Doctor of Philosophy (PhD) / A renewed focus on generalised pollinator systems has inspired a conceptual framework which highlights that spatial and temporal interactions among plants and their assemblage of pollinators can vary across the individual, population, regional and species levels. Pollination is clearly a dynamic interaction, varying in the number and interdependence of participants and the strength of the outcome of the interaction. Therefore, the role of variation in pollination is fundamental for understanding ecological dynamics of plant populations and is a major factor in the evolution and maintenance of generalised and specialised pollination systems. My study centred on these basic concepts by addressing the following questions: (1) How variable are pollinators in a generalised pollination system? To what degree do insect visitation rates and assemblage composition vary spatially among populations and temporally among flowering seasons? (2) How does variation in pollinators affect plant reproductive success? I chose to do this using a model system, Trachymene incisa subsp. incisa (Apiaceae), which is a widespread Australian herbaceous species with simple white flowers grouped into umbels that attract a high diversity of insect visitors. The Apiaceae are considered to be highly generalist in terms of pollination, due to their simple and uniform floral display and easily accessible floral rewards. Three populations of T. incisa located between 70 km and 210 km apart were studied over 2-3 years. The few studies investigating spatial and temporal variation simultaneously over geographic and yearly/seasonal scales indicate that there is a trend for more spatial than temporal variation in pollinators of generalist-pollinated plants. My study showed both spatial and temporal variation in assemblage composition among all populations and variation in insect visitation rates, in the form of a significant population by year interaction. However, removing ants from the analyses to restrict the assemblage to flying insects and the most likely pollinators, resulted in a significant difference in overall visitation rate between years but no difference in assemblage composition between the Myall Lakes and Tomago populations. These results indicate more temporal than spatial variation in the flying insect visitor assemblage of T. incisa. Foraging behaviour provides another source of variation in plant-pollinator interactions. Trachymene incisa exhibits umbels that function as either male or female at any one time and offer different floral rewards in each phase. For successful pollination, pollinators must visit both male and female umbels during a foraging trip. Insects showed both preferences and non-preferences for umbel phases in natural patches where the gender ratio was male biased. In contrast, insects showed no bias in visitation during a foraging trip or in time spent foraging on male and female umbels in experimental arrays where the gender ratio was equal. Pollinator assemblages consisting of a mixture of different pollinator types coupled with temporal variation in the assemblages of populations among years maintains generalisation at the population/local level. In addition, spatial variation in assemblages among populations maintains generalisation at the species level. Fire alters pollination in T. incisa by shifting the flowering season and reducing the abundance of flying insects. Therefore, fire plays an important role in maintaining spatial and temporal variation in this fire-prone system. Although insect pollinators are important in determining the mating opportunities of 90% of flowering plant species worldwide, few studies have looked at the effects of variation in pollinator assemblages on plant reproductive success and mating. In T. incisa, high insect visitation rates do not guarantee high plant reproductive success, indicating that the quality of visit is more important than the rate of visitation. This is shown by comparing the Agnes Banks and Myall Lakes populations in 2003: Agnes Banks received the highest visitation rate from an assemblage dominated by ants but produced the lowest reproductive output, and Myall Lakes received the lowest visitation rate by an assemblage dominated by a native bee and produced the highest seedling emergence. Interestingly, populations with different assemblage composition can produce similar percentage seed set per umbel. However, similar percentage seed set did not result in similar percentage seedling emergence. Differences among years in reproductive output (total seed production) were due to differences in umbel production (reproductive effort) and proportion of umbels with seeds, and not seed set per umbel. Trachymene incisa is self-compatible and suffers weak to intermediate levels of inbreeding depression through early stages of the life cycle when seeds are self-pollinated and biparentally inbred. Floral phenology, in the form of synchronous protandry, plays an important role in avoiding self-pollination within umbels and reducing the chance of geitonogamous pollination between umbels on the same plant. Although pollinators can increase the rate of inbreeding in T. incisa by foraging on both male and female phase umbels on the same plant or closely related plants, most consecutive insect movements were between plants not located adjacent to each other. This indicates that inbreeding is mostly avoided and that T. incisa is a predominantly outcrossing species, although further genetic analyses are required to confirm this hypothesis. A new conceptual understanding has emerged from the key empirical results in the study of this model generalised pollination system. The large differences among populations and between years indicate that populations are not equally serviced by pollinators and are not equally generalist. Insect visitation rates varied significantly throughout the day, highlighting that sampling of pollinators at one time will result in an inaccurate estimate and usually underestimate the degree of generalisation. The visitor assemblage is not equivalent to the pollinator assemblage, although non-pollinating floral visitors are likely to influence the overall effectiveness of the pollinator assemblage. Given the high degree of variation in both the number of pollinator species and number of pollinator types, I have constructed a model which includes the degree of ecological and functional specialisation of a plant species on pollinators and the variation encountered across different levels of plant organisation. This model describes the ecological or current state of plant species and their pollinators, as well as presenting the patterns of generalisation across a range of populations, which is critical for understanding the evolution and maintenance of the system. In-depth examination of pollination systems is required in order to understand the range of strategies utilised by plants and their pollinators, and I advocate a complete floral visitor assemblage approach to future studies in pollination ecology. In particular, future studies should focus on the role of introduced pollinators in altering generalised plant-pollinator systems and the contribution of non-pollinating floral visitors to pollinator assemblage effectiveness. Comparative studies involving plants with highly conserved floral displays, such as those in the genus Trachymene and in the Apiaceae, will be useful for investigating the dynamics of generalised pollination systems across a range of widespread and restricted species.

