A interação entre plantas e seus visitantes florais forma uma rede de interação complexa. Essa rede de interação possui propriedades emergentes que caracterizam a estrutura da rede. A composição, a riqueza e o papel das espécies nas redes de interação podem variar espaço-temporalmente e levar a alteração na estrutura das redes. A estrutura também pode variar de acordo com o grau de dependência entre os pares de espécies. É esperado que interações com elevado grau de dependência mútua variem menos no tempo e no espaço. A interação entre as plantas produtoras de óleo floral e as abelhas coletoras de óleo é de alta dependência mútua, já que as abelhas necessitam do óleo para alimentar as larvas e construir ninhos e são polinizadores mais importantes dessas plantas. O alimento larval destas abelhas é composto por pólen, óleo e néctar. A quantidade de alimento disponível para as larvas pode influenciar o tamanho dos adultos quando emergirem. As espécies de abelhas coletoras de óleo são solitárias, ou seja, não existe contato entre as gerações. Assim uma abelha recém-emergida deve ser capaz de encontrar as fontes de óleo, seguindo algum sinal de atração, que poderia ser através de odores voláteis. No presente trabalho analisamos se a estrutura da rede de interação formada pelas plantas produtoras de óleo floral e seus visitantes coletores de óleo varia espacialmente e temporalmente. Para a análise temporal coletamos dados da interação por dois anos em Itirapina, São Paulo. No estudo de variação espacial comparamos as redes de interação em cinco áreas de cerrado no interior de São Paulo. Em ambos os estudos usamos métricas de redes complexas. Além disso, estudamos a relação entre tamanho de célula de cria e quantidade de alimento disponível para as larvas e tamanho das abelhas adultas ao emergirem, para uma espécie de abelha coletora de óleo: Tetrapedia diversipes. Para isso medimos abelhas e suas células de cria e relacionamos estas medidas. Também manipulamos a quantidade de alimento disponível para as larvas e medimos os adultos emergidos. Por fim, procuramos odores voláteis em Byrsonima intermedia. Coletamos odores voláteis com a técnica de dynamic headspace. Analisamos as coletas com GCMS. A estrutura da rede de interação entre as plantas produtoras de óleo floral e as abelhas coletoras de óleo variou espaço-temporalmente. A análise temporal mostrou que o grau de especialização da rede foi fortemente alterado entre os anos de estudo e que a abundância foi um fator importante para explicar a ocorrência das interações. As espécies que tiveram suas abundâncias alteradas entre os anos também tiveram alteração no número de interações. Na análise espacial observamos que as interações e as espécies compartilhadas entre os locais de estudo foram aquelas com maior grau de interação (generalista). Além das interações e espécies generalistas se manterem nas cinco localidades, as espécies generalistas sustentam as espécies localmente raras. Byrsonima intermedia foi considerada uma espécie super-generalista nas cinco redes analisadas e 77% das interações compartilhadas envolveram esta espécie. A morfometria dos indivíduos de Tetrapedia diversipes estudados foi positivamente correlacionada com o volume da célula de cria e a quantidade de alimento. Quando relacionamos o volume das células de cria com as medidas corpóreas das abelhas verificamos que as abelhas maiores emergiram em células maiores. No entanto essa relação foi fraca. Após o experimento de manipulação de alimento verificamos que as abelhas emergidas de células sem manipulação foram maiores que as abelhas que tiveram o alimento retirado, e que entre as abelhas manipuladas aquelas que receberam mais alimento foram maiores. Nós não encontramos odores voláteis em flores de Byrsonima intermedia. / The plant pollinator interaction forms a complex network. These networks have emergent properties that characterize the structure of network. The number, identity and species role of the network may vary spatio-temporally and lead to changes in the structure of the networks. Moreover, the structure may vary according to the dependence degree of the interacting pairs. It is expected that interactions with a high mutual dependence degree are more stable. The interaction between oil producing plants and oil collecting bees is highly mutual dependence, since bees need oil to feed the larvae and are pollinators of these plants. The larval diet consists of pollen and oil/nectar. The amount of food available to the larvae may influence the adults size. The oil collecting bees are solitary, i.e., there is no contact between generations. Thus, a naive emerged bee should be able to find the oil plants, using some attractive cues that could be volatile odor. In this work we want to answer if the network structure of the interaction between oil producing plants and oil collecting bees varies spatially and temporally. For the