L'étude des bactéries pathogènes opportunistes d'origine environnementale ayant des modes de vie variés, libre et autonome ou contraint à une niche spécifique représentée par l'hôte, présente un intérêt dans la compréhension de l'adaptation des bactéries à leurs hôtes et de l'apparition de nouveaux pathogènes. Le genre Aeromonas regroupe des bactéries communes des milieux aquatiques, principalement des eaux douces. Elles sont capables d'entretenir différents types de relations avec leurs hôtes (parasitisme/symbiose) et peuvent être hébergées par un large spectre d'organismes. Chez l'homme, elles sont la cause d'une large variété d'infections (gastroentérite, bactériémie, infection de la peau et des tissus mous, etc.) mais les difficultés d'identification des souches et une taxonomie confuse engendrent une méconnaissance de la pathogénicité réelle des différentes espèces décrites.Le but de ce travail était d'étudier les mécanismes d'évolution génomique et génétique à l'origine de la remarquable capacité d'adaptation des Aeromonas à leurs hôtes, notamment à l'homme. Une analyse comparative de la diversité génétique et génomique d'une large collection de 195 souches représentative des différentes espèces du genre et d'origines variées (humaine, animale et environnementale) a été menée. La diversité génétique a été appréhendée au moyen d'une approche multilocus incluant l'étude des séquences de 7 gènes de ménage (dnaK, gltA, gyrB, radA, rpoB, tsf, zipA). En parallèle, nous avons étudié la variabilité des copies multiples du gène rrs en explorant leur diversité génétique par une méthode d'électrophorèse en condition dénaturante (PCR-TTGE) et la variabilité du nombre et de la répartition des opérons rrn dans le chromosome de ces bactéries par électrophorèse en champ pulsé.Ces différentes approches nous ont permis de mettre en évidence : i) une diversité très élevée des 7 gènes de ménage analysés ainsi que l'existence de transferts latéraux, ii) l'existence de sous-groupes de souches adaptées à un hôte ou à une localisation anatomique particulière, iii) un nombre important d'opérons rrn (8 à 11), iv) l'existence de profils de distribution chromosomique des opérons rrn spécifique d'espèce ou de groupes d'espèces proches, v) une forte proportion (41,5%) des souches présentant une hétérogénéité de séquences des différentes copies du gène rrs. Nos résultats montrent également la valeur taxonomique de l'étude de la diversité génétique et génomique à l'aide des approches proposées au sein du genre Aeromonas.Nous montrons que : i) l'ARN ribosomique 16S est un marqueur informatif pour étudier les modes d'évolution et conduire des études de taxonomie mixte et consensuelle dans le genre Aeromonas à condition d'étudier la diversité de ses multiples copies, ii) A. caviae présente des caractéristiques génétiques particulières témoignant d'un processus d'adaptation en cours à une niche écologique que nous supposons être l'intestin humain. Nos résultats supportent également un mode d'évolution des bactéries du genre Aeromonas dit en complexes d'espèces accompagné de phénomènes de spéciation pouvant en partie expliquer les difficultés rencontrées pour établir une taxonomie claire du genre Aeromonas. / Abstract :Studying opportunistic pathogenic bacteria with an environmental origin and a wide variety of lifestyles, either free-living or host-adapted, is useful to improve the understanding of bacterial adaptation to hosts and the emergence of novel pathogens. The genus Aeromonas groups water-living bacteria, mainly in freshwater. They are able to support several types of relations with their hosts (parasitism/ symbiosis) and are harbored by a large spectrum of hosts. In human, they are involved in a wide range of infections (gastroenteritis, bacteraemia, wound and soft tissue infection, etc.) but difficulties in identifying strains and a confused taxonomy results in incomplete knowledge of the real strain pathogenicity of each described species.The aim of this work was to study the mechanisms of genomic and genetic evolution related to the outstanding ability of Aeromonas adaptation to host, including human. We led a comparative analysis of the genetic and genomic diversity on a large strain collection (195 strains) representative of the species of the genus and from various sources (human, animal, environmental). We studied the genetic diversity using a 7 housekeeping gene multilocus strain analysis (dnaK, gltA, gyrB, radA, rpoB, tsf, zipA). We also described the variability in the i) rrs multiple gene copies using a PRC-TTGE method and ii) the number and distribution of the rrn operons within the chromosome using a pulse field gel electrophoresis. Our results also showed the taxonomic value of the study of genetic and genomic diversity using the approaches proposed in the genus Aeromonas.These various approaches enabled us to highlight: i) a high genetic diversity in the housekeeping genes together with horizontal gene transfers events, ii) some clusters that were either host-adapted or adapted to particular anatomical locations, iii) a high number of rrn operons (from 8 to 11), iv) the presence of patterns of rrn operon that were either species-specific or specific to groups of closely related species, v) a high frequency (41,5%) of strains harboring sequence heterogeneities between rrs copies. We showed that: i) 16 rRNA is a valuable marker for studying the modes of evolution of aeromonads and the taxonomy within the genus Aeromonas provided that multiple copy diversity is taken into account, ii) A. caviae displays particular genetic characteristic that suggested an ongoing process of adaptation to a niche that we supposed to be human digestive tract. Our results also support an evolution mode in complex of species with some speciation process that could at least in part explain difficulties for determining a clarified taxonomy within the genus Aeromonas.

