Fundação de Amparo a Pesquisa do Estado de Minas Gerais / The distyly is a floral polymorphism controlled genetically, where the populations present individuals with pin flowers, that possess long styles and short stamens, and individuals with thrum flowers, that possess short styles and long stamens. Their populations tend to be isoplethyc and anomalies in the distribution of pin and thrum flowers in the populations have usually been associated with the habitat fragmentation and environmental disturbances. Besides morphological characteristics of styles/stigmas anthers/pollen grains, these flowers can also present secondary differences in their morphology. Heterostylous plants usually possess a self-incompatibility system, where fruit-set occurs only after cross pollination between morphs (legitimate cross pollination). Palicourea rigida is a typically distylous species of the Rubiaceae, a family that contains the largest number of distylous species. The species is widely distributed in the Brazilian Cerrados and it is pollinated by hummingbirds. The objective of the present work was to study different populations from P. rigida comparing the distribution of pin vs. thrum individuals and other characteristics associated to heterostyly in order to verify if there were asymmetries between floral morphs and differences among areas that could be associated to environmental conditions or recent changes. The work was carried out in the Parque Estadual da Serra Caldas Novas - PESCAN (GO) and in RPPN of the Clube Caça e Pesca Itororó de Uberlândia - CCPIU (MG) where populations of P. rigida were studied in 2005 and 2006. Density, height and pin/thrum ratio were assessed for flowering individuals in both areas. Flower morphs were investigated for differences in floral morphology, nectar production, reproductive success and site of self incompatibility reactions. In spite of differences in density and individuals height, both populations were isoplethyc, with ratio between floral morphs not significantly different from 1:1. The populations were different probably due to their conservation histories, but such differences do not seem to have affected other characteristics associated to distyly. The anisoplethy was observed for populations of the species in Brasília and for other species of Palicourea. Besides the reciprocal herkogamy, with differences marked between pin/thrum styles and stamens, there were differences in the size of the anthers, stigmas and corolla that were larger in thrum flowers. The nectar production varied in volume and concentration in the two populations, but there was not a pattern that could be associated to each one of the floral morphs. There were no differences in reproductive success, which were high for the two morphs and for both populations, in spite of the species to be remarkably self-incompatible. There were differences in the site of incompatibility barriers between floral morphs that corroborated previous observations for the species and for many other species of Rubiaceae. The incompatibility reaction was more marked and interrupted pollen tube growth in the stigma of thrum flowers, while it was more variable and occurred along the style in pin flowers. All the pistils of flowers collected from both floral morphs in the study areas showed a profusion of pollen grains and pollen tubes, which indicates high pollination efficiency. The main floral visitor and pollinator, in both studied areas, was the hummingbird Eupetomena macroura. The high fruit-set indicates that the pollinators transported enough compatible pollen grains between floral morphs, although they commonly presented territorial behavior. The reproductive success was considered high, specially when compared with other self-incompatible species of the Cerrado. / A distilia é um polimorfismo floral controlado geneticamente, onde as populações apresentam indivíduos com flores longistilas, que possuem estiletes longos e estames curtos, e indivíduos com flores brevistilas, que possuem estiletes curtos e estames longos. Suas populações tendem a ser isopléticas, ou seja, com indivíduos dos dois morfos na proporção 1:1, e as anomalias referentes à distribuição dos morfos nas populações têm sido normalmente associadas à fragmentação de habitats e perturbações ambientais. Além das características morfológicas de estiletes/estigmas e anteras/grãos-de-pólen, estas flores podem ainda apresentar diferenças secundárias na morfologia. Plantas heterostílicas geralmente possuem um sistema de auto-incompatibilidade, onde só há formação de frutos nos cruzamentos entre morfos. Palicourea rigida é uma espécie tipicamente distílica de Rubiaceae, família que contém o maior número de espécies distílicas registradas. A espécie é amplamente distribuída nos cerrados brasileiros e é polinizada por beija-flores. O objetivo do presente trabalho foi estudar diferentes populações de P. rigida comparando a distribuição dos morfos florais