Cyclotides are plant proteins with a unique topology, defined as the cyclic cystine knot motif. The motif endows cyclotides with exceptional chemical and biological stability. They also exhibit a wide range of biological activities including insecticidal, cytotoxic, anti-HIV and antimicrobial effects. Hence, cyclotides have become potential candidates in the development of peptide-based drugs; either as scaffolds to stabilize susceptible peptide sequences or as drugs by their own right. In this thesis, important parameters that could be inputs toward this development have been tuned. An extraction protocol that can be extended to industrial scale production of the cyclotides from natural sources was developed; accordingly, a single maceration with hydroalcoholic solutions of medium polarity represented an optimum extraction method. Moreover, it was shown that investigating the cyclotide content of cyclotide-bearing plants from diverse environments is a promising approach for extending the knowledge of both structural and biological diversity of these proteins. Five novel cyclotides with new sequence diversity were isolated and characterized from a violet that grows on Ethiopian highlands at an altitude of 3400 m. One of the areas where the cyclotide framework has attracted interest is the development of stable antimicrobial peptides. A stability study was carried out to determine the stability of the cyclotide framework in a cocktail of bacterial proteases and serum where the native forms of tested cyclotides exhibited high stability profile. Understanding the modes of cyclotide-cell interaction is certainly an important factor for the potential development of cyclotide-based drugs. Cellular studies were carried out using the comet assay and microautoradiography. A bell-shaped dose response curve was obtained for the DNA damaging effect of the cyclotides in the comet assay, which was the first toxicological assay of its kind on this class of proteins. The microautoradiography study revealed that the cyclotides penetrate into the cells even at cytotoxic concentrations. From previous reports, it was known that the cyclotides interact with membranes; the cellular studies in this thesis added to this knowledge by clearly demonstrating that these proteins have multiple modes of action.

Un dispositif agronomique pérennisé entre 1995 et 1999 avec des systèmes de culture de maïs à couvertures végétales a été installé dans la zone écologique à forêt semi-décidue de Côte d'Ivoire. L'offre en N d'origine organique est étudiée dans le cadre de systèmes de culture de maïs, avec une durée de jachère de 6 mois (SC6MJ) et 18 mois (SC18MJ). Les jachères sont occupées soit par Chromolaena odorata (subspontanée), soit par Pueraria phaseoloides (introduite). Ces quatre systèmes sont comparés à deux témoins, un sans Chromolaena pour les SC6MJ et un avec brûlis de Chromolaena pour les SC18MJ. Les systèmes à couvertures végétales sont gérés en couvertures vivantes avec des herbicides et leur pérennité est assurée par une reprise de croissance des espèces avant la récolte de la céréale. Les systèmes à SC6MJ bénéficient d'une fertilisation modérée en phosphore. L'étude réalisée pose comme hypothèses de départ que (i) les déterminants de l'offre à court terme (cycle cultural) et à moyen terme (effets cumulés) dépendront principalement des quantités et de la qualité des litières de surface d'origine des jachères (ii) ces effets se concentreront à la surface du sol. Le potentiel de minéralisation des litières au laboratoire est corrélé avec leur rapport C/N, plus faible pour Pueraria (C/N entre 20 à 23) que pour Chromolaena (C/N entre 25 à 43). En 1998 la vitesse de disparition des litières suivie avec des " sacs de décomposition " est beaucoup plus rapide qu'en 1999 en relation au début du cycle de décomposition avec le régime pluviométrique. Les deux années la quantité de litières présentes après la coupe détermine le taux de décomposition à moyen terme des litières. Ces dynamiques, modélisées sur deux années, permettent de distinguer un compartiment de litières à disparition rapide et un compartiment à disparition lente. La comparaison des pertes en C dans des " sacs de décomposition " à maille différente montre que l'activité de la mésofaune et de la macrofaune du sol pourrait être responsable de la disparition de 50 % des litières au début du processus de décomposition et de 35% à la fin du processus de décomposition. Leur action de fragmentation et d'incorporation au sol des litières (meules des termites champignonnistes situées à la surface du sol) permet une disparition comparativement plus rapide des litières de Chromolaena que celles de Pueraria. L'application des modèles aux données obtenues sur les mobilisations en N des jachères en relation avec les besoins de la culture en N indique (i) un risque " d'asynchronie " en début de cycle (ii) une offre insuffisante en fin de cycle pour les SC6MJ (iii) peu de différenciation entre l'offre de Chromolaena et celle de Pueraria. L'offre en N in situ du sol en surface (0-10 cm) en 1998 et 1999, estimée pendant des périodes de 90 jours durant le cycle du maïs par des mesures et des incubations de sol, est corrélée pour les SC6MJ avec les mobilisations en N du maïs dans les parties aériennes. C'est le traitement avec Pueraria qui les deux années satisfait le