<p>In this thesis, I combined comparative and experimental approaches to examine selection on reproductive traits and population differentiation in the insect-pollinated, self-incompatible, perennial herb <i>Primula farinosa</i>. More specifically, I (1) determined whether the effects of floral display and interactions with pollinators and seed predators, and plant reproductive success were frequency-dependent and affected by surrounding vegetation context, (2) examined the consequences of intermittent drought years on population dynamics using numerical simulations based on demographic data collected over seven years, (3) analyzed among-population differentiation in flowering phenology and reproductive allocation, and its relationship to soil-depth at the site of origin. </p><p>A field experiment suggested that conspicuous plants facilitate inconspicuous plants in terms of pollinator attraction, and that the facilitation effect is contingent on the height of the surrounding vegetation. Further experiments revealed that both mutualistic and antagonistic interactions can result in frequency-dependent selection on floral display. Among inconspicuous plants, both fruit initiation, and damage from seed predators increased with the proportion of the conspicuous morph. The relative strength of these effects, and therefore their net outcome on the relationship between morph ratio and seed production varied among years. </p><p>I combined information on vital rates and their relation to environmental conditions in simulations to predict future population viability in changing environments. Simulated stochastic population growth rate decreased with increasing frequency of drought years. </p><p>Reproductive allocation varied significantly among populations both in the field and in a common-garden experiment, but was correlated with soil depth at the site of origin only in the field. The results suggest that among-population variation in reproductive effort in the field mainly reflects plastic responses to environmental conditions, and that this plasticity may be adaptive. The common-garden experiment suggested that the study populations have diverged genetically in flowering time.</p>

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.

Made available in DSpace on 2014-12-17T15:49:13Z (GMT). No. of bitstreams: 1 PamelaLR_DISSERT.pdf: 3477786 bytes, checksum: e908a438292b6fd69e56fc50da98d37c (MD5) Previous issue date: 2013-03-11 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Vriesea minarum is a rupiculous bromeliad species, with naturally fragmented populations, restricted to the Iron Quadrangle, Minas Gerais, Brazil. It is a threatened species, which is suffering from habitat loss due to the growth of cities and mining activities. The knowledge of genetic variability in plant populations is one of the main branches of conservation genetics, linking genetic data to conservation strategies while the knowledge about plant reproductive biology can aid in understanding key aspects of their life story, as well as in the comprehension of their distribution and survival strategies. Thus, the study of diversity, richness, and genetic structure, as well as the reproductive biology of populations of V. minarum can contribute to the development of conservation actions. Chapter 1 presents the transferability of 14 microsatellite loci for V. minarum. Among the results of this chapter, we highlight the successful transferability of 10 microsatellite loci described for other species of Bromeliaceae, all of which are polymorphic. In Chapter 2, we present the genetic analyses of 12 populations of V. minarum that are distributed throughout the Iron Quadrangle. We used the 10 microsatellite loci tested in Chapter 1. The results show a low population structuring (Fst = 0.088), but with different values of genetic richness (mean = 2.566) and gene diversity (mean = 0.635) for all populations; and a high inbreeding coefficient (Gis = 0.376). These may be the result of pollinators action and/or efficient seed dispersal, thus allowing a high connectivity among populations of naturally fragmented outcrops. The reproductive biology and floral morphology of a population of V. minarum, located in the Parque Estadual da Serra do Rola-Mo?a, are studied in Chapter 3. This reserve is the only public environmental protection area where the species occurs. As a result of field experiments and observations, we found that the species has its flowering period from January to March, with flowers that last for two days and that it has a mixed pollination syndrome. It is primarily alogamous, but also has the capacity to be self-ferilized. It is expected that data obtained in chapters 1, 2 and 3 serve as basis for other studies with species from the ferruginous rocky fields, since until now, to our knowledge, there are no other survey of endemic species from the Iron Quadrangle, seeking to merge the genetic knowledge, with the data of the reproductive biology, with the ultimate aim of biodiversity conservation. Considering the great habitat loss for the species by mining, it becomes crucial to analyze the creation of new protected areas for its conservation / Vriesea minarum ? uma esp?cie de brom?lias rup?cola, com popula??es naturalmente fragmentadas, restrita a regi?o do Quadril?tero Ferr?fero, em Minas Gerais, Brasil. ? uma esp?cie amea?ada, que est? sofrendo com a perda de habitat devido ao crescimento das cidades e ?s atividades de minera??o. O conhecimento da variabilidade gen?tica em popula??es de plantas ? um dos principais ramos de gen?tica da conserva??o, associando dados gen?ticos para as estrat?gias de conserva??o, enquanto que o conhecimento sobre a biologia reprodutiva pode ajudar na compreens?o de aspectos fundamentais da hist?ria de vida, distribui??o e estrat?gias de sobreviv?ncia das plantas. Assim, o estudo da diversidade, riqueza, estrutura gen?tica e biologia reprodutiva das popula??es de V. minarum podem contribuir para o desenvolvimento de a??es de conserva??o. O cap?tulo 1 apresenta a transferabilidade de 14 loci de microssat?lites para V. minarum. Entre os resultados desse cap?tulo, destaca-se o sucesso da transferabilidade de 10 loci de microssat?lites descritos para outras esp?cies de Bromeliaceae, sendo todos eles polim?rficos. No cap?tulo 2, ? apresentada a gen?tica de 12 popula??es de V. minarum que se distribuem por todo o Quadril?tero Ferr?fero. Os resultados obtidos mostram pouca estrutura??o entre as popula??es (Fst = 0,088), mas com diferentes valores de riqueza (m?dia = 2.566) e diversidade gen?tica (m?dia = 0.635) para todas as popula??es; o coeficiente de endogamia foi alto (Gis = 0.376). Estes dados podem ser resultado da a??o de polinizadores e/ou dispers?o de sementes eficientes, j? que as popula??es s?o naturalmente fragmentadas. No cap?tulo 3, ? estudada a biologia reprodutiva e morfologia floral de uma popula??o de V. minarum, situada no Parque Estadual da Serra do Rola-Mo?a, Minas Gerais. Como resultado, foi poss?vel identificar que a esp?cie possui flora??o de janeiro a mar?o; com flores que duram dois dias; s?ndrome mista de poliniza??o; sendo primariamente al?gama, mas tamb?m tem capacidade para ser auto fecundada. Espera-se que dados obtidos nos cap?tulos 1, 2 e 3 sirvam como base para outros estudos com esp?cies de campos rupestres ferruginosos, j? que at? o presente momento, n?o ? de nosso conhecimento a exist?ncia de registros de outras pesquisas feitas com esp?cies end?micas do Quadril?tero Ferr?fero, que busquem conciliar o entendimento da gen?tica, com os dados da biologia reprodutiva, tendo como alvo a conserva??o da biodiversidade neste h?bitat altamente amea?ado pela minera??o. Torna-se crucial uma an?lise cuidadosa para a cria??o de novas ?reas de prote??o, para conserva??o das popula??es da esp?cie