e as características associadas à heterostilia para verificar se existem assimetrias entre os morfos e diferenças entre as áreas que pudessem ser associadas às condições ou mudanças ambientais. O trabalho foi realizado no Parque Estadual da Serra de Caldas Novas - PESCAN (GO) e na RPPN do Clube Caça e Pesca Itororó de Uberlândia - CCPIU (MG) onde foram avaliadas populações de P. rigida nos anos de 2005 e 2006. As populações foram caracterizadas quanto à densidade, altura e razão entre morfos dos indivíduos floridos. Quanto aos morfos florais, foram investigadas diferenças na morfometria, produção de néctar, sucesso reprodutivo e reações de incompatibilidade. Apesar de diferirem em densidade e altura dos indivíduos, ambas as populações foram isopléticas, com razão entre morfos não significativamente diferente de 1:1. As populações se mostraram distintas provavelmente devido à suas histórias de conservação, mas tais diferenças não parecem ter afetado as características associadas à distilia. A anisopletia foi observada para populações da espécie em Brasília e para outras espécies de Palicourea. Além da hercogamia recíproca, com diferenças marcadas entre estiletes e estames dos dois morfos, ainda houve diferenças no tamanho das anteras, dos estigmas e da corola que foram maiores em flores brevistilas. A produção de néctar variou em volume e concentração nas duas populações, mas não houve um padrão que pudesse ser associado a cada um dos morfos. Não houve diferenças quanto ao sucesso reprodutivo, que foi alto para os dois morfos e em ambas as populações, apesar da espécie ser marcadamente auto-incompatível. O sucesso reprodutivo foi considerado alto, sobretudo quando comparado com outras espécies auto-incompatíveis do Cerrado. Todos os pistilos de flores coletadas ao acaso dos dois morfos nas áreas de estudo mostraram grãos-de-pólen e tubos polínicos em profusão, o que indica alta eficiência de polinização. Houve diferença nos locais de formação das barreiras de incompatibilidade entre os morfos que corroboraram as observações anteriores para a espécie e o encontrado para várias outras Rubiaceae. De uma maneira geral a reação de incompatibilidade foi mais marcada e interrompeu o crescimento dos tubos polínicos no estigma do morfo brevistilo, enquanto foi mais variável e ocorreu ao longo do estilete no morfo longistilo. O principal visitante floral e polinizador, em ambas as áreas estudadas, foi o beija-flor Eupetomena macroura. A alta produtividade de frutos indica que os polinizadores efetivamente transportaram quantidades suficientes de grãos-de-pólen compatíveis entre os morfos, apesar de comumente apresentarem comportamento territorial. / Mestre em Ecologia e Conservação de Recursos Naturais

O presente trabalho teve por objetivos avaliar o sistema reprodutivo de etnovariedades de batata-doce, mais especificamente, determinar quais genótipos são autocompatíveis e quais são compatíveis ou incompatíveis entre si. O sistema reprodutivo foi avaliado em 13 etnovariedades de batata-doce, através da frutificação e da observação in situ dos tubos polínicos no pistilo através da técnica de epifluorescência com azul de anilina, em flores submetidas a polinização controlada. Teste de viabilidade dos grãos de pólen foi realizado usando os corantes carmim acético e cloreto de tetrazólio em quatro horários diferentes de coleta (6:00; 8:00; 10:00 e 12:00 h). Visando avaliar alguma associação entre a incompatibilidade e a morfologia floral, foram tomadas medidas de todas as peças florais. A frutificação ocorreu apenas nas flores submetidas à polinização cruzada. Do total de 78 cruzamentos realizado, entre as 13 etnovariedades, 46,15% desses é que ocorreu a frutificação. Tubos polínicos nos pistilos foram verificados nas flores autopolinizadas (em 38,46% das etnovariedades) e nas flores submetidas à polinização cruzada (em 85,90% dos cruzamentos). Esses resultados revelam que a batata-doce possui sistema de reprodução sexual mista. A maioria das etnovariedades apresentou taxa de grãos de pólen viáveis acima de 80% quando corados com carmim, mas quando corados com tetrazólio essa taxa foi abaixo de 70%. Entretanto, a variação na viabilidade do pólen nos quatros horários de coleta foi quase inalterada, em ambos corantes. Todas as variáveis florais mensuradas mostraram-se diferenças significativas no nível de 5% de probabilidade, o que revela polimorfismo floral. Quatro padrões bem definido em relação à posição do estigma e da antera mais alta foi estabelecido; estigma bem acima antera, estigma um pouco acima da antera, estigma no mesmo nível da antera e estigma mais abaixo da antera. Mas, neuma associação foi verificada entre a morfologia floral e a incompatibilidade, os cruzamentos ocorreram entre todos os morfotipos. / The objective of this work was to study the reproductive system of sweet potato landraces - specifically, to determine which genotypes are either self-compatible or selfincompatible and which