Vitis vinifera L. est l'une des premières espèces fruitières à avoir été domestiquées. Sous l'effet de la sélection humaine, cette espèce a suivi une évolution agro-morphologique conduisant à une importante diversité répartie en deux morphotypes principaux selon l'usage des raisins (cuve vs table). L'objectif de cette thèse a été de mieux comprendre la structuration et l'origine de la diversité génétique de la vigne domestique au travers de l'étude de marqueurs moléculaires neutres (nucléaires et chloroplastiques) et de gènes codant pour des caractères d'intérêt agronomique (couleur des baies et architecture des grappes). Une meilleure connaissance des ressources génétiques de la vigne est en effet nécessaire pour leur gestion optimisée et leur utilisation appropriée dans de nouveaux programmes d'amélioration. Une étude de parenté basée sur l'analyse de 20 microsatellites nucléaires a d'abord été menée sur 2344 cultivars de la collection INRA du Domaine de Vassal. Elle a permis de préciser l'ascendance directe de plus de 800 cultivars et de révéler les géniteurs clés. A l'aide de ces mêmes marqueurs, une étude de la structuration génétique du compartiment cultivé a ensuite mis en évidence quatre grands groupes de diversité reliés à l'usage des fruits, la géographie et l'histoire de la viticulture. Ces premiers résultats ont été utilisés pour constituer un échantillon de travail de 595 génotypes comprenant i) des cultivars (subsp. vinifera, syn. sativa) représentatifs de la diversité neutre et de catégories historiques préalablement définies et ii) des représentants du compartiment sauvage (subsp. sylvestris). Les résultats de l'étude de la diversité de l'ADN chloroplastique sont compatibles avec l'existence d'un centre primaire de domestication oriental et de centres secondaires répartis sur le pourtour méditerranéen. Le polymorphisme de séquence (SNP et INDEL) a ensuite été exploré pour trois gènes associés à des caractères d'intérêt agronomique. L'analyse de la diversité des gènes VvMybA1 et VvMybA3, associés à la couleur des baies, a permis de préciser l'histoire de ce trait et sa diversification sous l'effet de la sélection artificielle. L'analyse du polymorphisme du gène VvTFL1A, associé à l'architecture des grappes, a montré une structuration différente principalement en relation avec l'usage des fruits. L'ensemble des résultats a permis de mettre en évidence certaines variétés ou groupes de variétés occupant une position originale dans l'histoire de la vigne cultivée depuis sa domestication. / Vitis vinifera L. is one of the first fruit species ever domesticated. Under human selection, this species underwent a morphological and agronomical evolution leading to an extensive diversity and to two distinct morphotypes according to the use of grapes (wine vs. table). The objective of this PhD thesis was to better understand the structure and origin of cultivated grapevine genetic diversity studying neutral (nuclear and chloroplastidial) molecular markers and genes encoding traits of agronomic interest (berry colour and bunch architecture). A better knowledge of grapevine genetic resources is indeed needed for their optimized management and appropriate use in new breeding programmes. A parentage study based on 20 nuclear microsatellites markers was first performed on 2344 cultivars held in the INRA “Domaine de Vassal” repository. This work allowed us to reveal the direct ascent of more than 800 cultivars and to uncover key genitors. Then, a study of the cultivated pool genetic structure was performed using the same markers. The four diversity groups found are related to use of fruits, geography and viticulture history. These first results were used to build a working sample of 595 genotypes that included i) cultivars (subsp. vinifera, syn. sativa) representative of both neutral markers diversity and previously defined historical categories and ii) representatives of the wild compartment (subsp. sylvestris). The results of chloroplastidial DNA diversity study are consistent with the existence of an eastern primary domestication centre with secondary centres distributed on the periphery of the Mediterranean sea. Sequence polymorphism (SNP and INDEL) was then explored in three genes associated with traits of agronomic interest. Diversity analysis of VvMybA1 and VvMybA3 genes associated with berry colour allowed us to better understand the diversification of this trait under artificial selection. Analysis of VvTFAL1A polymorphism, associated to bunch architecture, showed a different structuration mainly related to the use of fruits. All these results highlighted specific cultivars or groups of cultivars which hold an original position in the history of cultivated grapevine since its domestication.