temporal analysis we carried out field work for two years at Itirapina, São Paulo. In the study of spatial variation we compare interaction networks in five areas of cerrado. In both studies we use metrics of complex networks. Furthermore, we studied the relation between size of adult bees (Tetrapedia diversipes) and brood cell sizes and the amount of food available for the larvae. We measured the bees and their brood cells and relate it. Also we manipulated the amount of food available for the larvae and we took measures of the adults. Finally, we search for volatile odors in Byrsonima intermedia. We collect volatile odors with the technique of dynamic headspace. We analyzed the samples with GCMS. The network structure of interaction between oil producing plants and oil collecting bees varied spatio-temporally. The temporal analysis showed that the specialization degree of the network changed between the years and that the abundance has a key role to explain the occurrence of interactions. Species that had changed their abundances between years were also changed in the number of interactions. Spatial analysis found out that the interactions and species ubiquitous among the study sites were those that had the greatest interaction degree (generalist specie). Moreover, generalist species maintain the locally rare species. Byrsonima intermedia was regarded a super-generalist specie in the five networks analyzed and 77% of ubiquitous interactions involved this species. The morphometry of Tetrapedia diversipes individuals was positively correlated with the volume of the brood cells and the amount of food. When relating the volume of brood cells with measures of bees we found that larger bees emerged in larger cells. However this relationship was weak. The experiment of handling food showed that bees emerged from control cells were larger than bees that had the food removed, and between the manipulated bees those who received more food became bigger. We did not detect volatile odors in flowers Byrsonima intermedia.

Les recherches menées au cours des dernières années ont permis de mettre en évidence les nombreuses fonctions écologiques des composés organiques volatils (COVs) émis par les plantes. Cette thèse a eu pour objectif d'utiliser ces connaissances dans un cadre de protection des cultures. Nous avons essayé de poser les bases d'une stratégie de type « Push-Pull » contre la mouche du chou (Delia radicum) en utilisant des sources d'odeurs synthétiques (diffuseurs de COVs) ou des sources d'odeurs naturelles (plantes compagnes). A partir d'observations menées en plein champ nous avons sélectionné plusieurs brassicacées présentant des taux d'infestations contrastés vis-à-vis de la mouche du chou. Des expérimentations de laboratoires combinant des études comportementales et des analyses chromatographiques ont permis (i) d'établir un lien entre l'infestation et l'attractivité de certaines plantes (ii) d'identifier de nouveaux COVs potentiellement impliqués dans les prises de décisions comportementales de D. radicum. En parallèle, des études de terrain ont permis de tester l'efficacité de diffuseurs de COVs synthétiques au sein de parcelles expérimentales, que ce soit (i) pour favoriser le recrutement d'ennemis naturels (ii) attirer (composante « Pull ») ou repousser (composante « Push ») la mouche du chou. Enfin, l'utilisation de plantes pièges concentrant le ravageur semble être intéressante pour modifier la répartition de la mouche du chou à l'échelle de la parcelle en vue de protéger des cultures d'intérêt telles que le brocoli. / Volatile organic compounds (VOCs) emitted by plants are used by phytophagous, predatory and parasitoid insects to adjust behavioral decisions in complex environments. We aimed at using these ecological functions of VOCs in crop protection. We developed the first steps of a "Push-Pull" strategy toward the cabbage root fly (Delia radicum) with both dispensers of synthetic VOCs and natural blends of VOCs released by plants. We conducted a field study to select brassicaceous plants exhibiting contrasted levels of infestation toward the cabbage root fly. Laboratory bioassays combining behavioral observations and gas chromatographic analyses allowed to (i) investigate the relation between plant infestation and plant attractiveness (ii) identify novel VOCs involved in behavioral decisions of the cabbage root fly. Parallel field experiments were used to test the potential of dispensers of synthetic VOCs to (i) enhance natural enemies recruitment (ii) modify cabbage root fly behavior, attracting ("Pull" component) or repelling the pest ("Push" component). Finally, we demonstrated that attractive plants are promising for a trap crop approach enabling to redistribute cabbage root fly density at the field scale in order to protect main crop such as broccoli.

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No description available.

