Symbiotic nitrogen fixation occurs in plants harboring nitrogen-fixing bacteria within the plant tissue. The most widely studied association is between the legumes and rhizobia. In this relationship the plant (legumes) provides the bacteria (rhizobia) with reduced carbon derived from photosynthesis in exchange for reduced atmospheric nitrogen. This allows the plant to survive in soil, which is low in available of nitrogen. Rhizobia infect and enter plant root and reside in organs known as nodules. In the nodules the bacteria fix atmospheric nitrogen. The association between the legume, Medicago truncatula and the bacteria Sinorhizobium meliloti, has been studied in detail. Medicago mutants that have defects in nodulation help us understand the process of nitrogen fixation better. One such mutant is the Mtnip-1. Mtnip-1 plants respond to S. meliloti by producing abnormal nodules in which numerous aberrant infection threads are produced, with very rare rhizobial release into host plant cells. The mutant plant Mtnip-1 has an abnormal defense-like response in root nodules as well as defects in lateral root development. Three alleles of the Mtnip/latd mutants, Mtnip-1, Mtlatd and Mtnip-3 show different degrees of severity in their phenotype. Phylogenetic analysis showed that MtNIP/LATD encodes a protein belonging to the NRT1(PTR) family of nitrate, peptide, dicarboxylate and phytohprmone transporters. Experiments with Mtnip/latd mutants demonstrats a defective nitrate response associated with low (250 μM) external nitrate concentration rather than high (5 mM) nitrate concentration. This suggests that the mutants have defective nitrate transport. To test if MtNIP/LATD was a nitrate transporter, Xenopus laevis oocytes and Arabidopsis thaliana mutant plants Atchl1-5, defective in a major nitrate transporter AtNRT1.1(CHL1), were used as surrogate expression systems. Heterologous expression of MtNIP/LATD in X. laevis oocytes and Atchl1-5 mutant plants conferred on them the ability to take up nitrate from external media with high affinity, thus demonstrating that MtNIP/LATD was a high affinity nitrate transporter. Km for MtNIP/LATD was determined to be approximately160 μM in the X. laevis system and 113 μM in the Arabidopsis Atchl1-5 mutant lines thus supporting the previous observation of MtNIP/LATD being a high affinity nitrate transporter. X. laevis expressing the mutant Mtnip-1 and Mtlatd, were unable to transport nitrate. However X. laevis oocytes, expressing the less severe mutant allele Mtnip-3 were able to transport nitrate suggesting another role of the Mtnip/latd besides high affinity nitrate transport. Experimental evidence suggested that MtNIP/LATD might transport another substrate beside nitrate. MtNIP/LATD levels are regulated by phytohormones. Experiments performed with ABA (abscisic acid), IAA (indole acetic acid) and histidine as substrates in X. laevis system show that the MtNIP/LATD mRNA injected oocytes efflux IAA but do not transport histidine or ABA. When wild type A17 and mutant Mtnip-1 and Mtnip-3 plants, grown in the presence of different sources of nitrogen were screened in herbicide chlorate, a structural analog of nitrate, the A17 and Mtnip-3 mutant showed levels of susceptibility that was different from mutant Mtnip-1 lines. Evidence suggested that the amount of chlorate transported into the plants were regulated by the C:N status of the A17 and Mtnip-3 plants. This regulation was missing in the Mtnip-1 lines thus suggesting a sensor function of MtNIP/LATD gene.

