La longévité correspond à la capacité des graines à tolérer la dessiccation et d'être capable de germer sans perte de vigueur après conservation. Elle est acquise au cours de la maturation, après le remplissage des graines et l'acquisition de la tolérance à la dessiccation. Le but de cette thèse est d'analyser les caractéristiques physiologiques, biochimiques et moléculaires lors de la maturation des graines de Medicago truncatula, et d'identifier les mécanismes ou molécules qui corrèlent avec l'acquisition de la longévité. Pendant la maturation, la longévité augmente d'un facteur 30 entre 28 et 44 jours après pollinisation. Une analyse de l'acquisition de la longévité des graines sur plusieurs récoltes et des mesures biochimiques (teneur en sucres solubles, activité des systèmes antioxydants) montre une très forte corrélation entre la longévité et la teneur en saccharose(R²=-0.91) et stachyose(R²=0.92). Une étude protéomique a permis de suivre l'abondance de 38 spots qui correspondent à 16 protéines de type'Late Embryogenesis Abundant'(LEA). Seulement quatre polypeptides corrèlent avec la longévité(CapLEA,D113.II, D34.III et EM) et représentent 45% de l'intensité totale des protéines LEA. L'abondance de la plupart des polypeptides LEA augmente pendant la dernière phase de séchage de graines. La caractérisation de mutants abi5, un facteur de transcription, connu pour sa régulation de gènes LEA, montre que, contrairement à Arabidopsis, ABI5 chez M. truncatula joue un rôle clé dans la dormance et l'accumulation d'oligosaccharides. De surcroit, les graines de mutants Tnt1 d'abi5 ont une longévité deux fois moindre, et présentent une diminution des protéines d'EM, EM6 et D34.

Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Two experiments were conducted to find diversity in plant responses to arbuscular mycorrhizal (AM) fungi: positively and negatively/non-responsive combinations. Medic (Medicago truncatula L.) tomato (Solanum esculentum L.), leek (Allium porrum L.) and cucumber (Cucumis sativus L.) were inoculated with Glomus intraradices, Gigaspora margarita and Scutellospora calospora. It was found that soil differences critically affect plant responses to AM fungi. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1283734 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2007

Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Two experiments were conducted to find diversity in plant responses to arbuscular mycorrhizal (AM) fungi: positively and negatively/non-responsive combinations. Medic (Medicago truncatula L.) tomato (Solanum esculentum L.), leek (Allium porrum L.) and cucumber (Cucumis sativus L.) were inoculated with Glomus intraradices, Gigaspora margarita and Scutellospora calospora. It was found that soil differences critically affect plant responses to AM fungi. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1283734 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2007

Made available in DSpace on 2014-06-11T19:31:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2001-06-27Bitstream added on 2014-06-13T20:02:33Z : No. of bitstreams: 1 campos_at_dr_botfca.pdf: 1232453 bytes, checksum: 23f42c8e84fcf5f7b59eb8f0187628d3 (MD5) / O objetivo deste trabalho foi proceder ao levantamento e caracterização do consumo de energia nas atividades relativas à implantação, condução, processos de fenação e armazenamento, assim como ao balanço energético global de duas forrageiras diferentes, sendo uma gramínea, o coast-cross, e a outra uma leguminosa, a alfafa, inseridas em um Sistema Intensivo de Produção de Leite, localizado na região de Coronel Pacheco, MG. Para o desenvolvimento do estudo, foram utilizados dados coletados pela Embrapa Gado de Leite, em uma média de sete anos, considerando a produção de um hectare para cada cultura, relacionando o consumo dos diversos insumos envolvidos nos processos produtivos e o emprego de máquinas, equipamentos e instalações necessários à condução tecnificada da atividade. Para o levantamento dos diversos fatores componentes da matriz, foram utilizados coeficientes energéticos apresentados por autores especializados. Para ambas culturas utilizadas para estudo, coast-cross e alfafa, a energia direta, relativa aos insumos, teve maior participação que a indireta, sendo os percentuais encontrados de 94,64 e 5,31, respectivamente, para o coast-cross e 93,05 e 6,95, respectivamente, para a alfafa. O principal consumidor de energia indireta, para ambas forrageiras, foi o trator, que aciona todos implementos, seguido pelo equipamento de irrigação. O galpão de feno utilizado em conjunto pelas culturas, apesar do elevado consumo energético desprendido em sua construção, possui uma elevada vida útil, com o armazenamento considerado em 24 horas por dia, o que faz com que sua participação ponderada pela Demanda Específica de Energia seja reduzida drasticamente. O processo de fenação do coast-cross apresentou maior eficiência energética que o da alfafa, com valores de 4,28 para o primeiro e 3,19 para a segunda... / The aim of this work was to carry out a survey and characterization of the energy comsumption in the activities related to introduce, development, hay processing and storage, as well as global energy inputs of two different forage, one of them is grass, the coast-cross, and the other is leguminous, alfalfa, inserted in an Intensive System of Milk production, located in the Coronel Pacheco MG-Brasil region. In order to develop the study it was used data colected by Embrapa Gado de Leite, an average of seven years, considering the production of 1 ha of each culture, linking the consumption of various inputs involved in the productive process and the use of machines, equipments and building that are necessary to conduct the activity in a technique way. To carry on the survey and the various components factors of the matrix, it was used energetic coefficient introduced by specialized researchers. For both cultures used in the study, coast-cross and alfalfa, the direct energy, related to inputs had bigger participation, being the percentages found 94.64 and 5.31 respectively, to the coast-cross, and 93.05 and 6.95, respectively, to the alfalfa. The main consumer of indirect energy, for both forage, was the tractor, which turns on all the implements, followed by an irrigation system. The warehouse hay used together by the cultures, despite the high energetic consuption used in its construction, the warehouse hay has a long useful life, so what concerns to the storage, considered within 24 h per day, what makes its participation slightly ponderous by the specific demand of energy, and this process reduces its participation in a drastic way. The hay process coast-cross presented energetic efficiency as much bigger as the alfalfa, values such as 4.28 to the first one and 3.19 to the second one. Most of the energy used was that one derived from fossil resources, diesel...(Complete abstract click electronic access below)