En 1996, la FAO a reconnu le rôle des agriculteurs dans la construction des ressources génétiques. Dans ce contexte, l'objectif général du projet PLANTADIV, dans lequel s'inscrit cette thèse, est d'appréhender comment les facteurs anthropiques et biologiques interagissent et façonnent la diversité génétique des plantes cultivées dans le bassin du lac Tchad. Dans cette région, les populations ont mis au point une innovation agricole originale : l'utilisation en contre-saison de terres argileuses inondées pour repiquer du sorgho. Les variétés de sorgho repiqué sont capables de puiser dans les réserves hydriques du sol pour accomplir leur cycle végétatif en saison sèche sans autre apport d'eau. Le sorgho repiqué connaît un large développement dans la région depuis la moitié du XXe siècle. Le projet de thèse se focalise sur l'estimation de la diversité génétique des sorghos repiqués et sur les mécanismes biologiques et génétiques qui ont pu contribuer à sa structuration intra et inter variétale. Nous avons également entrepris de retracer l'histoire évolutive des sorghos repiqués en nous appuyant essentiellement sur des outils de génétique des populations pour discuter les hypothèses géographiques et historiques existantes. Cette étude a mis en évidence qu'au moins deux événements de dessaisonalisation avaient eu lieu à partir de sorghos pluviaux provenant de deux groupes génétiques différents. Les sorghos repiqués sont plus différenciés que les sorghos pluviaux. Ceci peut s'expliquer en partie par les pratiques paysannes : les sorghos pluviaux sont échangés plutôt via les amis, la famille et les voisins, alors que les sorghos repiqués le sont plutôt via les marchés. De par les nombreux échanges de semence entre les différentes populations humaines dans cette région, l'étude phylogénétique n'a pas permis de mettre en évidence le(s) lieu(x) d'origine de dessaisonalisation.La seconde partie de ce travail a été consacré à la biologie de la reproduction du sorgho repiqué. Les méthodes de calcul indirectes et directes ont montrées que le sorgho repiqué est, comme le sorgho pluvial, préférentiellement autofécondé. Le taux moyen chez le sorgho repiqué est cependant plus faible (1,8%) que pour le sorgho pluvial (12%). De même, les variations d'allofécondation entre des panicules d'un même type nommé semblent plus faibles que pour le sorgho pluvial. La dernière partie de la thèse est consacrée aux impacts des pratiques agricoles sur la structuration de la diversité génétique des sorghos repiqués à une échelle locale. Les analyses génétiques ont montré, que ce soit pour le village de Djongdong ou celui de Bouzar, situés à l'extrême Nord du Cameroun, que pour un agriculteur donné, chaque type nommé correspond à une entité génétique. De plus, un même morphotype chez différents agriculteurs correspond aussi à une entité génétique. Les modes de gestion des semences et les pratiques culturales ont été analysés, elles influencent peu la structuration de la diversité génétique du sorgho repiqué. / In 1996, FAO has recognized the role of farmers in building and managing genetic resources. This work is part of the project PLANTADIV which main objective is to understand how biological and anthropogenic factors interact and shape diversity of cultivated plants in the Lake Chad Basin. In this region, people have developed an original agricultural innovation: the use in dry-season of flooded clay soils for transplanting sorghum. Transplanted sorghum varieties are able to tap into soil moisture reserves to complete their growth cycle in the dry-season without any water supply. Transplanted sorghum cultivation undertook a large development in the region since the middle of the XX century.The thesis project focuses on the estimation of the genetic diversity of planted sorghum and on biological and genetic mechanisms that may have contributed to its structuration both within and between landraces. We also undertook to trace the evolutionary history of planted sorghum by relying primarily on population genetics approaches to elaborate over geographical and historical hypotheses.This study revealed that at least two events of deseasonalization occurred from rain- sorghum pools from two different genetic groups. Differentiation of dry-season sorghum is stronger than that of rain-sorghum. This may be partially due to social practices: rainy sorghum are mainly exchanged through friends, families and neighbors as planted sorghum seeds are often obtain from markets. Extensive seed exchange between different human populations across the region may have blurred the geographical pattern of the genetic diversity, not allowing us to identify potential sites for deseasonalization.The second part of this work is devoted to the reproductive biology of dry-season sorghum. Direct and indirect estimation methods have shown that dry-season sorghum is, as rain sorghum, preferably selfing. Average level of out crossing is nevertheless lower in dry-season sorghum (1.8%) than it is in rain-sorghum (12%). Within landraces, variations are also smaller for dry-season sorghum than for rain-sorghum.The last part of the thesis is devoted to the impacts of agricultural practices on the structure of the genetic diversity of dry-season sorghum at a local scale. Genetic analyzes have shown that in both studied villages of Djongdong and Bouzar, located in the extreme north of Cameroon, each landrace named by a farmer corresponds to a genetic entity. In addition, the same morphological type among different farmers corresponds to a genetic entity. Modes of seed management and cultural practices were analyzed; they seem to have little influence on the structure of the genetic diversity of dry-season sorghum.

La daurade royale (Sparus aurata) est une espèce de sparidé dont l’aire de répartition s’étend des côtes du Sénégal à l’Irlande, et est commune en mer Méditerranée. Elle est considérée comme rare en limite nord de répartition (Manche, mer d’Irlande et mer du Nord). Depuis une dizaine d’années, l’abondance d’individus pêchés en limite nord ne cesse d’augmenter. Ce phénomène peut être expliqué par l’évolution septentrionale de son aire de répartition, en lien avec le réchauffement climatique. Aucune donnée écologique sur les populations de cette espèce en limite nord de répartition, devenues une ressource de pêche en Manche, n’est actuellement publiée dans la littérature. Les traits de vie de S. aurata ont été étudiés face à la modification spatio-temporelle de son aire de répartition, via l’analyse du régime alimentaire, de la croissance et des déplacements côtiers. D’autre part, l’étude a conduit à l’analyse de la structure de ces populations septentrionales par une approche multi-marqueur couplant la génétique et la microchimie des otolithes.L’analyse des contenus stomacaux des populations de S. aurata en limite nord de répartition confirme un régime opportuniste, avec une forte proportion d’espèces-proies du genre Mytilus. Malgré un régime alimentaire similaire, la croissance des individus est, quant à elle, plus faible que celle observée sur des individus de mer Méditerranée, ce phénomène étant conditionné par les paramètres du milieu (température, salinité). Les variations élémentaires des otolithes ont permis de caractériser les