are either compatible or incompatible among themselves. The reproductive system was studied in 13 landraces of sweet potato, by means of the fruit production and in situ observation of the pollen tubes in the pistil with the epifluorescence technique with aniline blue in hand-pollinated flowers. A viability test was performed using the dyes acetic carmine and tetrazolium chloride in fou different collecting times (6:00; 8:00; 10:00 and 12:00 h). In order to detect a possible association between incompatibility and flower morphology, several flower parts were measured. Fruit production occurred only in those flowers subjected to cross-pollination. Among 78 crosses performed between 13 landraces, 46.15% produced fruit. Pollen tubes were observed in the pistils of the self-pollinated flowers of 38.46% of the landraces and of the cross-pollinated flowers of 85.90% of the crosses. These results show that sweet potato has a complex sexual reproductive system. Most landraces had a pollen viability rate over 80% when dyed with carmine, but below 70% when dyed with tetrazolium. However, the variation in pollen viability among the four collecting times was insignificant with either dye. All the flower variables measured showed significant differences at 5% of probability, revealing floral polymorphism. Even within-plant morphological pollen polymorphism, which is rather rare, was found. Four well-defined patterns of the relative position of stigma and highest anther were established: stigma well above the anther; stigma slightly above the anther; stigma at the same level as the anther; and stigma below the anther. However, no association was found between floral morphology and incompatibility patterns. There were successful crosses between all morphotypes.

Les Wolbachia sont des α-Protéobactéries endocellulaires transmises maternellement et qui manipulent la reproduction des Arthropodes pour augmenter leur transmission. Chez le moustique Culex pipiens, Wolbachia induit l'incompatibilité cytoplasmique (IC) qui se traduit par une forte mortalité embryonnaire lors de croisements entre individus infectés par des souches incompatibles de Wolbachia. Ce moustique se caractérise par une forte diversité génétique de ses Wolbachia (nommées wPip) et par des patrons d'IC complexes. Nous avons examiné les mécanismes qui façonnent la dynamique de cette association symbiotique aux niveaux génomique, phénotypique et populationnel. Nous avons montré que les souches wPip ont une origine génétique commune récente et qu'elles s'organisent en groupes génétiques présentant une structuration géographique. Nous avons mis en évidence des évènements de recombinaison entre souches wPip qui pourraient jouer un rôle majeur dans la diversité génétique des Wolbachia et dans l'évolution rapide des patrons d'IC. En croisant des lignées de moustiques d'origines géographiques diverses et infectées par des souches de différents groupes génétiques, nous avons montré que les IC (i) évoluent très rapidement chez Cx. pipiens; (ii) sont contrôlées par plusieurs déterminants génétiques, et (iii) qu'il y a une relation entre les patrons d'IC et les groupes génétiques des Wolbachia. Dans les populations naturelles, il apparaît que les IC sont contre sélectionnées au sein d'une population mais qu'une zone de contact entre populations infectées par des souches incompatibles peut se maintenir de façon stable. / Wolbachia are maternally inherited endocellular α-Proteobacteria that manipulate the reproduction of Arthropods to promote their own transmission. In the mosquito Culex pipiens, Wolbachia induce cytoplasmic incompatibility (CI) which results in high embryonic mortality in crosses between mosquitoes infected with incompatible Wolbachia strains. This mosquito is characterized by high genetic diversity of its Wolbachia (referred as wPip strains) and by complex CI patterns. We examined mechanisms that shape the dynamics of this symbiotic association at genomic, phenotypic and field population levels to understand how it evolves. We showed that wPip strains have a unique and recent evolutionary origin and that their diversity clusters into distinct genetic groups with a geographic structure. We revealed the existence of extensive recombinations among wPip strains, which could influence their adaptive dynamics by creating new wPip strains and thus allow the rapid emergence of new CI patterns. The analysis of crossing relationships between mosquito lines from different geographic origins and infected with wPip strains belonging to different genetic groups showed that CIs (i) evolve rapidly in Cx. pipiens; (ii) are controlled by several genetic factors, and (iii) there is a significant relationship between CI patterns and genetic divergence of wPip strains. In field populations, it appears that CIs are selected against within a population but a contact zone between populations infected by incompatible Wolbachia strains can be stably maintained.