mieux les besoins en N de la culture et cette offre est comparable à celle des SC18MJ. L'offre en N varie peu selon les traitements des SC18MJ. Le régime pluviométrique intervient (i) à une échelle pluriannuelle, en déterminant, l'intensité du pic de minéralisation en N minéral du sol et les accumulations en N dans les parties aériennes des jachères (ii) à l'échelle de la campagne en interaction avec les litières en surface et l'activité des systèmes racinaires des couvertures végétales. La litière de Pueraria semble mieux conserver l'humidité du sol et le système racinaire de Chromolaena mobiliser de plus fortes quantités d'eau. En situations hydriques limitantes, la minéralisation nette est corrélée aux humidités de sol. Les litières semblent être à l'origine en 1998 de l'augmentation de la biomasse microbienne du sol en surface créant ainsi une source potentielle d'immobilisation temporaire de N. En 1998 l'offre en N estimée par minéralisation nette indique une meilleure synchronie avec les besoins de la culture malgré une pluviométrie plus abondante en début de cycle par rapport en 1999. Une étude utilisant l'abondance naturelle 15N permet d'estimer que la litière de Pueraria peut contribuer fortement à la nutrition azotée de la culture, de 30 à 41% en 1998 et de 64 à 87% en 1999 dans le cadre du système de culture à jachère de 6 mois. Les différenciations sur les matières organiques du sol (MOS) indiquent un début d'effet sur les stocks évalués sur 0-10 cm en moyenne à 2.34 t ha-1 N pour les SC6MJ et en moyenne à 2.83 t ha-1 N pour les SC18MJ. Pour les SC6MJ la différence obtenue entre les apports et les pertes sur quatre années suggère que Chromolaena conserve mieux les stocks en C et N que Pueraria. La comparaison avec une jachère forestière située hors dispositif indique que la disponibilité en matériaux facilement minéralisables dans nos agrosystèmes est (i) réduite en fin de saison des pluies (ii) diminuée par la pratique du brûlis (iii) peu augmentée par la pratique de la jachère. Les études sur des incubations de longue durée (106 et 85 jours) montrent des effets significatifs sur les modélisations faites par ajustement aux points expérimentaux sur les cinétiques de respiration et de minéralisation nette cumulées (exprimées en proportion par rapport au C et N total du sol). Elles permettent d'identifier d'importants facteurs de différenciation au niveau du compartiment des MOS à taux de renouvellement élevé. Pour les SC6JM ce compartiment représente une part plus importante avec Pueraria, certainement en relation directe avec des apports cumulés de plus grande qualité biochimique (rapport lignine / N). La durée de la jachère n'améliore pas la disponibilité en N facilement minéralisable indiquant ainsi un blocage de C et N au niveau du sol de nature physique et/ou biochimique.

The growing problem of antibiotic resistance and the lack of promising prospective antibiotics have forced us to search for new classes of antibiotics. Among the candidates to develop into future antibacterials are antimicrobial peptides (AMPs). These potent, broad spectrum compounds are important components of innate immunity of organism from all kingdoms of life. One such family of mini-proteins from plants is called cyclotides, whose members are defines by cyclic backbone and a cystine knot (CCK), which confers to them extreme stability in the face of biological, chemical and physical insults.     Some cyclotides possess Gram-negative specific antibacterial activity; the purpose of this thesis was to characterize how these molecules kill bacteria, and how bacteria would respond to treatment with cyclotides. For this purpose, Salmonella enterica and Escherichia coli mutants resistant to the cyclotides cycloviolacin O2 and cycloviolacin O19, respectively, were selected. These mutants were characterized by whole genome sequencing, genetic reconstitution, fitness measurements, and cross-resistance studies. These studies identified a number of genetic pathways for resistance development to cyclotides. These mutants displayed variable fitness profiles in laboratory growth media and in mice competition experiments, with some mutants possessing a fitness advantage in mice. Cross-resistance studies resulted in the identification of several cases of cross-resistance and collateral sensitivity between cyclotides and other AMPs/antibiotics.      Antimicrobial effects of cyclotides were assayed in different conditions and in bacterial organisms with different surface characteristics. In addition, immunolocalization experiments were performed to explore the biological distribution of cyclotides in plants and to determine the mechanism of action of cyclotides in bacteria, respectively. Antibodies raised against cyO2 were used for this purpose. Immunohistochemical techniques applied to plant cells, tissues and organs provided the information that cyclotides were distributed in all plant organs, and were found in tissues vulnerable to pathogen attack, and that cyclotides were stored in the vacuoles of plant cells. Immunogold staining of cyclotide treated cells of S. typhimurium, showed effects of cyclotide treatment on the cell envelope components as well as cytoplasm. A higher number of cyclotide molecules was associated with the cell envelope, but a considerable fraction of them penetrated into the cytoplasm.

