Traditional pollination syndromes group angiosperms into categories based on how floral traits impact the functional group of pollinator most associated with those traits. The concept, while well supported for specialist-pollinated plants, is a poor predictor of pollinator identity in generalist systems, such as those common to the family Asteraceae. One potential avenue for future refinement of the concept is the combination of large floral trait datasets, quantitative pollinator data, and phylogenetic comparative methods. Helianthus is a well-studied genus of North American aster whose species include the agriculturally significant H. annuus, which represents the third largest oilseed crop globally. The genus is primarily bee pollinated and, while much is known about traits that are correlated with bee attraction at short ranges common to agricultural and horticultural settings, there has been little research on long range visual and chemical attraction traits within the genus. Using data on display size and shape, ray color, floral volatile composition, and floret depth collected from Helianthus species grown in a common garden, mixed models were constructed to predict pollinator visitation as a function of floral traits. For four of seven pollinator response variables, there was at least one model that outperformed null models, and three of the four best models were multivariate. This work will inform future research of pollination syndromes within generalist systems such as those common to Asteraceae.

<p>Determining the genetic basis of adaptation has become a central focus of evolutionary biology, and the incorporation of increasingly sophisticated analytical tools from molecular biology has made identifying causal genes a practical reality. The work presented herein addresses the effects of pleiotropic constraint on evolutionary change at the level of individual genes and genetic networks. In the first chapter, I combine molecular phylogenetic analyses and direct assays of enzymatic function to determine the evolutionary processes following a gene duplication in the anthocyanin pathway. My results show that, prior to duplication, the DFR gene was constrained from functional improvement by its multiple enzymatic roles. Following duplication, this constraint was released and adaptive evolution proceeded along both paralog lineages. In the second chapter, I determine the molecular genetic basis of a flower color transition that is associated with change in pollinator attraction in morning glories. A regulatory change in a branching gene in the flavonoid biosynthetic pathway restricted flux down the cyanidin-producing branch, conferring nearly exclusive production of red pelargonidin pigment in flowers. I further demonstrate that this regulatory change was restricted to floral tissue, and that ancestral pathway flux predominates in vegetative tissues. I propose that deleterious pleiotropic effects prevented evolutionary change via enzymatic changes in the pathway due to the numerous essential products downstream of this branching point. Together, these two results show that evolutionary change may be constrained by the molecular genetic context in which prospective adaptive mutations occur.</p> / Dissertation

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O estudo da ecologia floral à fundamental para o entendimento das relaÃÃes entre flores e seus visitantes, assim como para entender o papel dessa interaÃÃo no sucesso reprodutivo e na manutenÃÃo da populaÃÃo vegetal. No presente estudo, investigou-se a morfologia, a biologia floral e o sistema de reproduÃÃo de Ipomoea bahiensis Willd., bem como as interaÃÃes entre suas flores e a guilda de visitantes florais. A investigaÃÃo teve como objetivo central entender tanto as relaÃÃes ecolÃgicas e evolutivas entre I. bahiensis e insetos antÃfilos quanto o sistema reprodutivo da planta. Para tanto, foram empregados mÃtodos para analisar a morfologia funcional das flores, a relaÃÃo da morfologia e biologia floral com os visitantes, o comportamento e frequÃncia desses animais, o requerimento polÃnico e a relaÃÃo fruto/flor e, por fim, foi quantificada a limitaÃÃo de pÃlen. Para incrementar o conhecimento sobre a ecologia floral, foram observados os visitantes de nectÃrios extranupciais e sua relaÃÃo com inimigos florais. A partir da anÃlise dos dados obtidos, observou-se que as flores de I. bahiensis possuem atributos predominantemente de psicofilia, contudo tÃm plasticidade no sistemas de polinizaÃÃo, podendo ser polinizadas por abelhas de pequeno porte. A dinÃmica do nÃctar juntamente com o perÃodo de receptividade do estigma procura, possivelmente, garantir a visitaÃÃo de borboletas nesse perÃodo. No entanto, a variaÃÃo na frequÃncia dos visitantes florais foi entendida a partir da disponibilidade de pÃlen, pois os principais visitantes foram abelhas coletoras de pÃlen. Os testes de polinizaÃÃo mostraram que o sistema de reproduÃÃo à xenÃgamo, dependente de vetores biÃticos e apresenta auto-incompatibilidade. TambÃm foi observado que a planta à limitada por pÃlen / The study of floral ecology is pivotal to understanding the relationships between flowers and their visitors, as well as to understand the role of this interaction in reproductive success and maintenance of plant population. In this study, it was investigated the morphology, floral biology and breeding system of Ipomoea bahiensis Willd. and the interactions between its flowers and the guild of floral visitors. The research aimed mainly to understand the ecological and evolutionary relationships between I. bahiensis and anthophilous insects. For this, methods were used to analyze the functional morphology of flowers, the relationship of floral morphology and biology with visitors, the frequency and behavior these animals, the requirement for pollen and the rate fruit/flower and, finally, was estimated pollen limitation. To increase knowledge about the floral ecology were observed extranupcial nectariesâs visitors and its relation to floral enemies. From the analysis of the data, it was observed that the flowers of I. bahiensis have attributes predominantly psycophily, however they present plasticity in pollination systems, and can also be pollinated by small bees. The dynamics of nectar herewith with the period of stigma receptivity seeks to ensure the visitation of butterflies in this period. However, the variation in the frequency of floral visitors was understood from the availability of pollen, because the main visitors were bees pollen collecting. Pollination tests showed that the reproductive system is xenogamous, dependent on biotic vectors and presents self-incompatibility. It was also observed that the plant is limited by pollen

