Conselho Nacional de Desenvolvimento Científico e Tecnológico / The rate of nitrogen (N) in grapevines in the Rio Grande do Sul (RS) State, Southern Brazil, is established considering N content in leaves or leaves petioles and yield expectation limits. Once N fertilization needs have been diagnosed, the N is applied in indefinite times. However, N fertilization recommendations don t consider the impact of N fertilization in the yield, the consequences on grape composition and the contribution of N of crop residues to mineral nutrition of grapevines. The objective of this work was to estimate the recovery and accumulation of N in grapevines and the effect in the yield and grape composition. Three studies were carried on. Study 1, recovery, distribution and accumulation of N fertilizer was studied, in Cabernet Sauvignon grapevines, located at Bento Gonçalves city, area of the Serra Gaúcha (experiment 1), and Santana do Livramento city, area of the Campanha (experiment 2), in RS, Southern Brazil. In the experiment 1 and 2 the plants were fertilized with 15N in four forms and, when grapes were mature, plants were fractionated, oven-dried, weighted and analysed for total N and 15N enrichment. Study 2, grape yield and composition of grapevines treated with different amounts of N were studied, in the vineyards of the Study 1. Grapes were collected when ripe and yield was evaluated. Must soluble solids, pH, total acidity, tartaric acid, malic acid, total phenols, anthocyanins and N ammonia were evaluated in part of the berries. The remaining berries were triturated and total N, P, K, Ca and Mg were evaluated. Study 3, decomposition and nutrient release by aerial organs of Lolium perenne (LP) and Trifolium repens (TR) (experiment 1), and grapevines uptake of the N derived from decomposing material (experiment 2) were studied in Chardonnay grapevines located at the experimental Station of the University of Bologna, Italy. In experiment 1, 2 g (DW) of 15N-labelled litter of LP and TR were placed into litter bags which were placed on the vineyard soil surface. Collection of bags LP and TR was performed 8 (June) and 16 (August) weeks from the deposition. Remaining biomass, 15N, N, C, P, K, Ca, Mg and S concentrations were determined. In experiment 2, 30 g plant-1 of dry litter of LP and TR were placed on the herbicided soil under the grape plants. Grapevine leaves were collected 8 and 16 weeks from litter deposition; after 16 weeks, one year (including bunches) and two years shoots were collected, oven-dried, weighted and analysed for total N and 15N enrichement. Results showed that the absorbed N is distributed and accumulated especially in leaves and that most of the N found in the plant was not derived from the fertilizers but from the organic matter already present in the soil. However, N fertilization increased grape yield in the Serra Gaúcha grapevines. The rate can be split 25% at the beginning of bud burst + 25% at bud burst + 25% at flowering + 25% at growth berries; 50% at the beginning of bud burst + 50% at bud burst and 50% at flowering + 50% at growth berries. Moreover, N fertilization altered the grape composition, especially decrease the amount of anthocyanins. LP and TR residues decomposed at the same rate, but nutrient release was different between the two species and varied according with mineral element. The amount of N that grapevines recovered from the decomposing litter was low and similar for LP and TR residues. / No Rio Grande do Sul (RS), a dose de nitrogênio (N) para a videira é definida a partir do seu teor total na folha inteira ou no pecíolo, e na expectativa de produção. Quando diagnosticada a necessidade de aplicação, recomenda-se o seu parcelamento em épocas incertas. Entretanto, essa recomendação não infere informações sobre o impacto do N aplicado na produção e na composição da uva. Também não se considera a contribuição do N dos resíduos de plantas de cobertura depositados na superfície do solo. O presente trabalho teve como objetivo estudar a recuperação e acumulação de N em videiras e seu efeito na produtividade e na composição da uva e do mosto. Foram conduzidos três estudos: Estudo 1, realizado para estimar a recuperação, distribuição e a acumulação de N aplicado ao solo pela videira. Esse foi composto por dois experimentos: Experimento 1, implantado em um vinhedo de Cabernet Sauvignon