Submitted by Irvana Coutinho (irvana@ufpa.br) on 2015-10-14T12:58:54Z No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_IsolamentoMicrossatelitesEspecies.pdf: 871091 bytes, checksum: fb70b27bf298c90182fbfacb17aa112b (MD5) / Approved for entry into archive by Ana Rosa Silva (arosa@ufpa.br) on 2015-10-15T13:46:18Z (GMT) No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_IsolamentoMicrossatelitesEspecies.pdf: 871091 bytes, checksum: fb70b27bf298c90182fbfacb17aa112b (MD5) / Made available in DSpace on 2015-10-15T13:46:18Z (GMT). No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_IsolamentoMicrossatelitesEspecies.pdf: 871091 bytes, checksum: fb70b27bf298c90182fbfacb17aa112b (MD5) Previous issue date: 2004 / Aborda isolamento de microssatélites de espécies madeireiras no contexto da sustentabilidade genética no manejo florestal com o objetivo de isolar e caracterizar uma bateria de marcadores microssatélites para espécies arbóreas da Floresta Amazônica: Jacaranda copaia, Bagassa guianensis e Dipteryx odorata. Estes estudos vem sendo realizados como parte do projeto Dendrogene, e as análises genéticas populacionais posteriores fornecerão subsídios para conservação e manejo sustentável destas espécies.

Ce travail porte sur la dispersion des graines par les fourmis et se divise en deux parties : l’une expérimentale (Chapitres 1, 2 & 3) et l’autre synthétique (Chapitres 4 & 5). L’approche expérimentale a consisté en une exploration en conditions de laboratoire des comportements des fourmis à deux étapes du processus de myrmécochorie: à la source de graines et dans le nid. Des graines des plantes myrmécochores Viola odorata et Chelidonium majus ont été présentées aux fourmis Lasius niger et Myrmica rubra. Chaque étape de la séquence myrmécochorique a généré une variabilité des comportements propre à chacun des quatre couples fourmis-graines. L’élaiosome n’attire pas les fourmis à distance. Les fourmis suivent toujours la même séquence de comportements : antennation, manipulation et prise des graines. Le nombre d’antennations et de manipulations avant la prise de graines peut être considéré comme un indicateur de l’« hésitation » des fourmis à prendre les graines. L’espèce à tendance carnivore Myrmica rubra a été plus rapide et efficace dans la prise de graines que l’espèce éleveuse de pucerons Lasius niger. Parallèlement, les fourmis ont moins antenné, moins manipulé et plus pris de graines de Chelidonium majus, ce qui montre un intérêt particulier pour cette espèce. Un jour après l’expérience, toutes les graines des deux espèces se trouvaient dans les déchets à l’extérieur du nid, avec au moins la moitié des élaiosomes consommés (Chapitre 1). Lors du passage des graines à l’intérieur du nid, les fourmis Myrmica rubra ont également montré une capacité à traiter les graines rapidement, en montrant une dynamique d’arrachage de l’élaiosome et de rejet des graines hors du nid plus rapide. Le taux d’arrachage de l’élaiosome a été influencé par l’espèce de graine, plus important pour les graines de Chelidonium majus. Nous avons montré qu’une proportion variable de graines rapportées au nid (moins de la moitié) étaient déposées directement au contact des larves, les autres étant traitées ailleurs par les ouvrières ou laissées temporairement à l’abandon dans le nid. Par ailleurs, les dynamiques de rejet des items hors du nid ont curieusement été peu influencées par l’espèce de graine. Pour une graine, le fait de ne plus avoir d’élaiosome diminue le nombre moyen d’ouvrières qui la contacte simultanément. Parallèlement, même si la réponse n’est pas de type « tout ou rien », l’absence d’élaiosome accroit aussi la probabilité qu’une graine a d’être rejetée. (Chapitre 2). Nous avons isolé expérimentalement le paramètre de dessiccation des graines afin mesurer son influence sur le taux de prises. La dessiccation progressive des graines réduit les taux de prises par les fourmis Myrmica rubra. La réhydratation des mêmes graines leur permet de retrouver une attractivité et donc une valeur fonctionnelle prolongée. Les graines de Viola odorata perdent presque toute attractivité après 4 jours de dessiccation et leur réhydratation ne rétabli que faiblement leur attractivité. A l’inverse, les graines de Chelidonium majus gardent un tiers de leur attractivité après un mois de dessiccation et leur réhydratation restaure presque entièrement leur attractivité (Chapitre 3). La synthèse bibliographique a permis de dresser un aperçu des principales caractéristiques des fourmis dispersatrices de graines myrmécochores. Certains traits « généralistes » rendent les rencontres entre fourmis et graines très probables : leur ubiquité et diversité taxonomique, leur régime alimentaire omnivore, et leur fourragement « diffus » et opportuniste. Les fourmis possèdent des traits qui les rendent uniques par rapport aux autres insectes : le fourragement au sol, la capacité