Made available in DSpace on 2014-08-20T13:59:08Z (GMT). No. of bitstreams: 1 tese_elisia_rodrigues_correa.pdf: 1900185 bytes, checksum: fcb8093a27a373ee431423cfbdbcb391 (MD5) Previous issue date: 2014-05-21 / Improving the yield is the primary aim for the macadamia breeding program, together with it the breeder has the constant challenge of trying to add others traits which will promote macadamia value. Among these stands out the study of pollination character, a factor that brings benefits to the production, since plants with this feature are independent of pollinating vectors, which are currently declining in population. Along with this feature is the search for a product (nut) with a nutritional value which will bring a beneficial to human health, for macadamia the goal is the development of oil with a good rate of monounsaturated fats. Faced with these challenges, the objective was to identify genotypes with greater production of nuts and to provide a high rate of self-pollination, as well to establish strategies to better characterize the profile of macadamia nut oils. For the self-fertility experiment, racemes from cultivated and wild macadamia tree were tagged and bagged for comparing self-pollination and open pollination. T1002.003, T108.002 and T1023.003 were the only wild genotypes which presents Final Nut Set from self-pollination. In the oil studies, three experiments were carried out. The first aiming to quantify the sample size of macadamia nuts for best characterizes the fatty acids oil profile. Macadamia nuts were harvested, dried, the oil were extracted and after analyzed with gas chromatography. The variance found among the nuts from the same tree is bigger than the variance among the genotypes conducting to the second experiment, aiming to understand the possible source of variability at macadamia oil profile and the better way for controlling it, twenty nuts from four tree of macadamia cultivar, A16, were harvested. However the variance among the nuts was not controlled too. A third experiment was developed for checking if it has a pollen influence at fatty acids profile in the macadamia oil. Nuts from the selection 11.1 from three source of pollen (open pollination, self-pollination and a cross with the genotype 268) were harvested. Overall, the result from the experiments shows that the pollen has an influence in the oil profile. The aim of the last experiment was to identify the best genotypes in a population of macadamia plants in relation to annual production of almonds, aimed establishing a base population for a breeding. The results obtained by mixed model methodology (REML / BLUP) and the correlation between genotype and phenotype demonstrate the importance of using BLUP predictors as a tool for breeding programs related to this case study. / Aliado a busca por plantas de macadâmia mais produtivas, o pesquisador possui o desafio constante de tentar associar outras características as quais possam agregar valor a noz e assessorar os mais variados aspectos da produção. Dentre estes se destaca o estudo do caractere polinização, considerando que a autopolinização beneficia a produção, uma vez que plantas com esta característica independem de vetores polinizadores, que estão em declínio populacional atualmente. Junto a essa característica é incessante a busca por um produto (noz) com maior valor nutricional à saúde humana, no caso da macadâmia, um óleo com boa taxa de gorduras monoinsaturadas. Frente a estes desafios o objetivo do trabalho foi de identificar genótipos com maior produção de nozes e que apresentassem alta taxa de autopolinização, além de estabelecer estratégias para melhor a caracterização do perfil dos óleos de macadâmia para a inserção do estudo da composição dos óleos no melhoramento de macadâmia. Para o experimento de autopolinização, racemos florais de cultivares e matérias nativos foram ensacados e acompanhados quanto ao desenvolvimento da noz. Outros racemos florais foram apenas identificados e acompanhados para posterior comparação da polinização aberta em relação a autopolinizada. Três genótipos oriundos do grupo da macadâmia nativa apresentaram autopolinização. Nos estudos com óleo, três ensaios foram conduzidos. O primeiro com o intuito de quantificar o tamanho da amostra de macadâmia para a melhor representatividade referente ao perfil do óleo. Foram colhidas nozes de quatro cultivares, duas plantas por cultivar e vinte nozes por planta. As nozes foram secas e o perfil do óleo extraído determinado por cromatografia gasosa. A variância encontrada entre as nozes provenientes da mesma planta foi maior do que entre os genótipos, conduzindo assim ao objetivo do segundo ensaio. Com o intuito de identificar se o estádio de desenvolvimento da noz interfere no perfil do óleo, vinte nozes de quatro plantas da cultivar A16 foram colhidas e avaliadas. No entanto, como a causa potencial do alto nível de variabilidade é desconhecido ou não pode ser explorada, acredita-se que única outra opção é trabalhar com um grande número de nozes. O terceiro ensaio desenvolvido dentro do âmbito do perfil do óleo foi o da influência da fonte do pólen. Nozes da seleção 11.1 oriundas de três fontes de pólen (polinização aberta, autopolinização, e cruzamento com o genótipo 268) foram colhidas. Foi verificado que a fonte do pólen influencia no perfil do óleo. O objetivo do último experimento foi identificar os melhores genótipos em uma população de plantas de macadâmia, com relação à produção anual de amêndoas, visando estabelecer uma população base para programa de melhoramento e aprimorar a propagação vegetativa no sistema de produção. Os resultados obtidos pela metodologia de modelos mistos (REML/BLUP) e a correlação entre genótipo e fenótipo demonstram a importância do uso de preditores BLUP como ferramenta para programas de melhoramento relacionados a este estudo de caso.

