Thesis (MSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Haliotis midae, or more commonly known as Perlemoen, is an abalone species found along the coast of South Africa. It is the only cultured abalone species in South Africa and has a high demand abroad. Due to its popularity as a seafood delicacy, illegal harvesting has taken its toll on Perlemoen numbers. This increases the need for sustainable farming efforts and efficient implementation of law enforcement practices against poachers. Abalone farms make use of a limited number of broodstock for breeding, so it is necessary to ensure that genetic effects such as inbreeding and bottlenecks do not interfere with the viability of the offspring. Research that focuses on the genetics of Perlemoen will greatly aid the farms to continue sustainable production of this species as well as enhance their breeding efficiency. This study focuses on the construction of a linkage map for H. midae that will allow the future identification of markers associated with genes important to production, such as growth and disease resistance. Identification of these genes will allow breeders to select genetically superior abalone that will be used for breeding programmes in which the phenotype of the offspring will be enhanced. For the construction of a linkage map it is necessary to have enough informative markers for mapping. In this study, gene-linked microsatellite markers were developed by screening a contig assembly of H. midae’s transcriptome. Ninety-eight primer pairs could be developed from the contigs and 60 loci produced amplification products. Twenty-six microsatellites were found to be polymorphic (27% success rate). In addition to these markers, 239 previously developed microsatellites and 48 gene-linked SNPs were used to develop sex-average and sex-specific linkage maps in four full-sib families consisting of approximately 100 offspring each. Of these markers 99 were informative in family DS1 (31% success rate), 81 in family DS2 (26%), 77 in family DS5 (24%) and 71 in family DS6 (23%). These markers were used for linkage analysis (LOD>3). The average number of linkage groups for the sex-average maps ranged from 17-19. The average genome length for these maps ranged from 700cM to 1100cM with an average marker spacing of 8cM. The sex-specific maps’ linkage groups ranged from 13-17 with an average genome length of 600cM to 1500cM. The average marker spacing was approximately 16cM. The integrated map was constructed by merging the sex-average maps. This map contained 25 linkage groups with an average genome length calculation of 1700cM and an average marker spacing of 9.3cM. The linkage maps created in this study are the first to utilize SNPs in H. midae. Further incorporation of SNPs into linkage maps will enhance the density. The maps created in this study are of medium-density (65%) and provide a link to the development of high-density linkage maps to facilitate associations of phenotypic traits to certain markers, to so that QTL mapping can be performed. This information can be used for marker-assisted selection to produce genetically superior abalone. / AFRIKAANSE OPSOMMING: Haliotis midae, of meer algemeen bekend as Perlemoen, is 'n klipkous spesie wat langs die kus van Suid-Afrika voorkom. Dit is die enigste gekweekte klipkous spesie in Suid-Afrika en het 'n hoë aanvraag in die buiteland. As gevolg van sy gewildheid as 'n seekos lekkerny, het onwettige stropery sy tol geneem op Perlemoen getalle. Hierdie verhoog die behoefte vir volhoubare boerdery pogings en doeltreffende implementering van wetstoepassing teen stropers. Perlemoenplase maak gebruik van 'n beperkte aantal broeidiere vir teling, dus is dit nodig om te verseker dat genetiese effekte soos inteling en genetiese bottelnekke nie inmeng met die lewensvatbaarheid van die nageslag nie. Navorsing wat fokus op die genetika van Perlemoen sal grootliks die plase steun om die volhoubare produksie van hierdie spesie voort te sit, sowel as hul teling doeltreffendheid te verbeter. Hierdie studie fokus op die ontwikkeling van 'n genetiese koppelingskaart vir H. midae, wat die toekomstige identifisering van die merkers wat verband hou met die gene wat belangrik is vir die produksie, soos groei en weerstand teen siektes sal verbeter. Identifisering van hierdie gene sal toelaat dat telers genetiese voortreflike Perlemoen kan kies vir teelprogramme waartydens die fenotipe van die nageslag sal verbeter word. Vir die ontwikkeling van 'n genetiese koppelingskaart is dit nodig om genoeg informatiewe merkers