à transporter de la nourriture, ainsi que la nidification. Certains traits des fourmis ont une influence considérable sur la dispersion des graines : leur taille, les préférences de régime alimentaire, la phénologie, la capacité d’apprentissage et la fréquence de déménagement des nids. Nous développons également l’hypothèse que la rapidité et l’efficacité du traitement des graines par les fourmis seraient une conséquence d’un comportement hygiénique des fourmis à tendance carnivore, habituées à gérer des proies périssables (Chapitre 4). Nous avons dressé pour la première fois une liste des espèces de plantes myrmécochores et potentiellement myrmécochores des régions d’Europe tempérée (260 spp.). Nous montrons que ces dernières sont majoritairement herbacées, et ont tendance à fleurir plus précocement que les autres espèces. La proportion de graines myrmécochores comprises entre 1 et 3 mm et entre 0,6 et 10 mg est plus importante que dans le reste de la flore (Chapitre 5). / This study concerns seed dispersal by ants and is divided in two parts: one experimental (chapters 1, 2 & 3), and one synthetic (chapters 4 & 5). Experimental work consisted in a series of laboratory experiments, in which ant behaviour was studied at two stages of the dispersal process: at the seed source and inside the nest. Seeds of the myrmecochorous plants Viola odorata and Chelidonium majus were presented to two ant species: Lasius niger and Myrmica rubra. Each stage of the myrmecochory sequence generated a variability of behaviours for each of the four ant-seed pairs. The elaiosomedo not attract seed at a distance. Ants followed always the same behavioural sequence: antennations, manipulations, and removal. The number of antennations and manipulations before removal can be considered as a measure of ants’ “hesitation” to remove seeds. The carnivorous species Myrmica rubra was faster and more efficient at taking seeds than the aphid-tending Lasius niger. At the same time, ants antennated and manipulated less Chelidonium majus seeds, which shows a particular interest for this seed species. One day after the experiment, all seeds of both species were located outside the nest in the refuse piles. At least half of their elaiosomes had been consumed (chapter 1). Inside the nest, Myrmica rubra also showed a great ability to treat seeds quickly, i.e. quicker dynamics of elaiosome removal and seed rejection outside the nest. Elaiosome removal rates were influenced by seed species (higher for Chelidonium majus). We showed that a variable proportion of seeds (less than half) was directly deposited in contact with larvae. The rest of the seeds were handled elsewhere by workers, or left temporarily unattended in the nest. Dynamics of seed rejection outside the nest were curiously little influenced by the seed species. For a seed, to loose the elaiosome decreased the number of workers manipulating it. At the same time, even if the rejection response is not automatic, the absence of elaiosome increases the probability for a seed to be rejected (Chapter 2). We isolated the desiccation parameter in order to measure its influence on seed removal rates. Progressive seed desiccation reduced Myrmica rubra removal rates. Rehydration of the same seeds restored their attractiveness, thereby prolonging their functional life. Viola odorata seeds lost almost all their attractiveness after 4 days of desiccation, and rehydration only restored a reduced part of their attractiveness. On the contrary, Chelidonium majus seeds kept one third of their attractiveness after one month of desiccation, and recovered almost all their attractiveness after rehydration (Chapter 3). The bibliographic review allowed us to compile an outline of the main features of seed-disperser ants. Some generalist features highly increase the probability that ants encounter seeds: their ubiquity and taxonomic diversity, omnivorous diet and their opportunistic “diffuse” foraging. Among insects, ants have unique traits that make them broad dispsersers: ground foraging, the ability to transport items, and nesting behaviour. Some other traits have a great influence on the seed dispersal system: the ant body size, their diet preferences, the phenology of the colony, the learning, and the frequence of nest relocation. We also develop the hypothesis according to which, handling efficiency of ants is a byproduct of hygienic behaviour of carnivorous oriented species, since they are used to manage perishable preys (Chapter 4). We compiled the first list of myrmecochorous and potentially myrmecochorous plants species of European temperate regions (260 spp.). We show that these plants are mainly herbaceous. They also tend to flower earlier than the whole flora. The proportions of myrmecochorous seeds having a size range between 1 and 3 mm, and a weight range between 0.6 and 10 mg are higher than in the rest of the flora (Chapter 5).