The Eurasian spruce bark beetle Dendroctonus micans is a major pest of spruce which is expanding its range in France, Turkey, England and Wales. Its monospecific predator Rhizophagus grandis has followed naturally the bark beetle into most areas and, since the 1960s, has also been mass-produced and successfully released within newly infested locations. In this scope, the development of an effective trapping method would be very useful to assess the bark-beetle presence at previously uninfested sites, or predator establishment after release or natural spread. We demonstrated the efficiency of oxygenated monoterpenes-baited kairomone traps to monitor R. grandis in various epidemiological conditions, including areas localized behind or at the limit of the pest’s distribution, or in areas where artificial releases were performed. Because the predator is strictly species-specific, another exciting possibility offered by the kairomone trapping is the indirect monitoring of the pest itself in areas of unknown status (e.g. areas under colonization, or considered as at risk at medium- term). R. grandis is also considered as one of the most valuable natural enemies to strike aggressive North-American Dendroctonus species. In this respect, R. grandis has been recently applied in a neo-classical biological program against the red turpentine beetle D. valens, which invaded China from North America in the late 1990’s. In laboratory tests conducted on pine logs in the laboratory, or on living pine trees in the field, we demonstrated that R. grandis adults can successfully enter and reproduce into D. valens galleries. In Europe, R. grandis is the only species regularly found in the brood systems of D. micans, where adults and larvae attack the gregarious larvae of their prey. In such enclosed systems, R. grandis’ functional response is therefore influenced by various interrelated components, such as the prey density, the predator density, or the prey distribution. Measuring the predator’s success in terms of larval survival and growth rates, we demonstrated the time spent by R. grandis larvae to wound and kill their prey to be the main factor limiting their development. This factor may be considerably influenced by the proportions of diseased, wounded or molting prey rise in the brood system, for instance as a result of an increase in prey density, or due to the presence of conspecific adults (which wound their prey but do not consume them entirely). Furthermore, our tests suggest that no cannibalism or noticeable intraspecific competition occurred between R. grandis larvae, whereas some lighter mode of competition probably took place. R. grandis also exhibits a reproductive numerical response to prey density, which mainly relies on the perception of chemical stimuli and inhibitors released in the bark beetle brood system. In the current study, we developed a non-destructive approach to follow the dynamics of volatile compound production, using sequential sample collection on SPME fibers. Our tests demonstrated that the larval activity of D. micans or D. valens strongly influences the release of some oxygenated monoterpenes. However, our attempts to correlate the relative quantities of some identified chemicals to offspring production were less successful as it concerns the identification of potential oviposition stimuli and inhibitors. The problematic rose by the progression of D. micans, as well as detailed results of each of the described above studies are discussed in the two published papers and the three manuscripts forming this thesis. Bringing all these studies together, several perspectives are also presented in the general discussion. / Ravageur des épicéas, Dendroctonus micans est toujours en voie d’extension en France, en Turquie, en Angleterre et au Pays de Galles. Dans la plupart de ces zones, le dendroctone est accompagné de manière naturelle par son prédateur monospécifique, Rhizophagus grandis. Depuis les années 1960, le prédateur a également fait l’objet d’une production de masse et de programmes de lâchers dans les zones d’arrivée récente du scolyte. Dans le cadre de la lutte biologique contre D. micans, les gestionnaires forestiers doivent donc estimer au plus tôt la présence du ravageur dans des zones jusque là indemnes, mais également vérifier l’établissement du prédateur par progression naturelle ou résultant d’introductions délibérées. Dans la présente étude, nous avons démontré l’efficacité de pièges d’interception appatés à l’aide de monoterpènes oxygénés pour la capture de R. grandis. Celle-ci s’est faite dans différentes conditions épidémiologiques, incluant notamment des zones situées en arrière du front de progression du scolyte et des zones où des lâchers artificiels ont été réalisés. Comme R. grandis est strictement inféodé au dendroctone, un autre avantage de la technique est la possibilité de réaliser un dépistage indirect du ravageur dans les zones où son statut est incertain (zones en cours de colonisation, ou considérées comme à risque à moyen terme). Par ailleurs, R. grandis