vir die kartering te hê. In hierdie studie, is geen-gekoppelde mikrosatelliet-merkers ontwikkel deur ‘contig’ data van H. midae se transkriptoom te ondersoek. Agt en negentig inleier pare kon ontwikkel word uit die ‘contigs’ en 60 loki kon ‘n amplifiseringsproduk lewer. Ses-en-twintig mikrosatelliete was polimorfies (27% suksessyfer). Bykomend tot hierdie ontwikkelde merkers is 239 voorheen ontwikkelde mikrosatelliete en 48 geen-gekoppelde SNPs gebruik om geslagsgemiddelde en geslagspesifieke koppelingskaarte in vier volsib families, wat uit ongeveer 100 nageslag elk bestaan, te ontwikkel. Van hierdie merkers was 99 informatief in familie DS1 (31%), 81 in die familie DS2 (26%), 77 in die familie DS5 (24%) en 71 in die familie DS6 (23%). Hierdie merkers is gebruik vir 'n koppelingsanalise (LOD>3). Die gemiddelde aantal koppelingsgroepe vir die geslagsgemiddelde kaarte het gewissel van 17-19. Die gemiddelde genoom lengte vir hierdie kaarte het gewissel van 700cM tot 1100cM met 'n gemiddelde merker spasiëring van 8cm. Die koppelingsgroepe van die geslagspesifieke kaarte het gewissel van 13-17 met 'n gemiddelde genoom lengte van 600cM tot 1500cM. Die gemiddelde merker spasiëring was ongeveer 16cm. Die geïntegreerde kaart is saamgestel deur die samesmelting van die geslagsgemiddelde kaarte. Die kaart toon 25 koppelingsgroepe met 'n gemiddelde berekende genoom lengte van 1700cM en' n gemiddelde merker spasiëring van 9.3cM. Die genetiese koppelingskaarte wat in hierdie studie ontwikkel is, is die eerste om SNPs te gebruik in H. midae. Verdere insluiting van SNPs in koppelingskaarte sal die digtheid verhoog. Die kaarte wat in hierdie studie ontwikkel is, is van medium digtheid (65%) en bied 'n stap nader aan die ontwikkeling van hoë digtheid koppelingskaarte om fenotipiese eienskappe met sekere merkers te assosieer, vir kwantitatiewe kenmerk lokus kartering. Hierdie inligting kan gebruik word vir merker bemiddelde seleksie om geneties verbeterde Perlemoen te produseer.

The evolutionary relevance of homeotic alterations for the origin of new taxonomic entities is still a controversial objective in plant sciences. In this context, the discovery of a floral homeotic variant of Capsella bursa-pastoris in natural populations offers the unique opportunity to elucidate the evolutionary significance of homeotic mutants in the wild. Since all petals are transformed into additional stamens, the variant was termed Stamenoid petals (Spe). In this thesis, a combination of ecological and molecular characterisation of the variant was performed, to improve the understanding of evolutionary processes in plant populations. Molecular markers were used to analyze genetic differentiation among known provenances and also within a large sympatric population of wild-type and homeotic mutant. The results clearly suggest a repeated evolution of the novel flower morphology. Furthermore, genetic analyses provided substantial evidence, that the two floral variants are well-defined into flower-type dependent sub-samples within one population. The evaluation of phenotypic traits elucidated that the homeotic variant is not hampered in fitness. In greenhouse and field experiments, a significant ecological differentiation in the onset of flowering was detected among variants. Finally, the novel floral phenotype shows a co-dominant inheritance, and a marker-assisted mapping approach exposed a single locus in a genetic map. In conclusion, the comprehensive study of ecological and molecular aspects indicates that the floral homeotic variant may be treated as an established taxonomic entity and proved the predicted role as model for evolutionary objectives. Since morphological alterations like Spe are discussed as a result of macroevolution, the homeotic variant of C. bursa-pastoris provides the opportunity to survey a (macro)evolutionary novelty in association with continuous micro-evolutionary adaptation