Made available in DSpace on 2014-08-20T14:22:07Z (GMT). No. of bitstreams: 1 Tese_ Marcelo_ Couto.pdf: 11730472 bytes, checksum: afd142b8d56cb58f0be9327a636499ea (MD5) Previous issue date: 2006-11-27 / Insufficient chilling accumulation limits the production of peach in subtropical areas. Comparatively this combined with the wide temperature fluctuation during bud differentiation, through the bloom period and fruit growth and development may cause crop losses. Problems with low productivity in southern Rio Grande do Sul often attributed to the mild winter conditions, however, the environmental conditions especially around bloom period to be critical for optimal yields. In subtropical áreas temperature oscillations may cause problems during fertilization with subsequent reduction of fruit set and production instability of some peach cultivars. To better understand the effect of high temperatures on flower differentiation, pre-blooming, blooming, fruit set and fruit development and growth observations were made during the stages of bud differentiation, leafing and blooming, which included pollen production and viability, floral morphology, fruit set and the fruit growth and development. Temperature elevation with the tree was obtained by bagging branches with transparent plastic or with transparent plastic bottles. Plants of two low chilling peach cultivars (200 to 300 hours): Granada and Maciel were used in the years of 2003, 2004 and 2005. Conditions of high temperatures during the period of bud differentiation of Granada and Maciel did not promote the formation of double pistils or any morphologic anomalies. Bagging shoots with transparent plastic bags and plastic bottles is a simple and economical way of increasing the temperature on the branches under field conditions. High temperatures conditions during pre-blooming period advanced and accelerated the flowers anthesis and pistil growth. Flower morphology, regarding pistil length and weight, was not influenced by temperature elevation under experimental conditions. High temperatures during the pre-blooming negatively influenced the fruit set of the peach cultivar Granada. The percentage of in vitro pollen germination was different among cultivars and incubation temperatures. Pollen viability of cultivars Esmeralda, Granada, Jade and Maciel, can be evaluated by in vitro germination in standard culture medium (10% of sucrose + 1% of agar, dissolved in distilled water), three hours after the inoculation, with incubation at 24°C and 28°C. Under high temperatures conditions during the stage I of fruit growth and development of peach cultivars Granada and Maciel, cell division may be accelerated since a larger number of cells was counted in the pericarp tissue. / As horas de frio necessárias às gemas vegetativas e floríferas, quando insuficientes, limitam a produção de pessegueiros nas zonas subtropicais e temperadas brasileiras. A redução na produtividade pode ocorrer também, devido a variações bruscas na temperatura durante os períodos de diferenciação das gemas, préfloração, floração, crescimento e desenvolvimento dos frutos desta espécie. Estas variações de temperatura podem ser uma das causas da instabilidade produtiva de algumas cultivares de pessegueiro. Muito dos problemas que se verificam, após a floração ou mesmo na colheita, como baixa produtividade, atribuídos muitas vezes, apenas às condições de inverno com pouco frio que ocorrem no sul do Rio Grande do Sul, podem ter origem em etapa anterior do desenvolvimento e, muito provavelmente, na diferenciação floral. Com os objetivos de estudar a influência de altas temperaturas sobre diferenciação de gemas, pré-floração, floração, frutificação efetiva, crescimento e desenvolvimento de frutos, foram realizadas observações no avanço dos estádios de diferenciação das gemas, épocas de brotação e floração, testes de viabilidade e produção de pólen. Foram, ainda, feitas observações na morfologia e biologia floral, frutificação efetiva, crescimento e desenvolvimento dos frutos. A elevação da temperatura foi proporcionada com a utilização de casas de plástico ou através do ensacamento dos ramos com mangas de plástico transparente ou garrafas de plástico. Foram utilizadas plantas de duas cultivares comerciais de pessegueiro de baixa necessidade de frio (200 a 300 horas): Granada e Maciel em 2003, 2004 e 2005. Não há a formação de pistilos duplos ou quaisquer anomalias morfológicas mesmo sob temperaturas maiores que 25°C durante o período de diferenciação das gemas das cultivares de pessegueiro Granada e Maciel. Há diferenças entre cultivares e entre temperaturas de incubação quanto à porcentagem de germinação do pólen in vitro. A viabilidade do pólen das cultivares de pessegueiro testadas (Esmeralda, Granada, Jade e Maciel) pode ser avaliada por germinação in vitro em meio de cultura padrão (10% de sacarose + 1% de agar, dissolvidos em água destilada), três horas após a inoculação, com incubação a 24°C e 28°C. Ensacamento de ramos com plástico transparente ou com garrafas de plástico, é uma forma simples e econômica de aumentar a temperatura junto aos ramos das plantas, em condições de campo, sendo mais efetivo que o uso de estufas de plástico. Conclui-se que em condições de temperaturas elevadas, durante a pré-floração, ocorre a antecipação da antese das flores para as cultivares Granada e Maciel. A morfologia das flores, em relação ao comprimento e ao peso dos pistilos, não é influenciada pela elevação da temperatura nas condições experimentais utilizadas. Temperaturas elevadas durante a pré-floração, influenciam negativamente na frutificação efetiva da cultivar de pessegueiro Granada. Em condições de temperaturas, elevadas durante o estádio I de crescimento e desenvolvimento dos frutos, para as cultivares de pessegueiro Granada e Maciel, há uma tendência de formação de um maior número de células do pericarpo, o que ampliaria o potencial da produção de frutos com maior tamanho.