<p>  </p> <p>Giant ragweed (<em>Ambrosia trifida</em> L.) is an annual broadleaf plant capable of emergence throughout the cropping season, opportune colonization of disturbed soil, rapid biomass accumulation, and a propensity to evolve mutations that endow resistance to herbicides, all of which contribute to giant ragweed being one of the most challenging weeds to control in row-crop production. Many soybean growers rely on acetolactate synthase (ALS)-inhibiting herbicides such as cloransulam for control of giant ragweed prior to its emergence, though the spread of biotypes resistant to ALS inhibitors can render these herbicides largely ineffective. Mesotrione inhibits the 4-hydroxyphenylpyruvate dioxygenase (HPPD) enzyme, and applications of this herbicide have recently been approved for use in mesotrione-resistant soybean varieties. Field experiments demonstrated that preemergence applications of mesotrione resulted in greater control of giant ragweed populations segregating for ALS-inhibitor resistance than several other commonly used herbicide combinations. Where mesotrione was applied, giant ragweed biomass was reduced by an average of 84% relative to the nontreated, while treatments without mesotrione increased biomass by an average of 34% by suppressing competition from other weed species. Additionally, both soil- and agar-based bioassays demonstrated that combinations of mesotrione and metribuzin can be synergistic for control of giant ragweed. </p> <p>Cloransulam was shown to result in strong selection for giant ragweed individuals with ALS-inhibitor resistance, increasing the proportion of resistant plants that emerged at one field site from 15% to greater than 90% after a single preemergence application. This selection pressure was reduced when mesotrione was co-applied with cloransulam. However, no herbicide combination, including sequential applications of non-ALS-inhibiting herbicides, consistently resulted in a resistance frequency similar to the baseline if an ALS inhibitor was applied preemergence. Resistance to cloransulam and other ALS inhibitors is expressed in giant ragweed plants possessing at least one mutant (Trp574Leu) <em>ALS</em> allele. The distribution of this allele in one field violated the Hardy-Weinberg Equilibrium, despite the fact that <em>ALS</em> is a nuclear gene and the Trp574Leu mutation does not incur a fitness penalty. We suspected that the inheritance of this mutation may be linked with a gene or genes responsible for self-incompatibility (SI) in giant ragweed, and that linkage drag was disrupting pollination in resistant plants. This research provided evidence that giant ragweed does possess SI, as greater pollen retention, pollen tube growth, and seed set were observed in cross-pollinated plants compared with self-pollinated plants. Non-Mendelian inheritance of the Trp574Leu mutation was documented in crosses between plants from three different giant ragweed populations, indicating that the mutant <em>ALS</em> allele may be linked with an SI allele common to many plants because of a shared resistant ancestor.  In crosses between plants from one population, production of resistant F1 seeds was 33% greater on average compared with the expectation under Mendelian inheritance. </p> <p>Collectively, this research demonstrated that mesotrione may become a highly effective tool for control of giant ragweed in soybean. Applications of mesotrione can also reduce the selection for an increased frequency of ALS inhibitor-resistant biotypes induced by cloransulam, though a more robust weed management strategy may be necessary to maintain the long-term viability of ALS inhibitors.  The need for sound weed management practices is underscored by the impact of the linkage of SI and <em>ALS</em> genes, which may be encouraging a more rapid spread of herbicide-resistance than was previously anticipated.</p>

No description available.