La symbiose mycorhizienne à arbuscules (SMA) est le résultat de l'interaction entre les racines de plus de 80% des familles de plantes terrestres et les champignons MA. Divers types de membranes jouent un rôle crucial dans la mise en place et le fonctionnement de la SMA chez l’hôte végétal. Si l’électrophorèse bidimensionnelle (2-DE) reste la méthode la plus couramment utilisée pour des analyses protéomiques quantitatives dans la SMA, elle résout difficilement les protéines membranaires en raison de leur hydrophobicité, leur précipitation au point isoélectrique (pI) et leur faible abondance comparativement aux protéines cytosoliques. Donc peu nombreuses sont les protéines membranaires identifiées comme étant régulées en réponse à la symbiose. Afin d’avoir accès à cette catégorie de protéines et contourner les défauts de la 2-DE, l’application de nouvelles méthodes permet de réaliser des analyses quantitatives avec marquage chimique (comme l’iTRAQ) ou non (label-free). Dans ce contexte, deux méthodes de protéomique quantitative, iTRAQ-OFFGEL-CL-SM/SM et « label-free » 1-DE-CL-SM/SM, sont adoptées dans ce travail visant à identifier et quantifier les variations d’accumulation des protéines microsomales de racines de Medicago truncatula inoculées par Rhizophagus irregularis, préalable indispensable à l’analyse de leur rôle fonctionnel dans la SMA. Un protocole d’extraction donnant accès à des fractions radiculaires enrichies en protéines microsomales nécessaires pour les analyses ultérieures est décrit dans cette étude. En plus de l’analyse quantitative du protéome membranaire en réponse à la SMA, une approche méthodologique a été mise en place afin d’étudier l’impact du marquage iTRAQ sur le pI des peptides. / Arbuscular mycorrhizas (AM) are widespread symbiotic associations between plant roots and AM fungi. Deep membrane alterations are the foremost morphological changes occurring in the host plant in response to AM symbiosis. Two-dimensional gel electrophoresis (2-DE) is the workhorse method in AM proteomics. Membrane proteins are under-represented in 2-DE because of their hydrophobicity, low abundance, and precipitation at their isoelectric point, thereby few are the identified membrane proteins involved in sustaining the AM symbiosis. Membrane proteomics is still challenging due to 2-DE related shortcomings, however latest trends and advancements in mass spectrometry (MS)-based quantitative proteomics offer enormous potential to monitor membrane protein change in abundance in large scale experiments. In the current work microsomal proteins of Medicago truncatula roots inoculated with Rhizophagus irregularis were, for the first time, scrutinised by state-of-the-art MS-based proteomic approaches iTRAQ-OFFGEL-LC-MS/MS and label-free 1-DE-LC-MS/MS. The applied workflows combine two novel proteomic procedures, label-based and -free, targeting an insight view on the membrane proteome changes in AM symbiosis. A subcellular fractionation method is herein described to access the total membrane-associated proteins with sufficient recovery and purity for their subsequent in-depth analysis. In addition to the biological gain by shedding the light on candidate AM-related membrane proteins, a methodological approach was carried out in the present work in order to elucidate the iTRAQ labelling impact on peptide isoelectric points.