The use of herbicides in cropping systems is routine in western Canada as is the practice of rotating crops between cereals, oilseeds and pulse crops. Often, herbicides that are appropriate one year in the crop rotation are not compatible with the following crop. Additionally, certain herbicides are designed to target certain enzyme pathways that can interfere with amino acid synthesis. These pathways also exist in the microbial community, including Rhizobium species. Rhizobia have a unique symbiotic relationship with legumes. In return for a carbon source, rhizobia not only fix atmospheric dinitrogen (N2) for the plant, but also can increase soil N reserves for the following year. With herbicides targeting amino acid synthesis in both plants and microbes, there is a possibility that N2 fixation may be inhibited by the application of certain herbicides.<p> This project was designed to examine possible negative effects of herbicide application on N2 fixation in field pea (Pisum sativum L.) and chickpea (Cicer arietinum L.). The study included field, growth chamber and laboratory experiments in which the effects of pre- and post-emergent herbicides, as well as herbicide residues in soil were examined.<p> In the field experiments, some early season measurements suggested that herbicide application had a negative impact on various growth and N2 fixation parameters. However, as the season progressed, plants recovered from early herbicide damage and N2 fixation ultimately was relatively unaffected. Growth chamber experiments similarly revealed that N2 fixation was largely unaffected by herbicide application when the application rates were relatively low (i.e., at rates intended to simulate partial herbicide breakdown, and thus lower than the recommended field rate). Although, N2 fixation was suppressed where high rates of herbicide (i.e., greater than recommended field rate) were applied, the efficiency of the rhizobia to fix N2, (i.e., the amount of N2 fixed per unit nodule mass), was unaffected. This along with a laboratory experiment which monitored growth of rhizobia in vitro, confirmed that rhizobia were not directly affected by the herbicides used in this study and that overall N2 fixation was not inhibited directly by the application of these herbicides. It was concluded that any negative impact on N2 fixation caused by herbicides used in this study, was related to the impact of the herbicide on crop growth, and was not due to any direct effects of the herbicide on the rhizobia.

The use of herbicides in cropping systems is routine in western Canada as is the practice of rotating crops between cereals, oilseeds and pulse crops. Often, herbicides that are appropriate one year in the crop rotation are not compatible with the following crop. Additionally, certain herbicides are designed to target certain enzyme pathways that can interfere with amino acid synthesis. These pathways also exist in the microbial community, including Rhizobium species. Rhizobia have a unique symbiotic relationship with legumes. In return for a carbon source, rhizobia not only fix atmospheric dinitrogen (N2) for the plant, but also can increase soil N reserves for the following year. With herbicides targeting amino acid synthesis in both plants and microbes, there is a possibility that N2 fixation may be inhibited by the application of certain herbicides.<p> This project was designed to examine possible negative effects of herbicide application on N2 fixation in field pea (Pisum sativum L.) and chickpea (Cicer arietinum L.). The study included field, growth chamber and laboratory experiments in which the effects of pre- and post-emergent herbicides, as well as herbicide residues in soil were examined.<p> In the field experiments, some early season measurements suggested that herbicide application had a negative impact on various growth and N2 fixation parameters. However, as the season progressed, plants recovered from early herbicide damage and N2 fixation ultimately was relatively unaffected. Growth chamber experiments similarly revealed that N2 fixation was largely unaffected by herbicide application when the application rates were relatively low (i.e., at rates intended to simulate partial herbicide breakdown, and thus lower than the recommended field rate). Although, N2 fixation was suppressed where high rates of herbicide (i.e., greater than recommended field rate) were applied, the efficiency of the rhizobia to fix N2, (i.e., the amount of N2 fixed per unit nodule mass), was unaffected. This along with a laboratory experiment which monitored growth of rhizobia in vitro, confirmed that rhizobia were not directly affected by the herbicides used in this study and that overall N2 fixation was not inhibited directly by the application of these herbicides. It was concluded that any negative impact on N2 fixation caused by herbicides used in this study, was related to the impact of the herbicide on crop growth, and was not due to any direct effects of the herbicide on the rhizobia.

Upwelling, atmospheric nitrogen (N2) fixation by cyanobacteria, and freshwater inputs from the Mississippi River system have been shown to stimulate new production by alleviating nitrogen (N) limitation in the northern Gulf of Mexico (GoM). Stable carbon (delta13C) and nitrogen (delta15N) isotopes were used to investigate whether these sources are utilized differentially by coastal and marine pelagic food webs. Particulate organic matter (POM), Trichodesmium, and zooplankton were collected from the Mississippi River plume and Loop Current (LC) which were detected using remote sensing data. Stable isotope values were used to separate coastal and marine water masses and environmental data (salinity, nutrient and pigment concentrations) allowed me to relate variability to the degree of freshwater influence. Published food web data from these two environments were then assessed to establish whether isotopic baseline shifts observed in our data occur at an ecosystem level. Isotope values of the POM and zooplankton were found to be significantly different between coastal and marine water masses. This was not the case for Trichodesmium whose isotope values were not significantly different between the two water masses. We found that marine water masses (sal > 35) exhibited silicate concentrations, cyanobacterial pigments and DIN: P that suggest an increased abundance of diazotrophs. In contrast, coastal water masses (sal < 35) exhibited increased diatom pigments and molar C:N indicating terrestrial sources fuel phytoplankton production. When published food web data were compared, we found producer and consumer delta15N values were enriched in the coastal compared to the marine environments. This work suggests that differences in delta15N values within my data set and published data reflect a shift in the use of biologically available N where higher trophic levels are sustained by diazotrophic activity in marine environments versus those supported by terrestrial sources in coastal ones. Food webs that have been constructed without considering Trichodesmium as a significant source of organic matter in the GoM should be reconsidered. By re-evaluating published data, this research gives insight into the early life ecology of larval fishes and works to help answer questions about the structure and function of pelagic food webs.

Le cycle océanique de l'azote est à l'origine de deux rétro-actions climatiques au sein du système terre. D'unepart, il participe au contrôle du réservoir d'azote fixé disponible au développement du phytoplancton et à lamodulation de la pompe biologique, un des mécanismes de séquestration du carbone anthropique. D'autre part,le cycle de l'azote produit un gaz à effet de serre et destructuer d'ozone, le protoxyde d'azote (N2O). L'évolutionfuture du cycle de l'azote sous l'influence du rechauffement climatique, de la déoxygénation et de l'acidificationdes océans reste une question ouverte. Les processus tels que la fixation d'azote, la dénitrification et laproduction de protoxyde d'azote seront modifiés sous l'influence conjuguée des ces trois stresseurs. Cesinteractions peuvent être évaluées grâce aux modèles globaux de biogéochimie marine. Nous utilisons NEMOPISCESet l'ensemble des modèles CMIP5 pour projeter les modifications des taux de fixation d'azote, denitrification, de production et des flux air-mer de N2O à l'horizon de 2100 en réponse au scénario 'business-asusual'.Les effets liés à l'action combinée du rechauffement climatique et de l'acidification des océans sur leréservoir d'azote fixé, la production primaire et la rétro-action sur le bilan radiatif sont également évalués danscette thèse.! / The marine nitrogen cycle is responsible for two climate feedbacks in the Earth System. Firstly, it modulates thefixed nitrogen pool available for phytoplankton growth and hence it modulates in part the strength of thebiological pump, one of the mechanisms contributing to the oceanic uptake of anthropogenic CO2. Secondly, thenitrogen cycle produces a powerful greenhouse gas and ozone (O3) depletion agent called nitrous oxide (N2O).Future changes of the nitrogen cycle in response to global warming, ocean deoxygenation and oceanacidification are largely unknown. Processes such as N2-fixation, nitrification, denitrification and N2Oproduction will experience changes under the simultaneous effect of these three stressors. Global oceanbiogeochemical models allow us to study such interactions. Using NEMO-PISCES and the CMIP5 modelensemble we project changes in year 2100 under the business-as-usual high CO2 emissions scenario in globalscale N2-fixation rates, nitrification rates, N2O production and N2O sea-to-air fluxes adding CO2 sensitivefunctions into the model parameterizations. Second order effects due to the combination of global warming intandem with ocean acidification on the fixed nitrogen pool, primary productivity and N2O radiative forcingfeedbacks are also evaluated in this thesis.