migrations côtières lors des premières années de vie des poissons avec un passage en mer en hiver et la fréquentation de zones côtières. Cela suggère la présence de zones de nourricerie le long du pourtour atlantique et de la Manche. Une variabilité inter-individuelle a été mise en évidence suggérant une plasticité comportementale des individus. Les approches de génétique, combinant l’emploi de marqueurs mitochondriaux et des microsatellites, et de microchimie des otolithes ont mis en évidence l’absence de structure au sein des populations en limite nord de répartition. Ce phénomène concorde avec la colonisation récente des populations. Cependant, une différenciation entre les individus échantillonnés le plus au sud et ceux en limite nord d’échantillonnage a été mise en évidence, suggérant peu de mélange génétique. Les allèles communs entre les individus échantillonnés évoquent une colonisation des individus de proche en proche depuis la mer Méditerranée. Des différences génétiques et de traits de vie ont été observées entre les individus échantillonnés en Manche, suggérant l’existence d’une barrière biogéographique au sein de cet environnement. La présence de conditions environnementales favorables constitue un atout majeur pour cette espèce prédatrice qui semble présenter une capacité d’acclimatation importante. L’ensemble des approches abordées dans le cadre de ces travaux ont permis d’apporter les premières données sur les traits de vie et la structure des populations d’une espèce à fort intérêt commercial dans une zone d’expansion récente en lien avec des modifications globales des conditions environnementales. / The gilthead seabream (Sparus aurata) is a sparidae species whose natural distribution stretches from Senegal to Ireland coasts, with its common habitat in the Mediterranean Sea. This species is still considered as rare in its northern limit of distribution. For a decade, the abundance of individuals caught in the northern limit (English Channel, Irish Sea and North Sea), has increased. This species is now a fishing resources in the English Channel. This phenomenon, in link with global warming, is explained by the northern expansion of its distribution range. No ecological data about this species in northern distribution range is currently published. Life history traits of S. aurata in relation to the spatio-temporal modification of its distribution range has been studied, through diet, growth and sea/coastal movement analysis. On the other hand, the population structure analysis of these northern populations has been done with a multi-marker approach coupling the genetics and otolith microchemistry.Analysis of the stomach contents of S. aurata at the northern range confirms an opportunistic diet with a high proportion of prey species from the genus Mytilus. The growth of individuals is lower than that observed on individuals in the Mediterranean Sea. This phenomenon is conditioned by environmental parameters (temperature, salinity) at the distribution range limit. Elemental composition of the otoliths allowed us to characterize the sea/coastal migrations during the first year of life, with a sea transition in winter and the occupation of coastal zones. A variability between individuals has been observed as a behavioral plasticity of individuals. This suggests the presence of nursery areas along the Atlantic and Channel Sea coasts. Otolith microchemistry and genetic approaches, combining the use of mitochondrial and microsatellites markers, have demonstrated the lack of structure within populations at the northern distribution range. This phenomenon matches with the recent population colonization. However, a differentiation was identified between the most southern individuals sampled and those at the northern sampling boundary, suggesting little genetic mixing. Common alleles between individuals suggest a “step by step” colonization of individuals from the Mediterranean Sea. Genetic and life history traits differences were observed between individuals sampled in the Channel Sea, suggesting a biogeographic barrier within this environment.Favorable environmental conditions are a major asset for this predatory species, which appears to have a huge acclimatization ability. All the various approaches discussed in this work have then made it possible to provide the first data on the life characteristics and the population structure of a species with a high commercial interest on the northern range of its distribution.

La diversité génétique d’une population porte en elle la trace de pratiques culturelles et est un témoignage de certains éléments passés. Ainsi, la langue, qui peut agir comme une barrière culturelle à la reproduction, et le système de parenté, qui va déterminer quand, où et avec qui les individus se reproduisent, vont influencer la diversité génétique. L’objectif de cette thèse est de tenter de retracer les histoires démographiques de populations d’Asie du Sud et du Sud-Est, à plusieurs échelles culturelles, de la famille linguistique Austro-Asiatique aux groupes de filiation en utilisant des données génétiques. Dans un premier temps, nous nous sommes intéressés à la famille linguistique Austro-Asiatique. Les langues appartenant à une même famille linguistique descendent d’une même langue « mère » et nous avons voulu savoir si en plus de cette parenté linguistique, les locuteurs de ces langues partageaient une parenté génétique. Nous avons étudié la diversité génétique autosomale de populations parlant des langues Austro-Asiatiques et nous les avons comparées à des populations voisines appartenant à d’autres familles linguistiques. Nous n’avons pas pu mettre en évidence de parenté génétique particulière entre les populations parlant des langues Austro-Asiatiques. Ces résultats excluent l’hypothèse d’une origine commune des populations parlant des langues Austro-Asiatique et favorisent l’hypothèse d’une diffusion culturelle des langues Austro-Asiatiques. Puis, nous avons étudié la parenté génétique au sein de groupes de descendance dans huit populations d’Asie du Sud-Est. Les individus appartenant au même groupe de filiation disent descendre d’un même ancêtre commun, paternel dans les populations patrilinéaires ou maternel dans les populations matrilinéaires. Nous avons voulu savoir si l’ancêtre commun de ces populations est mythique ou biologique. Nous nous sommes intéressés aux diversités génétiques du chromosome Y et de l’ADN mitochondrial de huit populations d’Asie du Sud-Est : quatre populations matrilinéaires et quatre populations patrilinéaires. Nos données montrent que des individus appartenant à un même clan matrilinéaire sont fortement apparentés génétiquement au niveau de leur lignée maternelle, visible au niveau de leur ADN mitochondrial. A l’inverse, les individus appartenant à un même clan patrilinéaire ne sont pas plus apparentés génétiquement entre eux que deux individus pris au hasard dans la population au niveau de leur lignée paternelle, visible au niveau du chromosome Y. Ces résultats témoignent de réalités différentes entre les populations patrilinéaires et matrilinéaires d’Asie du Sud-Est avec un ancêtre commun réel dans les populations matrilinéaires et un ancêtre commun mythique dans les clans patrilinéaires. Pris ensemble, ces travaux