Les souches œnologiques présentent une importante diversité dans leur besoin en azote, qui se traduit par des différences de capacité fermentaire. A l'heure actuelle, les mécanismes impliqués dans la variabilité des profils fermentaires, suite à un épuisement en azote dans le milieu, ne sont pas connus. L'identification de ces mécanismes serait un atout dans la compréhension des phénomènes conduisant aux fermentations problématiques et dans les reprises de fermentation. Afin d'identifier ces mécanismes, nous avons couplé une approche de physiologie et génomique classique, à une approche de génétique impliquant la recherche de QTL basée sur l'efficacité fermentaire en condition de carence en azote. Nous avons ainsi caractérisé cette différence de besoin en azote entre souches comme étant une variabilité dans la capacité à percevoir la carence en azote et à développer un programme de quiescence réduisant le flux d'énergie et augmentant un état de stress. Ces remaniements d'énergie se traduisant alors par des différences de capacités fermentaires. L'approche QTL a quant à elle permis de détecter 23 régions du génome potentiellement impliquées dans le maintien de la capacité fermentaire. Après analyse nous avons identifié 4 gènes dont les variations alléliques sont responsables des variations phénotypiques entre souches. L'utilisation de ces gènes pourrait permettre la conception de marqueurs génétiques, exploités pour la sélection de souches ayant de bonnes capacités fermentaires. Les données issues de cette approche QTL suggèrent une étroite corrélation entre les différences de réponse au stress par les souches et l'implication des mécanismes de perception et de signalisation de l'azote. Enfin, l'ensemble de nos travaux offre une nouvelle hypothèse, en désignant la voie TOR comme le mécanisme responsable de la variation des capacités fermentaires entre souches. / Oenological strains present an important diversity in nitrogen requirement, which result by difference in the fermentative performances. Nowadays, mechanisms involved in variability of fermentation profiles, result in nitrogen depletion in the medium, are not known. The identification of these mechanisms would be an advantage in the comprehension of phenomena leading to problematic fermentation and the fermentation restart. To identify these mechanisms, we have coupled a physiological and classical genomic approach with a genetic approach involving the QTL mapping based on the fermentation capacity in conditions of deficiency. We have characterized this difference in nitrogen requirement between strains as variability in the ability to sense nitrogen starvation and develop a quiescent program that reduces the flow of energy and increases its adaptation to stress. These energy rearrangements result in differences of fermentative performances. QTL approach allowed to detect 23 genome regions potentially involved in maintaining of the fermentative capacity. After analysis we have identified 4 genes for which allelic variations are responsible for the phenotypic variation between strains. The use of these genes may allow the design of genetic markers, exploited for the selection of strains with good fermentation capacity. The data from this QTL approach suggest a correlation between differences in stress response and the involving of mechanisms sensing and nitrogen signaling. Finally, all our work supplies a new hypothesis, pointing to the TOR pathway as the mechanism responsible of variation in fermentation capacity between strains.

Orientadores: Anete Pereira de Souza, Luciana Rossini Pinto / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-06T10:12:59Z (GMT). No. of bitstreams: 1 Teixeira_LauraHelenaMarcon_M.pdf: 776155 bytes, checksum: d0c316ee167c455455a616d2024d8eea (MD5) Previous issue date: 2006 / Resumo: A cana-de-açúcar (Saccharum spp.) está entre as espécies de maior importância econômica no mundo, constituindo uma das principais fontes de produção de açúcar e álcool. Apesar de o Brasil ocupar posição de destaque, como o maior produtor mundial, os níveis de produtividade são considerados baixos. A obtenção de novas variedades de cana-de-açúcar, mais produtivas e resistentes a pragas e doenças, é um processo que consome vários anos até o lançamento para o plantio comercial. O desenvolvimento de mapas de ligação pode contribuir significativamente para os programas de melhoramento, principalmente na localização de genes associados a características agronômicas de interesse. Os bancos de dados de seqüências expressas (ESTs-database) oferecem uma oportunidade para a construção de mapas funcionais, os quais servem de base para a estratégia de genes candidatos (Candidate-gene approach). O projeto de seqüenciamento de ESTs (SUCEST) do programa Genoma da FAPESP já identificou cerca de 40 mil clusters que representam os genes de cana-de-açúcar. Deste modo, o mapeamento de ESTs pode ser conduzido pela análise do polimorfismo no comprimento de restrição (RFLPs) utilizando os ESTs como sondas de hibridização. Tendo em vista os avanços que serão alcançados no melhoramento genético da cana-de-açúcar com a exploração das informações contidas nos bancos de dados de ESTs, este projeto teve como objetivo mapear ESTs relacionados a genes de interesse em cana-de-açúcar, utilizando-os como sondas em ensaios de RFLP em uma progênie derivada do cruzamento entre duas variedades précomerciais de cana-de-açúcar. Assim é importante ressaltar que o presente projeto complementa um programa de mapeamento genético molecular de duas variedades pré-comerciais de cana-de-açúcar, e outro, de mapeamento de QTL¿s associados a características de interesse agronômico, utilizando como marcadores as seqüências produzidas pelo projeto SUCEST / Abstract: Sugarcane (Saccharum spp.) is the species that has the greatest economic importance in the world, as it is one of the main sources of sugar