This study examined population genetics of greater sage-grouse (Centrocercus urophasianus) in Strawberry Valley, Utah located in the north-central part of the state. The Strawberry Valley population of sage-grouse experienced a severe population decline with estimates of abundance in 1998 less than 5% (~150 individuals) of similar estimates from the 1930s (>3,000 individuals). Given the population decline and reduced genetic diversity, recovery team partners translocated sage-grouse from four different populations into Strawberry Valley over 6 years (2003-2008). Translocations have been used as a strategy to increase both population size and genetic diversity in wildlife populations. We assessed whether genetic diversity increased following the translocation of sage-grouse into Strawberry Valley by looking at both nuclear and mitochondrial DNA indices. We observed an overall increase of 16 microsatellite alleles across the 15 loci studied (x̅ =1.04 alleles per locus increase, SE ± 0.25). Haplotype diversity increased from 4 to 5. Levels of genetic diversity increased for both nuclear and mitochondrial DNA (16% and 25% increases for allelic richness and haplotype diversity, respectively). These results show that translocations of greater sage grouse into a wild population can be an effective tool to increase not only population size but also genetic diversity.Second, we studied fitness-related traits and related them to genetic diversity indices in a population of greater sage-grouse in Strawberry Valley, Utah from 2005 to 2013. We captured 93 sage-grouse in Strawberry Valley and fitted them with a radio collar and drew and preserved blood. We monitored sage-grouse weekly, throughout each year. From blood, we extracted and amplified DNA with 15 microsatellite loci. We determined genetic diversity as multilocus heterozygosity and mean d2. To determine if there was a relationship between genetic diversity and survival, we used known-fate models in Program MARK. We also determined if there was a relationship between genetic diversity measures and nest initiation, nest success, clutch size, and number of eggs hatched using generalized linear models where reproductive measures were modeled as a function of genetic diversity. We found no significant relationship between mean d2 and microsatellite heterozygosity with measures of survival or reproductive fitness. Overall, these results suggest that the often-reported strong heterozygosity-fitness correlations detected in small, inbred populations do not reflect a general phenomenon of increasing individual survival and reproductive fitness with increasing heterozygosity.

Primers for 10 polymorphic DNA microsatellite loci were developed and characterized for the endangered oyster mussel Epioblasma capsaeformis from the Clinch River, TN. Microsatellite loci also were amplified for individuals collected from the following additional populations or species: (1) E. capsaeformis from Duck River, TN; (2) E. florentina walkeri from Indian Creek, upper Clinch River, VA; (3) E. florentina walkeri from Big South Fork Cumberland River, TN; and (4) E. torulosa rangiana from Allegheny River, PA. Allelic diversity ranged from 9-20 alleles/locus, and averaged 13.6/locus for all 5 populations investigated. Average expected heterozygosity (HE) per locus ranged from 0.78-0.92, and averaged 0.86. A genetic characterization of extant populations of E. capsaeformis and E. florentina walkeri was conducted to assess taxonomic validity and to resolve conservation issues related to recovery planning. These mussel species exhibit pronounced phenotypic variation, and are difficult to characterize phylogenetically using DNA sequences. Monophyletic lineages, congruent with phenotypic variation among species, were obtained only after extensive analysis of combined mitochondrial (1378 bp of 16S, cytochrome-b, ND1) and nuclear (515 bp of ITS-1) DNA sequences. In contrast, analysis of variation at 10 hyper variable DNA microsatellite loci showed moderate to highly divergent populations based on FST values, which ranged from 0.12-0.39. Quantitative genetic variation was observed in fish host specificity, with transformation success of glochidia of E. capsaeformis significantly greater (p<0.05) on the greenside darter Etheostoma blennioides, and that of E. f. walkeri significantly greater (p<0.05) on the fantail darter E. flabellare. Lengths of glochidia differed significantly (p<0.001) between species, with sizes ranging from 241-272 μm. Underwater photographs of mantle-pads and micro-lures of female mussels documented fixed phenotypic variation between species. The texture and color of the mantle-pad of E. capsaeformis is smooth and bluish-white, while that of E. f. walkeri is pustuled and brown, with tan mottling. Based on extensive molecular, morphological, and life