The use of nitrogen (N) fertilizers is both costly to farmers and contributes to environmental degradation. N applied to wheat accounts for 18% of N applied to farmland globally, making it a prime target for reducing and optimizing N application. Chapter I is a review on nitrogen use efficiency (NUE) in wheat, with emphasis on breeding efforts and genetic resources available to increase NUE. The concept of effective use of nitrogen (EUN) as yield per unit N applied as a measure of N use, is also introduced. Chapter II is a study using two bi-parental double haploid families to evaluate genetic variability of both the genetic main effects (intercept) and linear response to N (slope) and determine the feasibility of selection for EUN in wheat. Using cross validation, a genomic prediction accuracy of 0.68 for intercept and 0.50 for slope was found, indicating that EUN is under genetic control and can be selected for. The prospect of breeding for EUN under limited resources, i.e., using fewer N rates and fewer experimental plots, is also explored. It was found that two different N treatments can be used to produce accurate predictions of intercept and slope as high as 0.98 and 0.95, respectively. Chapter III uses the same population described in chapter II to further investigate feasibility of selection for EUN using a normalized difference vegetation index (NDVI) obtained from multi-spectral aerial images gathered throughout the growing season. Cumulative photosynthesis across the growing season was estimated by integration across the NDVI curve, and compared to grain yield estimates to determine the efficacy of aerial imaging to identify high EUN lines. NDVI values and the area under the NDVI curve were able to predict yield and had the strongest ability to predict yield in moderate to low N treatments, with R2 values as high as 0.81 and 0.78 respectively. / Master of Science / Chapter I is a review on nitrogen use efficiency (NUE) in wheat, with emphasis on breeding efforts and genetic resources available to increase NUE. The concept of effective use of nitrogen (EUN) defined as grain yield per unit N applied, is contrasted to NUE as a more economic breeding goal. Chapter II uses two bi-parental mapping populations to evaluate genetic variability of both the genetic main effects and the linear response to N and determine the feasibility of selection for EUN in wheat. The efficacy of genomic prediction for EUN is explored and the prospect of breeding for EUN under limited resources is also explored. Chapter III uses the same populations described in chapter II to further investigate the feasibility of selection for EUN using a normalized difference vegetation index (NDVI) obtained from multi-spectral aerial images gathered throughout the growing season. Cumulative photosynthesis across the growing season was estimated and found to be predictive of grain yield estimates at accuracies ranging from 0.31 to 0.78.

Parmi les Prunus, genre botanique d’intérêt économique important, l’abricotier (Prunusarmeniaca L.) est une culture emblématique du Bassin Méditerranéen. Il y est soumis à des contraintes biotiques importantes, parmi lesquelles le dépérissement bactérien, causé par Pseudomonas syringae (Psy), peut mener à des phénomènes de mortalité en verger au niveau des régions à hivers froids et humides. La mise en évidence de différences variétales en verger offre potentiellement des perspectives de contrôle de la maladie à travers le levier génétique. Aussi, ce travail de thèse avait pour principaux objectifs (i) d’identifier chez la plante des régions génomiques liées à la résistance partielle à la bactérie et (ii) d’étudier un plan factoriel d’interaction entre les diversités de la plante et de la bactérie (GxG) afin d’apprécier la généricité de la résistance et sa durabilité. Afin de répondre au premier objectif, deux approches complémentaires ont été mobilisées : une cartographie de QRLs (Quantitative Resistance Loci) sur quatre populations biparentales dont trois sont issues du croisement avec un géniteur commun ainsi qu’une analyse d’association sur une core-collection. Les données phénotypiques mobilisées correspondent à des symptômes issus d’inoculations contrôlées ainsi que des notes de mortalité obtenues suite à infection naturelle en verger. Ces deux approches (analyse de liaison et d’association) ont permis de mettre en évidence 22 QRLs de résistance, parmi lesquels seuls 2 QRLs sur les chromosomes 6 et 7 colocalisent entre les deux approches. Deux régions majeures détectées en étude d’association sur les chromosomes 5 et 6 se sont révélées être en déséquilibre de liaison et contrôlent près de 26 et 43% de la variation des symptômes. Deux mécanismes complémentaires reposant sur le blocage de l’infection de Psy et sur la limitation de la progression locale de la bactérie dans les tissus ont été mis en évidence à travers la détection de QRLs sur les chromosomes 3, 6, 8 d’une part et 1,4et 6 d’autre part. Le second objectif a été abordé grâce à une étude d’un plan factoriel d’interaction entre 20 accessions d’abricotier et 9 souches de Psy, échantillonnées d’après la connaissance de l’épidémiologie de la maladie en verger. L’analyse