na Embrapa Uva e Vinho, Bento Gonçalves (RS), e o experimento 2, instalado na Empresa Pernod Ricard Brasil/Almadén, Santana do Livramento (RS). Nos experimentos 1 e 2 as videiras foram submetidas à aplicação de uma dose de N com 3% de átomos de 15N, em quatro modos de parcelamento. Na maturação da uva, as plantas foram colhidas, fracionadas, secas, determinada a matéria seca (MS) e analisado os totais de N e 15N. O Estudo 2 foi realizado para avaliar a produtividade e a composição da uva e do mosto, em videiras submetidas à aplicação de N. O Estudo 2 foi formado por dois experimentos: Experimento 1, implantado no vinhedo do experimento 1 do Estudo 1, as videiras receberam cinco doses de N e o experimento 2, instalado no vinhedo do experimento 2 do Estudo 1, as videiras foram submetidas à aplicação de seis doses de N. Nos experimentos 1 e 2 a uva foi colhida, determinada a produção e no seu suco avaliado os sólidos solúveis totais, o pH, a acidez total, o ácido tartárico e málico, os polifenóis totais, as antocianinas e o N amoniacal. Além disso, na baga foram analisados os totais de N, P, K, Ca e Mg. O Estudo 3 foi realizado para determinar a contribuição de N derivado da decomposição de resíduos de azevém (Lolium perenne) e trevo branco (Trifolium repens) para a videira. Esse compreendeu dois experimentos instalados em vinhedo de Chardonnay, em Bologna, Itália. No experimento 1, os resíduos de azevém perene e de trevo branco foram colocados no interior de bolsas teladas e depositados no solo. Ao longo do ciclo das videiras as bolsas teladas foram coletadas, determinada a MS remanescente e analisado os totais de 15N, N, C, P, K, Ca, Mg e S. No experimento 2, os resíduos das espécies foram colocados na superfície do solo e na maturação da uva as videiras foram colhidas, fracionadas, secas, determinado a MS e analisado os totais N e 15N. Os resultados obtidos mostram que o N adicionado e absorvido no ano é distribuído e acumulado, preferencialmente, nas folhas, mas a maior parte do N presente na videira é derivada de outras formas, que não a do fertilizante. Entretanto, existe aumento na produção quando da aplicação de N na Serra Gaúcha, podendo a dose ser parcelada 25% no início da brotação+25% na brotação+25% na floração+25% no crescimento das bagas; 50% no início da brotação+50% na brotação e 50% na floração+50% no crescimento das bagas. A aplicação de N altera a composição da uva, especialmente, diminui a quantidade de antocianinas no mosto. Os resíduos de azevém perene e trevo branco depositados na superfície do solo são decompostos com a mesma taxa no ciclo vegetativo e produtivo da videira, porém liberam nutrientes em taxas diferentes. No entanto, isso tem pouca importância, pois as plantas recuperam porcentagens pequenas e iguais de N derivado dos resíduos das duas espécies.

La nutrition minérale des plantes dépend à la fois du développement et du fonctionnement de leur appareil racinaire, incluant l'absorption mais aussi la capacité des plantes à influencer les cycles des nutriments, notamment par l'exsudation. Le but de cette thèse est de lier les rétroactions plantes-sol impliquant les cycles des nutriments aux stratégies d'exploration racinaire. Dans la revue bibliographique, je recense des mécanismes d'interaction plantes-sol et leurs échelles spatiales et temporelles. En considérant, à l'échelle de la rhizosphère, les interactions directes entre racines et sol, je propose que la combinaison entre exsudation et absorption des nutriments mène à des synergies entre racines d'une même plante. Ma seconde hypothèse est celle de l'existence d'un compromis entre l'exploration du sol et son occupation (défini comme la capacité des plantes à influencer efficacement le cycle des nutriments). Dans un premier chapitre, je développe un modèle général de recyclage des nutriments afin de déterminer sous quelles conditions les plantes auraient intérêt à limiter leur exploration du sol. Je montre qu'une exploration limitée est une stratégie de nutrition efficace sous certaines conditions, dont l'existence de synergies entre racines et le fait d'être dans un sol pauvre en nutriment. Dans un deuxième chapitre, je mesure le patron d'exploration racinaire et évalue le recyclage de l'azote à l'aide des outils isotopiques, chez trois espèces de Poacées pérennes de la savane de Hwange (Zimbabwe). Cette étude de terrain