and alcohol production. Although Brazil is the biggest producer of this crop - participating with 25% of world production, the productivity levels are considered low. The acquisition of new sugarcane varieties, which are more productive and resistant to plagues and illnesses, is a lengthy process that takes years until the launching of the commercial crops. Linking map development can contribute significantly towards improvement programs, mainly in the localization of genes associated to the agronomical traits of interest. The expressed sequence tag (ESTs) database offers a chance for the construction of functional maps, which serve as a base for the Candidate-gene approach. The EST sequence project (SUCEST) of the FAPESP Genome program has already identified about forty thousand clusters that represent sugarcane genes. In this way, EST mapping can be led by the restriction fragment length polimorfism (RFLPs) analysis using the ESTs as hibridization probes. In view of the advances that will be reached in sugarcane genetic improvement with the exploration of the information contained in the EST database, the aim of this project is to map ESTs related to sugarcane interest genes using them as probes in the RFLP assays in a lineage derived from the crossing between two sugarcane commercial crosses. The present project complements a molecular genetic mapping program; and another QTL mapping, associates the agronomics traits, using the sequences produced for the SUCEST project as markers / Mestrado / Genetica Vegetal e Melhoramento / Mestre em Genética e Biologia Molecular

Orientadores: Claudio Lopes de Souza Junior, Anete Pereira de Souza / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-06T11:28:57Z (GMT). No. of bitstreams: 1 Lima_MilenadeLunaAlves_D.pdf: 1291219 bytes, checksum: a3a078e58517d0d64e9051437ce18432 (MD5) Previous issue date: 2006 / Resumo: A maior parte dos caracteres de importância agronômica e econômica do milho estão sob o controle de diversos locos gênicos, denominados locos de caracteres quantitativos (QTL). A possibilidade do uso de marcadores moleculares e o aperfeiçoamento dos modelos estatístico-genéticos possibilitaram o mapeamento desses locos gênicos que afetam tais caracteres. Pouco enfoque no estudo de mapeamento de QTL foi dado em populações derivadas do germoplasma do milho tropical, o qual possui uma base genética ampla com maior diversidade do que o germoplasma temperado. Da mesma forma, pouco se conhece sobre as interações dos QTL nos diferentes ambientes (QTL X E). Duzentos e cinqüenta e seis progênies F2:3, derivadas do cruzamento de duas linhagens de milho tropical, foram avaliadas em cinco ambientes. O mapa genético foi desenvolvido com 139 marcadores microssatélites, utilizando o programa MAPMAKER/EXP versão 3.0b. As análises de mapeamento de QTL e a detecção da interação QTL X E foram realizadas utilizando o procedimento JZmapQTL do programa Windows QTL-Cartographer versão 2.5, que se baseia na análise de mapeamento em ambientes múltiplos (mCIM). A extensão total do mapa genético foi de 1.858,61 cM com intervalo médio entre marcadores de 13,47 cM. Dezesseis QTL foram mapeados para produção de grãos, oito para espiga por planta, seis para acamamento, seis para altura de planta, nove para altura de espiga e dois para número de folhas. Os efeitos genéticos dos QTL mapeados apresentaram variação em sinal e magnitude, demonstrando que cada QTL contribui de forma particular para a expressão dos caracteres. A maioria destes QTL apresentou ação gênica sobredominante, e muitos deles também apresentaram significante interação QTL X E. Esses resultados forneceram dados para uma melhor compreensão da arquitetura genética do genoma do milho tropical. Estas informações podem ser utilizadas em programas de seleção assistida dessa espécie, utilizando marcadores moleculares, gerando mais eficiência nos programas brasileiros de melhoramento / Abstract: Most of important agricultural and economical traits in maize are under the control of several gene loci, named quantitative trait loci (QTL). The possibility of using molecular markers and the statistic-genetic models made possible the mapping of these gene loci that affect such traits. Little focus has been given to QTL mapping study in populations derived from tropical maize germplasm, which has a broad genetic base with greater variability than temperate maize germplasm. Also, not much is known about the interaction of QTL in various environments (QTL X E). Two-hundred and fifty-six F2:3 progenies, derived from a crossing between two tropical maize inbred lines, were evaluated in five environments. The genetic map was developed with 139 microsatellite markers, using the software MAPMAKER/EXP version 3.0b. The analyses of QTL mapping and the detection of QTL X E interaction were performed using the Windows QTL-Cartographer version 2.5, JZmapQTL procedure, which is based on multiple-environment joint analysis (mCIM). The genetic map spanned 1,858.61 cM in length with an internal average of 13.47 cM between markers. Sixteen QTL were mapped for grain yield, eight for ears per plant, six for plant lodging, six for plant height, nine for ear height and two for number of leaves. The genetic effects of the mapped QTL presented varied signal and magnitude, displaying that each QTL contributes in a particular way for trait expression. Most of these QTL displayed gene action overdominance, many of them with significant QTL X E interaction detected. These results provide data for a better comprehension of genetic architecture on tropical maize genome. This information can be used in the marker-breeding selection of this species, leading more efficiency to Brazilian breeding programs / Doutorado / Genetica Vegetal e Melhoramento / Doutor em Genetica e Biologia Molecular