statistique de ce dispositif mis en œuvre à la fois en verger et en laboratoire a démontré la prédominance de l’effet du facteur souche dans la variabilité des symptômes étudiés et la très faible importance du facteur d’interaction GxG, indiquant une potentielle généricité des facteurs de résistance et des perspectives favorables à leur durabilité en verger.Les résultats issus de cette thèse contribuent à offrir une meilleure compréhension des mécanismes de résistance partielle au dépérissement bactérien de l’abricotier et fournissent des marqueurs et haplotypes, potentiellement mobilisables dans le cadre de programmes d’innovation variétale. / Within the genus Prunus, that contains highly valuable species, apricot (Prunusarmeniaca L.) is an emblematic Mediterranean crop. But apricot cultivation is constrainedby many biotic stresses, among which bacterial canker caused by Pseudomonas syringae(Psy) is particularly severe and can lead to the death of the trees in regions with humidand cold winters. Differences of susceptibilities have been observed between cultivars inorchards and create opportunities for disease management through genetic improvement.This thesis aimed to (i) identify genetic determinants linked to partial resistance to thebacterium and to (ii) study a factorial interaction design between both diversities of theplant and the pathogen (GxG interaction) in order to assess resistance genericity anddurability. With regard to the first objective, two complementary approaches were used :QRL (Quantitative Resistance Loci) mapping over four biparental progenies, amongwhich three were obtained with a cross involving a common genitor, and a genome-wideassociation study on a core-collection. The phenotypic data mobilized in this work rely onsymptoms issued from controlled inoculations and on mortality notations followingnatural infections in the orchard. These approaches led to the detection of 22 QRLs amongwhich only 2 QRLs, located on chromosomes 6 and 7, co-localized between the twomethods. Two main regions detected in the association study, over the chromosomes 5and 6, appeared to be in linkage disequilibrium and controlled 26 and 43% of the variationof the symptoms. A complementarity between two mechanisms, one that involves blockingthe infection of Psy and the other that limits bacterial mobility in the tissues has beenrevealed through the detection of QRLs over chromosomes 3, 6, 8 for one mechanism and1,4, 6 for the other, respectively. The second objective was fulfilled with a study of afactorial interaction design between 20 apricot accessions and 9 Psy strains, which weresampled according to the previous knowledge of the disease epidemiology in the orchard.Statistical analyses of phenotypic data obtained both from the orchard and a laboratorytest showed a clear predominance of the strain effect on symptom variability and a weakimportance of the GxG interaction factor. This last result highlighted a potentialgenericity of the resistance factors and favorable perspectives of durability in the orchard.The results issued from this thesis contribute to a better understanding of the mechanismsunderlying partial resistance of apricot to bacterial canker. Moreover, it provide markersand haplotypes of interest which could be mobilized in breeding programs.

Genetic material of Lithuanian origin varieties, breeding lines and wild ecotypes representing P. pratense, P. bertolonii and P. phleoides species were studied for the most important agro-morphological and feeding value indicators as well as at the genetic level employing different biochemical–molecular markers. The water stress experiment clearly demonstrated the existence of different levels of water stress response in Phleum species, suggesting that P. phleoides might have evolved under conditions of limited water availability. “Blind” mapping by HRM was used successfully to map water stress response genes of timothy in the perennial ryegrass mapping population. In total, 12 putative water stress response genes were mapped in seven perennial ryegrass linkage groups. / Pasitelkus agromorfologinius, kokybės ir biocheminius – molekulinius metodus ištirtos lietuviškos kilmės pašarinių, žemaūgių bei stepinių motiejukų veislės, selekcinės linijos bei laukiniai ekotipai. Fiziologiniai sausros atsparumo tyrimų rezultatai įrodė, kad stepiniai motiejukai turi geriau išvystytą atsparumo sausrai mechanizmą. Pirmą kartą motiejukuose aptikti ekspresuojami atsparumo sausrai kandidatiniai genai HRM metodu sužymėti daugiametės svidrės genolapyje.