The genus Restrepia is well known to orchid enthusiasts but its micromorphology has not been described, and its pollination and breeding systems have not been investigated. The aim of this investigation was, therefore, to add to existing knowledge so that the resultant data could be used to facilitate ex situ conservation initiatives. A detailed electron microscopy study (SEM) of the floral organs was performed. This confirmed the structure of the dorsal sepal and lateral petal osmophores, their secretory nature together with that of the synsepal and the labellum. It was postulated how, by manipulating different labellar surface textures, the flower might use these ‘tactile guides’ to steer the insect (fly) through the flower. The cirrhi were postulated to help by destabilising the pollinator in flight, trapping it and bringing about pollination. The papillate structure of the calli was established and their optical properties investigated. Media comparison investigations established that Western medium supported the highest germination rates and, with the addition of banana supplement, the highest rates for seedling growth and development. This represented the first protocol for axenic germination of Restrepia in the literature (Millner et al., 2008) and provided a tested methodology for investigating breeding systems and producing Restrepia plant material for both scientific and horticultural purposes. Self-pollinations were found to produce fewer embryos compared to cross-pollinations. The operation of self-incompatibility (SI) was confirmed by the study of pollen tube growth which further confirmed the time interval between pollination and fertilisation. A time line from pollination/fertilisation to flowering was established. The type of SI in operation was best explained by gametophytic incompatibility. This demonstrated that it was possible to raise Restrepia hybrids and species from seed, by performing intraspecific crosses so helping to preserve them for posterity and relieve pressure on wild populations. Narrow endemic Restrepia species face combined threats from habitat loss, habitat degradation and problems of viable seed production due to the effects of SI and inbreeding depression (ID). Recently developed online resources, such as GeoCAT, were used to perform a Red List assessment in order to identify the degree of threat individual species faced, both globally and nationally. All species were classified as facing substantial levels of threat; although this was lessened for populations in protected habitats. Conservation is needed for cultivated collections as well as these wild populations by keeping alive existing knowledge and expertise in growing these species.

The research presented in this dissertation integrates data and theory to examine three important topics in biology. In the first chapter, I investigate genetic variation at two loci involved in a genetic incompatibility in the genus Xiphophorus. In this genus, hybrids develop a fatal melanoma due to the interaction of an oncogene and its repressor. Using the genetic variation data from each locus, I fit evolutionary models to test for coevolution between the oncogene and the repressor. The results of this study suggest that the evolutionary trajectory of a microsatellite element in the proximal promoter of the repressor locus is affected by the presence of the oncogene. This study significantly advances our understanding of how loci involved in both a genetic incompatibility and a genetically determined cancer evolve. Chapter two addresses the role polyploidy, or whole genome duplication, has played in generating flowering plant diversity. The question of whether polyploidy events facilitate diversification has received considerable attention among plant and evolutionary biologists. To address this question, I estimated the speciation and genome duplication rates for 60 genera of flowering plants. The results suggest that diploids, as opposed to polyploids, generate more species diversity. This study represents the broadest comparative analysis to date of the effect of polyploidy on flowering plant diversity. In the final chapter, I develop a computational method for designing disease surveillance networks. The method is a data-driven, geographic optimization of surveillance sites. Networks constructed using this method are predicted to significantly outperform existing networks, in terms of information quality, efficiency, and robustness. This work involved the coordinated efforts of researchers in biology, epidemiology, and operations research with public health decision makers. Together, the results of this dissertation demonstrate the utility of applying quantitative theory and statistical methods to data in order to address complex, biological processes. / text

