O nitrogênio é o nutriente exigido em maior quantidade pelo cafeeiro e o segundo mais exportado pela planta. Usualmente se aplica ureia no cafeeiro e o N desta fonte é rapidamente metabolizado e incorporado na forma de aminoácidos e amidas. A assimilação do nitrogênio é afetada por diversos micronutrientes, entre eles o níquel (Ni), por ser este um constituinte da enzima urease, responsável pela degradação da ureia. O entendimento da interação do Ni com o metabolismo do N e o processo de senescência no cafeeiro é importante para o uso eficiente do nitrogênio pelas plantas. Pouco se sabe a respeito da nutrição com Ni no metabolismo do N, na senescência foliar e na interferência na absorção e transferência de outros nutrientes. A presente pesquisa foi realizada para avaliar se a aplicação de Ni (i) interferiria na absorção e transferência de outros nutrientes, bem como na partição de biomassa do cafeeiro e; (ii) aumentaria a eficiência de uso do N, tanto pela maior degradação da ureia, via atividade da urease, quanto pelo aumento da redistribuição de reservas de nitrogênio, por meio do incremento do catabolismo da arginina; e (iii) se o fornecimento de Ni atrasaria a senescência foliar, pela diminuição da produção de etileno endógeno na planta, o que aumentaria a duração foliar. A aplicação de Ni reduziu a biomassa do cafeeiro somente no maior teor do elemento disponível no solo (105 mg DM-3), e essa redução na biomassa afetou a concentração e acúmulo, principalmente, dos macronutrientes (N, P, K, Ca e Mg). Com relação aos micronutrientes metálicos (Cu, Fe, Mn e Zn), o incremento gradual no teor de Ni disponível, reduziu ou elevou gradualmente a concentração desses micronutrientes no cafeeiro. Os coeficientes de eficiência de uso de N pelo cafeeiro foram afetados somente em razão do nível de N, sem alteração em função do Ni disponível no solo. A aplicação de Ni no solo até teores de 60 mg dm-3 não afetou o crescimento da planta de cafeeiro, mas aumentou a retenção foliar no nível deficiente de N. O Ni reduziu a biossíntese de etileno na planta, apesar das concentrações de MDA e prolina aumentarem com o teor disponível de Ni no solo na pré-antese. Com relação ao N, houve incremento na redistribuição de N dos órgãos de reserva (ramo) para atender a demanda de folha e fruto. / Nitrogen is the nutrient required in greatest quantity by the coffee and the second most exported within the plant. Usually, N is applied as urea to coffee and urea-N is rapidly metabolized and incorporated into amino acids and amides. The assimilation of N is affected by several micronutrients, including nickel (Ni). Ni is a constituent of urease, the sole enzyme responsible for degradation (and subsequent assimilation) of urea. Understanding Ni interaction with N metabolism and the aging process is important for the efficient use of nitrogen by coffee, and by plants in general. Little is known about Ni nutrition as it relates to N metabolism in leaf senescence and its possible interference in the absorption and transfer of other nutrients. The present study was conducted in coffee to assess whether the application of Ni (i) interferes with the absorption and transfer of other nutrients as well as biomass partitioning; (Ii) increases the nitrogen use efficiency, both for greater degradation of urea via urease activity, and increased redistribution of nitrogen reserves by increased catabolism of arginine; and (iii) delays leaf senescence by decreasing endogenous ethylene production, thereby increasing leaf duration. Ni application reduced biomass of coffee only at the highest level of soil availability (105 mg dm-3). This level also brought about the reduction in biomass concentration and accumulation of macronutrients (N, P, K, Ca and mg). With respect to mineral micronutrients (Cu, Fe, Mn and Zn), gradual increases in available Ni content, either reduced or gradually increased the concentration of these micronutrients. N-use efficiency ratios in coffee were affected only by variations in management of N, and not by changes in available soil Ni . Soil Ni applications to 60 mg dm-3 did not affect the growth of the coffee plant, but increased foliar retention under N limitation. Ni reduced plant ethylene biosynthesis, in spite of the concentrations of MDA and proline increasing rising with increasing soil Ni during pre-anthesis. As Ni levels rose, there was an increase in the redistribution of N of storage organs (branch) to meet the demand of leaf and fruit.