rappellent la manière dont différents processus culturels ont laissé des signatures génétiques sur les diversités génétiques uniparentales et autosomales et illustrent la manière dont le généticien des populations peut utiliser ces diversités génétiques pour retracer l’histoire démographique des populations. / The genetic diversity of a population carries with it traces of cultural practices and is a testimony of certain past events. Thus, language, which can be a cultural barrier to reproduction, and the kinship system, which will determine when, where and with whom individuals reproduce, will influence genetic diversity. The aim of this PhD thesis is to try to retrace the demographic histories of populations of South and Southeast Asia, at several cultural scales, from the Austroasiatic language family to descent groups using the genetic data. Firstly, we studied in the Austroasiatic language family. Languages belonging to the same linguistic family come from the same "mother" language and we wanted to know whether, in addition to this linguistic kinship, the speakers of these languages shared a genetic kinship. We have studied the autosomal genetic diversity of Austroasiatic speaking populations and compared them to neighboring populations belonging to other linguistic families. We could not highlight any particular genetic kinship between populations speaking Austroasiatic languages. These results exclude the hypothesis of a common origin of populations speaking Austroasiatic languages and favor the hypothesis of a cultural diffusion of Austroasiatic languages. Then, we investigated genetic kinship in descent groups in eight Southeast Asian populations. Individuals belonging to the same descent group claim to descend from a common paternal ancestor, in patrilineal populations, or from a common maternal ancestor, in matrilineal populations. We wanted to know if the common ancestor of these populations is mythical or biological. We investigated the genetic diversities of the Y chromosome and mitochondrial DNA of eight populations in Southeast Asia: four matrilineal populations and four patrilineal populations. Our data showed that individuals belonging to the same matrilineal clan are closely genetically related to their maternal line, visible in their mitochondrial DNA. Conversely, individuals belonging to the same patrilineal clan are no more genetically related to each other than two individuals randomly selected from the population in their paternal line, visible on the Y chromosome. These results reflect different realities between the patrilineal and matrilineal populations of Southeast Asia with a real common ancestor in matrilineal populations and a mythical common ancestor in the patrilineal clans. Together, these results showed how different cultural processes have left genetic signatures on uniparental and autosomal genetic diversities and illustrated how the population geneticist can use these genetic diversities to trace the populations' demographic history.

A batata-da-serra, Ipomoea serrana Sim.-Bianch. & L.V. Vasconcelos, é uma liana endêmica da Chapada Diamantina, Bahia, cuja raiz tuberosa e consumida por populações humanas ha muitos anos. Apesar da espécie, classificada como vulnerável pela IUCN (União Internacional para a Conservação da Natureza), estar submetida à pressão antrópica devido à exploração de raízes tuberosas, para uso na alimentação, são raros os estudos com a espécie, razão pela qual e de grande importância conhecer a diversidade e estrutura genética da espécie. Estudos sobre diversidade e estrutura genética a partir de marcadores moleculares são importantes por fornecerem dados sobre impactos da exploração antrópica sobre as populações, podendo oferecer subsídio para planos de manejo e conservação de espécie. Cinco populações da Chapada Diamantina, constituindo um total de 142 indivíduos, foram investigados com quatro iniciadores Inter Simple Sequence Repeats (ISSR), resultando em 34 bandas, das quais 25 foram polimórficas. A analise dos parâmetros genéticos mostrou que as populações apresentam variabilidade moderada, com 73,8% de bandas polimórficas, 0,264 de índice de diversidade de Nei e 0,389 de índice de Shannon (valores médios). A maior parte da variação ocorreu dentro das populações (77%), estimado pela analise de variância molecular (AMOVA), enquanto que a variação entre populações foi de 23%, o que corroborou os resultados de estruturação obtidos pelo programa Structure, analise de coordenadas principais (PCoA) e agrupamento estimado pelo método Neighbor-Joining, a partir do coeficiente de dissimilaridade de Jaccard. A analise Bayesiana separou os indivíduos em quatro grupos, sendo que as populações Andaraí e Capão foram alocadas em grupos distintos, enquanto as outras três populações compartilharam indivíduos distribuídos em outros dois grupos. Este estudo, por seu caráter pioneiro com relação aos marcadores moleculares, constituiu o primeiro passo para o conhecimento da diversidade genética da espécie. Estudos futuros poderão ampliar o conhecimento sobre a espécie podendo oferecer subsidio para a elaboração de um plano de manejo para esta espécie que tem sido explorada na região. / batata-da-serra, Ipomoea serrana Sim.-Bianchi. & L.V. Vasconcelos, is an endemic liana from the Chapada Diamantina, Bahia, whose tuberous roots have been consumed by human populations for many years. Although the species, classified as vulnerable by the IUCN (International Union for Conservation of Nature), is subject to anthropic pressure due to the exploration of tuberous roots for food consumption, few studies have been conducted on the species, which is why it is of great importance to know the diversity and genetic structure of the species. Studies on genetic diversity and structure with molecular markers are important for providing data on the impacts of anthropogenic exploitation and can be useful for the species management and conservation. Five populations of Chapada Diamantina, consisting a total of 142 individuals were studied with four ISSR primers, resulting in 34 bands, 25 of which were polymorphic. The genetic diversity analysis showed that populations have a moderate variability, with 73.8% of polymorphic bands, Nei\'s unbiased gene diversity (He) was 0.264; Shannon diversity index (I) was 0.389, average values. Most of the variation was within populations (77%), as estimated by the analysis of molecular variance (AMOVA), whereas the variation between populations was 23% of the total, which corroborated the results of program Structure, principal co-ordinate analysis (PcoA) and cluster analyses, using the Neighbor-Joining method, and the dissimilarity coefficient of Jaccard. The Bayesian analysis separated the individuals into four groups, with populations Andarai and Capao allocated into different groups, while the other three populations shared individuals in two other groups. Considering the pioneering characteristic of this study at the molecular level, it represents the first step towards the knowledge on the genetic diversity of the species. Future studies will increase the knowledge on the genetics of the species and may provide subsidy for the development of a management plan for a species that has been explored in the region.