L'identité des organes floraux est définie par l’expression de gènes homéotiques appartenant à la famille des MADS-box au début du développement floral. Un de ces gènes, AGAMOUS (AG), est responsable de l’identité des étamines et des carpelles chez Arabidopsis thaliana. Dans ce manuscrit, je tente de comprendre les propriétés spatiales et temporelles de l’expression d’AG en cherchant à connaître les mécanismes impliqués dans le bon établissement de la dynamique d’expression d’AG pendant les jeunes stades du développement floral.Je débute par développer un modèle de réaction-diffusion qui prend en compte la croissance de la fleur pendant les stades d’intérêt, ainsi que quelques facteurs de transcriptions clefs impliqués dans la régulation d’AG. Ensuite j’ai imagé en direct et en 4D la croissance des fleurs pour quantifier l’activation de l’expression d’AG de son initiation à son patron d’expression stable. Je montre que son expression se déroule en deux phases: une phase de faible expression, et une phase de forte expression. Bien que toutes les cellules du dôme central de la fleur présentent un profil d’activation d’AG similaire, le temps précis au cours du développement où AG est activé est différent pour chacunes d’entre elles et est à l’origine de la stochasticité du patron d’expression. Avec l’aide du modèle, je propose quatres nouvelles hypothèses relatives à la régulation d’AG :AG est capable de maintenir sa propre activation en se liant directement à son second intron au travers d’un complexe protéique contenant au moins deux molécule d'AG, créant ainsi un seuil d'auto-activation.AP2 influence la valeur de ce seuil, restreint l’expression d’AG dans le dôme central de la fleur et produit un retard dans l’activation complète d’AG.LFY et WUS sont nécessaire à l’accumulation des protéines d’AG dans les cellules pour pouvoir atteindre le seuil d’auto-activation et obtenir une expression complète d’AG.Le mouvement d’AG est nécessaire pour obtenir l’expression d’AG dans toutes les cellules du dôme central. Pour prouver ces hypothèses, j’ai réalisé différentes expériences. En premier, utilisant une expérience de FRET-FLIM dans les protoplastes, nous proposons qu’AG est capable de s’associer en homodimer dans les cellules végétales. Néanmoins, sur-exprimer AG pour aider les cellules à atteindre le seuil d’auto-activation plus tôt que dans la plante sauvage ne semble pas modifier la dynamique d’expression de l’AG endogène. En deuxième, j’ai testé le rôle précis de LFY au cours des différentes phases et transitions de la dynamique d’expression d’AG en mutant les sites d'interactions spécifiques pour LFY au sein des séquences de régulation d’AG. Ces mutations retardent l’expression l’expression d’AG et modifient légèrement son patron d’expression. Je montre que seulement d’important retards dans l’activation d’AG induit des modifications phénotypiques. Ensuite, pour tester le rôle de la répression par AP2 dans la dynamique d’expression d’AG, j’analyse le rapporteur d’AG dans le contexte d’un mutant fort d’ap2. Dans ce mutant, l’expression d’AG s’étend à une région plus large et le retard entre l’initiation de l’expression d’AG et la transition entre les phases de faible et forte expressions est diminué. Ces résultats correspondent aux simulations du modèle. Finalement, pour comprendre l’importance du mouvement d’AG d’une cellule à l’autre dans sa propre dynamique, je bloque cette capacité de bouger en utilisant un tag de localisation nucléaire. Bien que cela induit un retard dans l’activation de quelques cellules au stade 3 au moment où toutes les cellules du dôme centrale de la fleur expriment AG dans la plante sauvage, ce retard n’a pas d’effets visible sur le phénotype. / The identity of flower organs is defined by the expression of homeotic genes during early development that belongs to the MADS-box family. One of these genes, AGAMOUS (AG), is responsible for the identity of the stamens and the carpels in Arabidopsis thaliana. In this manuscript, I attempt to fully understand the spatial and temporal properties of AG expression by investigating the mechanisms underlying the proper establishment of AG expression dynamics during the early stages of flower development. I start by developing a reaction-diffusion model that takes into account the growth of the flower at the relevant stages, as well as the few key transcription factors involved in AG regulation. Next I used real-time 4D imaging on growing flowers to quantify the activation of AG expression from its onset to the stable pattern. I show that the AG expression occurs in two phases: a low-expression phase and a high-expression phase. Thus although all cells of the central dome of the flower present similar profiles of AG activation, the precise developmental time at which AG is activated is different in each case, and is the origin of the initial stochastic pattern. With the aid of the model, I also propose four new hypotheses to explain AG regulation: AG is able to maintain its own activation by directly binding its own second intron through a protein complex containing at least two molecules of AG leading to the creation of an auto-activation threshold.AP2 influences the value of this threshold, restraining AG expression to the central dome of the flower and producing a delay in complete AG activation.LFY and WUS are necessary to accumulate AG proteins in cells in order to reach the auto-activation threshold and obtain a full expression of AG.AG movement is necessary to obtain expression of AG in every cell of the central dome. To prove these hypotheses, I have carried out various experiments, using FRET-FLIM in protoplast cells, we suggest that AG is able to form homo-dimers in plant cells. However, overexpressing AG to help cells reach the auto-activation threshold earlier than in the wild-type does not appear to alter the endogenous AG dynamics of expression. Secondly, I test the precise role of LFY in the different phases and transitions in the AG expression dynamics by mutating specific interaction sites for LFY within AG regulatory sequences. These mutations appear to delay AG expression and slightly modify its pattern of expression. I show that only important delays in AG activation induce phenotypic differences. Then, to test the role of AP2 repression in AG expression dynamics, I analyse the AG reporter in the context of a strong ap2 mutant. In these mutants, AG expression spreads to a wider region and reduces the delay between the onset of AG expression and the transition from low- to high-expression. These results match with simulations of the model. Lastly, to understand the importance of AG cell-to-cell movement in AG dynamics, I block its ability to move using a nuclear localisation tag. Although this induces a delay in the activation of few cells at stage 3, when all cells of the central dome of the flower express AG in the WT. This delay has no visible effects on the phenotype.