Doctor of Philosophy (PhD) / A renewed focus on generalised pollinator systems has inspired a conceptual framework which highlights that spatial and temporal interactions among plants and their assemblage of pollinators can vary across the individual, population, regional and species levels. Pollination is clearly a dynamic interaction, varying in the number and interdependence of participants and the strength of the outcome of the interaction. Therefore, the role of variation in pollination is fundamental for understanding ecological dynamics of plant populations and is a major factor in the evolution and maintenance of generalised and specialised pollination systems. My study centred on these basic concepts by addressing the following questions: (1) How variable are pollinators in a generalised pollination system? To what degree do insect visitation rates and assemblage composition vary spatially among populations and temporally among flowering seasons? (2) How does variation in pollinators affect plant reproductive success? I chose to do this using a model system, Trachymene incisa subsp. incisa (Apiaceae), which is a widespread Australian herbaceous species with simple white flowers grouped into umbels that attract a high diversity of insect visitors. The Apiaceae are considered to be highly generalist in terms of pollination, due to their simple and uniform floral display and easily accessible floral rewards. Three populations of T. incisa located between 70 km and 210 km apart were studied over 2-3 years. The few studies investigating spatial and temporal variation simultaneously over geographic and yearly/seasonal scales indicate that there is a trend for more spatial than temporal variation in pollinators of generalist-pollinated plants. My study showed both spatial and temporal variation in assemblage composition among all populations and variation in insect visitation rates, in the form of a significant population by year interaction. However, removing ants from the analyses to restrict the assemblage to flying insects and the most likely pollinators, resulted in a significant difference in overall visitation rate between years but no difference in assemblage composition between the Myall Lakes and Tomago populations. These results indicate more temporal than spatial variation in the flying insect visitor assemblage of T. incisa. Foraging behaviour provides another source of variation in plant-pollinator interactions. Trachymene incisa exhibits umbels that function as either male or female at any one time and offer different floral rewards in each phase. For successful pollination, pollinators must visit both male and female umbels during a foraging trip. Insects showed both preferences and non-preferences for umbel phases in natural patches where the gender ratio was male biased. In contrast, insects showed no bias in visitation during a foraging trip or in time spent foraging on male and female umbels in experimental arrays where the gender ratio was equal. Pollinator assemblages consisting of a mixture of different pollinator types coupled with temporal variation in the assemblages of populations among years maintains generalisation at the population/local level. In addition, spatial variation in assemblages among populations maintains generalisation at the species level. Fire alters pollination in T. incisa by shifting the flowering season and reducing the abundance of flying insects. Therefore, fire plays an important role in maintaining spatial and temporal variation in this fire-prone system. Although insect pollinators are important in determining the mating opportunities of 90% of flowering plant species worldwide, few studies have looked at the effects of variation in pollinator assemblages on plant reproductive success and mating. In T. incisa, high insect visitation rates do not guarantee high plant reproductive success, indicating that the quality of visit is more important than the rate of visitation. This is shown by comparing the Agnes Banks and Myall Lakes populations in 2003: Agnes Banks received the highest visitation rate from an assemblage dominated by ants but produced the lowest reproductive output, and Myall Lakes received the lowest visitation rate by an assemblage dominated by a native bee and produced the highest seedling emergence. Interestingly, populations with different assemblage composition can produce similar percentage seed set per umbel. However, similar percentage seed set did not result in similar percentage seedling emergence. Differences among years in reproductive output (total seed production) were due to differences in umbel production (reproductive effort) and proportion of umbels with seeds, and not seed set per umbel. Trachymene incisa is self-compatible and suffers weak to intermediate levels of inbreeding depression through early stages of the life cycle when seeds are self-pollinated and biparentally inbred. Floral phenology, in the form of synchronous protandry, plays an important role in avoiding self-pollination within umbels and reducing the chance of geitonogamous pollination between umbels on the same plant. Although pollinators can increase the