Chromolaena odorata (L.) King and Robinson (chromolaena, triffid weed) (Asteraceae: Eupatorieae) is one of the most problematic weeds in the subtropical northeastern parts of South Africa. Calycomyza eupatorivora Spencer (Diptera: Agromyzidae) was introduced as a biological control agent for the control of this weed. No study has yet been done to quantify field populations of C. eupatorivora since its establishment in 2003. The aim of this study was therefore to measure aspects of the field population and laboratory behaviour of C. eupatorivora on C. odorata. The first objective was to determine the percentage leaf area mined by larvae of C. eupatorivora on C. odorata plants exposed to three densities of mated flies, and also to determine the number of mines produced by these different densities, and their distribution on the plant. It also attempts to determine the relationship between chromolaena leaf quality and usage by C. eupatorivora. The maximum percentage of leaf area damaged was 37.5% for one of the trials involving five pairs of flies. Mean percentage leaf area damaged was slightly higher with five (28.5%) than ten pairs (22.0%) of adults and was lowest with one pair (6.5%), but these differences were not significant. In relation to the mean number of mines per plant, five and ten pairs of flies caused slightly more mines than one pair. The other significantly different parameter was number of leaves mined per plant, which was higher for five pairs. Within a plant, C. eupatorivora probably selects a subset of leaves with certain chemical and physical characteristics for oviposition since certain leaves were left unmined while others received multiple eggs. Percentage water content did not differ between mined and unmined leaves, but clear patterns were shown by acid detergent lignin which was higher in unmined leaves and nonstructural carbohydrates which were much higher in mined leaves. It is likely that leaf age plays a role in its suitability. The second objective was to quantify C. eupatorivora infestation levels, by counting and examining larval leaf mines, on C. odorata in the field at four times ('seasons' - September, December, March and July) over a 12-month period, and at three study sites that each included two habitats, viz. open and shady. At each of these six sampling sites, line transects were laid out and plants/branches sampled along them. Both plant/branch height and the number of leaves increased between September and March, and plants in the open habitats were taller and had more leaves than those in the shaded habitats. At the third site, the shady habitat supported taller plants with more leaves compared to the same habitat at the other sites. There was a steep increase in the number of C. eupatorivora mines from December to March. The mean number of mines, both total and in relation to leaves available, was highest in March, and was higher in the shaded habitats compared to the open habitats. The mean number of mines per damaged leaf was slightly higher in December compared to the other seasons, and was also higher in the open than the shaded habitats. Mean larval mortality was high (70%) in September but decreased to 32% in December, and increased again in late summer. The overall levels of mining by C. eupatorivora were low, with less than 5% of leaves sampled having mines. Taken together, the laboratory and field trials suggest that C. eupatorivora is restricted to a subset of the leaves of C. odorata for its development; that the field population is unable to make full use of the resource of young, palatable leaves that develop in early- to mid-summer because it only becomes large in late summer; and that the high mortality rate of young larvae negatively affects both the population of the fly and the level of damage to the plant. Given that these results were obtained in an area where the population of C. eupatorivora is relatively high, it is unlikely that the fly is having anything more than a negligible effect on C. odorata in South Africa at present. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.