O nitrogênio é o nutriente exigido em maior quantidade pelo cafeeiro e o segundo mais exportado pela planta. Usualmente se aplica ureia no cafeeiro e o N desta fonte é rapidamente metabolizado e incorporado na forma de aminoácidos e amidas. A assimilação do nitrogênio é afetada por diversos micronutrientes, entre eles o níquel (Ni), por ser este um constituinte da enzima urease, responsável pela degradação da ureia. O entendimento da interação do Ni com o metabolismo do N e o processo de senescência no cafeeiro é importante para o uso eficiente do nitrogênio pelas plantas. Pouco se sabe a respeito da nutrição com Ni no metabolismo do N, na senescência foliar e na interferência na absorção e transferência de outros nutrientes. A presente pesquisa foi realizada para avaliar se a aplicação de Ni (i) interferiria na absorção e transferência de outros nutrientes, bem como na partição de biomassa do cafeeiro e; (ii) aumentaria a eficiência de uso do N, tanto pela maior degradação da ureia, via atividade da urease, quanto pelo aumento da redistribuição de reservas de nitrogênio, por meio do incremento do catabolismo da arginina; e (iii) se o fornecimento de Ni atrasaria a senescência foliar, pela diminuição da produção de etileno endógeno na planta, o que aumentaria a duração foliar. A aplicação de Ni reduziu a biomassa do cafeeiro somente no maior teor do elemento disponível no solo (105 mg DM-3), e essa redução na biomassa afetou a concentração e acúmulo, principalmente, dos macronutrientes (N, P, K, Ca e Mg). Com relação aos micronutrientes metálicos (Cu, Fe, Mn e Zn), o incremento gradual no teor de Ni disponível, reduziu ou elevou gradualmente a concentração desses micronutrientes no cafeeiro. Os coeficientes de eficiência de uso de N pelo cafeeiro foram afetados somente em razão do nível de N, sem alteração em função do Ni disponível no solo. A aplicação de Ni no solo até teores de 60 mg dm-3 não afetou o crescimento da planta de cafeeiro, mas aumentou a retenção foliar no nível deficiente de N. O Ni reduziu a biossíntese de etileno na planta, apesar das concentrações de MDA e prolina aumentarem com o teor disponível de Ni no solo na pré-antese. Com relação ao N, houve incremento na redistribuição de N dos órgãos de reserva (ramo) para atender a demanda de folha e fruto. / Nitrogen is the nutrient required in greatest quantity by the coffee and the second most exported within the plant. Usually, N is applied as urea to coffee and urea-N is rapidly metabolized and incorporated into amino acids and amides. The assimilation of N is affected by several micronutrients, including nickel (Ni). Ni is a constituent of urease, the sole enzyme responsible for degradation (and subsequent assimilation) of urea. Understanding Ni interaction with N metabolism and the aging process is important for the efficient use of nitrogen by coffee, and by plants in general. Little is known about Ni nutrition as it relates to N metabolism in leaf senescence and its possible interference in the absorption and transfer of other nutrients. The present study was conducted in coffee to assess whether the application of Ni (i) interferes with the absorption and transfer of other nutrients as well as biomass partitioning; (Ii) increases the nitrogen use efficiency, both for greater degradation of urea via urease activity, and increased redistribution of nitrogen reserves by increased catabolism of arginine; and (iii) delays leaf senescence by decreasing endogenous ethylene production, thereby increasing leaf duration. Ni application reduced biomass of coffee only at the highest level of soil availability (105 mg dm-3). This level also brought about the reduction in biomass concentration and accumulation of macronutrients (N, P, K, Ca and mg). With respect to mineral micronutrients (Cu, Fe, Mn and Zn), gradual increases in available Ni content, either reduced or gradually increased the concentration of these micronutrients. N-use efficiency ratios in coffee were affected only by variations in management of N, and not by changes in available soil Ni . Soil Ni applications to 60 mg dm-3 did not affect the growth of the coffee plant, but increased foliar retention under N limitation. Ni reduced plant ethylene biosynthesis, in spite of the concentrations of MDA and proline increasing rising with increasing soil Ni during pre-anthesis. As Ni levels rose, there was an increase in the redistribution of N of storage organs (branch) to meet the demand of leaf and fruit.

In durum wheat (<i>Triticum turgidum</i> L. var <i>durum</i>), grain protein concentration (GPC) and gluten quality are among the important factors influencing pasta-making quality. Semolina with high protein content produces pasta with increased tolerance to overcooking and greater cooked firmness. However, genetic improvement of GPC is difficult largely because of its negative correlation with grain yield, and a strong genotype x environment interaction. Therefore, identification of quantitative trait loci (QTL) for high GPC and the associated markers is a priority to enhance selection efficiency in breeding durum wheat for elevated GPC. At a physiological level, GPC is influenced by several factors including nitrogen remobilization from vegetative organs and direct post-anthesis nitrogen uptake (NUP) from the soil. Understanding the relationship between elevated GPC and nitrogen remobilization, and post-anthesis NUP will enable durum wheat breeders to develop varieties that not only produce high yield and high GPC, but also exhibit better nitrogen use efficiency. The objectives of this study were: (1) to identify and validate QTL for elevated GPC in two durum wheat populations; and (2) to determine if elevated GPC is due to more efficient nitrogen remobilization and/or greater post-anthesis NUP. A genetic map was constructed with SSR and DArT® markers in a doubled haploid population from the cross Strongfield x DT695, and GPC data were collected in replicated trials in six Canadian environments from 2002 to 2005. Two stable QTL for high GPC, QGpc.usw-B3 on chromosome 2B and QGpc.usw-A3 on 7A, were identified. Strongfield, the high GPC parent, contributed the alleles for elevated GPC at both QTL. These two QTL were not associated with variation in grain weight (seed size) or grain yield. QGpc.usw-A3 was validated in a second Strongfield-derived population as that QTL was significant in all six testing environments. Averaged over five locations, selection for QGpc.usw-A3 resulted in a +0.4% to +1.0% increase in GPC, with only small effects on yield in most environments. A physiological study of grain protein accumulation revealed that regardless of the growing condition, nitrogen remobilization was the major contributor for grain nitrogen in durum genotypes evaluated, accounting for an average of 84.3% of total GPC. This study confirmed that introgression of Gpc-B1 into Langdon resulted in increased GPC, and this GPC increase was due to higher N remobilization. Strongfield expressed greater N remobilization than DT695 and the semi-dwarf cultivar Commander, but N remobilization was not the determining factor for Strongfields elevated GPC. Strongfield expressed greater post-anthesis NUP than DT695. Similarly, a selection of six high-GPC doubled haploid (DH) lines from the cross DT695 x Strongfield expressed significantly greater post-anthesis NUP than six low-GPC DH selections, supporting the hypothesis that elevated GPC in Strongfield is derived from greater post-anthesis NUP. All six high-GPC DH selections carried the Strongfield allele at QGpc.usw-A3, suggesting this QTL maybe associated with post-anthesis NUP.

In durum wheat (<i>Triticum turgidum</i> L. var <i>durum</i>), grain protein concentration (GPC) and gluten quality are among the important factors influencing pasta-making quality. Semolina with high protein content produces pasta with increased tolerance to overcooking and greater cooked firmness. However, genetic improvement of GPC is difficult largely because of its negative correlation with grain yield, and a strong genotype x environment interaction. Therefore, identification of quantitative trait loci (QTL) for high GPC and the associated markers is a priority to enhance selection efficiency in breeding durum wheat for elevated GPC. At a physiological level, GPC is influenced by several factors including nitrogen remobilization from vegetative organs and direct post-anthesis nitrogen uptake (NUP) from the soil. Understanding the relationship between elevated GPC and nitrogen remobilization, and post-anthesis NUP will enable durum wheat breeders to develop varieties that not only produce high yield and high GPC, but also exhibit better nitrogen use efficiency. The objectives of this study were: (1) to identify and validate QTL for elevated GPC in two durum wheat populations; and (2) to determine if elevated GPC is due to more efficient nitrogen remobilization and/or greater post-anthesis NUP. A genetic map was constructed with SSR and DArT® markers in a doubled haploid population from the cross Strongfield x DT695, and GPC data were collected in replicated trials in six Canadian environments from 2002 to 2005. Two stable QTL for high GPC, QGpc.usw-B3 on chromosome 2B and QGpc.usw-A3 on 7A, were identified. Strongfield, the