Disease resistance is critical in soft red winter wheat (Triticum aestivum L.) cultivars. Leaf rust caused by Puccinia triticina Eriks and stripe rust caused by Puccinia striiformis Westend. f.sp. tritici Eriks. are destructive pathogens of wheat. From 2014 to 2015 phenotypic data was collected at diverse locations for resistance to leaf rust (North Carolina, Texas, and Virginia) and stripe rust (Arkansas, North Carolina, Georgia, Texas, and Virginia) in a Pioneer ‘25R47’ /‘Jamestown’ (P47/JT) population composed of 186 F5:9 recombinant inbred lines (RILs). Analysis of the P47/JT population identified two quantitative trait loci (QTL) for leaf rust resistance on chromosome 5B and two QTL for stripe rust resistance on chromosomes 3B and 6A. Phenotypic variation (%) explained by the putative leaf rust resistance QTL of Jamestown on 5B was as high as 22.1%. Variation explained by the putative stripe rust resistance QTL of Jamestown on 3B and 6A was as high as 11.1 and 14.3%, respectively. Jamestown is postulated to contain gene Lr18. Seedlings of 186 F5:9 recombinant inbred lines from the P47/JT population and 200 F2 seedlings from eight other crosses including Jamestown and/or the Lr18 host differential line RL6009 (Thatcher*6/Africa 43) were screened with P. triticina race TNRJJ. Genetic analysis of the populations was conducted to validate the presence of Lr18 in Jamestown. Results of linkage analysis identified SNP maker IWB41960 linked within 5 cM of gene Lr18 in all three populations. From 2016 to 2017 phenotypic data was collected at diverse locations for resistance to leaf rust (Illinois, North Carolina, and Virginia) in a ‘2013412’ (PI 667644) / VA10W-21 (PI 676295) population (412/21) composed of 157 doubled haploid (DH) lines. The 412/21 DH lines were genotyped via genotyping by sequence (GBS). Analysis of the 412/21 population identified one quantitative trait loci (QTL) region associated with adult plant resistance to leaf rust on chromosome 1B. Phenotypic variation (%) explained by the putative leaf rust resistance QTL of 2013412 on 1B was as high as 40.1%. Kompetitive allele-specific (KASP) markers KASP_S1B_8414614 and KASP_S1B_8566239 were developed as markers for use in marker assisted selection. / Ph. D.

Leaf rust and stripe rust of wheat (Triticum aestivum L.) are caused by the fungal pathogens Puccinia triticina, and Puccinia striiformis f.sp. tritici, respectively. In North America, the incorporation of adult-plant resistance (APR) genes into breeding lines has been an important strategy to achieve durable resistance to both diseases. Previously, the spring wheat cultivar AC Domain was reported to express an effective level of adult-plant resistance (APR) to leaf rust under field conditions. Early gene postulation work had suggested AC Domain might carry the APR gene Lr34 due to its phenotypic similarity to other Lr34 carrying lines. However, new gene specific markers have shown that AC Domain is not a carrier of Lr34. The objective of this research was to genetically localize the resistance in AC Domain, which is important because the cultivar has frequently been used as a parent in Canadian breeding programs, primarily for its value as a source of pre-harvest sprouting resistance. A mapping population of 185 doubled haploid (DH) lines derived from the cross ‘RL4452’ by ‘AC Domain’ was used for this study. RL4452 is a known carrier of Lr34. During 2011-2012, the DH population was evaluated in field leaf rust nurseries at Saskatoon, SK and Portage, MB and at a stripe rust nursery at Lethbridge, AB. Field results indicated that rust resistance in the mapping population was variable, with lines ranging from highly resistant, to highly susceptible. DH lines carrying Lr34 showed a high level of resistance to both diseases. Thus, the non-Lr34 carriers were genotyped using select SSR markers, and by an Illumina 9k Infinium iSelect SNP assay for subsequent quantitative trait loci (QTL) analysis. QTL analysis revealed that AC Domain donated a major resistance QTL located on chromosome 2BS, that mapped 46 cM proximal to markers linked to Lr16, and explained a significant portion of the leaf and stripe rust phenotypic variance in all test environments. In addition, this QTL was significantly associated with the expression leaf tip necrosis (LTN), reduction in area under the disease progress curve (AUDPC), and coefficient of infection (CI). In certain environments the interaction between the 2B QTL and Lr34 was additive resulting in a superior level of rust resistance. Indoor rust testing showed AC Domain was susceptible to both diseases at the seedling stage. Taken together these results suggest that the identified resistance in AC Domain is likely due to the presence of an APR gene, on chromosome 2BS.