rate of inbreeding in T. incisa by foraging on both male and female phase umbels on the same plant or closely related plants, most consecutive insect movements were between plants not located adjacent to each other. This indicates that inbreeding is mostly avoided and that T. incisa is a predominantly outcrossing species, although further genetic analyses are required to confirm this hypothesis. A new conceptual understanding has emerged from the key empirical results in the study of this model generalised pollination system. The large differences among populations and between years indicate that populations are not equally serviced by pollinators and are not equally generalist. Insect visitation rates varied significantly throughout the day, highlighting that sampling of pollinators at one time will result in an inaccurate estimate and usually underestimate the degree of generalisation. The visitor assemblage is not equivalent to the pollinator assemblage, although non-pollinating floral visitors are likely to influence the overall effectiveness of the pollinator assemblage. Given the high degree of variation in both the number of pollinator species and number of pollinator types, I have constructed a model which includes the degree of ecological and functional specialisation of a plant species on pollinators and the variation encountered across different levels of plant organisation. This model describes the ecological or current state of plant species and their pollinators, as well as presenting the patterns of generalisation across a range of populations, which is critical for understanding the evolution and maintenance of the system. In-depth examination of pollination systems is required in order to understand the range of strategies utilised by plants and their pollinators, and I advocate a complete floral visitor assemblage approach to future studies in pollination ecology. In particular, future studies should focus on the role of introduced pollinators in altering generalised plant-pollinator systems and the contribution of non-pollinating floral visitors to pollinator assemblage effectiveness. Comparative studies involving plants with highly conserved floral displays, such as those in the genus Trachymene and in the Apiaceae, will be useful for investigating the dynamics of generalised pollination systems across a range of widespread and restricted species.

Pentadesma butyracea Sabine est l’une des quatre espèces du genre Pentadesma endémique de l’Afrique. Elle est distribuée de la Sierra Léone au Gabon dans deux grands types d’habitats :les forêts denses humides discontinues du domaine guinéo-congolais (Haute- et Basse-Guinée) et le domaine soudanien du couloir sec du Dahomey (assimilé à une barrière à l’échange de gènes et d’espèces entre les deux blocs guinéo-congolais). Dans ce dernier, l’espèce se retrouve dans des galeries forestières et occupe une place capitale dans le développement socio-économique des communautés locales en raison des multiples biens et services que procurent ses produits (alimentation, médecine et pharmacopée traditionnelle, etc.). Cependant, des pressions d’origines multiples, telles que le ramassage des graines pour fabriquer du beurre, la fragmentation de l’habitat et sa destruction en faveur du maraîchage, les pratiques culturales inadaptées, les incendies, font peser de lourdes menaces sur l’espèce.<p>Le but de ce travail est d’acquérir les connaissances requises pour la conservation et la gestion durable des ressources génétiques de l’espèce. Trois objectifs ont été définis :(i) étudier la phylogéographie de l’espèce, (ii) étudier sa variabilité morphologique et génétique au Bénin et (iii) caractériser sa biologie de reproduction. En amont de ces travaux, nous avons développé onze marqueurs microsatellites nucléaires chez P. butyracea (chapitre 2). Ils ont été utilisés pour l’étude de la phylogéographie et la diversité génétique de P. butyracea (chapitres 3 et 5), ainsi que pour étudier la dépression de consanguinité et les paramètres de son système de reproduction (chapitre 7).<p>La caractérisation de la répartition spatiale des lignées génétiques de régions intergéniques de l’ADN chloroplastique et de l’ADN ribosomal (ITS) a détecté deux lignées génétiques allopatriques entre le Haut et le Bas-Guinéen, indiquant une forte différenciation génétique et un signal phylogéographique. L’analyse des microsatellites détecte trois pools géniques correspondant aux trois régions étudiées (Haute Guinée, Dahomey Gap et Basse Guinée). La diversité génétique est faible dans le Dahomey Gap, modérée dans le Haut-Guinéen et élevée dans le Bas-Guinéen. Ces résultats indiquent une séparation très ancienne des populations d’Afrique centrale et d’Afrique de l’ouest, alors que celles du Dahomey Gap pourraient résulter des forêts denses humides de l’Afrique de l’ouest lors de la période Holocène humide africaine. Dans ce couloir sec, les populations ont subi une forte dérive génétique, potentiellement due à des évènements de fondation. Au Bénin, deux groupes éco-morphologiques ont été détectés suivant un gradient nord-sud, contrastant avec deux pools géniques présentant une distribution