high GPC parent, contributed the alleles for elevated GPC at both QTL. These two QTL were not associated with variation in grain weight (seed size) or grain yield. QGpc.usw-A3 was validated in a second Strongfield-derived population as that QTL was significant in all six testing environments. Averaged over five locations, selection for QGpc.usw-A3 resulted in a +0.4% to +1.0% increase in GPC, with only small effects on yield in most environments. A physiological study of grain protein accumulation revealed that regardless of the growing condition, nitrogen remobilization was the major contributor for grain nitrogen in durum genotypes evaluated, accounting for an average of 84.3% of total GPC. This study confirmed that introgression of Gpc-B1 into Langdon resulted in increased GPC, and this GPC increase was due to higher N remobilization. Strongfield expressed greater N remobilization than DT695 and the semi-dwarf cultivar Commander, but N remobilization was not the determining factor for Strongfields elevated GPC. Strongfield expressed greater post-anthesis NUP than DT695. Similarly, a selection of six high-GPC doubled haploid (DH) lines from the cross DT695 x Strongfield expressed significantly greater post-anthesis NUP than six low-GPC DH selections, supporting the hypothesis that elevated GPC in Strongfield is derived from greater post-anthesis NUP. All six high-GPC DH selections carried the Strongfield allele at QGpc.usw-A3, suggesting this QTL maybe associated with post-anthesis NUP.

Master of Science / Department of Agronomy / Ignacio Ciampitti / Soybean [Glycine max (L.) Merr.] yields have continuously increased over time. Seed yields are determined by the genotype, environment, and management practices (G × E × M) interaction. Closing yield gaps require a continuous improvement in the use of the available resources, which must be attained via implementation of better management decisions. Linear relationships between seed yield and nitrogen (N) demand are reported in the scientific literature. Main sources of N to the plant are the biological N fixation (BNF) and the soil mineralization processes. On overall, only 50-60% of soybean N demand is met by the BNF process. An unanswered scientific knowledge is still related to the ability of the BNF to satisfy soybean N demand at varying yield levels. Seed N demand not met by N fixation plus soil mineral N, is then fulfilled by the remobilization of N from vegetative organs during the seed filling period. An early remobilization process reduces the photosynthetic activity (leaves) and can limit seed yield. The objectives of this project were to: i) study yield improvements and contribution of N via utilization of contrasting N conditions under historical and modern soybean genotypes, and ii) quantify main seed N sources during the seed filling period. For objective one, four field experiments were conducted during the 2016 and 2017 growing seasons in Kansas, United States (US) and Santa Fe Province, Argentina (ARG). Those experiments investigated twenty-one historical and modern soybean genotypes with release decades from 1980s to 2010s. As for objective two, three field experiments were conducted during the 2015 and 2016 growing seasons in Kansas, US, studying three soybean genotypes: non-roundup ready (RR), released in 1997; RR-1, released in 2009; and RR-2, released in 2014. Across all studies, seeds were inoculated and tested under three N management strategies: i) control without N application (Zero-N); ii) 56 kg N ha-1 applied at reproductive growth stages (Late-N); and iii) 670 kg ha-1 equally split at three timings (Full-N). As for yield improvements and N limitation, soybean yield improvements from the 1980s to 2010s were documented, representing 29% increases in the US and 21% in ARG. Regarding N management, the Full-N fertilization produced a 12% increase in seed yields in the US and 4% in ARG. As for main seed N sources in objective two, remobilization accounted for 59% of seed N demand, and was negatively related to new N uptake occurring during the seed filling period. Seed N demand for greater yields was dependent on both, N remobilization and new N uptake, while for lower yields, seed N demand was mainly supported by the N remobilization process. These results suggest that: a) high seed yields are somehow limited by the availability of N to express their potential, although the question about N application still remains to be fully investigated, as related to the timing and the environment by plant interactions that could promote a N limitation in soybeans; b) remobilization accounts for majority (59%) of N sourced to the seed, and c) high yielding soybean (modern genotypes) rely on diverse N sources: the N remobilization process plus new uptake of N.

L’orge (Hordeum vulgare L.) est l'une des céréales les plus importantes du monde et l’une des premières cultures domestiquées. Elle a été utilisée pendant des siècles pour l'alimentation humaine. Comme toutes les autres plantes, l'orge est dépendante de l'azote inorganique. L’efficacité de remobilisation de l'azote est donc très importante pour le remplissage des grains et pour la teneur en protéines du grain. L'objectif de ce travail est de donner une image du métabolisme des feuilles sénescence chez l'orge lorsque les plantes sont cultivées dans des conditions limitantes ou non en nitrates. Les analyses biochimiques, physiologiques et moléculaires de la sénescence des feuilles d'orge ont été réalisées. La gestion de l'azote pendant la sénescence des feuilles a été suivie par l'évolution des différents composés azotés au cours du vieillissement de la feuille. Une étude de profilage métabolique a été effectuée afin de déterminer les caractéristiques métaboliques de la sénescence des feuilles dans l'orge. En parallèle, les enzymes impliquées dans la remobilisation de l'azote ont été étudiées. Leurs activités et les niveaux de leurs transcripts ont été mesurés. Une attention particulière a été portée aux glutamine synthétases et asparagine synthétases et aux protéines de la machinerie de l'autophagie, processus connus pour jouer un rôle dans la remobilisation de l'azote pendant la sénescence des feuilles. A partir de toutes les données de séquences disponibles, ADNc, EST et séquences génomiques, cinq gènes codant pour les isoformes de glutamine synthétase cytosoliques (GS1), cinq gènes codant pour les isoformes d’asparagine synthétase (AS) isoformes et 19 gènes codant pour des protéines de la machinerie de l'autophagie ont été identifiés. Les expressions de tous les gènes identifiés ont été suivies au cours de la sénescence des feuilles et en fonction de l'alimentation en nitrates. La plupart de ces gènes sont sur-exprimés dans les feuilles sénescentes et de façon différentielle en fonction des conditions de nutrition. Toutes les données de séquences fournies par ce travail seront utiles à d'autres études translationelles et d'association génétique. / Barley (Hordeum vulgare L.) is one of the most important cereals in the world. It was one of the first domesticated crops and was used for centuries for human food. As all plants, barley has a fundamental dependence of inorganic nitrogen and nitrogen remobilization efficiency is very important for grain filling and grain protein content. The aim of this work was then to give a picture of the leaf-senescence metabolism in barley leaves when plants are grown under low or high nitrate conditions. Biochemical, physiological and molecular analyses of barley leaf senescence were performed. Nitrogen management during leaf senescence was monitored measuring changes in the different nitrogen pools during leaf ageing. In addition a large metabolite profiling study was performed in order to determine the metabolic hallmarks of leaf senescence in barley. In parallel enzymes involved in nitrogen remobilization were studied measuring their activity and the transcript levels of their coding genes. There was a special focus on glutamine synthetase and asparagine synthetase enzymes and for autophagy machinery that are known to play a role in nitrogen remobilisation during leaf senescence.From all the sequences data available, cDNA, EST and genomic sequences, we could identified five genes coding for cytosolic glutamine synthetase (GS1), five genes coding for asparagine synthetase (AS) and 19 genes coding for autophagy machinery proteins. Transcript levels of all the genes identified were monitored during leaf senescence and depending on nitrate nutrition. Most of these genes were over-expressed in senescing leaves and differentially expressed depending on nitrate conditions. In addition to the characterization of autophagy, GS1 and ASN genes, phylogenic and gene structures were analysed. All the sequences data provided by this work will be helpful to further translational and genetic association studies.

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.