The objective of this study was to describe the impacts of the density of Chromolaena odorata (chromolaena) on species composition in coastal grasslands and to investigate serial changes in the vegetation following the implementation of a burning programme. The thesis deals with key ecological concepts and issues, so a comprehensive literature review is included. Chromolaena invades coastal grasslands that are not burnt regularly (i.e. biennially). Grasslands that were not burnt for 30 years were seral to secondary forest. The successional pathway from open grassland to closed canopy forest varied according to soil type. Coastal grasslands on Glenrosa soils were characterised by savanna at an intermediate stage between the grassland and forest states. Shading ended the persistence of savanna species (e.g. Combretum molle, Dichrostachys cinerea and Heteropyxis natalensis) in forest, whereas forest precursors (e.g. Canthium inerme, Maytenus undata and Protorhus longifolia) only established where fire was absent. Chromolaena infestations were characterised by multi-stemmed adult plants of variable height (i.e. 1-3 m), depending on soil type. Regic sands did not support stratified woody vegetation and chromolaena infestations were self-supporting, reaching a maximum height of 1.5 m. Glenrosa soils supported tree communities and chromolaena reached more than 3 m in places. The density of chromolaena affected species composition in grasslands with moderate to dense stands (> 5 adult plants m ¯² or >50000 shrubs ha ¯¹). Chromolaena stands became monospecific when the number of adult plants exceeded 7 m ¯². Succession to forest also ceased once chromolaena became thicket-forming. Fire-induced mortality of the chromolaena depended on grass fuel loads. Grass cover of 30% (c. 1 000 kg ha ¯¹) was required to achieve 80% mortality of the parent infestation after the initial burn. Dense infestations could only be killed by running head-fires from adjacent grasslands into thickets. Under conditions where head-fires could not be used, infestations were slashed and burnt at the height of the dry season (July to August) to achieve an 80% kill rate. Seedlings were killed (99%) by annual burning in sparse (≤ 10000 shrubs ha ¯¹) to moderate < 50 000 shrubs ha ¯¹) infestations. The suppression of chromolaena and other alien species, establishing on bare ground after clearing dense infestations, required chemical control until grass cover was sufficient (i.e. 1 000 kg ha ¯¹) to effect uniform burning. Certain secondary alien invaders (e.g. Lantana camara, Psidium guajava and Solanum mauritianum) persisted by coppicing profusely after fire and herbicides need to be integrated into burning programmes when these species occur. Grasslands on regic sands (e.g. Ischaemum fasciculatum, Panicum dregeanum and Themeda triandra) were more resilient to the modifying effects of woody vegetation, than grasslands on Glenrosa soils. Grasslands on Glenrosa soils did not revert to an open state but persisted as ruderal savanna grassland (e.g. Eragrostis curvula, Hyparrhenia tamba and Cymbopogon validus) once fire-resistant tree species (e.g. Combretum molle and Heteropyxis natalensis) had established. Depending the objectives for land management and the vegetation's condition, coastal grasslands can be rehabilitated and managed in multiple states, i.e. grassland, savanna or forest communities. A state-and-transition model based on the empirical data recorded in the study is presented and shows chromolaena altering vegetation states from open grassland to chromolaena dominated thicket. The model illustrates chromolaena thickets as the dominant phase of a moist coastal forest/savanna succession, irrespective of soil type, in absence of appropriate land management practices (e.g. control burning and integrated control of alien vegetation). This model should aid in planning strategies for the control of chromolaena in subtropical grasslands in South Africa. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2000.

Les premières souches bactériennes résistantes aux antibiotiques sont connues depuis 70 ans et se sont multipliées ces dernières années posant un grave problème de santé publique. Parmi les nombreux types d'infections induites par ces bactéries, nous nous sommes intéressés aux infections intestinales qui peuvent dégénérer en maladies inflammatoires de l'intestin et cancers. Notre travail de thèse a consisté à proposer des outils thérapeutiques dans le traitement des pathologies intestinales infectieuses : des peptides antimicrobiens dérivés de la chromogranine A et des extraits de plantes de la médecine traditionnelle béninoise. La chromogranine A est une protéine libérée par les cellules nerveuses, neuroendocrines et immunitaires au cours d'un stress et maturée en peptides. Des peptides actifs contre quatre souches bactériennes pathogènes (Klebsiella oxytoca, Salmonella enterica, Shigella sonnei et Vibrio cholera non O1) ont été identifiés et l'interaction bactérie-peptide analysée. L'étude de la combinaison peptide-antibiotique montre que la cateslytine permet de réduire les doses d'antibiotiques nécessaires. Ensuite, nous avons étudié l'implication de deux peptides sur un modèle de cellules neuroendocrines, les cellules BON. La chromofungine provoque la stimulation des cellules BON en induisant un influx de calcium extracellulaire, tandis que la catestatine est capable de bloquer l'activité de la chromofungine.Après un screening des extraits de 14 plantes du Bénin, nous avons isolé deux molécules, la sinensétine et l'O-tétraméthyléther scutellaréine, responsables de l'activité antibactérienne de Chromolaena odorata contre les pathogènes étudiés.