Depuis les années 1950, l’intensification des pratiques agricoles concourt à une augmentation de la pression anthropique entraînant une raréfaction des espèces spécialistes des parcelles cultivées, ce qui contribue à l’érosion de la diversité biologique des agroécosystèmes. Parmi les espèces en déclin, on compte de nombreuses messicoles dont le bleuet (Centaurea cyanus L.), fleur emblématique des moissons. Or, cette espèce peut rendre des services écosystémiques comme hôte de prédateurs de ravageurs des cultures et ressource privilégiée de certains pollinisateurs qui justifieraient son maintien dans les champs. Nous étudions ici des facteurs biologiques qui pourraient potentiellement contribuer à son déclin : distribution spatiale, potentialité de croissance, diversité génétique des populations, survie des semences, système de reproduction. Nous avons montré, par deux campagnes de relevés, que C. cyanus n’est pas une espèce indicatrice de diversité floristique dans la parcelle cultivée. Toutefois, lorsque que le bleuet est rare dans une région, il est associé à d’autres messicoles peu fréquentes. Par contre, s’il est commun, il est associé aux zones ayant la plus forte diversité végétale. Une expérience de semis dans différents compartiments de l’agrosystème, en absence de traitements herbicides, a mis en évidence que la potentialité de croissance des bleuets est plus élevée dans le plein champ du blé que dans la moutarde et les interfaces blé/bordure. Cette croissance est limitée de manière variable par les communautés adventices présentes dans les compartiments hors champs (bordures). L’analyse de la diversité génétique à l’aide de marqueurs microsatellites de bleuets dans une petite zone agricole montre que les populations sont connectées par des flux de gènes importants. Les barrières écologiques telles que les chemins, semblent être des facteurs de structuration plus importants que la distance géographique séparant les populations. La répartition du bleuet dans le paysage agricole n’est donc pas aléatoire et apparaît dépendante de la fréquence de l’espèce dans la région ainsi que des différents éléments du paysage. Les caractéristiques du cycle biologique du bleuet ont été étudiées grâce à des expériences au champ et en serre. Nous avons montré que la longévité des akènes enfouis dans le sol chute rapidement après deux ans. Le cycle saisonnier de la dormance permet deux cohortes de levées (automne et printemps). L’étude du système de reproduction a permis de mettre en évidence que les pollinisateurs sont nécessaires pour la fécondation et que les populations sont majoritairement auto-incompatibles. Il existe cependant des individus pseudo auto-incompatibles mais leur fréquence n’est pas liée à la taille des populations ou à leur niveau d’isolement spatial. Par ailleurs, dès que le coefficient de consanguinité augmente dans les populations, la valeur phénotypique des individus baisse. La dépression de consanguinité s’exprime alors principalement pendant la phase de la germination. L’ensemble de ces caractéristiques biologiques peuvent s’avérer défavorables au bleuet dans les agroécosystèmes simplifiés actuels et ont pu entraîner son déclin dans certaines régions / Agroecosystems are currently experiencing high biodiversity loss, in particular among the plant species specifically adapted to this habitat. This decline results from cropping systems that have been intensified in Western Europe since the 1950s. The cornflower (Centaurea cyanus L.), considered as emblem of the flora associated with traditional cereals, appears as a species that may be at risk and should be monitored. Indeed, cornflower can serve as a host to predators of crop pests and is strongly attractive for the pollinators. This study examines the biological factors that could potentially cause the decline of this species: spatial distribution, potential for growth, population’s genetic diversity, seeds survival and mating system.We have shown that C. cyanus is not a biological indicator of the weed diversity of a field. However, when it is not frequent in a region, cornflower is associated with other rare segetal species. When it is common in a region, C. cyanus is present in areas with higher weed diversity. Sowing seeds in different components of the agrosystem, without herbicide application, shows that the growth of cornflower was higher in the centre of a wheat field than in mustard and the crop edge. Growth success in field margins was variably limited according to the weed communities in the field boundaries. The analysis of the genetic diversity using microarrays has shown that populations in a small agricultural area are connected by important genes flow. The ecological barriers as country road seem to be factors more determinant than geographical distance to structure and divide populations. Therefore, the cornflower distribution in the agricultural landscape is not random and appears to be linked to species frequency in the area and to landscape elements.The cornflower life cycle characteristics were studied through field and greenhouse experiments. We have shown that the longevity of achenes buried in the soil falls rapidly after two years. There is a seasonal dormancy cycle that allows the emergence of two seedling cohorts (fall and spring). Our study of the mating system highlights the fact that pollinators are required for fertilization and that cornflower is strongly self-incompatible. However, there are pseudo self-incompatible individuals, but their frequency is not related to the population size or the spatial isolation level. Individual fitness decreases as the inbreeding coefficient increases in a population. Inbreeding depression is mainly expressed during the germination stage. All these biological characteristics can be detrimental to the cornflower in current simplified agroecosystems and may have led to cornflower decline in some areas