Orientadores: Anete Pereira de Souza, Antonio Augusto Franco Garcia / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T23:03:56Z (GMT). No. of bitstreams: 1 Mancini_MelinaCristina_D.pdf: 10767402 bytes, checksum: b866f6316486301d3466116f8be3049a (MD5) Previous issue date: 2014 / Resumo: A crescente busca por variedades de cana-de-açúcar com maior produtividade e resistentes às principais doenças consiste em um importante objetivo para o sucesso de um programa de melhoramento. Assim, a utilização de marcadores moleculares na identificação de locos que controlam características quantitativas (QTLs ¿ Quantitative Trait Loci) vêm ganhando cada vez mais destaque no programas de melhoramento genético. A presente tese teve como objetivo contribuir para o conhecimento básico sobre a genética e a biologia molecular da cana-de-açúcar através da detecção de marcadores ligados a características quantitativas. Foi utilizado uma população de cana-de-açúcar contendo 240 indivíduos F1 derivada do cruzamento entre as variedades comerciais SP81-3250 e RB925345. Para detectar os QTLs foi necessário realizar estudos fenotípicos e genotípicos. Foram coletados dados fenotípicos para as características de produção (altura, diâmetro, número e peso dos colmos) e de qualidade (sólidos solúveis, teor de sacarose do caldo e do colmo, pureza do caldo, teor de fibra) por três anos (2011, 2012 e 2013) nos municípios de Araras e Ipaussu, estado de São Paulo. Através de modelos mistos foi estimada a média, matriz de variância e covariância (VCOV), herdabilidade e a correlação fenotípica entre as características. Os resultados apresentados mostraram um ótimo controle ambiental, com menor valor de herdabilidade para pureza (0,77), além de 30 correlações fenotípicas significativas, confirmando que estes dados podem ser utilizados na detecção dos QTLs. Os dados genotípicos foram obtidos através da análise das regiões contendo microssatélites e de variações genéticas de único nucleotídeo, pelos marcadores SSR (Simple Sequence Repeat) e SNP (Single Nucleotide Polymorphism), respectivamente. A genotipagem dos SNPs foi realizada por espectrometria de massa pela Plataforma Sequenom MassARRAY® (Sequenom Inc., San Diego, California, USA). A análise foi realizada utilizando o programa SuperMASSA, que possibilitou estimar a ploidia dos locos. Assim, as marcas SNPs foram utilizadas na detecção dos QTLs para as características de produção e de qualidade. Por regressão linear foram encontradas 17 evidências de associação de QTL entre diâmetro dos colmos (quatro evidências), número de colmos (uma evidência), peso dos colmos (uma evidência), conteúdo de sólidos solúveis (duas evidências), teor de sacarose do caldo (três evidências), pureza (duas evidências), toneladas de cana por hectare (duas evidências) e toneladas de Pol por hectare (duas evidências). A proporção da variação fenotípica explicada pelo genótipo variou de 1,6% a 11,1%. Todos os SNPs que apresentaram associações com as características mencionadas tiveram os níveis de ploidia variando de hexaploide a dodecaploide. Por correlação genotípica-fenotípica, foi detectado sete evidências de associação de QTL entre diâmetro dos colmos (uma evidência), conteúdo de sólidos solúveis (duas evidências), teor de sacarose da cana (uma evidência), teor de sacarose do caldo (duas evidências) e pureza (uma evidência). Os SNPs detectados com correlações genotípica-fenotípica significativas apresentaram níveis de ploidia variando tetradecaploide a icosaploide. As diferentes ploidias permitiu a detecção de QTLs em multi-dose e podem ser usadas como informações prévias sobre os prováveis QTLs, contribuindo para o avanço do conhecimento da genética da cana-de-açúcar / Abstract: The increasing search for sugarcane varieties with higher productivity and resistant to major diseases is an important goal for the success of Sugarcane Breeding Program. Thus, molecular markers can be used to identify Quantitative Trait Loci (QTLs) and have become a powerful tool in Breeding Programs. This thesis aimed to contribute for the basic knowledge of genetics and molecular biology in sugarcane through detection of markers linked to quantitative traits. Was used a sugarcane population consisted of 240 F1 individuals derived from a cross between SP81-3250 and RB925345. To detect the QTLs it was necessary to perform phenotypic and genotypic studies. The phenotypic data were made for cane yield (stalk diameter, stalk height, stalk number, stalk weight and tons of cane per hectare) and quality traits (soluble solid content, sucrose content, juice sucrose content, purity, fiber and tons of Pol per hectare) for three harvest years (2011, 2012 and 2013) in Araras and Ipaussu cities, located in the state of São Paulo. The average, variance and covariance matrix (VCOV), heritability and phenotypic correlation was estimated via mixed models. All results showed a great environmental control, the lowest heritability was purity (0.77), besides 30 significant phenotypic correlations. confirming that these data can be used for QTLs detection. The genotypic data were obtained analyzing the regions containing microsatellites and single nucleotide genetic variants, by the markers SSR (Simple Sequence Repeat) and SNP (Single Nucleotide Polymorphism), respectively. The SNPs genotyping were performed via mass spectrometry by Sequenom MassARRAY® platform (Sequenom Inc., San Diego, California, USA). The analysis was performed using the SuperMASSA software allowing to estimate the loci ploidy. The SNPs markers were used for QTL detection for cane yield and quality traits. By linear regression 17 QTL association evidences were found for stalk diameter (four evidences), stalk number (one evidence), stalk weight (one evidence), soluble solid content (two evidences), juice sucrose content (three evidences), purity (two evidences), tons of cane per hectare (two evidences) and tons of Pol per hectare (two evidences). The phenotypic variation explained by genotype ranged from 1.6% to 11.1%. The SNPs associated with the traits mentioned had ploidy levels ranging from hexaploid to dodecaploide. Via genotypic-phenotypic correlation, it was detected seven QTL evidence of association for stalk diameter (one evidence), soluble solid content (two evidences), sucrose content (one evidence), juice sucrose content (two evidences) and purity (one evidence). The SNPs detected significant genotypic-phenotypic correlations showed ploidy levels ranging from tetradecaploide to icosaploide. The different ploidies allowed the detection of QTLs in multi-dose and can be used as prior information about QTL mapping, contributing to the advancement of the sugarcane genetics knowledge / Doutorado / Genetica Vegetal e Melhoramento / Doutora em Genética e Biologia Molecular