Comme pour tous les organismes pluricellulaires, la croissance et le développement des plantes nécessitent une coordination de la production de cellules via la mitose et la différenciation cellulaire. La progression du cycle cellulaire est contrôlée par les complexes CDK-cycline. Les inhibiteurs de ces complexes, les CKIs, représentent d’excellents candidats pour réguler cet équilibre entre les processus de prolifération et différentiation cellulaires qui ont lieu au cours du développement. Afin de mettre en évidence le rôle d’intégrateurs potentiel des CKIs, le développement floral a été utilisé en tant que modèle.Grâce à l’utilisation de la qRT-PCR, nous avons montré que durant le développement floral d’Arabidopsis thaliana, un groupe restreint de CKIs était exprimé. Nous avons choisi de travailler sur les deux CKIs les plus exprimés, KRP6 et KRP7. Une caractérisation fine de leur profil d’expression durant le développement a été réalisée en utilisant des approches complémentaires telles que l’analyse de l’activité de leur promoteur, de la dynamique de leur transcrit, de leur expression protéique et de leur régulation post-traductionnelle.Jusqu’à présent, seules des approches ‘gain de fonction’ ont été utilisées pour étudier le rôle des CKIs chez les plantes. C’est pour cela que nous avons choisi des approches ‘perte de fonction’ pour analyser le rôle de KRP6 et de KRP7 au cours du développement floral. Ainsi, nous avons généré des doubles mutants d’insertion krp6-krp7, krp3-krp6, krp3-krp7, des triples mutants d’insertion krp3-krp6-krp7 et diverses lignées ARN interférence avec des promoteurs spécifiques. Malgré l’étude de ces nombreuses lignées, nous n’avons pas réussi à mettre en évidence des effets phénotypiques associés à l’absence de la fonction CKI au cours du développement floral. Ces résultats mettent en évidence la redondance fonctionnelle qui semble exister entre les KRPs, ainsi un quadruple mutant pourrait être nécessaire pour entrainer des modifications développementales. Afin de mieux comprendre cette fonction d’intégrateurs des KRPs au cours du développement floral, les partenaires de KRP6 et de KRP7 ont été recherchés. Des criblages double-hybride ont été réalisés afin d’identifier des ADNc, spécifiques du développement floral, codant des protéines capables d’interagir avec KRP6 et KRP7. De façon intéressante, mis à part les cyclines de type D, un nouveau type d’interaction a pu être mis en évidence. Un sous-groupe de la famille des rémorines est capable d’interagir avec KRP6 ou KRP7 en système double-hybride. Les rémorines sont des protéines spécifiques du règne végétal, associées à la membrane plasmique mais dont la fonction reste à clarifier. Une approche BiFC en protoplastes BY-2 a permis de confirmer l’existence de ce type d’interaction. De plus, l’influence des rémorines sur la localisation intracellulaire des KRPs a été étudiée. En présence de ces nouveaux partenaires, KRP7 est capable d’adopter une localisation nucléo-cytoplasmique.Enfin, des résultats récents ont montré que l’AMPK était capable de phosphoryler p27KIP1, l’homologue fonctionnel des KRPs chez les mammifères. Ces évènements de phosphorylation entrainent des modifications de sa localisation intracellulaire et de son activité inhibitrice vis-à-vis des complexes CDK-cycline. Après la réalisation d’analyses in silico ayant permis de prédire des sites putatifs de phosphorylation par SnRK1, l’homologue de l’AMPK chez A. thaliana, pour certains KRPs, la protéine KRP6 sous forme recombinante a été utilisée pour réaliser des essais kinase in vitro. Une phosphorylation de KRP6 est détectée en présence de la sous unité catalytique activée de SnRK1. Contrairement aux mammifères, cet évènement de phosphorylation entraine une altération de l’activité inhibitrice de KRP6 sans modification de sa localisation intracellulaire. Cette abolition de l’activité de KRP6 a été confirmée in