Made available in DSpace on 2014-08-20T14:33:07Z (GMT). No. of bitstreams: 1 Dissertacao_Marcelo_ de_ Queiroz_ Rocha.pdf: 725823 bytes, checksum: baf6fa097603da126ae27ec658aca762 (MD5) Previous issue date: 2009-03-27 / The use of a new production system demand to know the behavior of cultures and establish the most appropriate management. It is necessary to understand the relationships that control the operation of the plant, and then understand the way in which these relations are dazzling together to result in final income. The growth dynamic, the fenological characterization and yield components of the red cherry tomato crop (Lycopersicon esculentum var. cerasiforme), cultivated in hydroponic system during summer-autumn 2008 crop season, were studied through two experiments conducted in plastic greenhouse, in the Campus of the Universidade Federal de Pelotas, RS. The first experiment was carried out to study the effect of different levels of ionic concentration of the nutrient solution on growth and yield components of cherry tomato (number of fruits, fruit mean weight and fruit yield). This experimental factor was assessed in four different levels: standard nutrient solution, containing 100% of the concentration of nutrients recommended by "Japan Horticultural Experimental Station", corresponding to an initial electrical conductivity (ECi) of 2,3 dS m-1, nutrient solution with reductions of 25 and 50% and with increment of 25% in the concentration of nutrients in relation to the standard solution, corresponding to ECis of 1,3; 1,8 and 2,8 dS m-1, respectively. Another experimental factor in this study was the position of flower cluster and its influence on the yield components. The second experiment was conducted to verify the effect of low sink demand on the vegetative growth in plants cultivated with standard nutrient solution. Two treatments were established: low sink demand (no fruit on the plant by removing 11 all inflorescences) and high sink demand (by maintaining of inflorescences and allowing the fruit to the plant). In both experiments, from the data of dry matter and leaf area accumulated during the experimental period, the production and distribution of dry matter among the different organs of the plant and growth index were determined. The results obtained in the first experiment allow to concluded that: the vegetative stage comprised 30,9% of the crop cycle and the number of the flowers is not dependent on the inflorescence position on the main stem; the position of floral cluster in the plant does not affect the yield components number and weight of the fruit, with little influence on the production of fruits per cluster of cherry tomato; the change of the ion concentration of the nutrient solution (in a range from 1,3 to 2,8 dS m-1) does not affect the number of fruits per plant, but an EC higher than 2,3 dS m-1 causes a reduction in mean weight of the fruits; the leaf expansion, the fruit growth and yield of cherry tomato decrease when the ionic concentration of the nutrient solution increases in the range from 1,8 to 2,8 dS m-1 and when it is reduced from 1,8 to 1,3 dS m-1; the nutrient solution of 1,8 dS m-1 EC can be recommended to increase the fruit yield of cherry tomato in hydroponic system in the summer-autumn crop season. The results observed in the second experiment showed that the fruits corresponded to approximately 26% of the total dry matter of plants in which the fruits are allowed. Thus, in red cherry tomato plants, fruits were not the biggest sink of photoassimilates, since the leaves represented approximately 39% of total dry matter of plants which the fruits are allowed. Thus, the leaf fraction was characterized as the largest source and at the same time as the largest sink of photoassimilates. The fruits has competed more strongly with the leaves than the stems by photoassimilates, indicating that stem and leaves are not characterized as a single compartment for storage of photoassimilates. / O emprego de um novo sistema de produção demanda conhecer o comportamento das culturas e definir o manejo mais adequado. É necessário, portanto, conhecer as relações que regem o funcionamento da planta e, em seguida, compreender a forma segundo a qual todas essas relações se encandeiam entre si para resultar no rendimento final. A dinâmica do crescimento, a caracterização fenológica e os componentes do rendimento da cultura do tomate cereja vermelho (Lycopersicon esculentum var. cerasiforme), cultivado em sistema hidropônico durante o ciclo de verão-outono de 2008, foram estudados através de dois experimentos conduzidos em estufa plástica, no Campus da Universidade Federal de Pelotas, RS. O primeiro experimento foi realizado objetivando-se estudar o efeito de diferentes níveis de concentração iônica da solução nutritiva sobre o crescimento e os componentes do rendimento do tomateiro cereja (número de frutos, peso médio do fruto e produção de frutos). Este fator experimental foi avaliado em quatro diferentes níveis: solução nutritiva padrão, contendo 100% da concentração de nutrientes recomendada pela Japan Horticultural Experimental Station , correspondendo a uma condutividade elétrica inicial (CEi) de 2,3 dS m-1; soluções nutritivas com reduções de 25% e 50% e com incremento de 25% da concentração de nutrientes em relação à solução padrão, correspondendo a CEi de 1,3; 1,8 e 2,8 dS m-1, respectivamente. Outro fator experimental estudado neste experimento foi a posição do cacho floral e sua influência sobre os componentes do rendimento. As avaliações fenológicas foram realizadas em plantas conduzidas na solução padrão. O segundo experimento foi conduzido a fim de verificar o efeito da baixa demanda de drenos sobre o crescimento vegetativo em plantas cultivadas com solução 9 nutritiva padrão. Dois tratamentos foram estabelecidos: baixa demanda de drenos (ausência de frutos na planta através da remoção de todas as inflorescências) e alta demanda de drenos (através da permanência das inflorescências e permitindo-se a frutificação na planta). Em ambos os experimentos, a partir dos dados de matéria seca e da área foliar acumuladas ao longo do período experimental, determinou-se a produção e distribuição de matéria seca entre os diferentes órgãos aéreos da planta e os índices de crescimento. Através dos resultados obtidos no primeiro experimento conclui-se que: a fase vegetativa corresponde a 30,9% do ciclo de cultivo e o número de flores emitidas independe da posição da inflorescência na haste principal; a posição do cacho floral na planta não afeta os componentes do rendimento número e peso médio do fruto, exercendo pouca influência sobre a produção de frutos por cacho do tomateiro cereja; a variação da concentração iônica da solução nutritiva (na faixa entre 1,3 e 2,8 dS m-1) não afeta o número de frutos colhidos por planta mas uma CE superior a 2,3 dS m-1 provoca uma redução no peso médio do fruto; a expansão foliar, o crescimento de frutos e a produtividade do tomateiro cereja diminuem quando a concentração iônica da solução nutritiva aumenta no intervalo entre 1,8 e 2,8 dS m-1 e quando é reduzida de 1,8 para 1,3 dS m-1; a solução nutritiva com CE de 1,8 dS m-1 pode ser recomendada para aumentar a produtividade do tomateiro cereja em sistema hidropônico no ciclo de verão-outono. Os resultados observados no segundo experimento mostram que os frutos corresponderam à aproximadamente 26% da matéria seca total das plantas nas quais se permitiu a frutificação. Desta forma, no tomateiro cereja vermelho, os frutos não foram os maiores drenos de fotoassimilados, uma vez que as folhas representaram aproximadamente 39% da matéria seca total das plantas que frutificaram. Assim, a fração folhas caracterizou-se como a maior fonte e, ao mesmo tempo, como o maior dreno de fotoassimilados. Os frutos competem mais fortemente com as folhas do que com o caule pelos assimilados, indicando que caule e folhas não se caracterizam como um compartimento único de estocagem de fotoassimilados.