Manguezais são ecossistemas localizados na confluência de terra e mar, característicos de áreas tropicais e subtropicais, cobrindo cerca de 18,1 milhões de hectares do planeta. A grande biodiversidade encontrada nestes ambientes ressalta a importância da busca por conhecimentos à seu respeito, como o estudo sobre novos princípios ativos derivados de microrganismos endofíticos presentes nas plantas de manguezais. Desta forma, o propósito do presente trabalho foi determinar a diversidade genética da comunidade de fungos endofíticos presentes em folhas e ramos das principais espécies arbóreas de manguezais de Cananéia e Bertioga (situados no estado de São Paulo, Brasil), e avaliar o potencial biotecnológico destes fungos em relação à produção de antibióticos contra os patógenos humanos Staphylococcus aureus e Escherichia coli, e contra o fitopatógeno Xanthomonas axonopodis citri . Os resultados da primeira etapa do trabalho, que envolveu o isolamento e a caracterização de fungos endofíticos filamentosos, mostraram que a comunidade fúngica associada às plantas de manguezais é formada por pelo menos 35 gêneros diferentes, sendo que os gêneros mais frequentes foram Diaporthe, Fusarium, Trichoderma, Colletotrichum e Xylaria. Grande parte dos gêneros encontrados neste trabalho é de fungos de solo, indicando que eles estão adaptados às condições adversas dos manguezais. Os resultados mostraram que, dentre as linhagens produtoras de antibiótico, 29,41% pertencem ao gênero Diaporthe, o qual apresentou maior frequência na comunidade fúngica estudada. Após a avaliação de 344 fungos quanto ao potencial de atividade antimicrobiana, foi selecionada a linhagem 41.1(1) de D. phaseolorum, um endófito de folha de Laguncularia racemosa, para elucidação da estrutura química do seu antibiótico purificado. Por meio das técnicas de Ressonância Magnética Nuclear e de Espectrometria de Massas o antibiótico foi identificado como ácido 3-hidroxipropiônico o qual apresentou atividade frente aos patógenos humanos Staphylococcus aureus e Salmonella tiphy. A estrutura química deste antibiótico foi modificada por meio de reação química de esterificação de Fischer-Speier para avaliar a relação da estrutura química e atividade biológica deste composto. O produto final da reação química de esterificação do antibiótico ácido 3-hidroxipropiônico não apresentou atividade antimicrobiana, indicando que o grupo hidroxila removido na reação é importante na atividade farmacológica desse composto. Além disso, a linhagem 41.1(1) de D. phaseolorum foi transformada geneticamente pelo sistema Agrobacterium tumefaciens, visando a obtenção de transformantes deficientes para produção de antibiótico e, com isso, a identificação de genes relacionados com a via de biossíntese do antibiótico ácido 3-hidroxipropiônico. A análise das sequências que flanqueiam o T-DNA, obtidas por TAIL-PCR, mostraram que os genes interrompidos nos transformantes estão relacionados com proteínas de domínios conservados envolvidos com diferentes funções como: translação de proteínas, homeostase do íon orgânico Mg2+, transporte intracelular, migração, adesão e proliferação celular e outras funções celulares. A caracterização da biblioteca de agrotransformantes constitui uma ferramenta importante para o estudo da biologia molecular de fungos que produzem compostos bioativos por meio do seu metabolismo secundário. / Mangroves are ecosystems situated beyond land and sea. They are more frequently found in tropical and subtropical areas englobing around 18.1 millions of hectares in the planet. The great biodiversity found in these ecosystems shows the importance of researching them, including studies regarding new compounds derived from endophytic fungi that inhabit these ecosystems. Therefore, the goal of this study was to determine the genetic diversity of the fungal endophytic community found in leaves and branches of the main arboreal species from mangrove of Cananéia and Bertioga (situated in São Paulo state, Brazil), and to evaluate the biotechnological potential of these fungi concerning the production of antibiotics against the human pathogens Staphylococcus aureus and Escherichia coli, and against the phytopathogen Xanthomonas axonopodis citri . The results of the first part of this work, including the isolation and characterization of the filamentous endophytic fungi, showed that the mangrove fungal community is made up of at least 35 different genera, from which the most frequent are Diaporthe, Fusarium, Trichoderma, Colletotrichum and Xylaria. Most of the fungal genera found in this study come from soil, which suggests that they are adapted to the adverse conditions of mangroves. The results show that among the antibiotic-produncing strains, 29.41% belong to the genus Diaporthe, which was the most frequently found in the studied fungal community. After the analysis of 344 fungi regarding the antibiotic activity potential, a strain of D. phaseolorum (a leaf endophyte of Laguncularia racemosa) was selected to unveil the chemical structure of their purified antibiotic. The nuclear magnetic resonance and the mass spectrometry techniques allowed the identification of the antibiotic as 3-hidroxypropionic acid, which displayed activity against the pathogens Staphylococcus aureus and Salmonella tiphy. The chemical structure of this antibiotic was modifyed by the chemical reaction of Fischer-Speier sterification in order to evaluate the chemical structure and biological activity of this compound. The final product of the chemical reaction of 3-hidroxipropionic acid sterification had no antibiotic activity, which suggests that the hydroxil group removed from the reaction is important to the pharmachological activity of this compound. Additionally, the strain 41.1(1) of D. phaseolorum was genetically transformed by the Agrobacterium tumefaciens system, in order to generate antibioticdeficient transformants, which would help to identify genes related to the biosynthesis pathway of the 3- hidroxypropionic acid antibiotic. The TAIL-PCR analysis revealed that the interrupted genes in the tranformants are related to proteins from conserved domains involved in different functions such as protein translation, Mg2+ ion homeostasis, intracellular transport, migration, adhesion and cellular proliferation and other cellular functions. The characterization of the agrotransformants library is an important tool to unveiling the molecular biology of fungi that produce bioactive compounds by the secondary metabolism.

Os tripanosomas são parasitas hemoflagelados com importância médica para humanos e animais domésticos. Surtos de infecções agudas graves por T.vivax têm sido relatados em todo Brasil onde perdas econômicas e falhas reprodutivas são observadas. Neste trabalho, uma infecção experimental por T. vivax em ovinos confirmou sua transmissão transplacentária. Em relação à epidemiologia de T. vivax, para entender o papel dos jumentos em surtos no Semiárido Brasileiro, foram realizados ensaios parasitológicos e moleculares em infecções naturais e experimentais. Nossos resultados mostraram animais assintomáticos, portanto, podem atuar como potenciais reservatórios. Além disso, estudos filogenéticos revelaram uma diversidade genética inesperada em tripanossomas no Leste da África, especialmente em moscas tsé-tsé. Novos genótipos de T. vivax foram caracterizados e tripanossomas do subgênero Nanomonnas se mostraram prevalentes nestas infecções. Nesta abordagem, conseguimos identificar T. suis e novas espécies do subgênero Pycnomonas presentes em animais selvagens e domésticos. / Trypanosomes are hemoflagellates parasites with medical importance to humans and domestic animals. Outbreaks of severe acute infections by T. vivax have been reported throughout Brazil, where economic loss and reproductive failure are observed. In this study, an experimental infection by T. vivax in sheep confirmed the transplacental transmission of this parasite. Regarding T. vivax epidemiology, to understand the role played by donkeys in outbreaks in the Brazilian Semiarid, we performed parasitological and molecular assays in naturally and experimentally infections. Our findings show that infected donkeys are asymptomatic and, thus, are potential reservoirs. In addition, phylogenetic studies have revealed an unexpected genetic diversity in East Africa, especially in tsetse flies. New genotypes of T. vivax were characterized and trypanosomes of the subgenus Nanomonnas were prevalent in these infections. In this approach, we can identify T. suis and new species of the subgenus Pycnomonas present in wild and domestic animals.

A exploração de produtos não madeireiro em florestas nativas tem sido alternativa na composição da renda de comunidades locais. Dentre as diversas espécies exploradas da Mata Atlântica, a biriba - Eschweilera ovata (Cambess.) Miers - vem sendo intensamente utilizada para a confecção do berimbau. Disponibilizar informações que auxiliem em apontar diretrizes para adequadas formas de conservação genética e manejo desta espécie é de fundamental importância, tanto do ponto de vista ecológico como econômico e social. Com este objetivo, realizou-se o estudo do sistema reprodutivo e da estrutura ge nética de E. ovata, através da técnica de eletroforese de isoenzimas, em três áreas de ocorrência natural de populações da espécie sob diferentes graus de antropização, sendo uma explorada e outras duas sem exploração, localizadas próximas à cidade de Salvador - BA. Os resultados do sistema de reprodução, obtidos para duas populações, mostram que a espécie reproduz-se predominantemente por cruzamento, tendo $ t m variado de 98,5% a 99,9%, valor este superior à média apresentada pelas espécies arbóreas tropicais. Uma certa taxa de cruzamentos endogâmicos foi evidenciada nas populações estudadas, sendo praticamente 100% da endogamia gerada nas progênies resultado de cruzamento entre parentes na população natural, e 76,8% na população explorada, sendo o restante atribuído a autofecundação. O valor estimado para o coeficiente de coancestralidade dentro de progênies ( $qF ) variou de 0,191 a 0,211. As estimativas das correlações de autofecundação ( s rˆ ), foram relativamente baixas em ambas as populações e não diferentes estatisticamente entre si, (variando de 0,100 a 0,107). O número provável de árvores doadoras de polén foi extremamente baixo, tendo em média, dois indivíduos por árvore matriz. O tamanho efetivo de variância (Ne(v)) médio das populações foi de 2,13, sendo necessário assim, para reter o tamanho efetivo de 50, a coletas de cerca de 23 árvores nestas populações. O tamanho efetivo populacional foi próximo ao número de indivíduos amostrados. A divergência genética entre as populações de adultos ( $ q pA ), foi de apenas 2,5%,indicando que a maior parte da diversidade genética encontra-se distribuída dentro das populações (97,5%). A divergência entre populações, estimada com base nas progênies ( $ q pP ) foi ainda menor, de 1,4% (0,1 a 3,5 O número médio de alelos por locos foi de 2,14 nos adultos e de 2,41 nas progênies. A porcentagem de locos polimórficos nas árvores adultas foi de 85,4% e nas progênies de 81,8%. A heterozigosidade esperada segundo as expectativas do Equilíbrio de Hardy-Weinberg (EHW) foram altas e variaram de 0,354 a 0,431, e a heterozigosidade observada variou de 0,332 a 0,371, sendo, tanto na progênies como nos adultos das populações, menor do que a heterozigosidade esperada, indicando que há mais homozigotos nas populações que o esperado pelas proporções do EHW. A análise de autocorrelação espacial das árvores de E. ovata dentro da populações estudadas mostrou haver estruturação, sendo que quanto mais próximos os indivíduos maior é probabilidade deles serem parentes. Os resultados obtidos possibilitaram realizar inferências a respeito da conservação e manejo da espécie. / The exploitation of