Dans le cadre du changement climatique et de l'évolution de la réglementation concernant les intrants azotés, la culture du colza risque d'être fortement pénalisée dans la mesure où c'est une culture qui nécessite d'importants apports azotés pour atteindre son potentiel de rendement. Par ailleurs, comme chez le colza un déficit hydrique induit l'accumulation de certains composés azotés, il est vraisemblable que cela conduise au détournement d'une quantité importante d'azote vers les organes végétatifs aux dépens des organes reproducteurs et donc du rendement. Chez le colza, la réponse métabolique au déficit hydrique se traduit par une très forte accumulation de proline et dans une moindre mesure une augmentation de la teneur en GABA (acide γ-aminobutyrique), deux acides aminés connus chez la plupart des plantes pour leur réponse à de nombreux stress abiotiques. L'objectif de cette thèse est de déterminer comment le métabolisme de ces deux molécules contribue à l'allocation de l'azote au cours du développement de la plante en situation normale comme en condition de stress hydrique et/ou azoté. Pour répondre à cette question nous avons fait le choix de caractériser deux voies enzymatiques majeures impliquées dans le catabolisme de la proline et du GABA : la proline déshydrogénase (ProDH) et la GABA-transaminase (GABA-T) et d'évaluer l'impact de carences hydriques et/ou azotées sur ces voies. Cette étude nécessitait d'identifier au préalable les gènes codant ces enzymes afin d'aborder une approche fonctionnelle. Les résultats montrent l'existence de multiples copies de gènes ProDH et GABA-T dans le génome du colza. L'analyse de leurs profils d'expression suggère que des processus de sub-fonctionnalisation sont en cours conduisant à l'expression spécifique, de certaines copies en réponse aux stress, et d'autres dans les processus développementaux. La comparaison des profils métaboliques avec les profils spécifiques des transcrits a permis d'élaborer des hypothèses sur le rôle de ces voies dans la gestion de l'azote. L'étude conjointe des métabolismes de la proline et du GABA suggère l'existence de régulations connexes entre les deux. Enfin, l'utilisation de plantules a permis - d'approfondir la régulation des gènes étudiés à des stades précoces de développement - et de mettre en évidence les effets délétères de l'inhibition de la GABA-T par une approche pharmacologique. En conclusion ces résultats apportent des précisions sur la régulation de ces deux enzymes et fournissent des éléments de réponse quant au rôle fonctionnel des catabolismes de la proline et du GABA dans les processus de gestion de l'eau et de l'azote chez le colza. Ces travaux constituent donc une première étape dans une démarche de validation de ces gènes comme candidats pour des programmes d'amélioration du colza visant à sélectionner des génotypes mieux adaptés aux conditions environnementales futures. / In the context of climate change and recent regulation concerning nitrogen inputs, the oilseed rape yields may be severely decreased because its crop requires significant nitrogen supply to reach high yield performance. Moreover, as water deficit induces the accumulation of some nitrogen compounds in oilseed rape, it is likely that this could lead to diversion of significant amounts of nitrogen to the vegetative organs at the expense of the reproductive ones and therefore of the yield. In oilseed rape, the metabolic response to water deficit results in a very high proline accumulation and, to a lesser extent, an increased content of GABA (γ-aminobutyric acid), both these amino acids known for their response to many environmental stresses in most species. The objective of the work presented here was to determine how the metabolism of proline and GABA contributes to the nitrogen allocation during plant development under optimal conditions and under water stress and/or nitrogen depletion. To answer this question, we have chosen to characterize two major enzymatic pathways involved in the catabolism of proline and GABA, proline dehydrogenase (ProDH) and GABA transaminase (GABA-T), and assess the impact of water and/or nitrogen deficiency on these pathways. This study has required to preliminary identify the genes encoding these enzymes in order to initiate a functional approach. The results show the presence of multiple copies of ProDH and GABA-T genes in the oilseed rape genome. Analysis of their expression profiles suggests that sub-functionalization processes are occurring, leading to the specific expression of some copies in response to stress, and some in developmental processes. Comparison of metabolic profiles with specific profiles of transcripts allows us to hypothesize about the role of these pathways in management of nitrogen. The combined study of proline and GABA metabolisms suggests the existence of relationships between them. Finally, the use of seedlings allows - further studying the regulation of genes in the early stages of development - and highlighting the deleterious effects of the inhibition of GABA-T by a pharmacological approach. In conclusion these results supply information on the regulation of these two enzymes and provide answers about the functional roles of proline and GABA catabolisms in the management processes of water and nitrogen in oilseed rape. These works constitute a first step in validation process of these genes as putative candidates for oilseed rape breeding programs to select genotypes better adapted to future environmental conditions.