Poaceae (or Gramineae) belong to the grass family and is one of the largest families among flowering plants on land. They include some of the most important cereal crops such as rice (Oryza sativa), barley (Hordeum vulgare), wheat (Triticum aestivum), maize (Zea mays), and sorghum (Sorghum bicolor). The characteristic bushy appearance of grass plants, including cereal crops, is formed by the activities of axillary meristems (AMs) generated in the leaf axil. These give rise to tillers from the basal nodes which recapitulate secondary growth axis and AMs are formed during vegetative development. On transition to flowering the apical meristem transforming to an inflorescence meristem (IM) which produces branches from axillary meristem. These IM gives rise to branches that ultimately bear florets. Vegetative branching/tillering determines plant biomass and influences the number of inflorescences per plant. While inflorescence branching determines the number of florets and hence seeds. Thus the overall activity of axillary meristems plays a key role in determining plant architecture during both vegetative and reproductive stages. In Arabidopsis, research on the plant specific transcription factor LEAFY (LFY) has pioneered our understanding of its regulatory functions during transition from vegetative to reproductive development and its role in specifying a floral meristem (FM) identity to the newly arising lateral meristems. In the FM LFY activates other FM genes and genes for floral organ patterning transcription factors. LFY is strongly expressed throughout the young floral meristems from the earliest stages of specification but is completely absent from the IM (Weigel et al., 1992). LFY expression can also be detected at low levels in the newly emerging leaf primordia during the vegetative phase, and these levels gradually increase until the floral transition (Blazquez et al., 1997; Hempel et al., 1997). In rice, the LFY ortholog-RFL/APO2 is expressed predominantly in very young branching panicles/ inflorescence meristems (Kyozuka et al., 1998; Prasad et al., 2003) while in the vegetative phase RFL is expressed at axils of leaves (Rao et al., 2008). In rice FMs expression is restricted to primordia of lodicules, stamens, carpels and ovules (Ikeda-Kawakatsu et al., 2012). Knockdown of RFL activity or loss of function mutants show delayed flowering and poor panicle branching with reduced number of florets and lower fertility (Rao et al., 2008, Ikeda-Kawakatsu et al., 2012). In some genotypes reduced vegetative axillary branching is also compromised (Rao et al., 2008). On the other hand RFL overexpression leads to the early flowering, attributing a role as an activator for the transition of vegetative meristems to inflorescence meristems (Rao et al., 2008). Thus, RFL shows a distinct developmental expression profile, has unique mutant phenotypes as compared to Arabidopsis LFY thus indicating a divergence in functions. We have used various functional genomics approaches to investigate regulatory networks controlledby RFL in the vegetative axillary meristems and in branching panicles with florets. These regulatory effects influence tillering and panicle branching, thus contributing to rice plant architecture. RFL functions in axillary meristem Vegetative AMs are secondary shoot meristems whose outgrowth determines plant architecture. In rice, AMs form tillers from basal nodes and mutants with altered tillering reveal that an interplay between transcription factors and the phytohormones - auxin, strigolactone underpins this process. We probed the relationship between RFL and other factors that control AM development. Our findings indicate that the derangements in AM development that occur on RFL knockdown arise from its early effects during specification of these meristems and also later effects during their outgrowth of AM as a tiller. Overall, the derailments of both steps of AM development lead to reduced tillering in plants with reduced RFL activity. Our studies on the gene expression status for key transcription factor genes, genes for strigolactone pathway and for auxin transporters gave an insight on the interplay between RFL, LAX1 and strigolactone signalling. Expression levels of LAX1 and CUC genes, that encode transcription factors with AM specification functions, were modulated upon RFL knockdown and on induction of RFL:ΔGR fusion protein. Thus our findings imply a likely, direct activating role for RFL in AM development that acts in part, through attaining appropriate LAX1 expression levels. Our data place meristem specification transcription factors LAX1 and CUC downstream to RFL. Arabidopsis LFY has a predominant role in conferring floral meristem (FM) identity (Weigel et al., 1992; Wagner, 2009; Irish, 2010; Moyroud et al., 2010). Its functions in axillary meristems were not known until recently. The latter functions were uncovered with the new LFYHARA allele with only partial defects in floral meristem identity (Chahtane et al., 2013). This mutant allele showed LFY can promote growth of vegetative AMs through its direct target REGULATOR OF AXILLARY MERISTEMS1 (RAX1), a R2R3 myb domain factor (Chahtane et al., 2013). These functions for Arabidopsis LFY and RAX1 in AMs development are parallel to and redundant with the pathway regulated by LATERAL SUPPRESSOR (LAS) and REGULATOR OF AXILLARY MERISTEM FORMATION1 (ROX1) (Yang et al., 2012; Greb et al., 2003). Interestingly, ROX1 is orthologous to rice LAX1 and our data show LAX1 expression levels in rice panicles and in culms with vegetative AMs is dependent on the expression status of RFL. Thus, we speculate that as compared to Arabidopsis AM development, in rice the LFY-dependent and LFY-independent regulatory pathways for AMs development are closely linked. In Arabidopsis, CUC2 and CUC3 genes in addition to their role in shoot meristem formation and organ separation play a role in AM development possibly by defining a boundary for the emerging AM. These functions for the Arabidopsis CUC genes are routed through their effects on LAS and also by mechanisms independent of LAS (Hibara et al., 2006; Raman et al., 2008). These data show modulation in RFL activity using the inducible RFL:∆GR protein leads to corresponding expression changes in CUC1/CUC2 and CUC3 genes expression in culm tissues. Thus, during rice AM development the meristem functions of RFL and CUC genes are related. Consequent