montre un gradient d'hétérogénéité racinaire entre ces trois espèces. Les Poacées exprimant le patron d'exploration le plus hétérogène ont un cycle de l'azote plus lent, mais potentiellement plus efficace. Dans un dernier chapitre, je développe un modèle mécaniste à l'échelle de la rhizosphère, pour une plante absorbant le phosphore et contrôlant sa disponibilité par l'exsudation de citrate. Je montre que, selon l'échelle d'influence des racines en terme d'exsudation et d'abaissement de la concentration en phosphore, la combinaison de l'exsudation et de l'absorption mène soit à une compétition, soit à une facilitation entre les racines d'une même plante. En me plaçant à l'échelle du système racinaire, je montre que les pertes en phosphore sont limitées par une exploration limitée du sol. Ce dernier résultat va dans le sens du compromis exploration/occupation. Au cours de cette thèse, j'ai donc développé des approches complémentaires, mettant en jeu différents mécanismes et échelles d'interactions plantes-sol. Le fait que les racines ne se limitent pas à un rôle d'absorption, mais agissent activement sur les cycles de nutriments a mené à deux résultats originaux : la facilitation inter-racinaire et intra-plante, et le fait qu'une exploration limitée puisse être considérée comme une stratégie efficace de nutrition. Enfin, ce travail souligne l'importance d'intégrer les divers mécanismes d'interaction plantes-sol pour comprendre les stratégies de nutrition des plantes et mieux prédire leur impact sur les cycles de nutriments à l'échelle des écosystèmes. / Plant nutrition depends on complementary mechanisms : the development of root systems, root uptake and plant ability to control nutrient cycling, e.g. through exudation. The aim of this thesis is to link plant-soil feedbacks involving the cycling of nutrients and root foraging strategies. I first review the different mechanisms of plant influence on nutrient cycling within the soil and assess their respective scales. Considering the direct effect of roots on the soil at the scale of the rhizosphere, I hypothesize that the combination of absorption and exudation may lead to synergies between the roots of a plant. At the scale of the whole root system, I propose a second, heuristic hypothesis: the existence of a trade-off between soil exploration and soil occupation (defined as the ability of plants to influence efficiently nutrient cycling). In a first chapter, I develop a general model of nutrient cycling, to determine under which condition plants should limit the exploration of soil by their roots. I show that limited exploration is an efficient strategy under specific conditions, especially nutrient-poor soils and the existence of synergies between roots. In a second chapter, I characterize soil occupation and nitrogen cycling, by the use of isotopes ratios, in the plant-soil system of three perennial grasses of the savanna of Hwange (Zimbabwe). This field study shows a gradient of root heterogeneity among these grass species. Those showing the more heterogeneous root pattern have a slower but potentially more efficient nitrogen cycling. In a last chapter, I develop a numerical mechanistic model at the rhizosphere scale for a plant taking up phosphorus and increasing its availability through exudation of citrate. I show that, depending on the extent of root influence on soil by exudation and nutrient depletion, competition between roots as well as facilitation arise from the combination of root uptake and exudation. By upscaling rhizosphere processes to the root system, I show that phosphorus losses are minimized by a restricted soil exploration, which backs the hypothesis of a trade-off between soil exploration and occupation. Overall, I developed complementary approaches that took into account several mechanisms and scales of plant-soil interactions. Considering that root functions are not limited to nutrient uptake, but also involve their influence on nutrient cycling, lead to two novel results: the potential existence of intra-plant and inter-root facilitation, and limited soil exploration as an efficient foraging strategy. This work underlines the importance of accurately integrating the mechanisms of plant-soil interaction to assess their nutrient strategies and to predict their impact on nutrient cycling within ecosystems.

No description available.

