1 texto completo.pdf: 244644 bytes, checksum: 7bb131def78ba69016b2b6a8a2dd140f (MD5) Previous issue date: 2014-02-27 / Linkage disequilibrium (LD) is defined as the non-random association between alleles of different loci in a population and association mapping (MA) refers to a statistically significant association between molecular marker and phenotypic trait. The objectives of this study were: (1) estimate the linkage disequilibrium in populations of popcorn related by cycles of selection using SNP markers, (2) compare the populations in the degree of LD, (3) evaluate changes in allele frequencies and (4) identify significant associations between markers and quality related characteristics of popcorn. In total 465 samples were evaluated, with 354 samples belonging to the population 'Viçosa' and 111 samples belonging to the population eija- asm Improvement Program popcorn, Federal University of Viçosa. The populations were subjected to the following cycles of selection: Viçosa cycle 1 and cycle 1 Beija-Flor, obtained after one cycle of selection among and within half-sib families, Viçosa cycle 4, obtained after four cycles of selection among and within half-sib families, Viçosa cycle 2 full-sib families (FIC), obtained after two cycles of selection among and within full-sib families, Viçosa S4, obtained by selection of progeny S4. Ninety-six SNPs markers in properly selected QTL regions for quality, already identified above were used. The following characters were measured: capacity expansion (ml/g), grain density (g/ml), sphericity of grains and 100 grains weight (g). In comparison the Viçosa c0 used as the reference population for the highest mean values for LD linking group found in Viçosa c2 fic (D' = 0,8911; r 2 = 0,1905) as well as to related and unrelated SNPs (D' = 0,8911, r 2 = 0,1905) . Regarding the MA significant associations (p < 0.05) were found for all traits, with thirteen associations related to the feature expandability, twenty three with the sphericity of the grains, eight with the weight of 100 grains and seventeen density of the grains. / Desequilíbrio de ligação (LD) é definido como a associação não aleatória entre alelos de diferentes locos em uma população e mapeamento associativo (MA) refere-se à associação estatística significativa entre o marcador molecular e a característica fenotípica. Os objetivos deste trabalho foram: (1) estimar o desequilíbrio de ligação em populações de milho pipoca relacionadas por ciclos de seleção utilizando marcadores SNP, (2) comparar as populações quanto ao grau de LD, (3) avaliar alterações de frequências alélicas e (4) identificar associações significativas entre marcadores e características relacionadas à qualidade do milho pipoca. No total foram avaliadas 465 amostras, sendo 354 amostras pertencentes à - populações pertencentes ao germoplasma do Programa de Melhoramento de Milho-Pipoca da Universidade Federal de Viçosa. As populações foram submetidas aos seguintes ciclos de seleção: Viçosa ciclo 1 e Beija-Flor ciclo 1, obtidas após um ciclo de seleção entre e dentro de famílias de meios-irmãos, Viçosa ciclo 4, obtida após quatro ciclos de seleção entre e dentro de famílias de meios-irmãos, Viçosa ciclo 2 famílias de irmãos completos (FIC), obtidas após dois ciclos de seleção entre e dentro de famílias de irmãos completos, Viçosa S 4, obtida por seleção de progênies S4. Foram utilizados 96 marcadores SNP devidamente selecionados em regiões de QTL pré-identificadas para qualidade. Os seguintes caracteres foram mensurados: capacidade de expansão (ml/g), densidade dos grãos (g/ml), esfericidade dos grãos e peso de 100 grãos (g). Em comparação a Viçosa c0, utilizada como população de referência os maiores valores médios de LD por grupo de ligação foram encontrados em Viçosa c2 fic r2=0,1905). Na análise de MA foram encontradas associações significativas (p < 0,05) para todas as características avaliadas, sendo treze associações relacionadas com a característica capacidade de expansão, vinte e três com a esfericidade dos grãos, oito com o peso de 100 grãos e dezessete com a densidade dos grãos.