La bactérie responsable de chancre sur bois, Pseudomonas syringae pv. actinidiae (Psa), a causé trois épidémies depuis les années 1980 et se décline en trois biovars. La plus récente et dévastatrice (causée par Psa biovar 3), a été détectée pour la première fois en 2008 en Italie et s’est rapidement répandue dans la majorité des pays producteurs de kiwi, dont en France en 2010. Nous avons analysé la diversité de 280 souches de P. syringae isolées de kiwi en France. La caractérisation biologique et l’analyse phylogénétique des souches par MLSA ont révélé que les biovars 1, 2 et 3 appartenaient à une même lignée génétique, groupant également P. s. pv. theae. Les souches de biovar 4 constituent un ensemble de 4 lignées génétiques distinctes qui ont été rassemblées au sein d’un nouveau pathovar (Pseudomonas syringae pv. actinidifoliorum (Psaf)). Ces souches sont caractérisées par une pathogénie réduite (taches foliaires mais pas de chancre). Cette nouvelle classification permet une meilleure gestion des épidémies de chancre bactérien du kiwi. Le développement d’un schéma MLVA composé de 11 VNTRs a permis d’étudier la structuration génétique de populations de Psa biovar 3, de révéler de la diversité au sein de ce pathovar et d’identifier l’origine italienne de l’épidémie en France. Le séquençage du génome de cinq souches de Psaf et la comparaison de ces séquences avec celles d’autres génomes de Psa et Psaf, disponibles sur NCBI, a permis le développement d’un nouvel outil de détection par PCR temps réel, plus spécifique de chaque biovar de Psa et de Psaf. La MLVA et la PCR temps réel développées ici contribueront à l’amélioration de la surveillance de Psa dans le monde. / The causal agent of bacterial canker, Pseudomonas syringae pv. actinidiae (Psa),has been responsible ofthree epidemics since 1980’s.Psa is divided in three biovars. The most recent and severe outbreak (causedby Psa biovar 3) was detected for the first time in Italy in 2008. It has spread very quickly in the main kiwifruit producing countries, as in France in 2010. We analyzed the diversity of 280 strains of P. syringae isolated fromkiwifruit in France. The biological characterization and the phylogenetic analysis of the strains by MLSArevealed that the biovars 1, 2 and 3 belong to the same genetic lineage, which include P. s. pv. theae, as well.The biovar 4 strains, which are structured in 4 distinct genetic lineages, have been grouped in a new pathovar(Pseudomonas syringae pv. actinidifoliorum (Psaf)). These strains are characterized by a low virulence (onlyspots on leaves and no canker on wood). This new classification help with the management of the bacterialkiwifruit canker outbreaks. The development of an MLVA scheme composed of 11 VNTRs allowed to studythe genetic structuration of Psa biovar 3 populations, to reveal the diversity within this pathovar and to identifythe Italian origin of the epidemic in France. The genome sequencing of five Psaf strains and the comparisonbetween these sequences and those of Psa and Psaf genomes already available on NCBI, allowed thedevelopment of a new detection tool by real-time PCR, specific of each Psa biovar and of Psaf. The MLVA andthe real-time PCR based detection technique developed here will contribute to the improvement of the monitoringof kiwifruit bacterial canker around the world.

L'endocardite infectieuse (EI) est une complication rare mais gravissime de la bactériémie à Staphylococcus aureus. Bien que certains facteurs de risque liés à l'hôte aient été décrits, l'implication de facteurs bactériens dans la survenue de l'EI est encore inconnue. Ces travaux de thèse ont visé à chercher tout élément bactérien associé à l'EI. Les facteurs phénotypiques décrits ou supposés comme potentiellement impliqués dans l'EI ont été testés. En parallèle, les profils génotypiques des souches obtenus par puces ADN ont été analysés par différents outils statistiques. L'analyse statistique univariée n'a montré aucune différence significative entre souches d'EI et souches de bactériémie, suggérant un processus complexe et multifactoriel. En effet, l'analyse discriminante en composante principale appliquée sur les données de puces ADN a permis de mettre en évidence une distinction entre les deux groupes de souches, confirmée sur une collection indépendante de souches. De plus, une fonction linéaire simplifiée, basée sur seulement 8 marqueurs génétiques, a permis d'obtenir des performances similaires, sur la collection de souches initiale ainsi que la collection indépendante de validation. En dernier lieu, les souches d'EI et de bactériémie ont été comparées à partir de séquences du génome complet (n = 40 (20 EI, 20 bactériémies)). L'analyse statistique par analyse discriminante en composante principale réalisée sur ces données génomiques confirme une distinction possible entre les deux groupes de souches. Au total, ces travaux de thèse apportent la preuve de concept que les facteurs bactériens sont impliqués dans la survenue de l'EI au cours de bactériémie à S. aureus / Infective endocarditis (IE) is a severe condition complicating 10-25% of Staphylococcus aureus bacteremia. Although host-related IE risk factors have been identified, the involvement of bacterial features in IE complication is still unclear. This PhD work aimed to characterize strictly defined IE and bacteremia isolates and searched for discriminant features. Phenotypic traits previously reported or hypothesized to be involved in staphylococcal IE pathogenesis were tested. In parallel, the genotypic profiles of all isolates, obtained by microarray, were analyzed. No significant difference was observed between IE and bacteremia strains, regarding either phenotypic or genotypic univariate analyses, suggesting a multifactorial process. However, the discriminant analysis of principal components (DAPC), applied on microarray data, segregated IE and bacteremia isolates. The performance of this model was confirmed with an independent collection of IE and bacteremia isolates. Finally, a simple linear discriminant function based on a subset of 8 genetic markers retained valuable performance both in study collection and in the independent validation collection. At last, IE and bacteremia isolates were compared based on whole genome sequence data from a subset of 40 isolates. When applied to this dataset, DAPC confirmed a possible segregation between the two groups of isolates. All in all, this PhD work provides the proof of concept that bacterial characteristics may contribute to the occurrence of IE in patients with S. aureus bacteremia