to specification of AM the buds are kept dormant. Bud outgrowth is influenced by auxin and strigolactone signalling pathways. We investigated the transcript levels, in rice culms of genes involved in strigolactone biosynthesis and perception and found the strigolactone biosynthesis gene D10 and hormone perception gene are significantly upregulated in RFL knockdown plants. Further, bioassays were done for strigolactone levels, where we used arbuscular mycorrhiza colonization assay as an indicator for strigolactone levels in wild type plants and in RFL knockdown plants. These data validate higher strigolactone signalling in RFL knockdown plants. To probe the relationship between RFL and the strigolactone pathway we created plants knocked down for both RFL and D3. For comparison of the tillering phenotype of these double knockdown plants we created plants with D3 knockdown alone. We observed reduced tillering in plants with knockdown of both RFL and D3 as compared to the tiller number in plants with knockdown of D3 alone. These data suggest that RFL acts upstream to D3 of control bud outgrowth. As effects of strigolactones are influenced by auxin transport we studied expression of OsPIN1 and OsPIN3 in RFL knockdown plants. Their reduced expression was correlated with auxin deficiency phenotypes of the roots in RFL knockdown plants. These data in conjunction with observations on OsPIN3 the gene expression modulation by the induction of RFL:∆GR allow us to speculate on a relationship between RFL, auxin transport and strigolactones with regard to bud outgrowth. We propose that the low tillering phenotype of RFL knockdown plants arises from weakened PATS, consequent to low levels of PIN1 and PIN3, coupled with moderate increase in strigolactones. Taken together, our findings suggest functions for RFL during AM specification and tiller bud outgrowth. RFL functions in panicle branching Prior studies on phenotypes of RFL knockdown or loss of function mutants suggested roles for RFL in transition to flowering, inflorescence meristem development, emergence of lateral organs and floral organ development (Rao et al., 2008; Ikeda-Kawakatsu et al., 2012). It has been speculated that RFL acts to suppress the transition from inflorescence meristem to floral meristem through its interaction with APO1 (Ikeda-Kawakatsu et al., 2012). The downstream genes regulated by RFL in these processes have not yet been elucidated. To identify direct targets of RFL in developing panicles we adopted ChIP-seq coupled with studies on gene expression modulation on induction of RFL. For the former we raised polyclonal anti-sera and chromatin from branching panicles with few florets. For gene expression modulation studies, we created transgenics with a T-DNA construct where an artificial miRNA against 3’UTR specifically knocked endogenous RFL and the same T-DNA had a second expression cassette for generation of a chemically inducible RFL-ΔGR protein that is not targeted by amiR RFL. Our preliminary ChIP-seq data in the wild type panicle tissues hints that RFL binds to hundreds of loci across the genome thus providing first glimpse of direct targets of RFL in these tissues. These data, while preliminary, were manually curated to identify likely targets that function in flowering, we summarize here some key findings. Our study indicates a role of RFL in flowering transition by activating genes like OsSPL14 and OsPRMT6a. Recent studies indicate that OsSPL14 directly binds to the promoter of OsMADS56 or FTL1, the rice homologs of SOC1 and FT to promote flowering (Lu et al., 2013). As RFL knockdown plants show highly reduced expression of OsMADS50/SOC1 and for RFT1 (Rao et al., 2008), and we show here RFL can bind and induce OsSPL14 expression we suggest the RFL¬OsSPL14 module can contribute to the transition of the SAM to flowering. Further, OsSPL14 in the young panicles directly activates DENSE AND ERECT PANICLE1 (DEP1) to control panicle length (Lu et al., 2013). Thus RFL-OsSPL14-DEP1 module could explain the role of RFL in controlling panicle architecture (Rao et al., 2008; Ikeda-Kawakatsu et al., 2012). Thus RFL plays a role in floral transition and this function is conserved across several LFY homologs. Our data ChIP-seq in the wild type tissue and gene expression modulation studies in transgenics also give molecular evidences for the role of RFL in suppression of floral fate. The direct binding of RFL to OsMADS17, OsYABBY3, OsMADS58 and HD-ZIP-IV loci and the changes in their transcript levels on induction of RFL support this hypothesis. Once the transition from SAM to FM takes place, we speculate RFL represses the conversion of inflorescence branch meristems to floral fate by negatively regulating OsYABBY3, HD-ZIP class IV and OsMADS17 that can promote differentiation. These hypotheses indicate a diverged function for RFL in floral fate repression. Arabidopsis LFY is known to activate the expression of AGAMOUS (AG), whose orthologs in rice are OsMADS3 and OsMADS58. Our studies confirm conservation with regard to RFL binding to cis elements at OsMADS58 locus that is homologous to Arabidopsis AG. But importantly we show altered consequences of this binding on gene expression. We find RFL can suppress the expression of OsMADS58 which we speculate can promote a meristematic fate. Further, we also present the abnormal upregulation of floral organ fate genes on RFL downregulation. These data too indicate functions of RFL, are in part, distinct from the role of Arabidopsis LFY where it works in promoting floral meristem specification and development. These inferences are supported by our data that rice gene homologs for AP1, AP3 and SEP3 are not directly regulated by RFL, unlike their direct regulation by Arabidopsis LFY during flower development. We also report the expression levels of LAX1, FZP, OsIDS1 and OsMADS34 genes involved in meristem phase change and IM branching are RFL dependent. This is consistent with its role in the suppression of determinacy, thereby extending the IM activity for branch formation. But as yet we do not know if these effects are direct. Together, our data report direct targets of RFL that contribute to its functions in meristem regulation, flowering transition, and suppression of floral organ development. Overall, our preliminary data on RFL chromatin occupancy combined with our detailed studies on the modulation of gene expression provides evidence