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Two experiments were conducted with sheep to determine the nutritional value of 'Lathco' flatpea (Lathyrus sylvestris) hay. In experiment 1, a finishing trial was conducted with 50 group fed crossbred lambs (25 ewes and 25 wethers) fed diets consisting of 70% chopped forage and 30% ground corn grain for 70 d. Five pens of five wethers and five pens of five ewes were selffed diets in which the forage consisted of 100:0, 75:25, 50:50, 25:75, and 0:100 proportions of alfalfa (Medicago sativa L.) and flatpea hay. Feed efficiencies decreased linearly (P<.05) as flatpea increased in the diet. Linear decreases (P<.05) were obtained in kidney and pelvic fat, backfat, leg conformation, and yield grade with increased flatpea in the diet. Blood urea-N (BUN), ruminal pH and NH3-N increased (P<.005) as proportion of flatpea increased. Total volatile fatty acid (VFA) and acetate concentration decreased linearly (P<.005) and propionate, isovalerate, and valerate levels increased linearly (P<.01) with increased proportions of flatpea hay. In experiment 2, 30 wether lambs were fed diets consisting of 100:0, 75:25, 50:50, 25:75 and 0:100 proportions of chopped alfalfa and flatpea hays in a digestion trial. Neutral detergent fiber (NOE), acid detergent fiber (ADE), cellulose, hemicellulose and energy digestibilities decreased linearly (P<.01) with increased levels of flatpea hay. Nitrogen retention was not different between sheep fed the different diets. Apparent absorbtion and retention of Ca decreased linearly (P<.05) with increased level of flatpea hay Ruminal pH and NHa-N, and BUN increased linearly (P<.OOl) as flatpea was increased in the diet. Ruminal total VEA, acetate and acetate to propionate ratio decreased linearly (P<.001) and propionate, isobutyrate, isovalerate, and valerate levels increased linearly (P<.001) as level of flatpea increased. / Master of Science

Les interactions entre protéines sont un enjeu majeur en recherche. Hautement spécifiques, elles régulent l'organisation cellulaire, ainsi que les réponses aux stimuli extérieurs ; ce sont des cibles idéales des agents thérapeutiques. Diverses techniques d'étude sont disponibles, mais peu d'entre elles permettent d'analyser simultanément plusieurs interactions. L'objectif de ce travail est d'utiliser le double hybride bactérien pour découvrir de nouveaux partenaires de 2 régulateurs de la prolifération tumorale humaine, p21 et STAT3, et en parallèle pour déterminer la fonction de MtSAP1, impliquée dans la germination et la réponse aux stress abiotiques chez Medicago truncatula. L'analyse des partenaires potentiels de l'oncogène STAT3 et de l'inhibiteur p21 suggère l'existence de nouveaux mécanismes moléculaires de la tumorogenèse auxquels participerait STAT3, et permet d'envisager de nouvelles hypothèses quant au rôle de p21. Un nouveau complexe, p21/prohibitine2, est étudié mais sa fonction reste supposée : il pourrait agir comme corégulateur transcriptionnel et/ou réguler la protéolyse de p21. L'étude du gène MtSAP1 et des partenaires d'interactions suggèrent fortement que MtSAP1 serait induit par l'hypoxie : elle jouerait un rôle considérable dans la détoxification et la tolérance de la plante au stress. Ce travail, appuyé par la littérature, met en évidence un lien fonctionnel entre p21, STAT3 et MtSAP1 au cours de l'hypoxie dans les cellules cancéreuses humaines. La confirmation de cette hypothèse permettrait d'approfondir les mécanismes de protection cellulaire face à l'hypoxie, tant au sein des tumeurs humaines que lors de la tolérance de Medicago truncatula.