In der vorliegenden Arbeit wird die Darstellung neuer organischer Polyazide dokumentiert, die durch einfache nucleophile Substitution mittels NaN3 dargestellt werden können. Organische Azide mit der Formel RN3 können sich unter Stickstoff-Abspaltung in exothermen, teilweise explosionsartigen Reaktionen zersetzen. Sie sind daher prinzipiell als energiereiche Materialien (HEDM) für entsprechende Anwendungen geeignet. Die als Treibladungsmaterialien potentiell geeignetsten, handhabungssicheren, dendritischen Polyazide werden unter anderem mittels Thermogravimetrie und Differenzkalorimetrie analysiert. In einer neuen Synthesemethode können die wenig bekannten Heteroazidomethane aus Tris(azidomethyl)amin erzeugt werden. Von besonderem Interesse ist dabei die Synthese neuartiger Azidohalogenmethane. Diese können durch analytische Gas-Chromatographie charakterisiert und mittels präparativer Gas-Chromatographie isoliert werden. Durch die 1,3-dipolare Cycloaddition mit Cyclooctin konnten einige Heteroazidomethane zu Triazolen abgefangen und so einer Einkristall-Röntgen-Strukturanalyse zugeführt werden. Als letztes in der homologen Reihe der Azidomethane noch fehlendes Azid konnte Tetraazidomethan synthetisiert werden. Das Perazidomethan besitzt mit 93.3% den für organische Azide höchstmöglichen Stickstoffgehalt. Seine Existenz wurde bislang lediglich durch molekültheoretische Berechnungen nahegelegt. Die Synthese dieses homoleptischen Kohlenstoffazides gelang durch die Behandlung von Trichloracetonitril mit Natriumazid. Es ließ sich durch präparative GC als extrem explosive, farblose Flüssigkeit isolieren. Mit Hilfe der analytischen GC konnten sowohl der Siedepunkt als auch die Polarität von C(N3)4 abgeschätzt werden. C(N3)4 wird desweiteren durch IR, MS, 13C-NMR und 15N-NMR-Spektroskopie sowie durch Einkristall-Röntgen-Strukturanalysen seiner Abfangprodukte mit Cyclooctin charakterisiert. Mit Wasser zeigt C(N3)4 eine quantitative Hydrolyse unter Bildung von Carbonyldiazid. Durch Austauschprozesse mit Na15N3 konnte die mögliche Dissoziation von C(N3)4 nachgewiesen werden. Reaktionen von C(N3)4 mit Phosphinen führen zu Cyanamidderivaten, mit Norbornen sowie Norbornadien wurden über vielstufige Reaktionsmechanismen Aminotetraazole erhalten.

No description available.

Un nouveau modèle de biomembrane de type liposome a été développé à partir de lipides synthétisés comportant une unité biphényle sur leur chaînes sn2 et une chaîne aliphatique sn1 de longueur et insaturation variables. L’anisotropie de susceptibilité magnétique positive de ces molécules induit une déformation en oblate de ces liposomes dits « biphényles » dans le champ magnétique B0. Cette déformation spécifique a été caractérisée par RMN des solides 31P et 2H en faisant varier différents paramètres : l’intensité de B0, l'élasticité membranaire, la température et la taille des liposomes (Helfrich, 1973). Ces vésicules déformées ont pu être observées par microscopies optiques et électroniques et la rémanence de la déformation en dehors de B0 a pu être analysée par diffusion des rayons X aux petits angles (SAXS). Enfin, les premières applications des liposomes biphényles comme nouveau modèle de biomembrane pour analyser la structure et l’orientation (par RMN des solides 15N) de peptides ou protéines membranaires, ont été étudiées. / A new model of biomembrane (liposome) was developed from synthesized lipids containing a biphenyl unit on the sn2 aliphatic chain and possessing a sn1 aliphatic chain which varies in length and unsaturation. The positive magnetic susceptibility anisotropy of these molecules induces an oblate deformation of these «biphenyls » liposomes under the magnetic field B0. This particular deformation has been characterized by 31P and 2H solid state NMR by varying different parameters: the intensity of B0, the membrane elasticity, the temperature and the size of the liposomes (Helfrich, 1973). These deformed vesicles were observed by optical and electron microscopy and the remanence of the deformation outside B0 has been analyzed by Small angles X-ray scattering (SAXS).Finally, the first applications of biphenyls liposomes as new biomembrane model to analyze the structure and orientation of membrane proteins or peptides were studied by 15N solid state NMR