for targets and pathways unique to the rice RFL during inflorescence development. Comparative analysis of genes downstream to RFL in vegetative tillers Vs panicles Tillers and panicle branches arise from the axillary meristems at vegetative and reproductive stages, respectively, of a rice plant and overall contribute to the plant architecture. Some regulatory factors control branching in both these tissues - for example, MOC1 and LAX1. Mutants at these loci affect tillers and panicle branch development thus indicating common mechanisms control lateral branch primordia development (Li et al., 2003; Komatsu et al., 2003; Oikawa and Kyozuka, 2009). Knockdown of RFL activity or loss-of-function mutants cause significantly reduced panicle branching and in few instances, reduction in vegetative axillary branching (Rao et al., 2008; Ikeda- Kawakatsu et al., 2012). We took up the global expression profiling of RFL knockdown plants compared to wild type plants in the axillary meristem and branching panicle tissue. These data provide a useful list of potential targets of RFL in axillary meristem and branching panicle tissue. The comparative analysis of the genes affected in the two tissues indicates only a subset of genes is affected by RFL in both the vegetative axillary meristems and branching panicle. These genes include transcription factors (OsSPL14, Zn finger domain protein, and bHLH domain protein), hormone signalling molecules (GA2 ox9) and cell signalling (LRR protein) as a set of genes activated by RFL in both tissues. On the other hand, these comparative expression profiling studies also show distinct set of genes deregulated by RFL knockdown in these two tissues therefore implicating RFL functions have a tissue-specific context. The genes deregulated only in axillary meristem tissue only include D3- involved in the perception of strigolactone, OsMADS34 speculated to have a role in floral transition and RCN1 involved in transition to flowering. On the other hand, the genes – CUC1, OsMADS3, OsMADS58 involved in organ development and floral meristem determination were found to be deregulated only in panicle tissues of RFL knockdown plants. These data point towards presence of distinct mechanisms for the development of AMs as tillers versus the development of panicle axillary as rachis branches. Overall, these data implicate genes involved in transition to flowering, axillary meristem development and floral meristem development are controlled by RFL in different meristems to thereby control plant architecture and transition to flowering.

LFY of Arabidopsis is a member of a unique plant specific transcription factor family. It is involved in giving meristem a determinate floral fate by the activation of floral organ identity genes and preventing inflorescence meristem identity. RFL is a homolog of FLO/LFY in rice. Studies from our lab on rice RFL, based on the effects of knockdown or overexpression, showed its major functions are in timing the conversion of SAM to IM and to prevent the premature conversion of branch meristem to spikelets. Additionally roles in vegetative axillary meristem specification have been also been identified in laboratory. Here, we attempt to delineate molecular pathways directly regulated by RFL as a transcription factor controlling inflorescence and floral development in rice. Part I: Identification of global target genes bound by RFL in developing rice inflorescences We carried out ChIP sequencing of the DNA bound by RFL in panicles (01.-0.3cm stage) using anti-RFL antibody. DNA sequences in one library pool were analyses by the MACS algorithm (FDR<0.01), to find 8000 binding sites while the SPP algorithm identified 5000 enriched peaks. These mapped to 2500 or 2800 gene-associated loci respectively, 617 of which were common loci to both pipelines. Several RFL bound gene loci were homologs of Arabidopsis thaliana LFY gene targets. Such gene targets underscore conserved downstream targets for LFY-proteins in evolutionarily very distinct species. AtLFY is known to bind variants of CCANT/G cis element classified as primary, inflorescence or seedling type. We scanned for these three types of cis elements at 123 RFL bound genes with likely functions in flowering. For a few of these 123 rice loci we find one of these cis motifs (p-value<0.001) in RFL bound ChIP-seq data. To validate these targets of RFL, we adopted in vitro DNA-protein binding assays with bacterially purified RFL protein. We confirm RFL target interactions with some genes implicated in flowering time, others in photoperiod triggered flowering, circadian rhythm, gibberellin hormone pathway, inflorescence development and branching. The in vitro experiments hint different RFL-DNA binding properties as compared to Arabidopsis LFY. We report binding to sequences at rice gene loci that are unique targets. Part II: Pathways regulated by RFL for reproductive transition and panicle development To co-relate DNA binding of RFL to target loci with changes in their gene expression, expression studies were taken up for selected set of genes implicated in rice flowering transition and panicle architecture. To study in planta and tissue specific gene regulation by RFL we raised RFL dsRNAi transgenics. Comparative transcript analysis in these RFL partial knockdown lines and matched wild type tissues reveal that RFL is an activator for some genes and repressor for other gene targets. We also examined if the gene expression effects of RFL knockdown can be reversed by induced complementation with an RFL-GR protein. We raised transgenics plants with a T-DNA ubi:RFL-GR, 35S CaMV:amiR RFL for these experiments. In planta target gene transcript levels were assessed in various conditions conditions. These studies validate rice RFL as an activator of some panicle architecture genes. Part III: Analysis of endogenous RFL protein in WT rice tissues Studies in Arabidopsis and in petunia with LFY and AFL, respectively, implicate these some abnormal mobility as compared to their predicted molecular weight when overexpressed. We studied endogenous RFL protein abundance in planta, adopting western analysis with anti-RFL antibody. We consistently identify two prominent cross reacting bands in different tissues which can be also be pulled-down from whole nuclear extracts of panicle and axillary meristem tissues. We speculate on likely modifications and possible functions for the same.