history data, a population of E. capsaeformis from the Duck River, TN is described and proposed as a separate species, and a population of E. f. walkeri from the upper Clinch River, VA is described and proposed as a separate subspecies. Genetic management guidelines were developed to assess taxonomic status, genetic variation of donor-recipient populations targeted for augmentation, and field and laboratory protocols to maximize genetically effective population size, minimize genetic changes in captive-reared progeny, and prevent the release of juvenile mussels into non-native drainages. A pragmatic approach to species recovery is advocated; one that incorporates the principles of conservation genetics into breeding programs, but prioritizes the immediate demographic needs of critically endangered mussel species. / Master of Science

La présente étude nous a permis de développer un système rapporteur, basé sur le gène bactérien codant pour la β-glucuronidase (GUS), afin de mesurer l’instabilité des microsatellites chez Arabidopsis thaliana. Les microsatellites sont des séquences particulières du génome susceptibles à des mutations fréquentes qui se produisent au cours de la réplication. On a démontré dans ce projet une corrélation entre la longueur du microsatellite et son taux de mutation. De même, l’orientation du microsatellite influence son instabilité. Dans deux études ultérieures, nous avons également étudié l’implication des gènes de correction des mésappariements (système MMR, « Mismatch repair ») dans l’instabilité des microsatellites. Le système MMR corrige les erreurs survenues au cours de la réplication des microsatellites. Nous avons montré que l’inactivation des gènes AtPMS1 et AtMSH2 entraîne l’augmentation de l’instabilité des séquences répétées, d’où l’importance de ces gènes dans le maintien de la stabilité des microsatellites en particulier et du génome en général. / The present study allowed us to develop a reporter system based on a bacterial gene encoding for β-glucuronidase (GUS), in order to measure microsatellite instability in Arabidopsis thaliana. Microsatellites are particular regions of the genome undergoing frequent mutations during replication. A correlation between length of the repetitive tracts and microsatellite instability was demonstrated. Besides, the orientation of the microsatellite influenced its instability. In two later studies, we have also studied the effect of mismatch repair genes (MMR) on microsatellite instability. Mismatch repair system corrects errors that occur during the replication of microsatellites. We showed that inactivation of AtPMS1 and AtMSH2 genes increased tracts instability, hence the importance of these genes in the maintenance of microsatellite stability and genome stability in general.

The field of conservation genetics, in combination with non-invasive sampling, provides a powerful set of tools for investigating the conservation status and natural history of rare species that are otherwise difficult to study. A systematic literature review demonstrated that this is certainly the case for many forest associated antelope species, which are poorly studied and yet constitute some of the most heavily hunted wildlife in Africa. The aim of the present study was to use non-invasive sampling to investigate genetic patterns in forest antelope populations in the high-biodiversity Udzungwa Mountains, Tanzania, within the context of the conservation of these species and the wider ecosystem. Genetic information was derived from faecal samples collected across the Udzungwa landscape and assigned to five antelope species (N = 618, collected 2006-09). Faecal pellet length was measured for a subset of samples but statistical assignment to species by this method proved unreliable. Phylogenetic analysis using mitochondrial control region sequences unexpectedly revealed that Harvey’s duiker within the Udzungwas are paraphyletic with respect to sequences from a putative sister species from southern Africa. However, there was no corresponding pattern in the microsatellite dataset suggesting that these mitochondrial lineages do not represent contemporary genetic isolation. Instead, Harvey’s duiker nuclear variation is shaped both by isolation by distance, due to positive spatial autocorrelation at short distances, and clustering of distinct genotypes from western outlying forests. These forests also harbour the endangered Abbott’s duiker and therefore require effective conservation management. Despite being detected throughout the Udzungwas, genetic diversity in Abbott’s duiker was very low in comparison to other species. These results suggest several promising research directions but also have significant conservation implications that will be disseminated to the Tanzanian wildlife authorities and the wider conservation community.

The European earwig exhibits a remarkable male-dimorphism in forceps morphology that is associated with alternative reproductive tactics under the control of a conditional evolutionarily stable strategy. Populations on the small, rocky islands of the Farnes off the Northumberland coast are known to sustain populations with dramatically higher morph ratios than observed on the UK mainland. A survey conducted of island and mainland sites around the UK showed that the dimorphic populations of the Farnes are similar to other islands and that mainland populations generally exhibit low morph ratios. Additionally, a correlation between morph ratio and population density was found lending support to the hypothesis that the ESS thresholds that define the morph ratios have diverged through local adaptation. A set of seven microsatellite markers are presented that were developed from a Farne island population of F. auricularia with one additional, previously published locus. These eight markers exhibit genetic variability within and between populations and as such can potentially be applied at a range of scales, from broad-scale phylogeography to within population parentage studies. A phylogeographic study of the UK populations using these markers suggests a single postglacial colonisation from mainland Europe and give further support to the local adaptation hypothesis of ESS threshold evolution. A study of ejaculate size in F. auricularia showed that the males transfer free sperm at a steady rate and that the morphs do not differ in the number of sperm per ejaculate. Measurements of change in body-mass were found to be ineffective measures of ejaculate size, but that macrolabic males lost more weight during copula than brachylabic males. This may be the result of differential investment in accessory ejaculate components between the morphs, as a result of the differing risk of sperm competition.

La dispersion est un trait d'histoire de vie qui joue un rôle majeur dans le fonctionnement des populations naturelles. Comprendre ce phénomène et son évolution est aujourd'hui déterminant pour la gestion des populations dans des écosystèmes de plus en plus anthropisés. Cette étude s'est attachée à caractériser la dispersion et ses déterminants chez le campagnol des champs, Microtus avalis, dans un paysage agricole de l'Ouest de la France. L'instabilité spatio-temporelle des agroécosystèmes et la démographie cyclique de ce petit rongeur colonial en font un modèle exceptionnel pour aborder cette problématique. La constitution d'une banque de marqueurs microsatellites nous a permis d'utiliser des approches de génétique des populations et de génétique paysagère. Une seule entité génétiquement homogène couvre les 500 km² du site d'étude et le patron d'isolement par la distance qui caractérise cette « population » indique que seule la distance limite le flux génique dans ce paysage. Les variations des patrons génétiques au cours des cycles reflètent l'instabilité de la balance entre dérive génétique et dispersion et montrent que le flux génique est positivement lié à la densité. Les patrons de dispersion, différents entre mâles et femelles (dispersion biaisée vers les mâles), suggèrent que la dispersion n'est pas déterminée par les mêmes causes évolutives selon le sexe. La colonisation par les femelles des habitats temporaire (les cultures annuelles), particulièrement massive lors des pullulations, permet la cohésion spatiale de la population. Sa cohésion génétique est assurée par les migrations répétées des mâles entre colonies pour la reproduction. Ce fonctionnement est rendu possible par l'extrême rapidité cycle de vie du campagnol des champs qui compense l'instabilité du paysage. L'intégration des différents résultats de cette étude dans un modèle de dispersion couplé à un modèle dynamique de paysage devrait permettre d'évaluer l'impact de l'évolution de l'agriculture sur cette espèce emblématique de la biodiversité de ces paysages.

Le but de cette étude était d'utiliser les techniques moléculaires dans deux populations de chevreuil, pour estimer la variance du succès reproducteur des mâles et identifier les principaux déterminants de cette variance.<br /> Comme attendu du fait de son faible dimorphisme sexuel, la variance dans le succès reproducteur à vie des mâles, et donc l'opportunité pour la sélection sexuelle, sont faibles chez le chevreuil, suggérant un faible degré de polygynie.<br /> Les mâles peuvent se reproduire dès 2 ans, mais avec un succès reproducteur inférieur à celui des mâles plus âgés.<br /> Le succès reproducteur tend aussi à décroître après 8 ans.<br /> La masse corporelle, la taille des bois, et la taille du territoire des mâles, affectent positivement leur succès reproducteur, mais pas la qualité de l'habitat dans leur territoire.<br /> Cette étude apporte de nouveaux éléments sur l'évolution de la territorialité et la sélection sexuelle chez les ongulés sauvages.