planta. En effet, les phénotypes associés à la surexpression de KRP6 peuvent être atténués par la surexpression simultanée de la sous-unité catalytique de SnRK1. L’existence de ce lien entre KRP6 et SnRK1 met en évidence une relation directe entre l’homéostasie énergétique et la prolifération cellulaire. / As in all multicellular organisms, growth and development in plants require the coordination of cell production by division and cell differentiation. Progression through cell cycle is controlled by the kinase activity of CDK/cyclin complexes. Inhibitors of these complexes, CKIs, represent excellent candidates to regulate the balance between proliferation and differentiation processes during development. To get insight in the potential integrator role of CKIs, floral development was chosen as a developmental model. Using a real time quantitative PCR approach, we bring to light that during floral development of Arabidopsis thaliana, a restricted subset of CKIs was preferentially expressed. It was decided to focus our work on the two major expressed CKIs, KRP6 and KRP7. A better characterization of their expression patterns of during development was undertaken using complementary approaches such as promoter activity analysis, mRNA dynamics, protein expression and post-translational regulation analysis. Because until now ‘gain of function’ approaches have been largely applied to unravel the role of plant CKIs, our challenge was to detect a floral phenotype for KRP6 and KRP7 loss of function mutants, either using knock-out mutants or RNAi lines. We generated krp6-krp7, krp3-krp6, krp3-krp7 double mutants and krp3-krp6-krp7 triple mutant and also several RNAi lines with specifics promoters. Despite the study of these numerous lines, we were not able to highlight phenotypic effects associated with the absence of CKI function during floral development. All these results emphasis functional redundancy which appears to exist between all KRPs, thus quadruple mutant might be needed to provoke some developmental modification.In order to better understand the integrative function of KRPs during floral development, partners of KRP6 and KRP7 were assessed. Two-hybrid screens were performed to identify cDNAs from a “floral-buds-development” library encoding proteins that are able to interact with KRP6 and KRP7. Interestingly, apart from D-type cyclins, we brought to light a new type of interaction. Indeed, a sub-class of the remorin protein family was able to interact with KRP6 or KRP7 in yeast two-hybrid. Remorins are plant specific plasma membrane associated proteins with unknown function. A BiFC approach in BY-2 protoplasts allowed us to confirm remorins/KRP6-7 interactions. Furthermore, the influence of the presence of remorin proteins on KRP6/7 localisation was assessed. KRP7 is able to adopt a nucleo-cytoplasmic localisation in presence of its new partners.Finally, recent results have shown that AMPK is phosphorylating p27KIP1, KRPs functional counterpart in mammals. These phosphorylation events lead to changes in its cellular localisation and its inhibitory activity toward CDK-cyclin complexes. After in silico analysis aiming to predict potential AMPK Arabidopsis homologue SnRK1 phosphorylation sites within some KRPs protein sequences, recombinant KRP6 was used in order to perform in vitro kinase assays. Phosphorylation occurs efficiently on KRP6 when activated SnRK1 catalytic subunit is present. Furthermore, unlike in mammals, this phosphorylation event leads to an alteration of KRP6 inhibitory activity without modification of its cellular localisation. This abolition of KRP6 activity was confirmed by in planta analysis. Indeed, KRP6 overexpression phenotype can be attenuated by simultaneous SnRK1 catalytic subunit overexpression. The existence of this link between KRP6 and SnRK1 underscores a direct relationship between energy homeostasis and cell proliferation.