Orientadores: Anete Pereira de Souza, Antonio Augusto Franco Garcia / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Economia / Made available in DSpace on 2018-08-20T23:12:17Z (GMT). No. of bitstreams: 1 Souza_LiviaMourade_D.pdf: 2370923 bytes, checksum: b0c800b4425fab2c0c9fda9cbbb4798d (MD5) Previous issue date: 2012 / Resumo: A seringueira é uma espécie de cruzamento misto e com um longo ciclo de vida, o que dificulta a geração de linhagens endogâmicas e, por consequência, a construção e integração de mapas de ligação usando as metodologias convencionais. A exemplo do que tem sido feito em outras espécies vegetais que apresentam limitações para a obtenção de linhagens endogâmicas, os mapas genéticos existentes para seringueira foram construídos utilizando populações F1 (com diferentes tipos de segregação). A integração dos mapas obtidos para cada um dos genitores é possível com base em marcadores bi-parental, onde ambos os genitores são heterozigóticos podendo segregar nas proporções 1:1:1:1, 1:2:1 e 3:1 na progênie F1. Além disso, o uso de marcadores codominantes e multialélicos, como os microssatélites, acrescentam muito à construção do mapa, uma vez que permite a obtenção de estimativas de frequência de recombinação e da fase de ligação com um menor viés. Os microssatélites são marcadores moleculares consagrados, sendo eles a ferramenta de escolha no estudo de diversos organismos pela simplicidade de uso e de análise. Entretanto, para espécies que ainda não têm grande parte de seu genoma sequenciado, a obtenção de marcadores desse tipo passa pela necessidade de desenvolvimento via bibliotecas genômicas. Esse trabalho objetivou desenvolver, caracterizar e mapear microssatélites em Hevea brasiliensis para possibilitar estudos genético-moleculares da variação morfológica, identificação de regiões genômicas associadas a fenótipos de interesse, bem como análises genético-populacionais. Uma biblioteca genômica enriquecida em motivos repetitivos foi construída, a partir da qual foram desenvolvidos os microssatélites utilizados nesse trabalho. A utilização dos microssatélites de Hevea brasiliensis em amplificações heterólogas envolvendo seis espécies do gênero Hevea resultou em aproximadamente 98% de sucesso nas amplificações, sugerindo a existência de um complexo formado pelas diferentes espécies do gênero. Um mapa genético-molecular foi construído utilizando-se 284 marcadores microssatélites na análise de uma população segregante F1 com 270 indivíduos, a partir do cruzamento entre os genitores PB217 e PR255. Por meio da utilização do programa ONEMAP foi possível a construção de um mapa de ligação integrado que revelou 2840 cM de extensão, distribuídos em 23 grupos de ligação. O mapeamento de QTLs realizado utilizando metodologia de mapeamento por intervalo composto (CIM) detectou 24 QTLs para altura e circunferência das plantas entre as estações de verão e inverno. Este trabalho é pioneiro na construção de um mapa integrado para seringueira pelo fato dele ter sido construído unicamente com marcadores microssatélites, os quais são altamente informativos. Além disso, é também o primeiro estudo envolvendo análise de QTL para características relacionadas ao crescimento de plantas em seringueira, bem como é inédita a aplicação da metodologia CIM para população F1 segregante / Abstract: The rubber tree is an mixed crossing species with a long life cycle, which makes it difficult for the generation of inbred lines, and, therefore, the construction and integration of linkage maps by using conventional methodologies. Similar to what has been reported from other plant species that have limitations to obtain inbred lines, the genetic maps for rubber tree have been constructed by using F1 populations (with different types of segregation). The integration of maps obtained for each of the genitors is possible based on bi-parental markers, where both genitors are heterozygous, being able to segregate in ratios 1:1:1:1, 1:2:1 and 3:1 in the F1 progeny. Moreover, the use of co-dominant and multi-allele markers, such as microsatellites, adds a lot to the construction of maps, since it allows the obtainment of estimates of recombination frequency and linkage phase with less bias. The microsatellites are established molecular markers which are the tool of choice in the study of various organisms for their simplicity of use and analysis. Nevertheless, their application in species whose genomes have not yet been sequenced requires a prior development phase. The present study intended to develop, characterize and map microsatellites for the species Hevea brasiliensis so that initiatives concerning morphological variation, identification of genomic regions linked to phenotypes of interest, as well as population genetic analysis. A repetitive DNA-enriched library was constructed from which it was developed the microsatellites used in this work. The use of Hevea brasiliensis microsatellites for heterologous amplification in other six Hevea species was done with approximately 98% of success ratio, suggesting the existence of a complex formed by