Les plantes aromatiques font parties des richesses naturelles du patrimoine de la région Midi-Pyrénées mais apparaissent cependant de moins en moins valorisées. Dans le cadre de la recherche de nouveaux aromatisants naturels de type WONF, nous avons choisi d'adopter la voie de valorisation des plantes aromatiques médiévales de cette région car elles ont toujours fait l'objet de cueillette à l'état sauvage sans jamais être cultivées. Dans une première étape, une étude bibliographique a été réalisée, à partir d'anciens ouvrages botaniques et de bases de données scientifiques, afin d'identifier les plantes régionales traditionnellement utilisées par nos ancêtres pour leurs activités aromatisantes, culinaires ou médicinales. Nous avons sélectionné cinq plantes aromatiques : Achillea millefolium, Calamintha grandiflora, Tanacetum balsamita, Myrrhis odorata et Monarda didyma. Dans une deuxième étape, des études physico-chimiques, sensorielles et biologiques des fractions volatiles et non volatiles, obtenues après extractions des plantes médiévales sélectionnées, ont été réalisées pour juger de leurs activités aromatiques, antioxydantes et antibactériennes ainsi que de leur intérêt dans le cadre de nouvelles réglementations européennes. Les résultats de ces travaux montrent des activités odorantes, antioxydantes et antibactériennes significatives pour l'ensemble des plantes sélectionnées. Dans une troisième étape, du fait de la rareté du végétal étudié, nous nous sommes intéressés à deux voies analytiques d'évaluation rapide de la composition de l'huile essentielle native de ces plantes aromatiques ; premièrement au moyen d'un échantillonnage classique couplé à un détecteur à forte sensibilité comme la chromatographie bidimensionnelle et deuxièmement par un échantillonnage spécifique couplé à un détecteur classique pour lequel nous avons développé un appareillage novateur dénommé "Crushing Finger Device" (CFD). Ce dernier permet de mimer instrumentalement l'écrasement de la feuille entre les doigts et de piéger les composés volatils odorants émis en vue de leur analyse chromatographique ultérieure. Enfin, pour une application pré-industrielle, afin d'élargir le champ d'application de ces plantes aromatiques, des études d'infusion des plantes et d'aromatisation des huiles végétales par les huiles essentielles ont été mises en place. Durant le processus d'aromatisation de l'huile végétale, les volatils émis par la plante ou par l'huile essentielle donnent un goût et une saveur spécifique tout en délivrant une activité biologique notable. / Midi-Pyrenees region has a rich and unvalorized wild vegetal heritage, especially for aromatic plants. In the framework of the search for new natural flavorings e.g. WONF, we chose to adopt the way of valorization of the medieval aromatic plants of this region, because they have been always picked in a wild state without being ever grown. In a first step, a global bibliographical survey study based on historical books, library archived documents and computerized data banks, was made in order to identify regional plants which were used by our ancestors for their odorous, culinary or medicinal activities. We selected five aromatic plants, e.g. Achillea millefolium, Calamintha grandiflora, Tanacetum balsamita, Myrrhis odorata and Monarda didyma. In a second step, physico-chemical, sensory and biological studies of volatiles and non volatiles fractions, obtained by extractions of the selected medieval plants were carried out, to judge their aromatic, antioxidant and antibacterial activities and their interest within the framework of new european regulations. Theses results show a significant odorous, antioxidant and antimicrobial activities for all selected plants. In a third step, such plants were rare or their harvesting problematic, inducing a limited sampling. We decided to investigate two strategic approaches to perform a rapid evaluation of native essential oil, based either on a classic sampling with a specific/ultra sensitive detection such as Comprehensive GC, nor the use of a specific/high effective sampling system with a classic detection. This last methodology was based on a self designed apparatus “Crushing Finger Device” (CFD), allowing to copy the crushing of leaves between fingers and to trap the volatiles emitted to their later chromatographic analysis. Finally, in the aim to expand the use of aromatic plants, also to enlarge the assortment of oil products, a pre-industrial application were carried out. Infusion of plants and flavoring of vegetable oil by essential oil were studied. During processing of aromatized vegetable oil, volatiles from added aromatic plants or essential oil migrate to oil to give a specific aroma and a significant biological activity. Plantes aromatiques oubliées, Huile essentielle, Antioxydant, Antibactérien, Simulateur de froissement, Composés d’arômes, Achillea millefolium, Calamintha grandiflora, Tanacetum balsamita, Myrrhis odorata, Monarda didyma.