The Russian wheat aphid (RWA) (Diuraphis noxia Kurdjumov) is one of the important insect pests of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.) and other grasses. To date, there are four RWA biotypes identified in South Africa. The virulent biotypes emerged, partly due to climate change and new genetic variations within populations of RWA; hence there is a need to improve host-plant resistance, as an effective control measure. Bread wheat (Triticum aestivum L.) accession Cereal Introduction (CItr) 2401 is known to be resistant to all RWA biotypes worldwide. The goal of this study was to use a backcrossed near-isogenic line (NIL) BC5F5 mapping population, developed from a cross between CItr 2401 and susceptible Kavkaz, to identify and validate single nucleotide polymorphism (SNP) markers linked to the resistance phenotype in CItr 2401. This was achieved by (i) conducting a preliminary study that evaluated the suitability of simple sequence repeat (SSR) markers previously reported in literature for discriminating stacked RWA resistance genes and, (ii) employing SNP markers for the first time in a RWA resistance study as a future alternative to the widely used SSR markers. None of the tested SSR markers showed potential use in marker-assisted selection (MAS). The mapping population was phenotypically evaluated for RWA resistance using the four South African biotypes, viz. RWASA1, RWASA2, RWASA3 and RWASA4. Analysis of variance (ANOVA) showed significant (P<0.001) differences of genotypes after confirming the normality of residuals and homogeneity of variance. The Illumina iSelect 9,000 wheat SNP platform was used to genotype the two crossing parents and a selection of 24 NIL genotypes from the mapping population. Eight SNP markers found to be linked to the phenotype were converted to breeder-friendly and high-throughput Kompetitive allele-specific polymerase chain reaction (KASP) markers. The designed KASP markers were validated on the two crossing parents, the 24 NIL sent for SNP genotyping, on the mapping population and on the preliminary study genotypes for their effectiveness. The KASP assays developed in this study will be useful for stacking the RWA resistance from CItr 2401 with other Dn genes effective against the RWA. / Life and Consumer Sciences / M. Sc. (Life Sciences)

L’huître creuse, Crassostrea gigas, est une espèce dont la production aquacole représente un intérêt économique tant au niveau mondial qu’aux niveaux européen et français. Cependant, cette espèce subit des mortalités estivales enregistrées dès le début du 20ème siècle et, depuis 2008, ce phénomène s’est amplifié et menace essentiellement les huîtres juvéniles. La production aquacole d’huître creuse subit les conséquences de ces mortalités massives ; c’est pourquoi ce phénomène est étudié depuis de nombreuses années. En France, la bactérie Vibrio splendidus et le virus Ostreid Herpes Virus 1 (OsHV-1) sont le plus souvent associés aux épisodes de mortalités massives d’huîtres creuses juvéniles et il a été démontré que les individus sélectionnés pour leur résistance aux mortalités estivales étaient capables de ralentir l’augmentation de la charge virale en OsHV-1 dans leurs tissus puis de la faire régresser. Ces mêmes individus présenteraient également un effort reproducteur plus modeste que des individus sélectionnés pour leur sensibilité aux mortalités estivales. Ce travail de thèse s’inscrit dans ce contexte et a donc eu pour principal objectif d’améliorer la connaissance de l’architecture génétique de la reproduction de C. gigas en identifiant des régions du génome ou QTLs (Quantitative Trait Loci) impliquées dans l’effort reproducteur et de mettre en évidence d’éventuelles relations génétiques entre survie et reproduction, des QTLs impliqués dans la survie ayant déjà été détectés. Afin de caractériser l’effort reproducteur, il a été nécessaire de développer un ensemble de nouveaux outils. D’un point de vue biologique, 21 familles F2 ont été produites à partir des lignées sélectionnées pour leur réponse contrastée aux mortalités estivales. D’un point de vue moléculaire, de nouveaux marqueurs SNP (Single Nucleotide Polymorphism) ont été développés afin d’augmenter la densité de la carte génétique déjà disponible pour C. gigas. D’un point de vue technique, l’Imagerie par Résonance Magnétique (IRM) a permis d’observer la gamétogenèse de 300 individus d’une même famille F2 au cours de huit sessions réparties sur deux années alors que les études précédentes étaient limitées à une observation ponctuelle; les méthodes classiques d’observation de la gamétogenèse entrainant nécessairement la mort des animaux. Une forte corrélation a été mise en évidence entre les observations par