La présence de plantes accélère la décomposition de la matière organique du sol (MOS) au travers de l’apport de composés riches en énergie (rhizodépôts et litières) stimulant les microorganismes ; un phénomène appelé « rhizosphere priming effect » (RPE). Une augmentation de la photosynthèse, activité pourvoyeuse d'énergie rhizodéposée, pourrait augmenter le RPE et l’offre du sol en nutriments. Récemment, le modèle SYMPHONY couplant activités photosynthétiques et microbiennes du sol suggère un ajustement de l'offre du sol en nutriments (delta minéralisation-immobilisation) à la demande des plantes. Cependant, le rôle clé de la photosynthèse sur cet ajustement offre-demande reste à étudier expérimentalement.L’objectif général de la thèse est d'étudier le rôle des interactions des activités photosynthétiques et microbiennes du sol dans les régulations des flux de carbone (C) et d'azote (N) des écosystèmes. Trois écosystèmes types ont été étudiés : la prairie, la monoculture de blé et un nouveau système de culture (NSC) associant blé et plantes pérennes de la prairie. Nous émettons l’hypothèse que les plantes pérennes, via une activité photosynthétique pourvoyant les microorganismes en énergie tout au long de l'année, sont essentielles à l'ajustement offre-demande en N. De nombreux défis techniques ont été relevés afin de construire une plateforme expérimentale de 40 mésocosmes sous éclairage et température naturels. Cette plateforme permet de coupler marquage 13C des plantes, mesures continues des échanges de CO2, du RPE, de la production végétale, du stockage de C du sol, le taux de minéralisation-immobilisation d'N et du lessivage d'N. Ce dispositif nous a permis de déterminer la contribution du RPE dans les flux de C des écosystèmes comprenant la production nette de l’écosystème (NEP), la production primaire brute (GPP) et la respiration de l’écosystème (RE) exprimées en g C m-2 24h-1. Nous avons montré une relation positive linéaire entre (1) RPE et GPP et (2) RPE et biomasse aérienne (AGB) (g C m-2). A partir de ces relations, le RPE peut être prédit en utilisant les équations suivantes : (...). Nous montrons un ajustement offre-demande de l’N au cours des saisons : une forte activité photosynthétique (printemps) est liée à un RPE et un delta minéralisation-immobilisation d’N élevés alors qu’une faible activité photosynthétique (automne) est liée à un RPE et un delta minéralisation-immobilisation d’N faibles. Cet ajustement était observé dans la prairie et dans le NSC mais pas en monoculture de blé. Logiquement, la lixiviation d’N était importante en monoculture de blé alors qu’elle était quasi nulle en prairie et dans le NSC. Après deux années de maintien des trois écosystèmes types, la production aérienne totale du NSC était équivalente à la prairie, tous deux étant supérieurs d’environ un facteur deux à la monoculture de blé. Ces résultats confirment l’importance des plantes pérennes dans la synchronisation offre-demande de l’N. L’ensemble de ces investigations souligne l’importance des activités des plantes et des processus rhizosphériques dans la régulation des cycles CN des écosystèmes. Ces régulations pourront être étudiées in situ et à l'échelle globale grâce aux proxys de ces processus rhizosphériques (RPE, ajustement offre-demande) déterminés dans la thèse. Des activités photosynthétiques et rhizosphériques tout au long de l'année sont essentielles à l'ajustement offre-demande en nutriments conduisant à une forte production primaire, à la fermeture des cycles des nutriments et au stockage de MOS. Ces découvertes offrent l'opportunité de construire de nouveaux systèmes de culture, à l'image de l’association blé-plantes pérennes étudiée, à hautes performances agro-environnementales. / The presence of plants accelerates the decomposition of soil organic matter (SOM) through the supply of energy-rich compounds (rhizodeposits and litter) stimulating microorganisms; a phenomenon called rhizosphere priming effect (RPE). An increase of photosynthesis, supplying soil with rhizodeposited energy, could increase the RPE and soil nutrients offer. Recently, the SYMPHONY model coupling photosynthesis and soil microbial activities suggested an adjustment of the soil nutrient offer (delta mineralization-immobilization) to plant demand. However, the key role of photosynthesis in this offer-demand adjustment needs to be investigated experimentally.The general objective of the thesis is to study the role of interactions between photosynthesis and soil microbial activities in the regulation of carbon (C) and nitrogen (N) fluxes of ecosystems. Three ecosystem types were studied: grassland, wheat monoculture and a new cropping system (NSC) where wheat and perennial grassland species were intercropped. We hypothesize that perennial species, through a continuous photosynthetic activity supplying microorganisms with energy over the year, are essential for offer-demand adjustment.Many technical challenges were overcame to build an experimental platform of 40 mesocosms under natural light and temperature. This platform allows to couple 13C labeling of plants, continuous CO2-exchange measurements, RPE, plant production, soil C storage, N mineralization-immobilization turnover and N leaching.This experimental platform allowed us to determine the contribution of RPE to C fluxes of ecosystems including net ecosystem production (NEP), gross primary production (GPP) and