A família Meliaceae compreende cerca de 51 gêneros e 550 espécies distribuídas principalmente na região Neotropical. Incluídas nesta família, estão espécies de elevado interesse comercial para a produção de madeiras nobres. Há carência de informações quanto à biologia floral dessas espécies, que aliada a problemas silviculturas dificulta a elaboração de programas efetivos de melhoramento genético das espécies de interesse econômico da família. O presente trabalho visa contribuir para o entendimento do desenvolvimento floral na família Meliaceae, com destaque para sete espécies de interesse econômico e/ou ecológico: Cedrela fissilis L., Cedrela odorata L., Swietenia macrophylla R. A. King, Trichilia claussenii C. DC., Guarea guidonea (L.) Sleumer, Toona ciliata M. J. Roem e Melia azedarach L. Para os estudos morfo-anatômicos utilizaram-se técnicas de microscopia óptica e microscopia eletrônica de varredura. Analisou-se igualmente o padrão de expressão de genes do modelo ABC, relacionados ao desenvolvimento floral, via hibridização in situ. Os estudos morfo-anatômicos permitiram caracterizar o desenvolvimento dos primórdios dos órgãos florais e o estabelecimento de estágios arbitrários de desenvolvimento, auxiliando na caracterização da expressão gênica. Estes resultados permitiram ainda a identificação de flores funcionalmente femininas ou masculinas em S. macrophylla C. fissilis, C. odorata e T. ciliata, as quais apresentam dimorfismo sexual. O padrão de expressão dos homólogos dos genes: APETALA1 (AP1), APETALA3 (AP3) e AGAMOUS (AG) foram diferentes daqueles previstos pelo modelo ABC. A expressão dos homólogos de AP1 foi verificada em todo o meristema floral nos estágios iniciais de desenvolvimento, em todas as espécies estudadas. O sinal de expressão concentrou-se nos primórdios das sépalas e pétalas em estágios mais avançados do desenvolvimento, porém um fraco sinal de hibridização de AP1 foi verificado em todos os verticilos. A expressão dos homólogos de AP3 foi verificada nas regiões previstas pelo modelo ABC, correspondentes ao segundo e terceiro verticilos. Porém um fraco sinal de AP3 foi também observado no quarto verticilo, o que não é previsto pelo modelo teórico. A expressão dos homólogos de AG foi restrita à região central do meristema floral, correspondente ao terceiro e quarto verticilos, em todos os estágios do desenvolvimento floral de todas as espécies estudadas. Entretanto, sinais adicionais de hibridização dos homólogos de AG foram visualizados na região abaxial dos primórdios de sépalas e pétalas em C. fissilis, C. odorata e T. ciliata. Os estudos da expressão dos genes do modelo ABC em Meliaceae revelaram padrões de expressão que não concordam com o modelo teórico vigente do controle molecular da determinação da identidade dos órgãos florais._________________________________________________________________________________________ ABSTRACT: The Meliaceae family comprises approximately 51 genus and 550 species mainly distributed in the Neotropical region. In this family there are many species of commercial interest for the production of noble wood. The lack of information on the floral biology associated with silvicultural problems limits the elaboration of effective breeding programs with species of economic interest. The present work aims to contribute with the understanding of floral development in Meliaceae, focusing in seven species of economical or ecological interest: Cedrela fissilis L., Cedrela odorata L., Swietenia macrophylla R. A. King, Trichilia claussenii C. DC., Guarea guidonea (L.) Sleumer, Toona ciliata M. J. Roem e Melia azedarach L. Morpho-anatomical analyses were done by light and scannning electron microscopy. The expression pattern of the ABC model genes, which are related to floral development, was analyzed by in situ hybridization. The characterization of the development of floral organ primordia and the establishment of arbitrary stages of floral development was done and was important for the characterization of gene expression. These results allowed for the identification of functionally female and male flowers in Swietenia macrophylla, Cedrela fissilis, Cedrela odorata and Toona ciliata, characterizing sexual dimorphism. The patterns of gene expression of the homologous of APETALA 1, APETALA 3 and AGAMOUS were different from those predicted by the ABC model. AP1 expression was observed in the entire floral meristem in the initial stages of development, in all seven species. The expression signal was more concentrated in sepal and petal primordia in further stages of development, although a weak hybridization signal of AP1 was verified in all four whorls. The expression of AP3 homologs was observed in the second and third whorls, as predicted by the ABC model. However, a weak signal of AP3 was also observed in the forth whorl, which was not predicted by the ABC model. AG homolog expression was restricted to the central region of the floral meristem, corresponding to the third and forth whorls, in all stages of development of all seven species. However, additional hybridization signal of AG homologs were seen in the abaxial region of sepal and petal primordia in Cedrela fissilis, Cedrela odorata and Toona ciliata. The analyses of the ABC model genes in Meliaceae revealed that the expression patterns do not agree with the theoretical currently accepted model that determines the molecular control of the identity of the floral organs.