est également considéré comme un des meilleurs ennemis naturels potentiels pour lutter contre d’autres espèces de Dendroctonus aggressifs. Dans cette optique, R. grandis a été récemment utilisé dans un programme de lute biologique contre D. valens, ravageur invasif arrivé en Chine dans la fin des années 1990 en provenance d’Amérique du Nord. Nous avons démontré la capacité de R. grandis à s’introduire et à se reproduire dans les galeries de D. valens lors de tests de laboratoire, mais aussi sur des arbres vivants en pinèdes. En Europe, R. grandis est strictement inféodé aux galeries de D. micans, où larves et adultes du prédateur s’attaquent aux larves grégaires du scolyte. Dans ce système clos, la réponse fonctionelle de R. grandis est influencée par plusieurs facteurs étroitement corrélés, la densité de proies, la densité de prédateurs, et la distribution des proies. En mesurant l’efficacité de prédation de R. grandis en termes de survie des larves et de taux de croissance, nous avons démontré l’influence sur leur développement du temps passé par les larves à blesser et à tuer leurs proies. Ce facteur est par ailleurs fortement dépendant de la proportion de larves malades, blessées ou en cours de mue au sein du système ; une proportion qui peut augmenter en réponse à une augmentation de la densité de proies, ou lorsque des adultes sont présents (ceux-ci blessent les proies mais ne les consomment pas entièrement). Enfin, nos tests suggèrent qu’il n’existe que peu de cannibalisme ou de compétition intraspécifique entre larves de R. grandis, tandis que des modes de compétition moins importants prennent vraisemblablement place. R. grandis présente également une réponse numérique reproductive à la densité de proies disponibles, principalement basée sur la perception de stimuli et d’inhibiteurs présents dans les galeries du scolyte. Par la collecte de composés volatils présents dans ces systèmes à l’aide de fibres SPME, nous avons développé une méthode non-destructive pour suivre la dynamique de production des médiateurs chimiques. Nos tests ont démontré que l’activité des larves de D. micans ou D. valens influence fortement la dynamique de production de certains monoterpènes oxygénés. En revanche, il n’a pas été été possible de corréler les différents composés identifiés au nombre de larves de R. grandis présentes dans le système. La problématique soulevée par la progression de D. micans, de même que les résultats détaillés de chacune des études décrites ci-dessus sont discutés dans les deux papiers publiés et les trois manuscrits formant cette thèse. Les différentes perspectives apportées par ce travail sont également présentées dans la discussion générale.

Rhizophagus grandis is one of the rare examples of totally specific predators: it only preys upon the bark beetle Dendroctonus micans in its native range (Europe and Asia). In the course of coevolution in such predator-prey systems, prey are selected for avoidance and resistance mechanisms against predators. Previous laboratory trials suggested that R. grandis could lay more eggs in presence of exotic prey species with an ecology similar to that of D. micans, but which could not evolve resistances against this predator, such as the North American parasitic bark beetles D. punctatus and D. valens. The specificity of the association R. grandis-D. micans is thought to depend on chemical signals (kairomones) used by the predator to find a gallery of its prey and to adjust its oviposition to the number of prey larvae available inside. However, the exact nature of the chemical signals regulating R. grandis’ oviposition is still unknown. While these signals seem specific in nature, stimulation of the predator’s oviposition by exotic prey species suggests that specificity may be constrained by geography. Unlike most bark beetles, which kill living trees or feed on dying trees, D. micans completes its development on healthy trees as a true parasite. This niche is highly defended by tree toxicants (terpenes among others) to which both D. micans and R. grandis are resistant. Insects possess their own detoxification processes, but they may also rely on symbiotic microorganisms (bacteria and fungi) to contend with the specific constraints of their niche. In comparison to other bark beetles, microbial communities of parasitic bark beetles are yet poorly known, as in the case of D. micans and D. punctatus. Apart from detoxification, insect symbionts may provide nutritional supplementation, protection against pathogens, or components of the chemical communication, which affect the hosts as well as partners of the tritrophic relationship tree-bark beetle-natural enemies. The system R. grandis-D. micans is a unique opportunity to study several aspects of this coevolution which are poorly understood to date. The objectives of this thesis were:- (1) the comparison of the oviposition performances of R. grandis on the specific prey D. micans, and on the exotic prey D. punctatus and D. valens, in order to emphasize possible prey resistance mechanisms; - (2) the