Les Angiospermes forment un clade connu pour sa grande diversité d’espèces et une répartition inégale de cette diversité en son sein. Ce travail se concentre sur la reconstruction d’états ancestraux de caractères floraux sur les phylogénies dans le but d’étudier leur impact sur la diversification. En premier lieu, nous nous sommes focalisés sur deux innovations-clés potentielles chez les Proteaceae, la symétrie bilatérale du périanthe et la présence dans les hotspots à climat méditerranéen. L’utilisation sur une phylogénie de modèles de diversification dépendants des états de caractère a permis de montrer que le premier état caractère n’a pas d’impact significatif sur la diversification des espèces dans ce groupe, contrairement au second. Ensuite, nous avons reconstruit les états ancestraux de la symétrie du périanthe à l’échelle de toutes les Angiospermes, avec une stratégie d’échantillonnage permettant de refléter toutes les transitions de ce caractère. Nous avons mis en évidence un minimum de 130 origines de la symétrie bilatérale, suivies de 69 réversions vers la symétrie radiaire. Enfin, la même méthode a été étendue à l’étude de quatre autres caractères du périanthe. Différents modèles de reconstruction d’états ancestraux ont été utilisés sur une phylogénie de 1232 espèces pour examiner l’influence des changements de modèle sur les états ancestraux reconstruits. Nos résultats montrent que les changements de modèles de reconstruction ont une influence sur l’état de certains nœuds seulement, jamais sur l’ensemble. Les différents résultats trouvés pour la symétrie, caractère commun aux trois chapitres, révèlent que notre méthode d’échantillonnage présente l’inconvénient d’estimer des taux de transition trop élevés pour donner des résultats concluants avec la méthode du maximum de vraisemblance. Les résultats sont en revanche beaucoup moins biaisés lorsque l’on examine plusieurs caractères simultanément. / Angiosperms are a clade known for its great species diversity and the uneven distribution of this diversity among its lineages. This work focuses on the ancestral state reconstruction of floral characters on phylogenies in the purpose of studying their impact on diversification. We first focused on two potential key innovations in Proteaceae, bilateral perianth symmetry and presence in Mediterranean-climate hotspots. Using character state dependent diversification models, we found that the first character state did not have any significant impact on species diversification in this group, contrary to the second one. We then reconstructed ancestral states for perianth symmetry in angiosperms as a whole, using a sampling strategy aimed at capturing all of the transitions in this character. We found a minimum of 130 origins of bilateral symmetry, followed by 69 subsequent reversals to radial symmetry. Lastly, the same approach was extended to the study of four other perianth characters. Different ancestral state reconstruction models were used on a phylogenetic tree of 1232 species to test the influence of model changes on reconstructed ancestral states. Our results show that changes in reconstruction models have an impact on the inferred ancestral state of some nodes, but not all of them. The various results obtained for symmetry, a character shared among the three chapters, indicate that our sampling method has the drawback of estimating transition rates too high to give conclusive results with maximum likelihood. On the other hand, results are much less biased when several characters are examined simultaneously.

Alstroemeria is an important cut flower and its breeding has been developed focused on aesthetic characteristics and vase life longevity, but little is known about its scent. Five different genotypes were assessed including the non scented cultivars ‘Rebecca’ and ‘Samora’ and the scented cultivars, ‘Sweet Laura’, ‘Ajax’ and the species A. caryophyllaea. The scented Alstroemerias emitted the terpenoids: isocaryophyllene and ocimene as the major floral volatile compounds. Characterization of an Alstroemeria TPS (ALSTER) was based on four ESTs previously found in A. cv ‘Rebecca’. Rapid amplification of cDNA ends (RACE) was performed and the full length ORF was used for characterizations of the genomic organization and amino acid sequences (phylogenetic analysis). ALSTER genomic region contains five introns and six exons. This unique genomic organization classified ALSTER as a member of the class III with a merged 5-6 exon. The deduced amino acid sequence was classified into the subfamily TPS-b. A functional analysis showed enzymatic activity of ALSTER with geranyl diphosphate (GPP) and the monoterpene myrcene was the only product obtained. Gene expression evaluated through real time and semi q RT-PCR on eight different stages of development (SO to S7) showed high expression of ALSTER at around S2 - S4 in the scented Alstroemerias, coinciding with high scent emission perceived and also with the maturation of reproductive organs. Evaluations through surveys focused on level of liking of floral scent, were performed finding positive correlations between floral scent liking and floral appearance liking and between floral scent liking and floral scent intensity. Finally, 17 new lines of A. caryophyllaea were evaluated in terms of their morphology, phenology and productivity. Although none of them were suitable for the market because of their low productivity, short stems and small flowers, they were all scented and identified as promising starting points for breeding purposes.