Les plasmides pSci de Spiroplasma citri GII3 possèdent une structure mosaïque avec de nombreuses régions conservées. Des dérivés du plasmide pSci2 ont été produits par délétions successives et leur capacité de réplication a été évaluée. Le plus petit réplicon obtenu ne contient plus qu’une CDS (pE) et ses régions flanquantes. L’inactivation dans un vecteur navette du gène pE est suffisante pour abolir la réplication de ce plasmide dans S. citri. Des dérivés des pSci ont été introduits efficacement dans S. kunkelii et S. phoeniceum, deux spiroplasmes phytopathogènes pour lesquels aucun outil génétique n’était disponible jusqu’à présent. La stabilité des dérivés des pSci a également été évaluée par leur capacité à persister en l’absence de pression de sélection. L’instabilité ségrégationnelle des plasmides dans lesquels soj a été délété ou inactivé indique que la protéine de partition Soj/ParA est essentielle au maintien des plasmides pSci. L’incompatibilité sélective entre un plasmide pSci et ses dérivés a été exploitée pour produire une collection de souches possédant des profils plasmidiques différents. L’analyse des phénotypes d’acquisition et de transmission de plusieurs de ces mutants suggère que la CDS traG portée par le pSci6 est essentielle à la transmission de S. citri GII3. En revanche, les plasmides pSci1-5, codant les protéines adhésine-like ScARPs, ne sont indispensables ni à l’acquisition ni à la transmission. Dans le cadre de ce travail, plusieurs outils génétiques ont été adaptés aux mollicutes. Le promoteur Pxyl/tetO2 a été utilisé pour contrôler l’expression du gène de la spiraline chez S. citri et M. agalactiae. Enfin, un système de linéarisation de plasmide in vivo basé sur l’expression de l’endonucléase I-SceI a été utilisé pour l’élimination de plasmides pSci chez S. citri. / Plasmids pSci from Spiroplasma citri GII3 display a mosaic gene organization with highly conserved regions. Through successive deletions, various pSci2 derivatives were constructed and assessed for their ability to replicate. The smallest functional replicon consists of one single CDS (pE) and its flanking, intergenic regions. Furthermore, shuttle (S. citri/E. coli) plasmids, in which the pE gene was disrupted, failed to replicate in S. citri, suggesting that PE is the replication protein. S. citri plasmids were efficiently introduced into S. kunkelii and S. phoeniceum, two plant pathogenic spiroplasmas, the transformation of which had never been described before. Studying stability of various pSci-derived plasmids in the absence of selection pressure strongly suggests the occurrence of an active partition system involving the soj-like gene. Selective incompatibility between a given pSci plasmid and its derivatives was used to remove plasmids from the wild-type strain. As a result, a collection of S. citri GII3 mutants differing in their plasmid contents was produced. Experimental transmission of these mutants through injection to or ingestion by the leafhopper vector will provide new insights into the role of plasmid encoded determinants in the biology of S. citri. First data indicated that pSci6 traG is required for insect transmission of S. citri GII3. In contrast, pSci1-5, encoding adhesin-like proteins, are not essential for both transmission and acquisition. In the frame of this work, new genetic tools have been adapted for use in mollicutes. The tetracycline inducible promoter, Pxyl/tetO2, was used to control spiraline gene in S. citri and M. agalactiae. Also, an in vivo linearization system based on the expression of the I-SceI-endonuclease was used to remove pSci plasmids from S. citri.

This PhD. thesis is dedicated to the legal regulation of instruments presenting guarantees of the independence of the member of parliament during the performance of his/her functions. The aim of this thesis is in particular to present a comprehensive picture of the topic concerning protection of independent performance of the parliamentary mandate, evaluate the Czech legal regulation in force and, last but not the least, to find an answer to the question to what extent is the Czech legal regulation in force complete and effective as well as present possible alternatives de lege ferenda. Member of parliament must be independent with regard to all aspects of the exercise of his/her mandate. Independency of the member of parliament is necessary precondition for his/her ability to exercise the mandate in such a way, which will enable him/her to face external pressures, not only from the state bodies, but also from other subjects or phenomenons. Instruments protecting independent performance of the mandate also support the possibility of members of parliament to fulfil their role freely and as a result to constitute working parliament. Presented PhD. thesis deals with the topic of parliamentary immunity, particular focus is put on the part presenting protection of freedom of voting and freedom of...