identification of the chemical signals regulating R. grandis’ oviposition, and the evaluation of their role in the specificity of the association with D. micans, in presence of the exotic prey D. punctatus and D. valens; - (3) the characterization of the bacterial and fungal communities associated with non-aggressive populations of the bark beetles D. micans, D. punctatus and D. valens, and investigating how these microbial symbionts may affect bark beetle hosts in respect of their ecology as well as the tritrophic relationship tree-bark beetle-natural enemies. Our results show that R. grandis laid equivalent numbers of eggs with the native prey D. micans and with the exotic prey D. punctatus and D. valens, which could illustrate that R. grandis’ specificity is constrained by geography but which does not emphasize possible prey resistance mechanisms against the long-standing predator. We identified robust candidates to the stimulation and inhibition of R. grandis’ oviposition which should be confirmed in bioassays. Finally, we report that the bacterial and fungal communities of non-aggressive populations of D. micans, D. punctatus and D. valens are mainly composed of widespread environmental Enterobacteria and yeasts, and we discuss the various ways they may influence bark beetle hosts in respect of their life histories including the attacked trees and their natural enemies. Overall, this thesis illustrates the need to encompass all levels of complexity, from prey symbionts and semiochemicals to predators, to study systems like R. grandis-D. micans. / Rhizophagus grandis est un des rares exemples de prédateurs totalement spécifiques :présent en Europe et en Asie, il se nourrit exclusivement du scolyte Dendroctonus micans. Dans de telles associations, la coévolution prédateur-proie mène à la sélection de mécanismes d’évitement ou de résistance chez la proie, contre le prédateur. De précédentes observations en laboratoire ont suggéré que R. grandis pouvait pondre davantage d’œufs en présence de proies exotiques d’écologie comparable à celle de D. micans, mais qui n’auraient pu développer de résistance à l’encontre de ce prédateur, à l’instar des scolytes parasitiques nord-américains D. punctatus et D. valens. Il semble que la spécificité de l’association R. grandis-D. micans repose sur des signaux chimiques (kairomones) qui permettent au prédateur de repérer les galeries de sa proie et d’y ajuster le nombre d’œufs déposé à la quantité de larves de proies disponible. Néanmoins, on ignore encore la nature exacte des signaux régulant l’oviposition de R. grandis. Le fait que l’oviposition soit stimulée par des proies exotiques indique que la spécificité de cette association pourrait être limitée par des barrières géographiques. Contrairement à la plupart des scolytes qui tue des arbres ou s’attaque à des arbres mourants, D. micans se développe intégralement sur des arbres en bonne santé, en véritable parasite. Cette niche est fortement défendue par les composés toxiques de l’arbre (en particulier les terpènes), auxquels à la fois D. micans et R. grandis sont résistants. Certains insectes possèdent leurs propres processus de détoxification, mais ils peuvent également bénéficier de l’aide de microorganismes symbiotiques (bactéries et champignons). En comparaison avec les autres scolytes, les communautés microbiennes associées aux scolytes parasitiques est très peu documentée, comme dans le cas de D. micans et D. punctatus. En dehors de la détoxification, les symbiotes d’insectes peuvent contribuer à leur nutrition, les protéger contre des pathogènes, ou intervenir dans la communication chimique, ce qui affecte leurs hôtes comme les autres acteurs de la relation tritrophique arbre-scolyte-ennemi naturel. L’association R. grandis-D. micans est une opportunité unique d’étudier des aspects méconnus de la coévolution. Les objectifs de cette thèse étaient de :- (1) comparer le nombre d’œufs pondu par R. grandis sur sa proie spécifique, D. micans, et sur les proies exotiques D. punctatus et D. valens, afin de mettre en évidence de possibles résistances ;- (2) identifier les signaux chimiques qui régulent l’oviposition de R. grandis, et évaluer leur rôle dans la spécificité de l’association avec D. micans, en présence des proies exotiques D. punctatus et D. valens ;- (3) caractériser les communautés bactérienne et fongique associées aux populations parasitiques des scolytes D. micans, D. punctatus et D. valens, et investiguer comment ces microorganismes symbiotiques peuvent influencer leurs hôtes, selon leurs contraintes écologiques, ainsi que leurs ennemis naturels et arbres-hôtes. Nos résultats révèlent une oviposition équivalente de R. grandis en présence de la proie native D. micans et des proies exotiques D. punctatus et D. valens, ce qui illustre que la spécificité de cette association pourrait être limitée par des barrières géographiques mais ne met pas en évidence de possibles résistances à l’encontre du prédateur de longue date. Nous avons identifié des candidats robustes à la stimulation et à l’inhibition de la ponte de R. grandis, et leur rôle devrait être confirmé par des bioessais. Enfin, nous rapportons que la communauté microbienne associée aux populations parasitiques des scolytes D. micans, D. punctatus et D. valens est principalement constituée d’Entérobactéries et de levures répandues, et nous discutons des différentes façons dont ces symbiotes peuvent affecter leurs hôtes et autres acteurs de la relation tritrophique arbre-scolyte-ennemi naturel, selon leurs écologies respectives. Dans son ensemble, cette thèse souligne l’importance de considérer tous les niveaux de complexité biologique, des microorganismes associés aux proies jusqu’aux prédateurs, afin d’étudier des systèmes comme R. grandis-D. micans. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

The Eurasian spruce bark beetle Dendroctonus micans is a major pest of spruce which is expanding its range in France, Turkey, England and Wales. Its monospecific predator Rhizophagus grandis has followed naturally the bark beetle into most areas and, since the 1960s, has also been mass-produced and successfully released within newly infested locations. <p>In this scope, the development of an effective trapping method would be very useful to assess the bark-beetle presence at previously uninfested sites, or predator establishment after release or natural spread. We demonstrated the efficiency of oxygenated monoterpenes-baited kairomone traps to monitor R. grandis in various epidemiological conditions, including areas localized behind or at the limit of the pest’s distribution, or in areas where artificial releases were performed. Because the predator is strictly species-specific, another exciting possibility offered by the kairomone trapping is the indirect monitoring of the pest itself in areas of unknown status (e.g. areas under colonization, or considered as at risk at medium- term).<p>R. grandis is also considered as one of the most valuable natural enemies to strike aggressive North-American Dendroctonus species. In this respect, R. grandis has been recently applied in a neo-classical biological program against the red turpentine beetle D. valens, which invaded China from North America in the late 1990’s. In laboratory tests conducted on pine logs in the laboratory, or on living pine trees in the field, we demonstrated that R. grandis adults can successfully enter and reproduce into D. valens galleries. <p>In Europe, R. grandis is the only species regularly found in the brood systems of D. micans, where adults and larvae attack the gregarious larvae of their prey. In such enclosed systems, R. grandis’ functional response is therefore influenced by various interrelated components, such as the prey density, the predator density, or the prey distribution. Measuring the predator’s success in terms of larval survival and growth rates, we demonstrated the time spent by R. grandis larvae to wound and kill their prey to be the main factor limiting their development. This factor may be considerably influenced by the proportions of diseased, wounded or molting prey rise in the brood system, for instance as a result of an increase in prey density, or due to the presence of conspecific adults (which wound their prey but do not consume them entirely). Furthermore, our tests suggest that no cannibalism or noticeable intraspecific competition occurred between R. grandis larvae, whereas some lighter mode of competition probably took place. <p>R. grandis also exhibits a reproductive numerical response to prey density, which mainly relies on the perception of chemical stimuli and inhibitors released in the bark beetle brood system. In the current study, we developed a non-destructive approach to follow the dynamics of volatile compound production, using sequential sample collection on SPME fibers. Our tests demonstrated that the larval activity of D. micans or D. valens strongly influences the release of some oxygenated monoterpenes. However, our attempts to correlate the relative quantities of some identified chemicals to offspring production were less successful as it concerns the identification of potential oviposition stimuli and inhibitors. <p>The problematic rose by the progression of D. micans, as well as detailed results of each of the described above studies are discussed in the two published papers and the three manuscripts forming this thesis. Bringing all these studies together, several perspectives are also presented in the general discussion. <p>/<p>Ravageur des épicéas, Dendroctonus micans est toujours en voie d’extension en France, en Turquie, en Angleterre et au Pays de Galles. Dans la plupart de ces zones, le dendroctone est accompagné de manière naturelle par son prédateur monospécifique, Rhizophagus grandis. Depuis les années 1960, le prédateur a également fait l’objet d’une production de masse et de programmes de lâchers dans les zones d’arrivée récente du scolyte.