Thesis (PhD)--Stellenbosch University, 2013. / Pollinators play a fundamental role in floral evolution. They can exert selection on the flowers they visit in a plethora of different ways, ranging from innate floral preferences to differences in body size and shape and behavioural elements such as flower constancy and learning capacity. Since different pollinators exhibit differences in these characters, shifts between pollinating species are often considered the most likely drivers of floral diversification. While many lines of evidence support this claim, numerous angiosperms pollinated by a single species also exhibit floral variation. Throughout my thesis, I explore and investigate floral diversification in such species in the absence of pollinator shifts. In Chapter 2, I investigate variation in the preference of conspecific male and female pollinators for the floral traits of a sexually deceptive daisy that comprises distinct floral forms. I show that its pollinator exhibits gender-specific variation in floral preferences, and that some floral forms have specialized on the male pollinator. This chapter thus illustrates the importance of intraspecific variation in pollinator preference for floral diversification, an underappreciated mechanism in this field of research. The innate preferences of pollinators are likely to have a genetic basis, especially innate preferences that govern mate choice. Genetic structure within the pollinators of sexually deceptive plants, which mimic female insects to achieve pollination, may thus provide an important source of selection on the plants they pollinate. This depends on an association between genetic divergence and divergent mate preferences, and I explore this intriguing idea in Chapter 3. While pollinators associated with sexually deceptive floral forms did exhibit significant genetic structuring, male pollinators from different phylogeographic clades all exhibited preference for the same sexually deceptive floral form, thus rejecting this hypothesis. Another behavioural attribute of pollinators that may affect floral evolution, particularly in deceptive plant species, is learning ability. Studies on sexually deceptive orchids often report that male pollinators tend to avoid sexually deceptive flowers with experience. In Chapter 4, I systematically investigate learning abilities within male pollinators and the costs they suffer on sexually deceptive floral forms that vary in deceptiveness. Results reveal a positive relationship between the level of floral deceptiveness and the 4 associated mating costs that deceived males suffer. Pollinator learning, however, appears to occur only on the most deceptive floral forms, suggesting a link between the costs suffered to the occurrence of learning. In Chapter 4, I systematically investigate learning abilities within male pollinators and the costs they suffer on sexually deceptive floral forms that vary in deceptiveness. Results reveal a positive relationship between the level of floral deceptiveness and the associated mating costs that deceived males suffer. Pollinator learning, however, appears to occur only on the most deceptive floral forms, suggesting a link between the costs suffered to the occurrence of learning. In Chapter 5, I explore the importance of florivory damage in a polymorphic daisy. Studies on floral evolution often overlook the significance of florivorous visits and focus only on pollinator-mediated selection. I show that floral polymorphism is maintained by antagonistic selection exerted by pollinators and florivores on the same floral traits. Lastly, I focus on evolutionary history to explore similarity in the patterns of South African angiosperm evolution and the pollinator species used throughout my thesis. Molecular dating shows this pollinator exhibits broadly congruent evolutionary patterns to these angiosperms, indicative of a shared biogeography. Taken together, my thesis demonstrates the vast impact of floral visitors, in particular pollinating insects, on the evolution of floral form. / My research was funded by the National Research Foundation of South Africa (NRF) and personal funding was provided by a NRF Innovation scholarship and merit bursaries from the Botany and Zoology department at Stellenbosch University. A WhiteSci Travel Grant and financial support from Prof. Erik Svensson at Lund University also allowed me to present parts of my research at international conference.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Comprendre la diversité morphologique des fleurs passe par l'étude de son origine moléculaire et développementale et de ses conséquences fonctionnelles et écologiques. Le périanthe est composé d'organes stériles, sépales et pétales, qui jouent un rôle majeur dans le succès reproducteur des plantes pollinisées par les animaux du fait de leur fonction d'attraction.Cette thèse propose une approche multidisciplinaire visant à comprendre l'origine génétique et moléculaire de la diversité morphologique du périanthe et sa signification évolutive, à l'aide du modèle Nigella damascena L. Cette Renonculacée présente un dimorphisme spontané. La forme probablement ancestrale, trouvée en populations naturelles, a un périanthe bipartite composé de cinq sépales pétaloïdes et huit pétales nectarifères. Dans la forme variante, cultivée à des fins d'horticulture, les pétales sont remplacés par un nombre élevé d'organes allant d'une forme proche des sépales à une forme proche des étamines.La première partie de cette thèse