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Crop wild relatives (CWR) are an indispensable and at the same time threatened genetic resources for plant breeding. The study uses wild species related to celery to demonstrate how genetic resources of CWRs can be actively maintained in their natural surroundings (in-situ). Genetic reserves should be designated for long term conservation of selected occurrences. The study presents the selection procedure in detail, aiming at the identification of occurrences and sites suitable for the designation of genetic reserves, the spatial model of a genetic reserve and first practical results of the project. The overall aim of the project is the establishment of a nationwide network of genetic reserves for Apium graveolens, Helosciadium repens, H. nodiflorum and H. inundatum, the four wild celery species native to Germany. Helosciadum repens (Jacq.) W.D.J.Koch is threatened by genetic erosion due to a decline in population numbers and sizes. The loss of any population is an irretrievable loss of diversity and opportunity to enhance crops in the future. Genetic reserves are one way to conserve these populations and their genetic potential. Twenty-seven populations were selected for the analysis in a decision process based on site information. Microsatellites (SSR) were used to elucidate the genetic diversity of German populations. A cluster analysis was performed to see if the individuals form clusters of similarity. For that, a discriminate analysis of principal components (DAPC) was conducted, as the inbreeding index indicated a high number of inbreeding events in the populations and thus discordance with HWE (Hardy-Weinberg equilibrium). The analysis identified six genetic groups, which coincide well with the geographic origin of the analysed plants. The allelic richness (mean counts of alleles per individual per population) was higher in the southern populations compared to the northern ones. This North-South discrepancy was also visible as a high heterogeneity in the cluster assignments in the DAPC analysis. These differences in genetic diversity might be a result of the biogeographic history of Europe, especially the last glacial maximum. For the establishment of genetic reserves, two populations were considered as most important: The population that differs the most from the average genetic composition and the population that represents the average genetic composition of a population the best. The two extremes of differentiation were interpreted as such that the former has a specific adaptation to its local environment, and the latter represents all populations the best. DifferInt was used to analyse the SSR data and validate the differentiation of all populations compared to a pool of populations. However, SSRs are not capable of detecting adaptive traits. Populations were additionally chosen from different eco-geographic units (EGU), to increase the chance of capturing different traits. EGUs (Naturräume) are areas of specific abiotic and biotic features. These features may influence selection pressures and induce local adaptations. Based on site parameters and genetic data, 14 most appropriate wild populations (MAWP) were identified for genetic reserves establishment. For H. repens, two eco-forms are known and described in the literature. Besides their different habitats (terrestrial/semi-terrestrial and aquatic) they can be differentiated by morphological traits. Leave and stolon sizes and flowering behaviour differ significantly. Furthermore, the roots of the aquatic forms do not anchor in soil but on other aquatic plants, wood or roots of trees, while the terrestrial form exhibits a shallow root system network similar to other perennial species. To this end, no genetic analysis was conducted to clarify the phylogenetic status of the putative forms and authors avoided the usage of any specific noun rather than form. The SSR data from the previous study was evaluated, particularly with regards to the two forms. Additionally, an ISSR analysis was conducted, and the data was used to perform a PCA. There was no genetic clustering regarding the two forms neither in the SSR nor in the ISSR data. Nonetheless, the North-South discrepancy in the genetic diversity that was visible in the DAPC plot was confirmed in the PCA of the ISSR data. However, markers may fail to detect quantitative variation for adaptively important traits. As the most obvious difference in the two habitats was the water availability, the adaptation of both forms to drought stress was studied by measuring the relative water content of leaves, system water content and water loss during drought stress conditions. The stomatal index was measured for different water treatment levels. The results indicate that phenotypic plasticity rather than genotypic adaptation is responsible for different H. repens phenotypes.