ecosystem respiration (ER) expressed in g C m-2 24h-1. We found positive linear relationships between (1) RPE and GPP and (2) RPE and aboveground biomass (AGB) (g C m-2). Using these relationships, the RPE can be predicted with the following equations: (...).We show an adjustment of soil N-offer to plant N-demand across seasons: a high photosynthetic activity (spring) is linked to high RPE and delta mineralization-immobilization of N whereas a low photosynthetic activity (autumn) is linked to low RPE and delta mineralization-immobilization of N. This adjustment was observed in grassland and NSC but not in wheat monoculture. Consistently, N leaching was high in wheat monoculture while it was almost null in grassland and NSC. After two years of establishment of the three ecosystems, the total aboveground production of the NSC was equivalent to the grassland, each being about twice as high as the wheat monoculture. These results confirm the importance of perennial species in the offer-demand adjustment of N.Our findings underline the importance of plant activities and rhizosphere processes in the regulation of ecosystems C N cycles. Using the proxies of rhizosphere processes (RPE, offer-demand adjustment) provided in the thesis, further studies could investigate these regulations in situ and at the global scale. The presence of photosynthetic and rhizospheric activities over the year are essential for offer-demand adjustment of nutrients leading to high primary production, closing nutrient cycles and SOM storage. These findings offer the opportunity to build new cropping systems such as the wheat-perennial species studied, with high agro-environmental performances.

No cultivo do milho safra, os produtores em sistema de produção em plantio direto (SPD) antecipam a dose da adubação nitrogenada de cobertura, que seria feita nos estádios V3 e V6, na cultura da aveia durante a entressafra ou em présemeadura do milho. Não se conhece a eficiência destes sistemas de adubação do milho. Assim, esta pesquisa foi realizada com a finalidade de elucidar a dinâmica do nitrogênio (N) por meio da avaliação do seu aproveitamento nos compartimentos solo, milho e no resíduo da aveia, e determinar a eficiência de uso do fertilizante nitrogenado (EUFN). O experimento foi realizado nos municípios de Guarapuava-PR e Taquarituba-SP, com a aplicação de 180 kg ha-1 e 150 kg ha-1 de N, respectivamente. O delineamento experimental foi em blocos ao acaso com cinco sistemas de adubação e quatro repetições. Os sistemas de adubação foram: N no perfilhamento da aveia; N na pré-semeadura do milho; N na semeadura do milho; N em cobertura no estádio V3 do milho; e N em duas coberturas, nos estádios V3 e V6 do milho. Em ambos locais não houve diferença na produtividade de grãos, em razão do sistema de adubação, em Guarapuava a produtividade média foi de 14,2 Mg ha-1 de grãos, e em Taquarituba 10,6 Mg ha-1 de grãos. A antecipação do N na aveia reduziu a EUFN em até 41%, com maior dependência do N fornecido pelo solo, e foi o sistema de adubação que mais perdeu 15N-fertilizante, uma vez que não recuperou 50% do 15N aplicado. Os sistemas de adubação, em que o 15N foi aplicado no estádio V3 ou nos estádios V3 e V6 foram mais eficientes, com EUFN de 51% e 55%, nessa ordem. A avaliação da produção de grãos não serve para avaliar a eficiência de sistema de adubação. Os sistemas de adubação antecipado na cultura do milho não são eficientes no uso do N proveniente do fertilizante. / In maize crop, sidedress nitrogen fertilization in V3 or V6 phenological stages was moved to cultivated oat during off season or in pre-sowing by farmers that adopt the no-tillage maize system. However, the efficiency of this fertilization system is unknow. This research was designed to understand nitrogen (N) dynamic by evaluating the recovery in soil, maize plant and oat straw to calculate the N use efficiency (NUE) by maize crop. Maize crop fields experiments were conducted in Guarapuava, Paraná state and Taquarituba, São Paulo state, where fertilization rate was 180 kg ha-1 and 150 kg ha-1 of N respectively. Experimental desing was randomized block with four repetitions and five fertilization systems: N in oat tillering; N in pre-sowing corn, N in sowing corn; N sidedress in V3 and N sidedress in V3 and V6. Grain yield, with average 14,2 Mg ha-1 in Guarapuava and 10,6 Mg ha-1 in Taquarituba were not statiscally different. N fertilization in oat showed NUE 41% lower due to N soil contribution and high N loss with 50% 15N unaccounted. Fertilization systems which N-fertilizer was applied sidedress in V3 or V3 and V6 maize stage were more efficient with NUE 51% and 55%, respectively. Grain yield was not able to measure the efficiency of fertilization system. The anticipation of N fertilization in maize crop had low N use efficiency from N derived from fertilizer..