est consacrée à l'étude de l'origine développementale, génétique et moléculaire du dimorphisme, par la caractérisation détaillée de la morphologie florale et de son développement dans les deux morphes dans le cadre d'une approche gène candidat. Par analyse d'expression et validation fonctionnelle, nous avons montré que le gène NdAP3-3 est responsable de l'ensemble des aspects du dimorphisme floral de N. damascena, ce qui suggère que ce gène joue un rôle dans l'identité du pétale mais aussi dans l'architecture du méristème, potentiellement via la régulation du nombre d'organes et de la frontière entre périanthe et étamines.La seconde partie de cette thèse concerne l'impact du dimorphisme floral sur le mode de reproduction des deux morphes et leur maintien potentiel. Nous avons caractérisé les stratégies reproductives et la valeur sélective des deux morphes en conditions naturelles dans des populations expérimentales. Le variant sans pétale est peu visité par les pollinisateurs, et se reproduit majoritairement en autogamie. L'analyse de la vigueur de ses descendants suggère une dépression de consanguinité. Par ailleurs, dans notre matériel, il semble que l'allèle donnant le phénotype sans pétale soit lié à un allèle augmentant la valeur sélective. A la lumière de nos résultats, nous discutons les conditions du maintien de ce polymorphisme. / Understanding flower diversity requires on one hand the study of the molecular and developmental origin of floral architecture, and on the other the study of the functional and ecological consequences of flower morphology. A great deal of that diversity can be found at the perianth level which comprises the sepals and petals, sterile and versatile organs that play a major role in the reproductive success of animal pollinated flowering plants through their attractive characteristics.This thesis is the result of a multidisciplinary effort to understand the genetic and molecular origin as well as the evolutionary significance of perianth diversity, using the Nigella damascena L. as a model. This Ranunculaceae species presents a rare naturally occurring floral dimorphism affecting perianth architecture. The putatively ancestral form found in natural populations has a well differentiated bipartite perianth composed of five petaloid sepals and eight nectariferous petals, while the perianth in the alternative apetalous mutant, cultivated for horticultural purpose, has no petals and but is instead composed of numerous organs showing a continuum of forms from outer sepal-like to inner stamen-like.The first part of this thesis was dedicated to the study of the developmental, genetic and molecular origin of this dimorphism, via a detailed characterization of floral morphology and development in both morphs, which laid a foundation for the interpretation of the results of a candidate gene approach. Using expression analysis and functional validation we showed that NdAP3-3 is fully responsible for the complex N. damascena floral dimorphism, suggesting that it plays a role not only in petal identity but also in meristem patterning, possibly through the regulation of perianth organ number and perianth-stamen boundary.The second half of this thesis focused on the impact of the floral dimorphism on the reproduction mode and evolutionary maintenance of the two morphs. We assessed reproduction strategies and reproductive success in the two morphs by studying a polymorphic experimental population in natural conditions. The absence of petals in the mutant form was associated with a qualitative drop in pollinator visitation which resulted in a shift towards selfing. The study of their progeny suggests that selfing had a negative effect on the descendant’s vigor via inbreeding depression. Additionally, in our material, the allele responsible for the apetalous phenotype seems to be linked to a favorable allele increasing fitness. We discuss the mechanisms of the dimorphism maintenance in light of these results.

Lippia alba (mill) N. E. Br. ex Britt l Wilson - Verbenaceae - é relatada como uma planta medicinal com efeito fungitóxico para vários fungos fitopatogênicos e entre esses Colletotrilhum gloeosporioides (penz), causador da doença"queda de frutos jovens de citros". Testes realizados a partir de duas formas de L. alba - forma 1 (F1) e forma 2 (F2) mantidos em contato com gotas de suspensão de conídios de C. glocosponcides, sobre superfície de poliestireno durante diferentes períodos de tempo, demonstraram que substância(s) presente(s) em folhas secas das duas formas de L. alba altera(m) o padrão de germinação dos conídios, aumentado o comportamento e a largura do tubo germinativa e impedindo a formação do apressório. A não formação de apressórios pelos conídios foi também demonstrada a partir do contato de extratos acetônicos, etanólicos e hexânicos com gotas de suspensão de esporos em lâminas de poliestireno. Utilizando-se esses extratos, sobre meio BDA em placas de Petri, verificou-se também o efeito dos mesmos sobre o crescimento micelial de C. gloeosporioides. O extrato etanólico mostrou-se como e mais eficiente na redução do crescimento micelial, enquanto que o extrato aquoso não exibiu efeito algum nesse sentido. Através de bioensaios para frações fungitóxicas, pelo uso de placas de cromato-grafia delgada, demonstrou-se também o efeito fungitóxico dos extratos provenientes das duas formas de L.alba. Com o aparecimento de diferentes zonas de inibição do crescimento de L. gloeosporioides, comprovando-se o efeito fungitóxico de todos os extratos inclusive o aquoso, para as duas formas de L. alba. A quantificação de proteínas e carboidratos totais, bem como análise de macro e micronutrientes realizadas nas folhas secas das duas formas de L. alba não forneceram evidências para se efetuar correlações sobre o papel desses compostos na atividade fungistática apresentada / not available