est-ouest. <p>P. butyracea est une espèce auto-compatible majoritairement allogame. La corrélation de paternité est plus élevée aux niveaux intra-fruit vs. inter-fruits, et au sein d’une population de petite taille vs. de grande taille. Les principaux pollinisateurs au Bénin sont deux oiseaux (Cyanomitra verticalis, Cinnyris coccinigastrus) et trois abeilles (Apis mellifera, Meliponula togoensis, Hypotrigona sp.). La productivité totale en fruits augmente en fonction de l’âge de l’arbre et varie en fonction de l’année, atteignant un pic pour les arbres ayant un diamètre de 60-80 cm. Les graines sont récalcitrantes et ont une teneur en eau de 42.5 ± 2.9 %. <p>L’analyse des paramètres de reproduction et de diversité génétique, associés aux facteurs écogéographiques, nous a permis de proposer un échantillon de neuf populations représentatives de la diversité à l’échelle du Bénin, dans la perspective d’une conservation in situ. Le succès de celle-ci dépendra des efforts conjugués des communautés locales, de la recherche forestière et de la définition d’un cadre législatif par le politique pour la protection des habitats. La conservation ex situ est envisagée sous forme d’un verger rassemblant diverses origines, présentant l’intérêt supplémentaire de permettre d’étudier les contributions de la diversité génétique et de la plasticité phénotypique à la variation phénotypique. / Pentadesma butyracea Sabine is one of the four species of the endemic genus Pentadesma in Africa. The species is distributed from Sierra Leone to Gabon in two major types of habitats: the discontinuous and dense Guineo-Congolian rainforests (Upper and Lower Guinea) and the Sudanian domain of the dry corridor of Dahomey (considered as a barrier to the exchange of genes and species between Upper and Lower Guinea). In the latter, the species is found in gallery forests and plays a vital role in the socio-economic livelihood of local communities due to the various resources and services that provide its products (food, medicine and traditional, etc.). However, pressure from many sources including the collection of seeds to make butter, habitat fragmentation and its destruction for market gardening, inadequate agricultural practices, fires, are serious threats to the species.<p>The aim of this work was to acquire appropriate knowledge for the conservation and sustainable management of genetic resources of the species. Three objectives were defined (i) study the phylogeography of the species; (ii) evaluate its morphological and genetic variability in Benin; and (iii) characterize its reproductive biology. In a preliminary work, eleven nuclear microsatellite markers of P. butyracea were developed (Chapter 2). They were used for the study of phylogeography and genetic diversity of P. butyracea (chapters 3 and 5), and to study the inbreeding depression and parameters of its breeding system (Chapter 7).<p>The characterization of the genetic lineages and their spatial distribution using intergenic regions from chloroplast DNA and ribosomal DNA (ITS) region detected two allopatric genetic lineages between Upper and Lower Guinea, indicating a high genetic differentiation and a phylogeographic signal. Microsatellite markers allowed us to detect three genepools matching with the three studied regions (Upper Guinea, Dahomey-Gap and Lower Guinea). Genetic diversity was low in the Dahomey Gap, moderate in Upper Guinea and high in Lower Guinea. These results indicate an ancient separation of populations from Central and West Africa, while those from Dahomey Gap could originate West African rainforests (Upper Guinea) during the African humid Holocene period. In this dry corridor, populations experienced high genetic drift, possibly due to founding events. In Benin, two eco-morphological groups were detected following a north-south gradient, contrasting with two gene pools presenting an east-west distribution.<p>Pentadesma butyracea is a self-compatible, mainly allogamous species. The correlation of paternity was higher within-fruit vs. among-fruits, and in population of small size vs. large size. The main pollinators in Benin are two birds (Cyanomitra verticalis, Cinnyris coccinigastrus) and three bees (Apis mellifera, Meliponula togoensis, Hypotrigona sp.). Total productivity in fruit increases with tree age and varies yearly, reaching a peak for trees of 60-80 cm of diameter class. Seeds are recalcitrant (i.e. they cannot be conserved at low temperature), having a water content of 42.5 ± 2.9% at maturity.<p>The analysis of reproduction and genetics parameters, associated with eco-geographic factors, enabled us to select nine populations representative of the diversity in Benin, from the perspective of in situ conservation. The success of the latter will depend on combined efforts of local communities, forest research and an adequate legislative framework for the protection of habitats. Ex situ conservation is envisaged as an orchard assembling various origins, and would have the additional advantage of allowing to study the contribution of genetic diversity and phenotypic plasticity to phenotypic variation. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished