A cana-de-açúcar (Saccharum spp.) desempenha um papel de grande importância no cenário socioeconômico brasileiro, e representa 42% da matriz energética renovável do país. A expansão da área de cultivo da cana-de-açúcar para solos marginais e a necessidade de manutenção da alta produtividade dessa cultura tem levado à maior aplicação de fertilizantes a base de nitrogênio (N). Tal fato aliado à baixa responsividade da cana-de-açúcar a fertilizantes nitrogenados acarreta altos custos econômicos e ambientais. O amônio é a fonte preferencial de N para essa gramínea, sendo que pouco se conhece sobre a funcionalidade dos transportadores de NH4+ pertencentes à família gênica AMT (AMMONIUM TRANSPORTER). Neste contexto, é relevante esclarecer os mecanismos que influenciam na eficiência do uso de N (NUE), visando reduzir o impacto econômico e ambiental da aplicação dos fertilizantes nitrogenados nos sistemas agrícolas. Dessa forma, esse trabalho teve como objetivo a caracterização molecular e funcional de membros da subfamília AMT2 de cana-de-açúcar através de expressão heteróloga em mutantes de Saccharomyces cerevisiae (cepa 31019b) e Arabidopsis thaliana (qko), defectivos no transporte de amônio. As sequências gênicas e promotoras de ScAMT2;1 e ScAMT3;3A foram identificadas em biblioteca de BAC (bacterial artificial chromosome) de cana-de-açúcar (cultivar \'R570\'). Análises de expressão gênica de ScAMT2;1 e ScAMT3;3A em cana-de-açúcar demonstraram uma expressão preferencial em raízes e em folhas maduras, respectivamente, e que estes genes são regulados de maneira distinta entre si e entre os órgãos, de acordo com o desenvolvimento e com o status de N da planta. A complementação de levedura com os AMT2 de cana-de-açúcar demonstrou que estes genes restauram o crescimento do mutante, sendo que ScAMT2;1 permite maior absorção de amônio; porém o experimento não indicou sensibilidade dessas proteínas ao metilamônio (análogo tóxico ao amônio). Experimentos de localização da expressão órgão/tecido específico em arabidopsis selvagem \'Col-0\', utilizando os promotores de ScAMT2;1 ou ScAMT3;3A fusionados a GUS ou GFP, demonstraram que esses AMTs são preferencialmente expressos na região da endoderme/periciclo e vascular das células das raízes e região vascular da parte aérea, sendo regulados pela disponibilidade e fonte de N. Plantas de arabidopsis qko superexpressando ScAMT2;1, ScAMT3;3A ou transformadas com ScAMT2;1 dirigido por seu promotor endógeno, crescidas in vitro com amônio como fonte exclusiva de N, apresentaram um aumento significativo na produção de biomassa em relação a qko não transformada, principalmente para ScAMT2;1, indicando que essas proteínas são capazes de transportar amônio e complementar o mutante. Dados de influxo e acúmulo de 15N-amônio in vivo em raízes e parte aérea de plantas qko superexpressando ScAMT2;1 ou ScAMT3;3A demonstraram que ScAMT2;1 atua na absorção de amônio pelas raízes e provavelmente do carregamento do xilema, enquanto ScAMT3;3A está possivelmente envolvida na remobilização de amônio na parte aérea, podendo atuar aditivamente na absorção de NH4+ em raízes sob alto amônio. Esses resultados indicam que os transportadores ScAMT2;1 e ScAMT3;3A de cana-de-açúcar são funcionais, atuando com propriedades e funções distintas no transporte de amônio nessa gramínea e de acordo com a disponibilidade de N. / Sugarcane (Saccharum spp.) plays a major role in the Brazilian socio-economic scenario, and represents 42% of renewable energy sources in the country. The expansion of sugarcane cultivation to marginal lands and the requirement to maintain high yields have led to increased application of nitrogen (N) fertilizer. This fact, coupled with the low response of sugarcane to N fertilization, entails high economic and environmental costs. Ammonium is the preferred source of N by this grass; however, little is known about the functionality of NH4+ transporters belonging to the AMT gene family (AMMONIUM TRANSPORTER). In this context, it is important to clarify the mechanisms that affect the nitrogen use efficiency (NUE) in order to reduce the economic and environmental impact of the application of N fertilizers in agricultural systems. Therefore, this study aimed to conduct the molecular and functional characterization of members of the AMT2 subfamily from sugarcane by heterologous expression in mutants of Saccharomyces cerevisiae (strain 31019b) and Arabidopsis thaliana (qko), both defective in ammonium transport. Gene and regulatory region sequences of ScAMT2;1 and ScAMT3;3A were identified in a bacterial artificial chromosome (BAC) library of sugarcane (cultivar \'R570\'). Expression analysis of ScAMT2;1 and ScAMT3;3A in sugarcane showed a preferential expression in roots and mature leaves, respectively, and indicated a distinct expression pattern between genes and organs according to the ontogeny and the N status of the plant. The yeast complementation with AMT2 of sugarcane demonstrated that these genes restore the mutant growth, with ScAMT2;1 enabling higher ammonium absorption; however, the experiment did not indicate sensitivity to methylammonium (toxic ammonium analog). Arabidopsis wild type \'Col-0\' transformed with the promoter region of ScAMT2;1 or ScAMT3;3A directing the expression of GUS or GFP, demonstrated preferential expression in the endodermis/pericycle regions of roots and vascular region in shoots, being regulated by the availability and source of N. Arabidopsis qko overexpressing ScAMT2;1, ScAMT3;3A or transformed with ScAMT2;1 driven by its endogenous promoter, grown in vitro with ammonium as the sole source of nitrogen, showed a significant increase in biomass production compared to untransformed qko, especially for ScAMT2;1, indicating that these proteins are capable of transporting ammonium and complementing the mutant. Data of 15N-ammonium influx and accumulation in vivo in roots and shoots of qko plants overexpressing ScAMT2;1 or ScAMT3;3A showed that ScAMT2;1 acts in ammonium uptake by roots and probably in the xylem loading, while ScAMT3;3A is possibly involved in ammonium remobilization in shoots, and may act additively in the absorption of NH4+ in roots under high ammonium. These results indicate that ScAMT2;1 and ScAMT3;3A from sugarcane are functional, working with distinct properties and functions in ammonium transport according to the availability of N

Made available in DSpace on 2017-07-10T14:17:10Z (GMT). No. of bitstreams: 1 simas.pdf: 1840643 bytes, checksum: 95c5a43ded5855449c86bf6bf87f3e58 (MD5) Previous issue date: 2015-02-12 / The aim of this study was to analyze the effects of the physical exercise of climbing stairs in bone and cartilage histomorphometric parameters in rats submitted to a model of osteoporosis and immobilization. 36 Wistar rats were separated into six groups were used: G1, G2 and G3 groups were subjected to pseudo-oophorectomy; and G4, G5 and G6 to oophorectomy. After surgery, all groups remained 60-day rest and the rats G2, G3, G5 and G6 had the right hind limb (MPD) immobilized during 15 days, followed by remobilization same period, being free in the box to G2 and G5, and climb stairs in G3 and G6. At the end of the experiment, the rats were euthanized, their tibias removed bilaterally and submitted to histological routine. Morphometric analysis showed that there was a statistically significant decrease in area (p = 0.0178) and cortical thickness (p = 0.0024), thickness of articular cartilage (p = 0.0138) and epiphyseal plate were made (p = 0.0187), and the number of osteocytes (p <0.0001) and chondrocytes (p = 0.0006) as well as significant increase in the medullary canal (p = 0.0384) in immobilized limbs of ovariectomized rats. However, the stair climbing exercise was able to reverse the loss of bone cortex (area (F (5; 29) = 6.24, p = 0.0007) and thickness (F (5; 29) = 4.11 p = 0.0062)), cartilage (articular cartilage thickness (F (5; 29) = 13.88, p <0.0001) and epiphyseal plate (F (5; 29) = 14.72, p <0.0001)) and cellular (osteocytes (F (5; 29) = 14.55, p <0.0001) and chondrocytes (F (5; 29) = 10.16, p <0.0001)) resulting of oophorectomy associated with immobilization. It was also observed a significant decrease in trabecular thickness and area in the members subjected to immobilization, however, both as the free remobilization and climb stairs were able to recover from this loss. Morphological analysis of tibia articular cartilage, no changes in cell structure or organization of the rats not subjected to immobilization (G1 and G4) were verified by observing only considerable decrease in the thickness and number of chondrocytes in G4 (oophorectomized group) . In female rats that were free remobilization (G2 and G5), it was observed degeneration regions of articular cartilage with subchondral bone exposure, loss of cellular organization, discontinuity of tidemark, the presence of fissures and flocculation, as well as decreased number of chondrocytes. On the other hand, G2 female rats (pseudo-oophorectomized group, immobilized and free remobilization) showed some regions of granulation tissue (pannus). In rats subjected to exercise climb stairs (G3 and G6), there were repair signs of cartilaginous structures, the presence of clones and pannus. In G6 (oophorectomized group, remobilization and immobilized ladder), although it was observed subcortical invasion of blood vessels in the calcified zone, and increasing the amount of isogenous groups and the thickness of the calcified zone. From the results obtained in this study, it can be concluded that climbing stairs exercise was effective in the recovery process of bone and cartilage tissue damaged by immobilization on osteoporosis model by ovariectomy in rats. / O objetivo deste estudo foi analisar os efeitos do exercício físico de subida em escada sobre parâmetros histomorfométricos ósseos e cartilaginosos de ratas submetidas à um modelo de osteoporose e imobilização. Foram utilizadas 36 ratas Wistar separadas em seis grupos: G1, G2 e G3 submetidas à pseudo-ooforectomia; e G4, G5 e G6 à ooforectomia. Após a cirurgia, permaneceram 60 dias em repouso e as ratas de G2, G3, G5 e G6 tiveram o membro posterior direito (MPD) imobilizado por 15 dias, seguido pelo mesmo período em remobilização, sendo livres na caixa para G2 e G5, e de subida em escada para G3 e G6. Ao final do experimento, as ratas foram eutanasiadas, suas tíbias retiradas bilateralmente e submetidas à rotina histológica. Foram realizadas análises morfométricas, nas quais observou-se que houve diminuição estatisticamente significativa de área (p= 0,0178) e espessura cortical (p= 0,0024), espessura da cartilagem articular (p= 0,0138) e da placa epifisária (p= 0,0187), e do número de osteócitos (p < 0,0001) e condrócitos (p= 0,0006), como também aumento significativo do canal medular (p= 0,0384), em membros imobilizados de ratas ooforectomizadas. No entanto, o exercício de subida em escada foi capaz de reverter a perda óssea cortical (área (F (5;29) = 6,24; p= 0,0007) e espessura (F(5;29)=4,11; p= 0,0062)), cartilaginosa (espessura da cartilagem articular (F(5;29)=13,88; p<0,0001) e da placa epifisária (F(5;29)=14,72; p<0,0001)) e celular (osteócitos (F(5;29)=14,55; p<0,0001) e condrócitos (F(5;29)=10,16; p<0,0001)) decorrente da ooforectomia associada à imobilização. Observou-se também diminuição significativa de área e espessura trabecular nos membros submetidos à imobilização, contudo, tanto a remobilização livre como em escada foram capazes de recuperar essa perda. Nas análises morfológicas da cartilagem articular da tíbia, não foram verificadas mudanças de estrutura nem de organização celular das ratas não submetidas à imobilização (G1 e G4), observando-se apenas considerável diminuição na espessura e no número de condrócitos em G4 (grupo ooforectomizado). Nas ratas que ficaram em remobilização livre (G2 e G5), visualizou-se regiões de degeneração da cartilagem articular com exposição de osso subcondral, perda da organização celular, descontinuidade da tidemark, presença de fissuras e floculações, como também diminuição do número de condrócitos. No entanto em G2 (grupo pseudo-ooforectomizado, imobilizado e remobilização livre) algumas regiões apresentavam tecido de granulação (pannus). Nas ratas submetidas ao exercício de subida em escada (G3 e G6), havia sinais de reparação das estruturas cartilaginosas, com presença de clones e pannus. Em G6 (grupo ooforectomizado, imobilizado e remobilização em escada), observou-se ainda invasão de vasos sanguíneos subcorticais na zona calcificada, além de aumento da quantidade de grupos isógenos e da espessura da zona calcificada. A partir dos resultados obtidos neste estudo, pode-se concluir que o exercício de subida em escada mostrou-se efetivo no processo de recuperação dos tecidos ósseo e cartilaginoso danificados pela imobilização, em modelo de osteoporose por ooforectomia em ratas.

Orientador: Ciro Antonio Rosolem / Banca: Leonardo Theodoro Bull / Banca: Ederaldo José Chiavegato / Banca: Luiz Henrique Carvalho / Banca: Cassio Hamilton Abreu Junior / Resumo: O boro é um micronutriente de grande importância para o crescimento e desenvolvimento das plantas. Este elemento tem baixa mobilidade no floema e é reconhecidamente o micronutriente cuja deficiência é mais comum no algodoeiro. Partindo-se da hipótese de existe diferença de resposta ao B entre cultivares de algodão, este trabalho teve como objetivo, estudar a absorção e mobilidade do boro disponibilizado via solução nutritiva, em três cultivares de algodão, bem como, a conseqüência de uma eventual deficiência do nutriente na fotossíntese das plantas. O experimento foi conduzido em condições de casa de vegetação e as plantas foram cultivadas em vasos contendo solução nutritiva. Os tratamentos foram constituídos de três cultivares de algodão (FMT 701, DP 604BG e FMX 993), e cinco doses de boro na solução nutritiva (0,0 - 2,5 - 5,0 - 10,0 e 20,0 μmol L-1). O delineamento experimental utilizado foi em blocos casualizados com quatro repetições em esquema fatorial com 3 cultivares x 5 doses. Foram feitas avaliações visuais dos sintomas de deficiência, durante todo o experimento, sendo os sintomas anotados e fotografados. As avaliações fisiológicas foram feitas quando as plantas atingiram o estádio de emissão do primeiro botão floral e foram repetidas após uma e quatro semanas após este estádio. As avaliações fitotécnicas e nutricionais foram feitas em quatro semanas, com a primeira iniciando uma semana após a o estádio de emissão do primeiro botão floral. Os resultados obtidos neste experimento levaram a concluir que: A época de aparecimento e a intensidade de sintomas de deficiência de boro entre cultivares de algodão é diferente. A cultivar DP604BG é inicialmente menos exigente em B, porém, há necessidade de maior disponibilidade de boro no meio nutritivo para evitar o aparecimento de deficiência. O crescimento do algodoeiro... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Boron is an micronutrient of great importance for growth and development of plants. This element has low mobility in the phloem and is known to the micronutrient whose deficiency is more common in cotton. Based on the hypothesis that there is difference of sensitivity to B between cotton cultivars, this work aimed to study the uptake and mobility of boron available in nutrient solution at three cotton cultivars, as well as the consequence of boron deficiency in the photosynthesis of plants. The experiment was conducted under greenhouse conditions and plants were grown in plots containing nutrient solution. The treatments were constituted of three cotton cultivars (FMT 701, DP 604BG and FMX 993), and five doses of boron in the nutrient solution (0.0 - 2,5 - 5.0 to 10.0 and 20.0 mmol L -1). The experimental design used was the completely randomized with four replications in outline factorial 3 cultivars x 5 doses. Evaluations were made of visual deficiency symptoms during all of experiment period and the symptoms recorded and photographed. Physiological evaluations were made when the plants reached the season of first flower bud and were repeated after one and four weeks after this season. Agronomic parameters and nutritional available were made in four weeks, with the first available starting one week after the season of first stage bud. Time of appearance and severity of symptoms of boron deficiency between cotton cultivars is different. Cultivar DP604BG is initially less boron requiring, but there is need of great availability boron in the nutrient solution to prevent the appearance of deficiency. The cotton growth was loss by boron deficiency regardless of differences of symptoms appearance, without deference between cultivars. The boron deficiency affect the physiological activity of three cotton cultivars / Doutor

Le colza est une culture oléagineuse très exigeante en intrants azotés associée à une faible efficience d’usage de l’azote (EUA). Le défi majeur vise à améliorer le bilan agroenvironnemental du colza par une optimisation de l’EUA, notamment en condition où l’azote est limitant dans le sol. L’EUA est limitée par une faible efficience de remobilisation de l’azote (ERA) lors de la sénescence des feuilles. L’objectif de ce travail de thèse a consisté à rechercher, chez le colza, la topologie et l’orientation des attributs métaboliques associées à l’ERA pendant la sénescence foliaire.Les résultats montrent que le métabolome des feuilles évolue tout au long de leur développement végétatif, de leur croissance à leur chute. Il est spécifique à chaque étage foliaire et traduit des relations trophiques et environnementales particulières liées au positionnement des feuilles dans le couvert végétal. Ces spécificités sont associées à des variations de teneurs en glucides, d’acides aminés,de glucosinolates et de coumarines en lien étroit avec la régulation phytohormonale du développement foliaire et avec leur translocation dans le phloème. Le cas de la Proline a été plus particulièrement approfondi et l’activation de son catabolisme sous régulation circadienne dans les tissus sénescents a été mise en évidence. Une approche combinée de transcriptomique et de métabolomique a permis de démontrer une variabilité génotypique importante dans les processus de dégradation et de transport des protéines, glucides et acides aminés entre deux génotypes à forte ERA. De la même manière, des relati / Oilseed rape is a very demanding oleaginous crop for nitrogen inputs associated with a low nitrogen use efficiency (NUE). The main challenge to improve the agri-environmental balance of oilseed rape is to optimize the NUE, especially under nitrogen deprivation. The NUE is limited by a low nitrogen remobilization efficiency (NRE) during leaf senescence. The aim of this thesis was to define the metabolome topology and orientation associated with NRE during leaf senescence in oilseed rape.The results show that leaf metabolome dynamically evolves throughout their vegetative growth, until their fall. Metabolome was found specific to each leaf rank, reflecting the trophic and environmental relationships related to the leaf positioning in the canopy. These specificities are associated with variations in carbohydrates, amino acids, glucosinolates and coumarins contents in close connection with the phytohormonal regulation of leaf development and with their translocation in the phloem.In particular, the activation of Proline circadian-controlled catabolism in senescent tissues was demonstrated. Finally, significant variations in the degradation and transport of proteins, carbohydrates and amino acids between two highly efficient NRE genotypes were highlighted using a combined transcriptomic and metabolomic approach. Similarly, a close relationship has been described between the genes expression levels and the metabolic content involved to increase NRE under low nitrogen input.The results are discussed regarding nitrogen remobilization improvement and more generally nutrients i

Resumo: Com o objetivo de avaliar o efeito do aumento da densidade populacional sobre o metabolismo de C e N, a remobilização e exportação dos nutrientes e as características agronômicas das plantas de milho, conduziu-se um experimento nas condições de campo em Jaboticabal-SP (48°15'18''W e 21°15'22''S), na safra 2005/2006. Os tratamentos se basearam em cinco densidades populacionais do híbrido AG 9010 (30, 45, 60, 75 e 90 mil plantas por ha), com quatro repetições. A adubação de semeadura foi realizada com 320 kg ha-1 da fórmula 05-15-10 e, em cobertura foram aplicados 134 kg ha-1 de N na forma de sulfato de amônio. As avaliações foram realizadas nos estádios de pendoamento (VT) e de grão pastoso (R4), e na colheita final de grãos. O acúmulo de massa seca, dos metabólitos e as atividades enzimáticas do metabolismo de C e N (em R4) nos colmos, folhas e grãos por planta de milho diminuíram em função do aumento da densidade populacional, exceto a invertase solúvel nos grãos e sacarose sintase na folha. O estudo do metabolismo das plantas em condições de campo assemelhou-se aos de cultivo de grãos in vitro, quanto aos aminoácidos livres e a atividade da sacarose sintase. A elevação da densidade de plantas promoveu um maior redirecionamento de fotoassimilados para os grãos, uma redução do acúmulo dos macro e micronutrientes e da eficiência de remobilização de N e K, da parte vegetativa para os grãos, e aumento na eficiência do uso dos nutrientes N, P e K. O incremento na densidade populacional aumentou a altura das plantas e inserção da espiga, diminuiu o número de espigas por planta, o número de grãos por espiga, da massa de mil grãos e da massa de grãos por espiga. A produtividade de grãos e a exportação dos nutrientes aumentaram com o aumento da densidade populacional. / Abstract: With the objective of evaluating the effect of the population density increase on the C and N metabolism, the nutrients remobilization and exportation and the agronomic characteristics of corn plants, a field experiment was carried out, at Jaboticabal-SP (48°15'18''W e 21°15'22''S), in the agricultural year of 2005/2006. The plots were represented by five population densities of the hybrid AG 9010 (30, 45, 60, 75 and 90 thousand plants per hectare), with four replications. Sowing fertilization was performed with 320 kg ha-1 of the formula 05-15-10 and side dressing with 134 kg ha-1 of N, by the form of ammonium sulfate. The variables were measured on the stages of tassel (VT) and pasty grain (R4), and final grain harvest. The dry mass storage of the metabolites and enzymatic activities of C and N metabolism (in R4) in the stalks, leaves and corn plant grains deceased in function of the population density increase, except for the soluble invertase in the grains and sucrose syntax in the leaves. In relation to free aminoacids and sucrose syntax activity, the study of plant metabolism on field conditions was very similar to the studies of grains in vitro. The plant density increase promoted a bigger redirection of assimilates for the grains, reduction of the macro and micronutrients storage and of the N and K remobilization efficiency, from the vegetative parts to the grains, and increased the use efficiency of N, P and K nutrients. The enhance of the population density raised the plant height and insertion of the first ear, decreased the number of ears per plant, number of grains per ear, mass of a thousand grains and grain mass per ear. Grain yield and nutrient exportation rose with the population density increase. / Orientador: Jairo Osvaldo Cazetta / Coorientador: Domingos Fornasieri Filho / Banca: Aildson Pereira Duarte / Banca: Paulo Roberto de Camargo e Castro / Banca: Leandro Borges Lemos / Banca: Renato de Mello Prado / Doutor

The evidence of a flood wave passing through a catchment remains visible even for a long time after it occurs. The morphological update in the channel and floodplains, together with the processes related to the mass transport within the aquatic environment, can be regarded as flood event evidence. The advancement in hydroinformatics brought the development of numerical modeling as a tool for the solution of broad hydrological tasks. Thanks to the scenario modeling, flood events with interconnected processes can be explored in detail. This thesis is broadly focused on the mass transport initialization issue both in the polluted and clear middle-European water environments. The aim of the thesis is the evaluation of the principal issues connected with the mass transport initialization based on complex and integrated numerical modeling. The thesis brings original datasets resulting from several case studies. The aim of the thesis is also to bring a comparative study of methodological approaches evaluating the possibilities and limits regarding the accuracy of inputs vs. outputs and computational time requirements. This thesis also brings several useful comparisons and innovative solutions design. The mass transport initialization issue is solved in both balance and event-scale processed- based models....

Le colza d’hiver (Brassica napus) est cultivé pour l’huile contenue dans ses graines ainsi que pour les tourteaux qui sont une source de protéines pour l’alimentation animale. La culture de colza demande de forts apports d’azote et cette espèce est caractérisée par sa faible efficacité d’utilisation de l’azote. Une forte proportion de l’azote absorbé est restituée au sol lors de la chute précoce des feuilles au stade végétatif. L’amélioration de la remobilisation de l’azote est donc de première importance pour améliorer le rendement de cette culture tout en satisfaisant le besoin de réduction des intrants. La glutamine et l’asparagine jouent un rôle important dans le transport de l’azote au sein de la plante, notamment au cours de la sénescence foliaire. Les deux familles multigéniques des glutamine synthétases cytosoliques (GLN1) et des asparagine synthétases (ASN) assurent leur synthèse. Ce travail de thèse s’est intéressé à ces enzymes chez deux Brassicacées : le colza et Arabidopsis thaliana. Dans un premier temps, l’expression des gènes GLN1 a été étudiée chez Arabidopsis par une combinaison d’approches de biologie moléculaire, cellulaire et de cytologie. Les spécificités d’expression de chacun des cinq gènes d’Arabidopsis ont été mises en évidence. L’identification des gènes BnaGLN1 chez Brassica napus a permis une analyse de leur expression en fonction de l’âge des feuilles et de la disponibilité en azote. Les profils d’expression observés chez le colza se sont révélés similaires à ceux des gènes homologues d’Arabidopsis, amenant l’hypothèse d’une conservation des fonctions chez les deux espèces. Le rôle des gènes GLN1 d’Arabidopsis dans la remobilisation de l’azote vers les graines a été étudié grâce à un marquage ¹⁵N effectué sur des mutants simples. Le rôle des gènes GLN1 dans la remobilisation de l’azote des tissus végétatifs vers les tissus reproducteurs a été mis en évidence sans toutefois cibler spécifiquement une isoforme. L’étude de la famille ASN chez Arabidopsis a permis de mettre en évidence des profils d’expression spécifiques en fonction des organes, de l’âge des tissus et de la disponibilité en azote pour chacun des trois gènes. Le marquage ¹⁵N a également révélé une implication des gènes ASN1 et ASN2 dans la remobilisation de l’azote de la rosette vers les tissus reproducteurs. Les travaux présentés dans ce manuscrit sont une base pour de futures approches translationnelles vers le colza. / Winter oilseed rape (Brassica napus) is grown for its oil-rich seeds and for proteins, used in animal feed cake. It requires high nitrogen inputs due to the low efficiency of nitrogen utilization that characterizes this species. A large proportion of absorbed nitrogen is indeed returned to the soil when leaves fall. Improving nitrogen remobilization to promote seed filling is then required to improve yield and limit fertilizer use. Asparagine and glutamine are important amino acids for phloem translocation. This thesis focuses on the two multigenic families in charge of asparagine and glutamine synthesis: cytosolic glutamine synthetase (GLN1) and asparagine synthetase (ASN). Studies were performed on the two Brassicaceae, rapeseed and Arabidopsis thaliana. The GLN1 gene expressions were investigated in Arabidopsis by a combination of molecular biology and cytology. The five GLN1 genes are differentially expressed in Arabidopsis depending on ageing and nitrogen availability. The identified BnaGLN1 genes in Brassica napus also showed age and nitrogen dependent expressions. Interestingly, expression profiles were similar between homologous genes in Arabidopsis and rapeseed, suggesting that homologous genes share similar function in the two species. The role of Arabidopsis GLN1 genes for nitrogen remobilization to the seeds was monitored using ¹⁵N tracing experiments on individual mutants. The GLN1 genes play a role in the remobilization of nitrogen from the rosette leaves to the reproductive organs. However, their effect is weak and non-specific of one GS1 isoform. ASN genes also presented specific expression profiles depending on organs, age and nitrogen availability. The ¹⁵N tracing revealed that ASN1 and ASN2 are both involved in nitrogen remobilization from the rosette to the seeds. Our studies provide a basis for future translational approaches to improve oilseed rape.

The Kiruna mining district, located in the northern Norrbotten ore province, Sweden, is a geologically and economically important area, being the type-locality for Kiruna iron oxide-apatite (IOA) deposits and also host to a variety of other deposits including syngenetic stratiform exhalative Cu-(Fe-Zn) (Viscaria, Eastern Pahtohavare), epigenetic stratabound Cu ± Au (Pahtohavare), and iron oxide-copper-gold (IOCG, Rakkurijärvi) deposits. However, the timing of IOA versus IOCG within the tectonic evolution is in question based on structural investigations showing Cu- and Fe-sulfides occur in late-orogenic structures. Here we use an established tectonic framework to constrain mineral systems (tectonic/thermal drives, metal and ligand sources, fluid pathways, traps, remobilization mechanisms) related to the early and late phases of the Svecokarelian orogeny in the Kiruna mining district. U-Pb zircon geochronology of intrusions in the district indicates a thermal drive was present during the early phase of the Svecokarelian orogeny from ca. 1920-1865 Ma, however remains enigmatic for the late Svecokarelian orogeny. Zircon grains from a magnetite-ilmenite gabbro yielded an age of 1881 ± 8 Ma, coeval with the Kiirunavaara IOA deposit and suggested to represent an important generation of mafic magmatism related to the ore. Lithogeochemistry of early bimodal Svecokarelian intrusions in the district indicates a within-plate to active continental margin environment with a volcanic arc affinity, pointing to a back arc environment. Epsilon Ndi and 87Sr/86Sri values calculated from the U-Pb ages for the igneous intrusions were compared to samples of ore-related alteration from epigenetic Pahtohavare and Rakkurijärvi deposits, district greenstone, and Archean samples. Results show that each deposit sourced Sr and Nd from a variety of rocks suggesting broad fluid transport. Each deposit has a distinct Sr mixing trend suggesting they formed from different ore-forming fluids and pathways. This is supported by new structural data that constrain the folding event and the ore-related quartz-carbonate-sulfide veins in the Pahtohavare area to a late orogenic timing, compared to the early orogenic timing of Rakkurijärvi. Sulfide trace element and sulfur isotope data from structurally constrained ores within the tectonic framework also record distinct characteristics between early and late deposits. However, remobilization of early Pahtohavare sulfides associated to an increase in Co content and heavier sulfur isotope compositions is recorded. The results of this study illustrate that using a structural framework approach to constrain the ingredients of mineral systems is a powerful strategy for interpreting ore deposit processes in tectonically complex terrains where both IOA and IOCG deposits occur.

Les HAP font partie de la classe des polluants organiques persistants dans les sols. Dans le cadre de la réhabilitation d'anciennes friches industrielles, il est nécessaire de comprendre l'origine de leur forte rétention et de connaître leurs possibilités de re-mobilisation à l'eau. Pour cela, une évaluation du potentiel de re-mobilisation à l'eau de matrices industrielles polluées a été réalisée sur 6 mois à l'aide de réacteurs fermés dont la solution est renouvelée mensuellement. Des expériences d'adsorption/désorption du 14C-fluoranthène sur ces mêmes matrices ainsi que sur une terre agricole ont confirmé que le fluoranthène s'adsorbe par interaction hydrophobe. L'affinité exceptionnelle des HAP pour les matrices présentant une pollution amassée est associée à leur re-mobilisation limitée par le faible contact de l'eau avec les surfaces de ces matrices hydrophobes. Notre étude met également en évidence la désorption plus lente de HAP "natifs" par rapport à des HAP ajoutés récemment, traduisant une modification du comportement de HAP au cours du temps. Afin de comprendre les mécanismes impliqués dans ce "vieillissement" au sein de terres industrielles, une expérience d'incubation de petites colonnes de sol, contaminées par du 14C-fluoranthène, a été conduite sur 6 mois. Dans les matrices à pollution amassée, le fluoranthène reste sous une forme extractible. Dans les matrices présentant une pollution moins importante et non amassée, le devenir des résidus du 14C-fluoranthène est contrôlé par l'activité de la microflore. Il est montré en particulier que les produits de dégradation sont en effet plus mobiles et peuvent pénétrer dans la porosité et y être séquestrés / PAH are persistent organic pollutants in soils. In the framework of industrial waste lands rehabilitation, one needs to understand the reasons of their strong retention and to assess their potential for water remobilization. For these purposes, the remobilization potential of industrial polluted matrices was assessed during 6 months in closed reactors with a monthly renewal of the solution. Adsorption / desorption experiments with 14C-fluoranthene on the same matrices and an agricultural soils confirmed the adsorption of fluoranthene by hydrophobic interaction. The PAH dramatic affinity for matrices containing pure organic phases is combined with their hindered remobilization due to the low surface contact with water of these hydrophobic matrices. Our work also pointed out desorption differences between native and a lab-added PAH, indicating modifications of PAH behaviour in soils. In order to understand the mechanisms involved in this "ageing" in industrial soils, an incubation experiment was conducted during 6 months with small soil columns spiked with 14C-fluoranthene. In matrices containing pure organic phase, fluoranthene remained in an extractable state. In less polluted matrices containing a diffuse pollution, fluoranthene fate was controlled by microflora activity. Our results demonstrated in particular that degradation products that are more mobile than the parent compound can diffuse in the porosity and be sequestered within

Tomato (Solanum lycopersicum L.) consumption has increased every year due to the fruit attractiveness, several utilizations, and beneficial effects for human health. However, tomato fruits can accumulate a Cd concentration that exceeds the safety threshold for human consumption of vegetables, even when plants are grown in soil with acceptable Cd level. Cd is a non-essential, hazardous element to biological systems, triggering several diseases in humans. In plants, Cd disturbs the antioxidant machinery, changes the nutritional status, and impairs the photoassimilate production and/or partitioning, hence reducing fruit yield and quality. However, distinct tomato accessions can present contrasting tolerance degree to Cd toxicity, as detected by our group in previous studies. The use of these accessions is a powerful approach to identify strategies employed by plants to cope with Cdinduced challenges, and the acknowledgement of such strategies can be potentially used in breeding and biotechnological programs to improve fruit yield and quality in crops that were cultivated in contaminated fields. The set of studies that compose the present thesis aimed (i) to identify the main mechanisms for the contrasting tolerance degree to Cd-induced toxicity in tomato accessions after short and long-term Cd exposure; (ii) to evaluate the relationship among tolerance degree and fruits attributes in plants that were grown in Cd-containing soil, and (iii) to determine the transgenerational effects of Cd-induced stress. In the first experiment, nine tomato accessions with a varied tolerance degree, which was based on biomass accumulation, to Cd exposure were grown in hydroponic solution containing CdCl2 35 &mu;M for 6 days. Avoidance of high Mg concentration in roots was identified as a plant strategy to mitigate Cd toxicity by preventing formation of root hairs. Regarding the mode of action of Cd toxicity, Mn excess in leaves, in addition to the high Cd concentration per se, seems to be coupled to leaf damages that are enhanced by the increased Zn and B concentrations in the photosynthetic tissues. In the second experiment, tolerant (Yoshimatsu) and sensitive (Tropic Two Orders) genotypes were grown in Cd-containing soil, in order to