Projet de recherche réalisé avec Bernard Angers comme directeur de maîtrise, Denis Réale en tant que co-directeur et grâce à la collaboration active d'Emmanuel Milot. / La région métropolitaine de Montréal est formée de nombreuses îles à la jonction du fleuve Saint-Laurent et de la rivière des Outaouais, isolant ainsi les populations insulaires en fonction de distances respectives ainsi que des courants. Ce système offre un contexte idéal pour évaluer l’effet de la perte d’habitat liée à la pression d'urbanisation dans un paysage métropolitain insulaire ou en situation d’archipel. La présente étude a pour objectif de comparer l’effet de la perte d’habitat sur la diversité génétique de deux serpents très distincts, Storeria dekayi et Thamnophis sirtalis. Des analyses réalisées à l’aide de marqueurs microsatellites révèlent une plus importante structure génétique entre les populations de S. dekayi (FST=0,19) qu’entre celles de T. sirtalis (FST=0,07) dans la région montréalaise. Chez les deux espèces étudiées, la majorité des populations des habitats réduits présente une richesse allélique moyenne comparable à celle observée dans les habitats plus vastes. Néanmoins, certaines populations présentent des réponses différentes, dont des traces de goulots d’étranglement, une perte de richesse allélique ou encore une importante modification des fréquences alléliques. Au niveau régional, les résultats présentent une importante perte de diversité génétique chez les couleuvres se trouvant sur le continent alors que les populations insulaires de la région montréalaise constituent désormais un réservoir de diversité génétique. Les résultats observés auprès des populations insulaires démontrent que les effets de la perte d’habitat peuvent s’avérer très spécifiques à chaque situation et que la détection de traces génétiques d’un tel phénomène peut nécessiter un contexte logistique très particulier. Un nombre croissant de publications reportent une absence de signature génétique suite à la perte d’habitat chez des oiseaux et des mammifères. Il s’agit de la première étude témoignant de ce phénomène chez les reptiles. Une note est fournie en annexe à l’intention des gestionnaires au sujet de la conservation de la couleuvre brune, S. dekayi. / The Montreal metropolitan community includes numerous islands located at the confluence of the Saint-Lawrence and Ottawa Rivers. In such a fragmented landscape, dispersal of animals is limited by the distance between islands as well as the currents. This system offers an ideal context for the study of the effects of habitat loss on the genetic diversity of animal populations located on islands or archipelagos. This study seeks to assess the effects of habitat area by comparing the organization of genetic diversity of two highly distinct snake species, Storeria dekayi and Thamnophis sirtalis. Analysis realized with microsatellite markers reveals a much stronger genetic organisation in S. dekayi (FST=0.19) than in T. sirtalis (FST=0.07) in the Montreal area. For both studied species, most populations found in reduced habitats showed similar genetic diversity to what was observed in larger habitats. Nevertheless, some populations showed different responses to the loss of habitat, including traces of genetic bottlenecks, a loss in mean allelic richness or an important alteration of their allelic frequencies. This study also reveals an important loss of genetic diversity in the continental snake populations. At the regional scale, the results reveal an important loss of genetic diversity in the continental snake populations and that the insular populations of the Montreal area now constitute a reservoir of the remnant genetic diversity. Moreover, this study not only demonstrates that the genetic response to habitat loss can be very case-specific, but also that to detect traces of such a phenomenon can require a very particular framework. A growing number of publications based on birds and mammals have reported the absence of a genetic signature following a habitat loss. This is the first study to report this phenomenon in reptiles. A note intended for managers is provided about the conservation of the Dekay’s brown snake, S. dekayi.