La maîtrise des qualités physiologique (performance germinative) et sanitaire des semences (vection d'agents pathogènes) constitue un verrou scientifique et technologique majeur dans le cadre d'une agriculture raisonnée et mondialisée. La qualité physiologique repose notamment sur la tolérance à la dessiccation, la dormance et l'aptitude à la conservation qui s'acquièrent pendant le développement de la graine. Cependant les mécanismes régulateurs de ces processus restent mal compris. Chez Medicago truncatula, par une approche transcriptomique nous avons montré que la signalisation par l'acide abscissique participe via le facteur de transcription ABI5 à la régulation de la survie à l'état sec et à la dormance. ABI5 joue un rôle prépondérant dans les phases du développement allant de la fin de la maturation et à la levée. La transmission à et par les semences est l'un des principaux moyens de survie et de dissémination des bactéries phytopathogènes telles que Xanthomonas. Les éléments du dialogue moléculaire qui pourrait s'établir entre ces bactéries et la graine sont cependant méconnus. Après avoir établi que la contamination est plus efficace en situation compatible [Xanthomonas alfalfae subsp. alfalfae] qu'en situation incompatible [X. campestris pv. campestris, Xcc], nous démontrons l'existence d'un dialogue moléculaire entre Xcc et la graine en développement. La réponse transcriptionelle de la graine contaminée par Xcc et la réduction de leur poids suggèrent un trade-off entre l'activation des défenses basales et son développement. Cette thèse apporte des éléments permettant de suggérer que les qualités sanitaire et physiologique sont liées.

Pas de résumé en français / In plants, long distance transport of sugars from photosynthetic source leaves to sink organs comprises different crucial steps depending on the species and organ types. Sucrose, the main carbohydrate for long distance transport is synthesized in the mesophyll and then loaded into the phloem. After long distance transport through the phloem vessels, sucrose is finally unloaded towards sink organs. Alternatively, sugar can also be transferred to non‐plant sinks and plant colonization by heterotrophic organisms increases the sink strength and creates an additional sugar demand for the host plant. These sugar fluxes are coordinated by transport systems. Main sugar transporters in plants comprise sucrose (SUTs) and monosaccharide (MSTs) transporters which constitute key components for carbon partitioning at the whole plant level and in interactions with fungi. Although complete SUTs and MSTs gene families have been identified from the reference Dicot Arabidopsis thaliana and Monocot rice (Oriza sativa), sugar transporter families of the leguminous plant Medicago truncatula, which represents a widely used model for studying plant-fungal interactions in arbuscular mycorrhiza (AM), have not yet been investigated.With the recent completion of the M. truncatula genome sequencing as well as the release of transcriptomic databases, monosaccharide and sucrose transporter families of M. truncatula were identified and now comprise 62 MtMSTs and 6 MtSUTs. I focused on the study of the newly identified MtSUTs at a full family scale; phylogenetic analyses showed that the 6 members of the MtSUT family distributed in all three Dicotyledonous SUT clades; they were named upon phylogenetic grouping into particular clades: MtSUT1-1, MtSUT1-2, MtSUT1-3, MtSUT2, MtSUT4-1 and MtSUT4-2. Functional analyses by yeast complementation and expression profiles obtained by quantitative RT-PCR revealed that MtSUT1-1 and MtSUT4-1 are H+/sucrose symporters and represent key members of the MtSUT family. Conservation of transport capacity between orthologous leguminous proteins, expression profiles and subcellular localization compared to previously characterized plant SUTs indicate that MtSUT1-1 is the main protein involved in phloem loading in source leaves whilst MtSUT4-1 mediates vacuolar sucrose export for remobilization of intracellular reserve.The AM symbiosis between plants and fungi from the phylum Glomeromycota is characterized by trophic exchanges between the two partners. The fungus supplies the autotrophic host with nutrients and thereby promotes plant growth. In return, the host plant provides photosynthate (sugars) to the heterotrophic symbiont. Here, sugar fluxes from plant source leaves towards colonized sink roots in the association between the model leguminous plant M. truncatula and the arbuscular mycorrhizal fungus (AMF) Glomus intraradices were investigated. Sugar transporter candidates from both the plant and fungal partners presenting differential expression profiles using available transcriptomic tools were pinpointed. Gene expression profiles of MtSUTs and sugar quantification analyses upon high and low phosphorus nutrient supply and inoculation by the AMF suggest a mycorrhiza-driven stronger sink in AM roots with a fine-tuning regulation of MtSUT gene expression. Conserved regulation patterns were observed for orthologous SUTs in response to colonization by glomeromycotan fungi.In parallel, a non-targeted strategy consisting in the development of a M. truncatula - G. intraradices expression library suitable for yeast functional complementation and screening of symbiotic marker genes, similar to the approach that led to the identification of the first glomeromycotan hexose transporter (GpMST1), has been developed in this study. [...]