Les phytoplasmes sont des bactéries phytopathogènes, sans paroi, qui appartenant à la classe des Mollicutes. Ils ne peuvent pas etre cultivés in vitro et sont limités à des tubes du phloème. Ils provoquent des centaines de maladies chez de nombreuses espèces végétales dans le monde entier, ce qui conduit à des pertes de récolte importantes. Les phytoplasmes sont transmis naturellement par des insectes suceurs de sève dans laquelle ils se multiplient. Ils induisent des symptômes graves, notamment le jaunissement, la croissance limitée, déclin, ainsi que des anomalies des fleurs et des fruits. L'infection par le phytoplasme du stolbur, en particulier, affect fortement la morphologie florale. Dans la tomate, deux isolats différents du phytoplasme du stolbur, nommé C et PO, induisent des symptômes différents. La tomate infectée par le phytoplasme du stolbur PO montrent des malformations florale telles que les sépales hypertrophiés, les pétales et les étamines avortées ce qui conduit à la stérilité. En revanche, la tomate infecté par le phytoplasme du stolbur C ont de petites feuilles de tomate en retrait, mais les fleurs presque normale, et produisent des fruits. Nous avons précédemment montré que SlDEF, un gène impliqué dans la formation des pétales est réprimé dans des plantes de tomate infectée par le stolbur phytoplasme PO. Toutefois, l'expression de son facteur de transcription, codée par le gène FA, est resté stable ou voir légèrement augmentée. Nous avons donc émis l'hypothèse que la répression de SlDEF pourrait être dû à une méthylation de l'ADN. Pour tester cette hypothèse, nous avons étudié l'expression des gènes de méthylases et de déméthylases. Ils étaient en général réprimés dans les tomates infectées par le phytoplasme du stolbur PO, ce qui était en accord avec l'hypothèse De plus, nous avons étudié les voies de défense activée chez les tomates infectées par le phytoplasme du stolbur. Pour se défendre, les plantes utilisées des molécules de signalisation comme l'acide salicylique (SA), l'acide jasmonique (JA) et d'éthylène (ET). Nous avons étudié l'expression de 21 gènes de défense dépendants SA / JA / ET, des gènes de biosynthèse et les facteurs de transcription chez les tomates infectées par les phytoplasmes du stolbur C et PO. Nous avons également étudié l'effet de la pré-activation des voies de SA et JA sur la production des symptômes. Nos résultats montrent clairement que les voies de défense ont été activées différemment dans les tomates infectés par le phytoplasme du stolbur C et PO. En effet, les voies de défense dépendantes de SA, ET et JA ont été activées chez les tomates infectées par le phytoplasme du stolbur C alors que seulement les voies dépendantes SA et ET ont été activés dans les tomates infectées par stolbur PO . En outre, la pré-activation de la voie de défense dépendante SA par l'application de BTH modifie légèrement l'évolution des symptômes de maladies causées par le phytoplasme du stolbur PO / Phytoplasma are cell wall-less, phytopathogenic bacteria belonging to the class Mollicutes. They have not been cultured in vitro and are restricted to the phloem sieve tubes. They cause hundreds of diseases in many plant species worldwide, resulting in important crop losses. Phytoplasmas are naturally transmitted by sap-sucking insects in which they multiply. They induce severe symptoms including yellowing, restricted growth, decline, as well as major flowers and fruits abnormalities.The stolbur phytoplasma infection, in particular, has been reported to strongly affect floral morphology. In tomato, two different isolates of stolbur phytoplasma, named C and PO, induce different symptoms. The stolbur PO phytoplasma-infected plants show abnormal flower development such as hypertrophied sepals, and aborted petals and stamens leading to sterility. In contrast, stolbur C phytoplasma-infected tomato have small indented leaves but nearly normal flowers, and produce fruits. We have previously shown that SlDEF, one gene involved in petal formation, was repressed in stolbur PO phytoplasma-infected tomato. However, the expression of its transcription factor, encoded by the gene FA, was unchanged or slightly up-regulated. So we hypothesized that SlDEF repression could be due to DNA methylation. To test this hypothesis, we studied the expression of DNA methylases and demethylases genes. They were in general down-regulated in stolbur PO infected tomato, which was in agreement with the hypothesis. However, the regulation of SlDEF expression could not be firmly correlated to the DNA methylation status of its promoter region. In addition, we studied the plant defense pathways activated in stolbur phytoplasma-infected tomato. To defend themselves, plants used signalling molecules like Salicylic acid (SA), Jasmonic acid (JA) and Ethylene (ET). We studied the expression of 21 SA/JA/ET regulated defense and biosynthesis genes including transcription factors in stolbur C and PO phytoplasma-infected tomato as compared to healthy ones. We also studied the effect of pre-activation of SA and JA mediated defense pathways on symptom production. Our results clearly showed that defense pathways were activated differently in stolbur C and PO phytoplasma-infected tomato. Indeed, SA ET and JA dependant pathways were activated in stolbur C-infected tomato while only SA and ET dependant pathways were activated in stolbur PO-infected plants. In addition, pre-activation of SA-dependent defense pathway by application of BTH slightly modify the evolution of disease symptoms caused by stolbur PO phytoplasma whereas no effect was observed after treatment with an analogue of JA.