Specialization on insect and animal pollinators is thought to be the driving force for the evolution of floral traits. Specifically in the New World (NW), the oil-bee pollination syndrome has led to the convergence of floral characters in two distantly related families of core eudicots, Malpighiaceae and Krameriaceae. Both families display a flag-like structure that establishes a zygomorphic flower and floral oil rewards in epithelial elaiophores. These traits work concomitantly to attract and reward female oil-bees that help fertilize these flowers and in return receive oils. The underlying genetics of floral zygomorphy were studied in several clades of core eudicots, which implicated CYCLOIDEA2-(CYC2-)like genes of the TCP gene family to play a role in the establishment and maintenance of this trait. In Malpighiaceae, previous work demonstrated that two CYC2-like genes, CYC2A and CYC2B, are expressed during development correlated with establishing zygomorphy in flowers of NW Malpighiaceae. In this thesis work, I investigated the underlying developmental and genetic basis for the establishment of non-homologous and yet functionally similar traits in the oil-bee pollinated flowers of Malpighiaceae and Krameriaceae. In Chapter 1, I investigated the modification of a conserved CYC2 genetic program in the Old World (OW) acridocarpoids of Malpighiaceae following a major biogeographic disjunction. And in Chapter 2, I studied the floral ontogeny and genetic basis of floral zygomorphy development in Krameria lanceolata Torrey of Krameriaceae. This thesis work sheds light on the significance of the interdisciplinary study of floral symmetry evolution through comparative pollination ecology, development, and genetics.

La convergence, soit l'acquisition indépendante de phénotypes similaires, est un aspect intéressant de la diversité qui peut fournir des informations importantes sur la nature du changement évolutif. Dans les systèmes végétaux, les syndromes de pollinisation – combinaisons de traits floraux adaptés à leurs pollinisateurs – constituent de bons exemples de convergence se produisant sur les fleurs. Nous avons utilisé une approche globale incluant la morphologie cellulaire et la transcriptomique pour analyser la convergence de formes florales de deux syndromes de pollinisation trouvés dans les genres frères non Gesneria et Rhytidophyllum (Gesneriaceae), un groupe antillais qui contient environ 81 espèces avec différentes morphologies et stratégies de pollinisation variables dans leur degré de spécialisation écologique. Il a déjà été démontré que la forme des fleurs joue un rôle important dans l’évolution de ce groupe, qui présente de nombreuses transitions entre les stratégies de pollinisation. Nous avons testé la présence de convergence dans les forms de cellules de la corolle et dans l’expression des gènes de la corolle en utilisant (1) une analyse pour mesurer la forme des cellules de pétales matures à l’aide d’un modèle phylogénétique mixte et (2) une approche transcriptomique comparative combinant l'expression différentielle des gènes (DESEq2) et l'analyse de co-expression (WGCNA) de gènes exprimés dans certaines regions précises des pétales. Toutes les analyses ont pris en compte les relations phylogénétiques entre les espèces. Nous avons trouvé une anisotropie cellulaire convergente se produisant dans les régions distales des pétales au sein des espèces du même syndrome (forme). Nous avons également constaté une plus grande similarité dans les modèles d'expression génique entre les espèces d’un même syndrome qu'entre les espèces apparentées et avons produit une liste de 203 gènes potentiellement associés aux formes de fleurs convergentes. La convergence morphologique florale observée dans les syndromes de pollinisation des espèces étudiées se retrouve tant au niveau cellulaire qu'au niveau de l'expression. Les résultats présentés ici amplifient les informations de base sur la famille des Gesneriaceae pour les études futures sur la convergence et la forme florale dans le groupe. / Convergence, the independent acquisition of similar phenotypes, is an important aspect of diversity that can provide valuable insights about the nature of evolutionary change. In plants, pollination syndromes - combinations of floral traits adapted to their pollinators - make good examples of convergence occurring on flowers. We used a comprehensive approach that includes cell morphology and transcriptomics to analyze the floral shape convergence of two pollination syndromes found in the sister genera Gesneria and Rhytidophyllum (Gesneriaceae), an Antillean group that contains approximately 81 species with different morphologies and pollination strategies varying in their degree of ecological specialization. Flower shape has already been found to play an important role in the evolution of this group, which shows many transitions between pollination strategies. We tested convergence in the corolla cell shapes and in gene expression for the pollination syndromes using (1) cell measurement statistical analysis (Phylogenetic Mixed Model) of mature petals and (2) a comparative transcriptomic approach that combined differential gene expression (DESEq2) and co-expression analysis (WGCNA) in genes expressed in specific regions of the petals. All analyses took the phylogenetic relationships of the species into account. We found convergent cellular anisotropy occurring in the distal regions of the petals within species of the same syndrome (form). We also found greater similarity in gene expression patterns occurring among species of the same syndromes than between more closely related species and produced a list of 203 genes potentially associated with convergent flower forms. The floral morphological convergence observed in the pollination syndromes of the investigated species is paralleled both at the cellular and expression levels. The results shown here amplify the background information of the Gesneriaceae family for future studies of convergence and floral form in the group.