different species. A molecular genetic map was constructed using 284 microsatellite markers in the analysis of an F1 segregating population with 270 individuals from a cross between the parents PB217 and PR255. By using the program ONEMAP it was possible the construction of an integrated linkage map that revealed 2840 cm in length, divided into 23 linkage groups. The QTL mapping methodology was performed using composite interval mapping (CIM) and detected 24 QTLs for plant height and circumference between summer and winter. This work is a pioneer in building an integrated map for rubber because it was built only with microsatellite markers, which are highly informative. Furthermore, it is also the first one involving the analysis of QTL for characteristics related to the growth of rubber tree plants, as well as novel application of the CIM methodology for this type of population / Doutorado / Genetica Vegetal e Melhoramento / Doutor em Genetica e Biologia Molecular

La pourriture brune des fruits (BR), causée par les champignons du genre Monilinia, est une maladie courante qui peut provoquer jusqu’à 40% de pertes de récolte chez la pêche. Toutes les pêches cultivées sont plus ou moins sensibles à la moniliose. Aucune alternative aux traitements chimiques n’est disponible, ce qui rend nécessaire les applications de fongicides jusqu'au stade pré-récolte, qui sont préjudiciables pour l'environnement et peuvent laisser des résidus sur les fruits. Une revue de la littérature compile les connaissances disponibles sur le couple pêcher-monilioses.Le but de cette étude est d'étudier les facteurs de résistance du fruit à M. laxa à différents stades de croissance chez la pêche et de déterminer leur contrôle génétique.Nous avons focalisé d’abord sur quelques cultivars pour étudier l'évolution de la sensibilité des fruits à M. laxa au cours de leur développement en relation avec les caractéristiques structurales et biochimiques des fruits (conductance cuticulaire du fruit, micro-fissures de l’épiderme et composés de surface des fruits). Certains composés ont été détectés pour la première fois chez la pêche. Les résultats ont confirmé que lors de la phase I les fruits immatures sont sensibles à la moniliose. Au stade de durcissement du noyau, les fruits sont résistants, la conductance cuticulaire faible et les niveaux de composés de surface présentaient un pic de teneurs. A l’approche de la maturité, les fruits sont sensibles de nouveau. Au stade I, nous avons exploré le rôle des stomates et de la conductance du fruit immature en relation avec la sensibilité à M. laxa. Une centaine de génotypes d'une descendance interspécifique de pêchers appelée BC2 a été caractérisée par une infection au laboratoire, un suivi de pertes transpiratoires des fruits et une estimation de la densité de stomates (uniquement pour les nectarines). Des symptômes inattendus (une ‘tache claire’ qui ne progresse pas) ont été observés. La conductance cuticulaire était significativement liée à la probabilité d'infection, mais le nombre de stomates n’a montré aucun effet sur la probabilité d'infection. Des QTL contrôlant la résistance des fruits à la moniliose, à la conductance cuticulaire et au nombre de stomates ont été identifiés et des co-localisations observées.A maturité, le contrôle génétique de la résistance à la moniliose et des composés biochimiques de l'épiderme des fruits a été étudié. Pendant trois ans, les fruits de la BC2 ont été infectés avec une suspension de spores de champignon selon deux modalités. La BC2 a affiché une forte variabilité de résistance à la moniliose. Malgré une faible stabilité entre les années, des génotypes à haut niveau de résistance ont été identifiés. De plus en 2015, nous avons exploré la variation des composés de l'épiderme des fruits au sein de la BC2. Les composés phénoliques, les terpènes et dérivés ont été quantifiés par HPLC. La relation entre la résistance à la moniliose et la présence et / ou les niveaux de certains composés de l'épiderme et le contrôle génétique de ces composés ont été étudiés.La moniliose des fruits de pêche est un problème complexe qui est encore loin d'être résolu. Des progrès ont été accomplis dans la connaissance des caractéristiques structurales et biochimiques impliquées dans la résistance et des régions du génome qui pourraient conférer une certaine tolérance à la maladie ont été détectées. Des travaux supplémentaires sont nécessaires pour développer des marqueurs moléculaires pour la sélection assistée par marqueurs. Les résultats obtenus suggèrent que des solutions pour l'avenir résident dans l’association de cultivars tolérants _ moins sensibles aux micro-fissures et à haute teneur en composés épidermiques potentiellement inhibiteurs du développement du champignon _ avec des pratiques culturales réduisant les risques de fissuration des fruits et d'apparition de conditions climatiques favorables à la propagation de la moniliose. / Brown rot (BR) in peach fruit caused by the fungus Monilinia spp. is a common disease that can provoke as much as 30 to 40% losses of crop. Currently, all cultivated peaches are more or less sensitive to BR. No other alternative than chemical treatment is available, hence fungicide applications are required until pre-harvest. Such applications are damaging the environment and may let residues in fruits. A review of literature was accomplished to compile the knowledge scattered in the literature from many years. The aim of this study is to investigate the factors of resistance of the fruit to M. laxa at different stages of fruit growth and their genetic control by studying contrasted genotypes and an interspecific peach progeny. The first focus was made on few cultivars to study the evolution of sensibility of fruits to M. laxa during their development in relation with structural and biochemical characteristics of the fruit, e.g. cuticular conductance, micro-cracks and fruit surface compounds. Some compounds were detected for the first time on peach fruit. The results confirmed that during the stage I immature fruits are susceptible to BR. Fruit cuticular conductance was high probably due to high density of stomata and thin cuticule in formation. In contrary, at pit hardening stage fruits were resistant, cuticular conductance was low and the levels of surface compounds exhibit a peak. When maturity approaches, fruit become susceptible again. With rapid development of the fruit during this stage, the surface compounds were diluted and micro-cracks often appear which resulted in high cuticular conductance. At stage I we explored the different physical characteristics of the immature fruit in relation with susceptibility to M. laxa. A hundred of individuals of an interspecific peach progeny called BC2 were characterized through laboratory infection, monitoring of fruit transpiratory losses and estimating stomata density (only for nectarines). Unexpected symptoms (not progressing ‘clear spot’) were observed. The cuticular conductance was significantly linked to the likelihood of infection, but the stomata number had no effect on the likelihood of infection. QTL controlling fruit resistance to BR, cuticular conductance and stomata number have been identified and some co-locations observed. At maturity stage we investigated the genetic control of BR resistance together with biochemical compounds of fruit epidermis. For three years, mature fruits from the BC2 progeny were infected with two modalities of infection: spray until runoff in the orchard to measure infection probability and drop in the laboratory conditions in order to observe the characters of beginning, progression and speed of infection. The BC2 progeny displayed high variability for BR resistance. Despite low stability between years, genotypes with high level of resistance were identified. In addition in 2015, we explored the variation in epidermis compounds of fruit within the BC2 progeny. Phenolic compounds, terpenoids and derivatives were quantified by HPLC. The relationship between BR resistance and presence and/or levels of certain epidermis compounds and the genetic control of these compounds were investigated. BR of peach fruit is a complex problem which is still far from resolved. Progress has been made in the knowledge of structural and biochemical characteristics involved in BR resistance and regions of the genome that could confer certain disease tolerance have been detected. Further work is needed to develop molecular markers for marker assisted selection. The results obtained suggest that solutions for the future lie in associations of tolerant cultivars _ less susceptible to micro-cracks and with high content of epidermis compounds potential inhibitor of the fungus development _ with cultural practices reducing both risks of fruit cracking and occurrence of micro-climatic conditions favorable to BR spread and sporulation.

Evaluer l'effet du climat sur la croissance des arbres forestiers, et notamment leur capacité de production de biomasse en situation de contrainte hydrique, passe par la quantification du niveau de plasticité phénotypique des individus et de la diversité génétique des populations au sein de l'espèce étudiée. Le pin maritime, plante pérenne d'intérêt économique majeur en Aquitaine, largement étudiée au niveau génétique et écophysiologique, est un excellent modèle biologique pour mener à bien ce type d'étude. Dans le cadre de cette thèse, nous avons étudié la variabilité phénotypique de nombreux caractères liés à la croissance du pin maritime au jeune âge, en fonction de contraintes abiotiques essentiellement liées à la disponibilité en eau. Nous avons par ailleurs documenté l'architecture génétique de ces traits en termes de nombre, position sur une carte génétique et effet des gènes majeurs (QTL) qui contrôlent une partie de cette variabilité phénotypique.Dans le cadre de travaux de cette thèse, nous avons utilisé deux dispositifs expérimentaux: l'un composé de trois familles F1 issues de croisements contrôlés de parents d'origines géographiques contrastées (Corse, Landes et Maroc), l'autre d'une famille F2 issue de l'autofécondation d'un hybride Corse x Landes. Le premier essai comprenait 1500 individus semés en 2007 sur lesquels la hauteur et le diamètre ont été mesurés annuellement de 2010 à 2014. Nous avons également noté l'évolution de la phénologie du débourrement apical pendant deux années consécutives (2012 et 2013). Enfin, la dynamique de la croissance radiale de 239 génotypes (répartis sur deux des trois familles) a été suivie en continu pendant trois années (2011-2013) grâce à un dispositif unique de capteurs de déplacement continu (microdendromètres). Le second essai comprenait 500 arbres semés en 1998 et dont le carottage au niveau du tronc début 2011 a permis d’établir le profil microdensitométrique sur 7 années consécutives (2004-2010). En parallèle, le génotypage des descendants des 4 croisements a été réalisé grâce à l'aide de biopuces à ADN, ce qui a permis de construire des cartes génétiques. L'analyse conjointe de l'information phénotypique et génotypique a permis d'identifier des QTL pour l'ensemble des caractères et d'étudier leur stabilité en fonction des conditions environnementales et du fond génétique.Cette étude a montré que le débourrement est variable en années en fonction des contraintes de températures et du fond génétique. Tout comme la croissance primaire et secondaire, le débourrement est contrôlé par de nombreux QTL à effets modérés qui varient en fonction de l’environnement climatique et du fond génétique. Le suivi de la dynamique saisonnière de la formation du bois a également montré une interaction QTL x environnement révélant que la densité du bois est régulée par différents gènes ou le même jeu de gènes régulé de façon différentielle en fonction du climat. Enfin, La dernière partie de cette étude a permis de mettre en évidence, pour la première fois, la variabilité intra-spécifique des fluctuations journalières du tronc et son interaction avec des variables environnementales. [...] / Evaluating the impact of climate change on current plantations supposes the evaluation of their phenotypic plasticity and their genotypic diversity within the species, under abiotic pressure. Maritime pine is a perennial species of major economical interest in the french Aquitaine region. Wildly studied genetically and ecophysiologically, maritime pine is a very good biological model to see that type of study to the end. In this thesis, we intend to study various traits related to maritime pine growth under a biotic constraints, according to the following approaches: (1) evalutation of the phenotypic variability and (2)dissection of the genetic architecture of the traits (number, location and effects of QTLs). The comparisonbetween envrionmental and phenotypic data will allow us to appreciate the phenotypic pasticity of individuals. Afterwards, studying the genetic architecture of these traits and its variability according to the genetic background of individuals and environmental conditions will allow us to assess the stability ofdetected QTLs.We used 4 progenies of maritime pines: 3 controlled crosses of parents originated from contrasted ecotypes (Corsica, Landes and Morocco) and 1 controlled cross from a second generation of self-pollination (F2). Micro-cores were extracted from the individuals of the F2 population andmicrodensity profiles were established trough 7 consecutive years. Total height and diameter of eachindividual were measured once a year on the 3 others crosses, from 2010 to 2014. Dynamics of apical budburst was also followed on the same individuals in 2012 and 2013. Finally, dynamics of radial growth were monitored on a sub-sample of 239 individuals (spread in 2 of the 3 controlled crosses) during 3 yearsthanks to a unique device of microdendrometers.At the same time, all individuals (form the 4 crosses) were genotyped with several DNA bioarraysof molecular markers, allowing the building of genetic maps. The confrontation of phenotypic and genotypic data enabled to identify genome are as involved in the genetic architecture behind the traitsand to study their stability according to environmental conditions and the genetic background of individuals.This study showed that bud burst varies from year to year, depending on the conditions oftemperature and of the genetic background of individuals. Same way as growth, bud burst is controlled bymany QTLs of moderate effect, varying according to climatic conditions and the genetic background of individuals. The monitoring of seasonal dynamics of wood formation also showed a QTL x environment interaction revealing that wood density is regulated by different genes or the same set of genes,differentially regulated in response to the climate. The last part of the study puts forwards, for the firsttime, the variability of radius daily fluctuations within a full-sib family and its interaction with environmental variables. [...]

Les bases moléculaires responsables des propriétés technologiques des levures œnologiques sont pas ou peu connues. Or, ces connaissances sont nécessaires pour mettre en œuvre des stratégies d'amélioration génétique des capacités fermentaires ou de l'impact organoleptique des levures par croisements et une meilleure exploitation de la biodiversité. Ces travaux de thèse ont visé à l'identification des bases génétiques de plusieurs propriétés d'intérêt, telles que la production de sulfites, la capacité fermentaire en milieu carencé en azote ou la formation d'esters. Une approche de recherche de QTL a été mise en œuvre en s'appuyant sur la création d'une population de ségrégants méiotiques issus du croisement de deux souches œnologiques aux caractéristiques contrastées. Une caractérisation des phénotypes des ségrégants associé à leur génotypage a permis d'identifier des QTL pour les différents traits étudiés. L'implication d'une région du génome dans le contrôle de la voie d'assimilation des sulfates a été démontrée. Ce locus est responsable de la variation phénotypique de plusieurs métabolites de cette voie et de métabolites indirectement connectés. D'autres QTL ont également pu être mis en évidence pour les capacités fermentaires sur un milieu carencé en azote et la production de différents composés volatils. Leur analyse a permis d'identifier plusieurs gènes candidats dont les fonctions sont liées directement ou indirectement aux phénotypes étudiés. / The molecular basis of technological properties of wine yeasts are not or poorly known. However, this knowledge is required for the improvement of the fermentation capacity and of the organoleptic impact of wine yeasts by breeding strategies and a better exploitation of the yeast biodiversity. This thesis aimed at identifying the genetic bases of several traits of interest such as: sulfite production, fermentation ability in low nitrogen medium or esters formation. A QTL mapping approach has been implemented from a population of meiotic segregants derived from the cross of two strains with contrasting oenological characteristics. Phenotyping of the a population of meiotic segregants was combined with a genotyping to identify QTL for these traits. The involvement of a region of the genome in the control of sulphate assimilation pathway has been demonstrated. This locus is responsible for the phenotypic variation of several metabolites of this pathway and indirectly connected metabolites. Other QTLs have also been demonstrated for the fermentation capacities on a nitrogen limiting medium and production of various volatile compounds. Their analysis revealed several candidate genes whose functions are related directly or indirectly to the studied phenotypes.

Notre étude a porté sur le contrôle génétique des comportements sociaux ou de peur et leur relation avec les caractères de production au sein d’un croisement entre deux lignées de cailles sélectionnées de façon divergente sur la motivation sociale. Les résultats alertent sur un possible effet délétère de la sélection pour une plus forte productivité sur la sociabilité et la réactivité émotionnelle des animaux. Dans le même temps, ils indiquent des synergies possibles entre une plus forte motivation sociale et la précocité de la ponte, ou une plus faible réactivité émotionnelle et une production d’œufs plus importante. L’étude a permis la construction de la première carte génétique de moyenne densité à l’aide de marqueurs SNP chez la caille. Les analyses de liaison ont révélé un total de 45 QTLs dont 23 pour les caractères comportementaux et 22 pour ceux de production. Ce sont pour les critères de motivation sociale que les QTLs sont les plus nombreux (15). Certaines régions contrôlent à la fois la réactivité émotionnelle et le poids ou la sociabilité et l’âge au premier œuf. Plusieurs gènes candidats en lien avec la sociabilité ont été suggérés. / Our study focused on the genetic control of social and fear behaviors and their relationships with production traits in a second generation crossing between two lines of quail divergently selected for their social reinstatement behavior. The results warn us about a possible deleterious effect of the selection for higher productivity on the animal’s sociability and emotional reactivity. At the same time, they also indicate possible synergies between a stronger social motivation and a precocious laying onset, or a lower emotional reactivity toward a novel object and a higher egg production. The study allowed us to construct the first genetic map of medium density using SNP markers in the quail. Linkage analyses reveal a total of 45 QTLs with 23 linked to behavioral traits and 22 to the production traits. Most of the behavioral QTLs were linked to the social motivation (15). Also, some regions control both emotional reactivity and weight or sociability and the age at first egg. At this stage of the study, several candidate genes related to sociability were suggested.

Dezoito galinhas de uma população experimental utilizada em um cruzamento recíproco entre as linhagens de frangos de corte (TT) e de postura (CC) foram sequenciadas pela tecnologia de nova geração na plataforma Illumina com uma cobertura média de 10X. A descoberta de variantes genéticas foi realizada em uma região de locos de característica quantitativa (Quantitative Trait Locus, QTL), associado anteriormente com peso e percentagem de gordura abdominal no cromossomo 3 da galinha (GGA3), entre os marcadores microssatélites LEI0161 e ADL0371 (33,595,706-42,632,651 pb). O programa SAMtools foi utilizado na identificação de 136.054 SNPs únicos e 15.496 INDELs únicas nos 18 animais sequenciados e após a filtragem das mutações, 92.518 SNPs únicos e 9.298 INDELs únicas foram mantidas. Uma lista de 77 genes foi analisada buscando genes relacionados ao metabolismo de lipídios. Variantes localizadas na região codificante (386 SNPs e 15 INDELs) foram identificadas e associadas com vias metabólicas importantes. Variantes nos genes LOC771163, EGLN1, GNPAT, FAM120B, THBS2 e GGPS1 foram identificadas e podem ser responsáveis pela associação do QTL com a deposição de gordura na carcaça em galinhas. / Eighteen chickens from a parental generation used in a reciprocal cross with broiler and layer lines were sequenced by new generation technology with an average of 10-fold coverage. The DNA sequencing was performed by Illumina next generation platform. The genetic variants discovery was performed in a quantitative trait loci (QTL) region which was previously associated with abdominal fat weight and percentage in chicken chromosome 3 (GGA3) between the microsatellite markers LEI0161 and ADL0371 (33,595,706-42,632,651 bp). SAMtools software was used to detect 136,054 unique SNPs and 15,496 unique INDELs for the 18 chickens, and after quality filtration 92,518 unique SNPs and 9,298 unique INDELs were retained. One list of 77 genes was analised and genes related to lipid metabolism were searched. Variants located in coding region (386 SNPs and 15 INDELs) were identified and associated with important metabolic pathways. Loss of functional variants in the genes LOC771163, EGLN1, GNPAT, FAM120B, THBS2 and GGPS1 may be responsible for the QTL associated with fat deposition in chicken.