Myrtaceae possui cerca de 4.600-5.800 espécies, distribuídas em áreas tropicais e subtropicais. Myrteae é uma das tribos mais ricas da família, com ca. de 2.500 espécies. Estudos sobre desenvolvimento e evolução de flor em Myrteae têm mostrado a condição homoplástica de caracteres anteriormente considerados diagnósticos, culminando em um melhor entendimento taxonômico. O hiper-diverso gênero Eugenia, com ca. de 1.050 espécies, está incluído nessa tribo e tem se mostrado morfologicamente homogêneo. Contudo, gêneros previamente segregados de Eugenia pelo grau de fusão do cálice são fontes de controvérsia taxonômica. A filogenia molecular baseada em cinco regiões de DNA reconstruiu que o cálice fundido evoluiu independentemente diversas vezes em Eugenia. A investigação morfológica da fusão do cálice revelou cinco padrões de desenvolvimento, além da condição dos lobos livres. Embora os padrões de desenvolvimento sejam homoplásticos, o sinal filogenético indicou que eles podem ser utilizados para complementar caracteres e sustentar clados. O padrão membranissépalo é uma exceção, sendo exclusivo e diagnóstico de um clado Amazônico. Espécies tradicionalmente reconhecidas com seis pétalas são interpretadas como padrão petaloide, isto é, quatro pétalas são precedidas por duas sépalas internas e petaloides, e duas sépalas externas e fundidas entre si. Padrão longohipanto é reconhecido como uma condição extremamente rara em Eugenia, em que o hipanto se alonga estende, bem como os verticilos estaminais, resultando em estames curvos no botão em vez de eretos. Mudanças nomenclaturais e uma recircunscrição e revisão taxonômica de Eugenia sect. Schizocalomyrtus derivam destes resultados. Dez espécies são descritas e comentadas em suas distribuições, sua conservação e taxonomia. A inflorescência de Eugenia também é analisada sob uma abordagem integrativa, que revela arranjos previamente ignorados como relevantes para a sistemática e taxonomia. Sete padrões foram descritos, e cinco deles são considerados para uma análise evolutiva. O padrão auxotélico foi recuperado em linhagens que primeiro divergiram. Isso sugere que o seu ancestral hipotético é similar ao padrão auxotélico. O padrão fasciculiforme é amplamente distribuído na seção de maior riqueza, Eugenia sect. Umbellatae, indicando que altas taxas de diversificação podem estar relacionadas com a aquisição desse padrão. Este estudo também fornece novas hipóteses para a evolução dos arranjos da inflorescência, indicando que flexibilidade no padrão racemoso de ramificação é, provavelmente, a inovação chave que promoveu a diversificação de Eugenia na região Neotropical / Myrtaceae has about 4600-5800 species distributed in tropical and subtropical areas. Myrteae is one of the most species rich tribe in Myrtaceae with ca. 2500 species. Studies on Myrteae flower development and evolution have shown the homoplastic nature of characters previously considered diagnostic that are now culminating in much-improved taxonomic understanding. The hyper-diverse Eugenia with ca. 1050 species is nested in this tribe, and have been shown as morphologically homogeneous. However, genera previously segregated from Eugenia by the degree of calyx fusion are source of controversy. Molecular phylogeny based on five DNA regions reconstructed fused calyx as evolved several times independently in Eugenia. Morphological assessment of the calyx fusion revealed five development patterns besides the standard condition of the free lobes. Although the development patterns are homoplastic, phylogenetic signal indicates that they can be used in combination to complementary characters to support clades. Membranisepalous pattern is an exception, recovered as exclusive allowing to be used to diagnose an Amazonian clade. Traditionally six-petal species are newly interpreted as petaloid pattern, i.e. four petals are followed by two internal petal-like sepals, and two external fused sepals. Longohypanthium pattern is recognized as an extremely rare condition in Eugenia which hypanthium extends as the stamens whorl resulting in a display of curved stamens in the bud instead the standard straight. Nomenclatural changings and a taxonomic revision of Eugenia sect. Schizocalomyrtus newly circumscribed follow these results. Ten species are detailed described and commented under distribution, conservation, and taxonomy. The inflorescence of Eugenia was also analysed under an integrative approach that revealed arrangements previously ignored as relevant for systematic and taxonomy. Seven patterns were described but five are regarded to an evolutionary assessment. Auxotelic pattern was recovered in the early lineages. It suggests that a hypothetical ancestral is similar to the auxotelic pattern. Fasciculiform pattern is widely found in the most speciose Eugenia sect. Umbellatae, indicating that high rates of diversification may be related to the acquisition of this pattern. This study also provides insights in the evolution of inflorescence arrangement by indicating that flexibility in the racemose branching pattern is likely the innovative key that promoted the diversification of Eugenia in the Neotropics