Les Truffes sont des champignons ascomycètes ectomycorhiziens appartenant à la famille des Tuberaceae et plus précisément au genre Tuber. On dénombre à ce jour plus d’une trentaine d’espèces de Tuber en Europe. Lors de ce travail de thèse, nous nous sommes plus précisément focalisés sur le modèle Tuber aestivum-uncinatum. Cette truffe communément appelée « Truffe de Bourgogne » présente un intérêt à la fois gastronomique et culturel.La première partie de ce travail de thèse a porté sur la clarification du statut taxonomique de la truffe de Bourgogne (Tuber uncinatum). Pour cela, nous avons utilisé une approche multi-marqueurs combinant des marqueurs génétiques couramment utilisés à l’échelle interspécifique. Nos analyses ont montré que les deux taxons Tuber aestivum (la truffe d’été) et Tuber uncinatum sont conspécifiques. Durant la deuxième partie, nous nous sommes intéressés à la diversité génétique de Tuber aestivum. Pour cela, nous avons tout d’abord développé des marqueurs microsatellites spécifiques par une approche de « direct shotgun pyrosequencing ». Cette méthode a permis le développement de 15 marqueurs microsatellites polymorphes. Nous les avons ensuite utilisés pour génotyper des individus provenant de différentes localisations en Europe. Nous avons pu identifier quatre sous populations différenciées qui ne correspondent pas, pour la majorité, à une répartition géographique. Cependant, un des clusters se différencie des autres à la fois par sa situation géographique (sud de la France) et ses caractéristiques génétiques (présence d’allèles rares). Ces résultats préliminaires pourraient indiquer l’existence d’un écotype particulier attaché à une écologie méridionale, Tuber aestivum sensu stricto.Nous nous sommes ensuite intéressés, dans la troisième partie de ce travail de thèse à la diversité aromatique de Tuber aestivum à l’échelle locale. Les résultats obtenus permettent de mettre en évidence l’existence d’une différenciation modérée entre les individus issus d’une truffière naturelle et les individus issus d’une truffière plantée. D’une saison de récolte à l’autre, une stabilité génotypique a été observée. Au niveau aromatique, seuls les composés C8 semblent être liés aux génotypes.Dans la dernière partie, nous nous sommes intéressés à l’analyse de données de récolte sur plus de trente ans au sein d’une truffière plantée de noisetiers inoculés initialement par Tuber melanosporum. Grâce à des analyses statistiques simples, nous avons pu noter les fluctuations tant en quantité qu’en poids des truffes récoltées suivant les saisons et les arbres truffiers. Il apparait que le remplacement de Tuber melanosporum par Tuber aestivum s’est fait de manière très rapide (trois ans). La disparition de Tuber melanosporum peut probablement être expliquée par la fermeture de la canopée des noisetiers, Tuber melanosporum n’appréciant pas un ombrage excessif / Truffles are ectomycorrhizal Ascomycota fungi belonging to the Tuberaceae family and more specifically to the Tuber genus. More than thirty Tuber species are currently described in Europe. In this thesis, we specifically focused on the Tuber aestivum-uncinatum model. This truffle is commonly called "Burgundy Truffle" and has a gastronomic and cultural interest.The first part of this thesis focused on the taxonomic status of the Burgundy truffle (Tuber uncinatum). For this, we used a multi-marker approach combining several genetic markers commonly used at the interspecific scale. Our analyses showed that the two taxa, Tuber aestivum (summer truffle) and Tuber uncinatum are conspecific.In the second part, we addressed the genetic diversity of Tuber aestivum. To do this, we firstly developed specific microsatellite markers by "direct shotgun pyrosequencing". This method has allowed the development of 15 polymorphic microsatellite markers. Then, we used those markers to genotype individuals from different European locations. We have identified four differentiated subpopulations that not correspond, for the majority, to a geographical distribution. However, one cluster differs from the others by its location (south of France) and its genetic characteristics (presence of rare alleles). These preliminary results may indicate the existence of a particular ecotype attached to a southern ecology: Tuber aestivum sensu stricto.We were then interested, in the third part of this thesis, to the aromatic diversity of Tuber aestivum at a local scale. Our results highlight the existence of a moderate differentiation between individuals from a natural truffle orchard and individuals from planted orchard. From one season to another, genotypic stability was observed. Only C8 volatile organic compounds seem to be related to the genotypes.In the last part, we analyzed harvesting data, over more than thirty years, from an hazelnut truffle orchard initially inoculated by Tuber melanosporum. Through simple statistical analyzes, we noted changes in both quantity and weight of truffles harvested according to the seasons and hazelnut trees. It appears that Tuber aestivum rapidly replaced Tuber melanosporum (in three years). The disappearance of Tuber melanosporum can probably be explained by the canopy closure; Tuber melanosporum not appreciating excessive shading.