Orientador: Leonor Patricia Cerdeira Morellato / Resumo: A visão é o principal sentido utilizado pelos polinizadores para forragear, no qual a cor das flores é a primeira característica de atração. A percepção dos sinais de cores depende da coloração do segundo plano das flores, composto principalmente pelas folhas. Diferenças na composição das espécies e na estrutura da vegetação, e também na sazonalidade climática, podem interferir na cor do segundo plano de folhas em diferentes vegetações e estações, interferindo como os polinizadores percebem a flor. Portanto, a diversidade de cores das flores e os padrões de floração de uma comunidade podem estar relacionados com a composição de polinizadores e condições ambientais. Desta maneira, utilizando comunidades vegetais e considerando o sistema visual das abelhas, nós descrevemos e comparamos a diversidade de cores das flores e seus sinais, dando importância para a cor do segundo plano de folhas em duas vegetações sazonais tropicais e uma vegetação sazonal temperada. Em seguida, para vincular os sinais florais com a sazonalidade, nós analisamos a importância das síndromes de polinização levantadas, o padrão de floração, a influência da cor do segundo plano de folhas no padrão de cor das flores entre as estações. Nós encontramos diferenças na diversidade de cores das flores e confirmamos a influência da coloração do segundo plano de folhas, juntamente com a estrutura da vegetação e a intensidade da sazonalidade nos sinais florais exibidos em diferentes vegetações sazonais, de acordo co... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Vision is the main sense used to forage by pollinators being the flower colour the primary feature of attraction. Colour signals perception depend on the flower background colouration, mainly composed by leaves. Differences in species composition and vegetation structure, and also the seasonality, may change the leaf-background colouration of different vegetations and seasons, interfering how the pollinators perceive a flower. Therefore, flower colour diversity and flowering patterns of a community may be related to the pollinators’ composition and environmental conditions. Using a community level-approach and according to bee visual system, we described and compared the flower colour diversity and the signals of a temperate and two tropical seasonal vegetations, considering their leaf background colouration. Then, to link flower signals to seasonality and using the cerrado sensu stricto as a model of seasonal vegetation, we analysed the importance of the surveyed pollination syndromes, the community flowering pattern, flowering patterns according to the colour of flowers and the influence of seasonal changes in the background colouration in the flower colour signals between seasons. We found differences in flower colour diversity and confirmed the influence of the leaf- background colouration, along with the vegetation structure and seasonality intensity, in flower colour signals displayed in different seasonal vegetations according to the colour vision of bees. Higher value... (Complete abstract click electronic access below) / Mestre

Les plantes présentent une incroyable diversité de tailles, formes et couleurs, étroitement liée à certaines de leurs fonctions biologiques telles que la photosynthèse, la reproduction, etc. De ce fait, la façon dont ces organismes multicellulaires acquièrent des formes complexes est une question clé en biologie du développement. La morphologie des organes végétaux résulte en effet de la modulation, à l’échelle cellulaire, de patrons d’expression génétique, de croissance et de différenciation. Bien que la morphogénèse ait été largement étudiée d’un point de vue moléculaire, nous ne savons toujours pas comment ces réseaux génétiques sont traduits en formes biologiques. Le but de ce projet de recherche est donc d’étudier le développement des organes latéraux (feuilles juvéniles, feuilles caulinaires et organes floraux, id sépales, pétales et anthères) chez l’espèce modèle Arabidopsis thaliana. Afin d’approcher la question du rôle des interactions complexes entre cellules et organes lors du développement, nous nous intéressons à la variabilité entre les organes, mais aussi à la variabilité cellulaire intrinsèque de chaque organe. Nous avons donc testé (1) si la diversité de formes observées chez les organes latéraux résulte de modulations d’un programme développemental commun; (2) si la croissance et le développement des organes latéraux est un phénomène stochastique ou dépend de mécanismes sous-jacents spécifiques. Pour ce faire, nous utilisons une approche multidisciplinaire basée sur la génétique, la microscopie confocale et l’analyse d’images 3D pour extraire les patrons de croissance inhérents aux différents organes. Les résultats de la première étude (Chapitre 2) montrent que la forme des organes dépend de l’équilibre entre croissance et différentiation, dont la régulation précise permet l'acquisition de fonctions hautement spécialisées. La feuille caulinaire, par exemple, présente un retard de différenciation qui permet une activité morphogénétique prolongée et une redistribution de la croissance. À travers la suppression transitoire de la croissance lors des premiers stades de développement, la trajectoire développementale de la feuille caulinaire permet sa double fonction, à la fois protectrice et photosynthétique.\par La deuxième étude (Chapitre 3), quant-à-elle, s’intéresse aux comportements des cellules individuelles, dont la croissance, bien que contrôlée par des informations positionnelles, est souvent hétérogène. Cette variabilité résulte de la différenciation de cellules spécialisés, les stomates, qui suivent un programme de développement spécifique. Le comportement autonome de ces cellules, asynchrone, est la principale source de variabilité dans des tissus dont la croissance est autrement homogènes. Dans l’ensemble, cette thèse a permis de mettre en lumière l’importance de la temporalité lors du développement des organes végétaux. Que ce soit à l’échelle de l’organe, du tissu ou de la cellule, la modulation et la synchronisation de la croissance et de la différentiation sont nécessaires à l’acquisition des formes stéréotypiques des organes et à leur complexité fonctionnelle. / Plants display an incredible diversity of sizes, shapes, and colors, closely linked to some of their biological functions, such as photosynthesis, reproduction, etc. How these multicellular organisms acquire complex shapes is, therefore, a key question in developmental biology. The morphology of plant organs results from cell-level modulation of patterns of gene expression, growth, and differentiation. Although morphogenesis has been extensively studied from a molecular point of view, how genetic networks are translated into biological forms is still unclear. Thus, the aim of this research project is to study the development of lateral organs (rosette leaves, cauline leaves, and floral organs, i.e. sepals, petals, and anthers) in the model species Arabidopsis thaliana. To address the question of the role of complex cell-organ interactions during development, we are interested not only in variability between organs but also in the intrinsic cellular variability of each organ. We, therefore, tested (1) whether the diversity of shapes observed in lateral organs results from modulations of a common developmental program; (2) whether the growth and development of lateral organs is a stochastic phenomenon or depends on specific underlying mechanisms. To this end, we are using a multidisciplinary approach based on genetics, confocal microscopy, and 3D image analysis to extract the growth patterns inherent in the different organs. The results of the first study (Chapter 2) show that organ shape depends on the balance between growth and differentiation, which fine regulation enables the acquisition of highly specialized functions. The cauline leaf, for example, shows a delay in differentiation that allows for prolonged morphogenetic activity and growth redistribution. Through the transient growth suppression at early stages, the cauline leaf developmental trajectory allows for its dual function, from protection to photosynthesis. The second study (Chapter 3) focuses on the behavior of individual cells, whose growth, although controlled by positional information, is often heterogeneous. This variability results from the differentiation of specialized cells, the stomata, which follow a specific developmental program. The autonomous, asynchronous behavior of these cells is the main source of variability in tissues whose growth is otherwise homogeneous. Overall, this thesis has shed light on the importance of timing in plant organ development. Whether at the organ, tissue, or cell level, modulation and synchronization of growth and differentiation are necessary for the acquisition of stereotypic organ shapes and functional complexity.