<p>Dans le cadre de la lutte biologique contre D. micans, les gestionnaires forestiers doivent donc estimer au plus tôt la présence du ravageur dans des zones jusque là indemnes, mais également vérifier l’établissement du prédateur par progression naturelle ou résultant d’introductions délibérées. Dans la présente étude, nous avons démontré l’efficacité de pièges d’interception appatés à l’aide de monoterpènes oxygénés pour la capture de R. grandis. Celle-ci s’est faite dans différentes conditions épidémiologiques, incluant notamment des zones situées en arrière du front de progression du scolyte et des zones où des lâchers artificiels ont été réalisés. Comme R. grandis est strictement inféodé au dendroctone, un autre avantage de la technique est la possibilité de réaliser un dépistage indirect du ravageur dans les zones où son statut est incertain (zones en cours de colonisation, ou considérées comme à risque à moyen terme).<p>Par ailleurs, R. grandis est également considéré comme un des meilleurs ennemis naturels potentiels pour lutter contre d’autres espèces de Dendroctonus aggressifs. Dans cette optique, R. grandis a été récemment utilisé dans un programme de lute biologique contre D. valens, ravageur invasif arrivé en Chine dans la fin des années 1990 en provenance d’Amérique du Nord. Nous avons démontré la capacité de R. grandis à s’introduire et à se reproduire dans les galeries de D. valens lors de tests de laboratoire, mais aussi sur des arbres vivants en pinèdes. <p>En Europe, R. grandis est strictement inféodé aux galeries de D. micans, où larves et adultes du prédateur s’attaquent aux larves grégaires du scolyte. Dans ce système clos, la réponse fonctionelle de R. grandis est influencée par plusieurs facteurs étroitement corrélés, la densité de proies, la densité de prédateurs, et la distribution des proies. En mesurant l’efficacité de prédation de R. grandis en termes de survie des larves et de taux de croissance, nous avons démontré l’influence sur leur développement du temps passé par les larves à blesser et à tuer leurs proies. Ce facteur est par ailleurs fortement dépendant de la proportion de larves malades, blessées ou en cours de mue au sein du système ;une proportion qui peut augmenter en réponse à une augmentation de la densité de proies, ou lorsque des adultes sont présents (ceux-ci blessent les proies mais ne les consomment pas entièrement). Enfin, nos tests suggèrent qu’il n’existe que peu de cannibalisme ou de compétition intraspécifique entre larves de R. grandis, tandis que des modes de compétition moins importants prennent vraisemblablement place.<p>R. grandis présente également une réponse numérique reproductive à la densité de proies disponibles, principalement basée sur la perception de stimuli et d’inhibiteurs présents dans les galeries du scolyte. Par la collecte de composés volatils présents dans ces systèmes à l’aide de fibres SPME, nous avons développé une méthode non-destructive pour suivre la dynamique de production des médiateurs chimiques. Nos tests ont démontré que l’activité des larves de D. micans ou D. valens influence fortement la dynamique de production de certains monoterpènes oxygénés. En revanche, il n’a pas été été possible de corréler les différents composés identifiés au nombre de larves de R. grandis présentes dans le système. <p>La problématique soulevée par la progression de D. micans, de même que les résultats détaillés de chacune des études décrites ci-dessus sont discutés dans les deux papiers publiés et les trois manuscrits formant cette thèse. Les différentes perspectives apportées par ce travail sont également présentées dans la discussion générale.<p> / Doctorat en Sciences agronomiques et ingénierie biologique / info:eu-repo/semantics/nonPublished

Metabolomics is the methodical scientific study of biochemical processes associated with the metabolome—which comprises the entire collection of metabolites in any biological entity. Metabolome changes occur as a result of modifications in the genome and proteome, and are, therefore, directly related to cellular phenotype. Thus, metabolomic analysis is capable of providing a snapshot of cellular physiology. Untargeted metabolomics is an impartial, all-inclusive approach for detecting as many metabolites as possible without a priori knowledge of their identity. Hence, it is a valuable exploratory tool capable of providing extensive chemical information for discovery and hypothesis-generation regarding biochemical processes. A history of metabolomics and advances in the field corresponding to improved analytical technologies are described in Chapter 1 of this dissertation. Additionally, Chapter 1 introduces the analytical workflows involved in untargeted metabolomics research to provide a foundation for Chapters 2 – 5. Part I of this dissertation which encompasses Chapters 2 – 3 describes the utilization of mass spectrometry (MS)-based untargeted metabolomic analysis to acquire new insight into cancer detection. There is a knowledge deficit regarding the biochemical processes of the origin and proliferative molecular mechanisms of many types of cancer which has also led to a shortage of sensitive and specific biomarkers. Chapter 2 describes the development of