Resumo: O presente trabalho teve como objetivo produzir ferramentas e informações úteis para a conservação e exploração racional de Casearia sylvestris Sw. (Salicaceae), uma espécie que produz diterpenos clerodânicos de grande importância farmacológica (casearinas), e que é explorada por extrativismo. Tal objetivo foi alcançado através do desenvolvimento de marcadores microssatélites específicos para C. sylvestris e de um estudo da diversidade genética e química existente entre e dentro de populações do Estado de São Paulo. Tradicionalmente são reconhecidas duas variedades em C. sylvestris (var. sylvestris e var. lingua), o que é motivo de debate devido à existência de formas intermediárias. Este trabalho objetivou, adicionalmente, contribuir com argumentos genéticos para esta discussão. Foi construída uma biblioteca enriquecida em microssatélites, a partir da qual obtiveram-se e validaram-se dez pares de iniciadores (primers) microssatélites específicos para C. sylvestris. Estes pares de iniciadores foram utilizados para o estudo da estrutura genética de populações de C. sylvestris através da amostragem de 376 indivíduos em nove populações distribuídas em quatro ecossistemas (Floresta Ombrófila Densa, Floresta Estacional Semidecidual, Cerrado e ecótonos). As duas variedades foram amostradas de acordo com sua distribuição nestes ecossistemas. A genotipagem dos indivíduos para os locos amostrados foi realizada através de eletroforese em gel de acrilamida lido a 700 e 800 nm por um seqüenciador IR2-DNA Analyser (LI-COR). Os dados foram analisados através de abordagens frequentistas, bayesianas e baseadas na teoria de coalescência, utilizando-se diversos programas computacionais. Para o estudo da diversidade química, as mesmas populações foram amostradas, selecionando-se 12 indivíduos por população, totalizando 108 indivíduos. Adicionalmente, foram... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This work aimed obtaining tools and information for the conservation and rational exploitation of Casearia sylvestris Sw. (Salicaceae), a tree species which produces casearins, pharmacologically important clerodane diterpenes. This goal was achieved through the development of a set of polymorphic microsatellite markers, and through the study of chemical and genetic diversity in populations of C. sylvestris from São Paulo State. Also, we aimed contributing for the debate on the existence of two varieties within this species (var. sylvestris e var. lingua). A genomic library was constructed and 10 primer pairs were obtained. Those primers were utilized for a population genetic structure analysis, in which 376 individuals from nine populations distributed on four different ecosystems (Evergreen Atlantic Forest, Semideciduous Atlantic Forest, Cerrado and ecotones) were sampled. The two varieties were sampled according to its distribution among these populations. Genotyping was performed at 700 and 800 nm by electrophoresis on an IR2-DNA Analyser (LI-COR). The data were analyzed through frequentist, Bayesian and coalescence-based approaches, through the use of several softwares. Chemical diversity was studied by sampling in the same populations (12 individuals per population, i.e. 108 individuals). Also, cuttings of these individuals were prepared, aiming to verify its' chemical compounds after a year of green-house cultivation. Cuttings' rooting was problematic and a methodology was developed. Only 46 cuttings survived. Casearins from these 154 individuals (108 + 46) were extracted and analyzed by HPLC. Genetic analysis results suggests a partial genome duplication, as more than two alleles for the same locus were observed in 8% of var. sylvestris individuals and in 70% of var. lingua individuals. Additional studies are necessary to verify the hypothesis of partial genome duplication... (Complete abstract click electronic access below) / Orientador: Marcos Aparecido Gimenes / Coorientador: Alberto José Cavalheiro / Banca: Roseli B. Torres / Banca: Maria Imaculada Zucchi / Banca: Giancarlo C. X. Oliveira / Doutor