Das Biopolymer-produzierende Knallgasbakterium Ralstonia eutropha H16 gilt mit seinem außergewöhnlichen Stoffwechsel als vielversprechender Produktionsstamm für die weiße Biotechnologie. Es wächst auf einer Vielzahl organischer Substrate sowie chemolithoautotroph mit H2 und CO2 als einzige Energie- bzw. Kohlenstoffquelle. Unter anaeroben Bedingungen ist es zudem zur Denitrifikation befähigt. In dieser Arbeit wurde das Proteinprofil von R. eutropha unter chemolithoautotrophen sowie anaeroben Bedingungen mittels GeLC-MS/MS untersucht. Beide Proteomstudien offenbarten, dass die Nutzung unterschiedlicher Elektronendonoren bzw. -akzeptoren mit zahlreichen Veränderungen im Proteinbestand der Zellen einherging. Hierbei waren neben Proteinen metabolischer und Transportprozesse auch jene der Zellbewegung betroffen. Die Ergebnisse stellen im Vergleich zu vorangegangenen Studien den bisher umfassendsten Überblick zum Proteinbestand beim H2-basierten sowie anaeroben Wachstum in R. eutropha dar. Von besonderer Bedeutung war dabei das Einbinden der Analyse der Membran als Ort wichtiger Energie- und Transportprozesse. Besonderes Interesse galt einem unter H2/CO2-Bedingungen abundanten Zweikomponentensystem. Sequenzvergleiche zeigten Ähnlichkeit zum Regulationssystem der Katabolitrepression des Biphenylabbaus in Acidovorax sp. KKS102. Die Deletion des Response-Regulator-Gens führte zu vielfältigen Wachstumseffekten auf Substraten wie Fructose, Glycerin sowie auf H2/CO2. Der pleiotrope Phänotyp sowie die Ergebnisse von Genexpressionsstudien und der Suche nach Regulator-Bindestellen lassen eine globale Rolle des Systems im Energie- und/oder Kohlenstoffmetabolismus von R. eutropha H16 annehmen. Histidin-Kinase und Response Regulator wurden in GloS bzw. GloR umbenannt. Die vorliegende Arbeit zeigt eindrucksvoll das Potential der Proteomik als Teil der funktionellen Genomik für den Anstoß neuer Forschungsansätze zur Evaluierung des biotechnologischen Potentials von Mikroorganismen. / Due to its remarkable metabolism the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16 is ranked as a promising production strain for white biotechnology. It grows on a wide range of organic substrates as well as lithoautotrophically on H2 and CO2 as sole energy and carbon source, respectively. Under anaerobic conditions it thrives by denitrification. This thesis focused on characterizing the protein profiles of lithoautotrophically and anaerobically grown R. eutropha cells. Proteome analyses revealed an extensive protein repertoire adapting the organism to alternative electron donors and acceptors, respectively. Changes concerned proteins involved in metabolic and transport processes as well as in cell movement. Compared to previous studies the results reported here offer the most comprehensive proteomic survey regarding the H2-based as well as anaerobic lifestyle of R. eutropha so far. In this context analyzing the cell membrane as a place for a number of energy, transport and signal transduction processes was of particular importance. Special interest aroused the identification of a two-component system upregulated on H2/CO2. Sequence analysis offered high similarity to the regulatory system for catabolite control of biphenyl degradation in Acidovorax sp. KKS102. Deletion of the response regulator gene led to versatile growth effects on substrates such as fructose and glycerol as well as H2/CO2. This pleiotrophic phenotype as well as the results of gene expression studies and the search for regulator binding sites suggests that the two-component system is a global player in energy and/or carbon metabolism in R. eutropha and possibly other bacteria. Thus, histidine kinase and response regulator have been renamed GloS/R. Since their characterization was initiated by proteomic data this study impressively elucidates the power of functional genomics in terms of revealing new research approaches to evaluate the biotechnological use of microbes.