an in vitro diagnostic multivariate index assay (IVDMIA) for prostate cancer (PCa) prediction based on ultra performance liquid chromatography-mass spectrometry (UPLC-MS) metabolic profiling of blood serum samples from 64 PCa patients and 50 healthy individuals. A panel of 40 metabolic spectral features was found to be differential with 92.1% sensitivity, 94.3% specificity, and 93.0% accuracy. The performance of the IVDMIA was higher than the prevalent prostate-specific antigen blood test, thus, highlighting that a combination of multiple discriminant features yields higher predictive power for PCa detection than the univariate analysis of a single marker. Chapter 3 describes two approaches that were taken to investigate metabolic patterns for early detection of ovarian cancer (OC). First, Dicer-Pten double knockout (DKO) mice that phenocopy many of the features of metastatic high-grade serous carcinoma (HGSC) observed in women were studied. Using UPLC-MS, serum samples from 14 early-stage tumor DKO mice and 11 controls were analyzed. Iterative multivariate classification selected 18 metabolites that, when considered as a panel, yielded 100% accuracy, sensitivity, and specificity for early-stage HGSC detection. In the second approach, serum metabolic phenotypes of an early-stage OC pilot patient cohort were characterized. Serum samples were collected from 24 early-stage OC patients and 40 healthy women, and subsequently analyzed using UPLC-MS. Multivariate statistical analysis employing support vector machine learning methods and recursive feature elimination selected a panel of metabolites that differentiated between age-matched samples with 100% cross-validated accuracy, sensitivity, and specificity. This small pilot study demonstrated that metabolic phenotypes may be useful for detecting early-stage OC and, thus, supports conducting larger, more comprehensive studies. Many challenges exist in the field of untargeted metabolomics. Part II of this dissertation which encompasses Chapters 4 – 5 focuses on two specific challenges. While metabolomic data may be used to generate hypothesis concerning biological processes, determining causal relationships within metabolic networks with only metabolomic data is impractical. Proteins play major roles in these networks; therefore, pairing metabolomic information with that acquired from proteomics gives a more comprehensive snapshot of perturbations to metabolic pathways. Chapter 4 describes the integration of MS- and NMR-based metabolomics with proteomics analyses to investigate the role of chemically mediated ecological interactions between Karenia brevis and two diatom competitors, Asterionellopsis glacialis and Thalassiosira pseudonana. This integrated systems biology approach showed that K. brevis allelopathy distinctively perturbed the metabolisms of these two competitors. A. glacialis had a more robust metabolic response to K. brevis allelopathy which may be a result of its repeated exposure to K. brevis blooms in the Gulf of Mexico. However, K. brevis allelopathy disrupted energy metabolism and obstructed cellular protection mechanisms including altering cell membrane components, inhibiting osmoregulation, and increasing oxidative stress in T. pseudonana. This work represents the first instance of metabolites and proteins measured simultaneously to understand the effects of allelopathy or in fact any form of competition. Chromatography is traditionally coupled to MS for untargeted metabolomics studies. While coupling chromatography to MS greatly enhances metabolome analysis due to the orthogonality of the techniques, the lengthy analysis times pose challenges for large metabolomics studies. Consequently, there is still a need for developing higher throughput MS approaches. A rapid metabolic fingerprinting method that utilizes a new transmission mode direct analysis in real time (TM-DART) ambient sampling technique is presented in Chapter 5. The optimization of TM-DART parameters directly affecting metabolite desorption and ionization, such as sample position and ionizing gas desorption temperature, was critical in achieving high sensitivity and detecting a broad mass range of metabolites. In terms of reproducibility, TM-DART compared favorably with traditional probe mode DART analysis, with coefficients of variation as low as 16%. TM-DART MS proved to be a powerful analytical technique for rapid metabolome analysis of human blood sera and was adapted for exhaled breath condensate (EBC) analysis. To determine the feasibility of utilizing TM-DART for metabolomics investigations, TM-DART was interfaced with traveling wave ion mobility spectrometry (TWIMS) time-of-flight (TOF) MS for the analysis of EBC samples from cystic fibrosis patients and healthy controls. TM-DART-TWIMS-TOF MS was able to successfully detect cystic fibrosis in this small sample cohort, thereby, demonstrating it can be employed for probing metabolome changes. Finally, in Chapter 6, a perspective on the presented work is provided along with goals on which future studies may focus.