non-timber resources from tropical forest has been an alternative income to local human communities. Among the tree species which are exploited in the Atlantic forest, the “biriba" – Eschweilera ovata (Cambess.) Miers – has been used to manufacture the arc of “berimbau". The genetic conservation and the sustainable management of E. ovata populations are necessary to guarantee the longterm exploitation of this resource in an ecological, as well economic and social point of view. Adequate management strategies can be planned by getting information about the population genetic structure and mating system of the species. The aim of this study was to evaluate the mating system and the population genetic structure of E. ovata, using the allozyme electrophoresis technique. Three populations under different levels of human action in an area of natural occurrence of E. ovata near Salvador, Bahia state, were chosen. The results of the evaluation of the mating system of E. ovata, obtained for two population, indicate that the species reproduces itself through outcrossing. The outcrossing rate ($ t m ) varied from 98.5% to 99.9%. These values are higher than that average estimates observed in tropical tree species. Inbred crosses were observed in the progenies. Almost 100% of the inbreeding observed in the non-disturbed population and 76.8% in the exploited population was due to crossing among related individuals. Selffertilization was also observed. The estimated value of the coefficient of the coancestrality among the progenies ( $qF ) varied from 0.191 to 0.211. The estimates of self- fertilization correlation ( s rˆ ) were low and statistically similar and non significant in both populations (the values varied from 0.100 to 0.107). The expected number of pollen donors was extremely low, having an average of two pollen donors by mother-tree. The average effective number of variance (Ne(v)) was 2.13. With this estimate was possible to conclude that it is necessary to collect about 23 adult trees in these populations to keep an effective size of 50. The effective population size was similar to the number of individuals sampled. The genetic divergence among the adult populations ( $ q pA ) was 2.5%, showing the genetic diversity is mainly distributed within populations (97.5%). The genetic divergence among the populations obtained from the progenies ( $ q pP ) was also lower, 1.4% (0.1 to 3.5%). The average number of alleles by loci was 2.14 in the adult populations and 2.41 in the progenies. The percentage of polymorphic loci was 85.4% in the adults and 81.8% in the progenies. The expected heterozigosity varied form 0.354 to 0.431 and the observed heterozigosity varied from 0.332 to 0.371. The values of observed heterozigosity were inferior to the expected heterozigosity, indicating that there are more homozygous individuals than expected by the Hardy Weinberg Equilibrium. The spatial autocorrelation analysis of E. ovata shows that there is intrapopulational genetic structure in the studied populations of adult individuals. Closer individuals show a higher probability of being genetic related. The results obtained in this study are helpful to infer about the genetic conservation and the management of the species.

Clostridium perfringens causa enterite necrótica aviária devido à produção de toxinas que lesam o intestino. Neste estudo, de 94 amostras nove apresentaram C. perfringens, totalizando 22 isolados. Todos exceto um isolado, possuíram os genes nanI (95%) e/ou nanJ (81%), e 19/22, mostraram atividade neuraminidase em hemácias de frango. A atividade hemaglutinante foi observada em poucos isolados (26%). Todos os isolados foram plc positivos (toxina α), sendo classificados como tipo A. Sete isolados (31,8%) abrigaram o gene tpeL que codifica a toxina TpeL. Isolados tpeL+ mostraram efeito citotóxico característico da ação desta toxina. Alguns isolados mostraram capacidade de aderir e invadir células Vero. A maioria dos isolados foi resistente à sulfaquinoxalina (100%), cefalexina (95%) e eritromicina (95%) e sensíveis (100%) à cefoxitina, amoxicilina, enrofloxacina, amoxicilina-ácido clavulânico, penicilina-estreptomicina, cloranfenicol e metronidazol. Todos os isolados foram agrupados geneticamente em sete clusters, apresentando se como um grupo heterogêneo. / Clostridium perfringens cause avian necrotic enteritis due to production of toxins that damage the intestine. In this study, nine out of 94 samples had C. perfringens, totaling 22 isolates. All the isolates with exception of one, possessed the genes nanI (95 %) and/or nanJ (81 %), and 19/22 showed neuraminidase activity in chicken erythrocytes. The hemagglutinating activity was observed in a few isolates (26 %). All isolates were plc positive (toxin α) being classified as type A. Seven isolates (31.8%) harbored tpeL gene encoding the toxin TpeL. TpeL + isolates showed characteristic cytotoxic effect of the action of this toxin. Some isolates showed ability to adhere and invade Hep-2 cells. Most of the isolates were resistant sulphaquinoxaline (100%), cephalexin (95%) and erythromycin (95%) and sensitivity (100%) to cefoxitin, amoxicillin, enrofloxacin, amoxicillin - clavulanic acid, penicillin -streptomycin, chloramphenicol and metronidazole. All isolates were genetically grouped into seven clusters, presenting itself as a heterogeneous group.