No cultivo do milho safra, os produtores em sistema de produção em plantio direto (SPD) antecipam a dose da adubação nitrogenada de cobertura, que seria feita nos estádios V3 e V6, na cultura da aveia durante a entressafra ou em présemeadura do milho. Não se conhece a eficiência destes sistemas de adubação do milho. Assim, esta pesquisa foi realizada com a finalidade de elucidar a dinâmica do nitrogênio (N) por meio da avaliação do seu aproveitamento nos compartimentos solo, milho e no resíduo da aveia, e determinar a eficiência de uso do fertilizante nitrogenado (EUFN). O experimento foi realizado nos municípios de Guarapuava-PR e Taquarituba-SP, com a aplicação de 180 kg ha-1 e 150 kg ha-1 de N, respectivamente. O delineamento experimental foi em blocos ao acaso com cinco sistemas de adubação e quatro repetições. Os sistemas de adubação foram: N no perfilhamento da aveia; N na pré-semeadura do milho; N na semeadura do milho; N em cobertura no estádio V3 do milho; e N em duas coberturas, nos estádios V3 e V6 do milho. Em ambos locais não houve diferença na produtividade de grãos, em razão do sistema de adubação, em Guarapuava a produtividade média foi de 14,2 Mg ha-1 de grãos, e em Taquarituba 10,6 Mg ha-1 de grãos. A antecipação do N na aveia reduziu a EUFN em até 41%, com maior dependência do N fornecido pelo solo, e foi o sistema de adubação que mais perdeu 15N-fertilizante, uma vez que não recuperou 50% do 15N aplicado. Os sistemas de adubação, em que o 15N foi aplicado no estádio V3 ou nos estádios V3 e V6 foram mais eficientes, com EUFN de 51% e 55%, nessa ordem. A avaliação da produção de grãos não serve para avaliar a eficiência de sistema de adubação. Os sistemas de adubação antecipado na cultura do milho não são eficientes no uso do N proveniente do fertilizante. / In maize crop, sidedress nitrogen fertilization in V3 or V6 phenological stages was moved to cultivated oat during off season or in pre-sowing by farmers that adopt the no-tillage maize system. However, the efficiency of this fertilization system is unknow. This research was designed to understand nitrogen (N) dynamic by evaluating the recovery in soil, maize plant and oat straw to calculate the N use efficiency (NUE) by maize crop. Maize crop fields experiments were conducted in Guarapuava, Paraná state and Taquarituba, São Paulo state, where fertilization rate was 180 kg ha-1 and 150 kg ha-1 of N respectively. Experimental desing was randomized block with four repetitions and five fertilization systems: N in oat tillering; N in pre-sowing corn, N in sowing corn; N sidedress in V3 and N sidedress in V3 and V6. Grain yield, with average 14,2 Mg ha-1 in Guarapuava and 10,6 Mg ha-1 in Taquarituba were not statiscally different. N fertilization in oat showed NUE 41% lower due to N soil contribution and high N loss with 50% 15N unaccounted. Fertilization systems which N-fertilizer was applied sidedress in V3 or V3 and V6 maize stage were more efficient with NUE 51% and 55%, respectively. Grain yield was not able to measure the efficiency of fertilization system. The anticipation of N fertilization in maize crop had low N use efficiency from N derived from fertilizer..