evaluated production parameters. After plant exposure to Cd, the tolerant genotype presented an increased fruit diameter, height and weight, when compared to the control plants. In both cultivars, Cd concentration varied according to the following descending order: roots = leaf blade > (floral receptacle, peduncle and sepals) > stem = fruit peel = fruit pulp. Moreover, data suggested that floral receptacle and its related-structures acted as a barrier to the Cd transportation to the fruits, but it was not enough to avoid Cd reaching the fruits. Furthermore, Cd exposure provoked remarkable reductions in the Mg concentration in roots of sensitive and tolerant genotypes, revealing that both tomato cultivars are able to employ this mechanism for plant acclimation to long-term Cd exposure. Considering such information, it is possible that, under the short-term Cd exposure, tolerant accessions activate this mechanism either early or faster than sensitive genotypes. In addition, positive transgenerational effects on seed germination and vigor of the tolerant genotype were triggered by the plant-mother cultivation in Cd-containing media, despite of the increased chromosomal abnormality. This work reported new insights about the effects of Cd exposure on tomato development, tolerance mechanisms, fruit quality and yield of tomato, as well as Cd distribution in the plants. / O consumo de tomate (Solanum lycopersicum L.) tem aumentado a cada ano devido a atratividade dos frutos, suas diversas utilizações e efeitos benéficos para a saúde humana. No entanto, os frutos de tomate podem acumular uma concentração de cádmio (Cd) que excede o limiar de segurança para o consumo humano, mesmo quando as plantas são cultivadas em solo com níveis aceitáveis de Cd. Cádmio e um elemento não-essencial, extremamente perigoso para os sistemas biológicos, desencadeando varias doenças em seres humanos. Nas plantas, o Cd perturba a maquinaria antioxidante, altera o estado nutricional e prejudica a produção e /ou o particionamento de fotoassimilados, frequentemente reduzindo a produtividade e qualidade de frutos. No entanto, diferentes acessos de tomateiros podem apresentar contrastantes graus de tolerância a toxicidade gerada pela exposição ao Cd, como detectado em estudos anteriores de nosso grupo. O uso desses acessos e uma abordagem poderosa para identificar as estratégias empregadas pelas plantas para lidar com os desafios induzidos pelo Cd; e o conhecimento de tais estratégias pode ser potencialmente utilizado em programas biotecnológicos e de melhoramento genético. Deste modo, o conjunto de estudos que compõem a presente tese objetivou (i) identificar os principais mecanismos que suportam o grau de tolerância contrastante a toxicidade induzida por Cd em acessos de tomate após exposição a curto e longo prazos a este metal pesado; (ii) avaliar a relação entre o grau de tolerância e os atributos físico-químico de frutos oriundos de tomateiros cultivados em solo contendo Cd, e (iii) determinar os efeitos transgeracionais do estresse induzido por Cd. No primeiro experimento, nove acessos de tomateiro com graus variados de tolerância a exposição ao Cd, baseado na acumulação de biomassa, foram cultivados em solução hidropônica contendo 35 &mu;M de CdCl2 durante 6 dias. O impedimento de elevada concentração de magnésio (Mg) em raízes foi identificado como possível estratégia da planta para mitigar a toxicidade de Cd, por meio da evitação da formação de pelos radiculares. Em relação ao modo de ação da toxicidade induzida por Cd, o excesso de Mn, em adição a elevada concentração de Cd, parece estar acoplado aos danos foliares que são acentuados ainda mais pelas altas concentrações de zinco (Zn) e boro (B) nos tecidos fotossintéticos de plantas sob exposição ao Cd. No segundo experimento, os genótipos tolerantes (Yoshimatsu) e sensíveis (Tropic Two Orders) foram cultivados em solo contendo Cd, a fim de avaliar os parâmetros de produção. O genótipo tolerante apresentou frutos com maior diâmetro, altura e peso após o cultivo em solo contendo Cd, quando comparado as plantas controle. Em ambas as cultivares, a concentração de Cd variou de acordo com a seguinte ordem descendente: raízes = folíolos> (receptáculo floral, pedúnculo e sépalas) > caule = casca de fruta = polpa de fruta. Alem disso, dados sugerem que o receptáculo floral e suas estruturas atuaram como uma barreira ao transporte de Cd para os frutos, entretanto, ela não foi suficiente para evitar que o Cd atingisse os frutos. Em adição, a exposição ao Cd provocou notáveis reduções na concentração de Mg nas raízes de genótipos sensíveis e tolerantes, revelando que a aclimatação das plantas depende do baixo status de Mg em tecidos radiculares. Desde que ambas as cultivares são capazes de empregar este mecanismo, os dados sugerem que, durante a exposição a curto prazo ao Cd, acessos tolerantes são capazes de ativa-lo ou mais cedo ou mais rápido do que acessos sensíveis. Ademais, efeitos transgeracionais positivos na germinação e vigor das sementes do genótipo tolerante foram desencadeados pelo cultivo planta-mãe em solo com Cd, apesar do aumento de anormalidades cromossômicas. Este trabalho reportou novos conhecimentos sobre os efeitos da exposição ao Cd sobre o desenvolvimento do tomateiro, mecanismos de tolerância, qualidade e rendimento de frutos, bem como a distribuição de Cd dentro da planta.