IRM et par la méthode classique de l’histologie. En plus de l’estimation du rapport gonado-somatique (indice traditionnellement utilisé pour caractériser l’effort reproducteur), l’IRM a également permis d’observer des variations individuelles de la cinétique de la gamétogenèse ainsi que des différences entre les mâles et les femelles; le sexe étant identifiable sur les images obtenues par IRM. Parallèlement, 300 individus de deux autres familles F2 ont été sacrifiés pour estimer le rapport gonado-somatique par histologie. Cette approche a ainsi permis de détecter des QTLs impliqués dans de nombreux traits concernant la gamétogenèse. Des individus provenant des trois familles F2 caractérisés pour l’effort reproducteur ont été caractérisés pour la survie à un épisode de mortalités estivales. Cette étude a permis de détecter des QTLs impliqués dans le caractère « survie ». Ces QTLs correspondent, pour certains, à ceux détectés au cours d’une étude précédente. De plus, ces QTLs sont parfois colocalisés avec des QTLs impliqués dans l’effort reproducteur. Bien que la reproduction de l’huître creuse soit un caractère complexe à suivre, les nouveaux outils utilisés au cours de ce travail de thèse ont permis d’acquérir de nouvelles connaissances. Le séquençage du génome complet de Crassostrea gigas ainsi que les nouvelles méthodes de séquençage pourront peut-être permettre d’affiner les régions QTLs détectées. / The Pacific oyster, Crassostrea gigas, is a major aquacultured species whose production represents an economic interest at worldwide, european and french levels. However, this species undergoes summer mortalities recorded from the beginning of the 20th century and, since 2008, this phenomenon increased and threatens mainly juvenile oysters. Aquaculture production of oysters suffers consequences of mass mortalities, that’s why this phenomenon has been studied for many years. In France, the bacterium Vibrio splendidus and the Ostreid virus Herpes Virus 1 (OsHV-1) are often associated with mass mortality outbreaks of juveniles oysters and it was demonstrated that selected individuals for resistance to summer mortality were able to slow the increasing in viral load OsHV-1 in their tissues and then to decline it. These same individuals also present a lighter reproductive effort than individuals selected for their sensitivity to summer mortality. In this context, this study aimed to improve the knowledge of genetic architecture of reproduction of C. gigas by identifying some regions of the genome called QTLs (Quantitative Trait Loci) involved in reproductive effort and highlighting possible genetic relationships between reproduction and survival; QTLs involved in survival being already detected. To characterize the reproductive effort, it was necessary to develop a set of new tools. From a biological point of view, 21 F2 families were produced from lines selected for their contrasting response to summer mortality. From a molecular point of view, new SNPs (Single NucleotidePolymorphism) were developed to increase density of the genetic map already available for C. gigas. On a technical point of view, Magnetic Resonance Imaging (MRI) allowed to observe the gametogenesis of 300 individuals of the same family F2 during eight sessions over two years while previous studies were limited to a one-time observation because of the conventional methods of observation of gametogenesis leading necessarily to the death of the animals. A strong correlation was found between observations by MRI and observations by the conventional method of histology. In addition to the estimation of gonadic index (index traditionally used to characterize there productive effort), MRI also revealed individual variations in kinetics of gametogenesis and differences between males and females, the sex being identifiable on MRI images. In parallel, 300 individuals from two F2 families were sacrificed to estimate the gonadic index by histology. This approach enabled the detection of QTLs involved in many gametogenesis traits. Individuals from the three families characterized for F2 reproductive effort were characterized for survival during a summer mortality outbreak. This study was able to detect QTLs involved in the trait "survival". These QTLs correspond to some of those detected in a previous study. In addition, these QTLs are often collocated with QTLs involved in reproductive effort. Although there production of the Pacific oyster is a complex trait to follow, the new tools used in this thesis allowed acquiring new knowledges. The sequencing of genome of Crassostrea gigas and Next-Generation Sequencing technologies may be able to help to refine the detected QTL regions.

No description available.