Philosophiae Doctor - PhD / Growth of symbiotic legume hosts is P limited, because of the high energetic requirements associated with N2 fixation. Attempts to overcome P deficiency in soils where legumes are grown involve addition of P-based fertilisers. However, these are produced from fmite, non-renewable resources that could be exhausted in the next 50-80 years. For this and other prudent reasons, viable alternatives are sought that include producing genetically enhanced plants with better P use efficiency (PUE). There exist some inter- and intraspecific genetic variation for associated traits of PUE in various legumes and these will have to be exploited to realize the development of P efficient cultivars. With the advent of sophisticated molecular tools, good progress has been made to understand the molecular response of some common physiological and morphological functions observed under LP. The research aims here were to investigate the energy costs and the alternative metabolic routes associated with C and N metabolism under LP in legumes, which is very scant in literature. We also investigated the recovery responses of nodulated roots upon P alleviation. Consequently, improvement strategies to produce legume varieties for better adaptation in poor P soils are envisaged. We have demonstrated varying degrees of sensitivity between the amide and ureide legume systems being investigated under short-term LP. The species-specific responses were ascribed to differences related to the agro-climatic origins, nodule morphologies and the type of N containing export product of the different legume types. These different responses also underscore possible different regulatory mechanisms under LP. Lupins were probed further, because of its apparent tolerance to P deficiency. Lupin nodules had between 3 to 5-fold higher Pj concentrations compared with soybeans under LP and HP, respectively. The maintenance of Pj levels, as oppose to a decline in the total P pool, is discussed in relation to its role in maintaining N2 fixation in lupins. Under LP, an effective Pj recycling mechanism in nodules is proposed to occur via the induction of the PEPc- MDH-ME route. This route also enhanced the capacity of root nodules to procure high malate concentrations that are used to fuel bacteroid respiration and N2 fixation. Two distinctly different cMDH proteins, one corresponding to HP and another corresponding to LP, were identified. The high malate concentrations reported here are speculated to have arisen through LP-induced cMDH. Metabolically available Pj decline developed gradually as P deficiency progressed. This coincided with a 15% decline in the %Ndfa. Moreover, under prolonged P deficiency the disproportionate synthesis of organic acids, most notably malate, that occurred at the expense of amino acids was proposed to account for this decline. The recovery in response to alleviation from LP involved alterations in the allocation of respiratory costs to growth and nutrient acquisition. Under LP, smaller nodules were formed and nodule metabolism revolved around accentuating PUE. Thus, there is considerable potential for improvement of P efficiency in legumes through manipulation of root: shoot partitioning.

Philosophiae Doctor - PhD / The tripartite symbiosis between Phaseolus vulgaris, arbuscular-mycorrhiza and the nodule bacteria, Rhizobia have been the focus of many studies ranging over a number of decades, however these studies have failed to answer certain questions relating the role of the symbionts in regard to host nutrition and the subsequent influence of these symbionts on the host C- economy. There is little doubt over the synergistic benefits involved in the dual inoculation of bean plants, as well as the resultant C-costs of maintaining the 2 symbionts, yet the specific contribution of the individual symbionts to the hosts overall nutrient and C-economy remain to be clarified. Thus the aim of this thesis is to help clarify these points by determining the symbiont induced photosynthetic, respiratory and nutritional changes taking place in the host. This was achieved by a series of experiments in which nodulated bean plants were split into two categories-those with and without AM colonized roots. These plants were then exposed to a range of growing conditions, including hi and low P, and a series of N treatments, ranging from zero N through to 3 mM NH/. Under these differing nutrient conditions growth, photosynthetic, respiratory, nutrient and amino acid responses were monitored, thus allowing for the determination of the symbionts influence on the host and the hosts reliance on the respective symbionts. Host reliance was noted most strongly under nutrient limiting conditions. Under low P treatment AM was the dominant symbiont as far as host C was concerned, allowing for the early establishment of the AM, thus ensuring the uptake of P for both host and nodule development. High P affected AM colonization to a greater extent than it did nodule dry weight and conversely the addition of N~ + led to a greater decrease in nodule dry weight than it did AM colonization. In spite of this decline, AM benefited the host by improving host N nutrition and relieving N-feedback inhibition of the export amino acid asparagine on BNF. These AM induced benefits did come at a cost to the host though, the dual inoculated plants had higher below ground respiratory costs and subsequently higher photosynthetic rates to compensate for the increased demand for C. The higher photosynthetic rates associated with dual inoculation were as a result of symbiont induced sink stimulation and not due to the improved nutrition of the host, as shown by the photosynthetic and nutrient response ratios. However, the respiratory costs associated with the uptake of soil nutrients were lower in AM colonized roots, thus showing an increased efficiency in nutrient gain by AM colonized roots. This improvement in host N nutrition as a result of AM colonization, coupled with the lower respiratory costs of AM nutrition led to the conclusion that under certain growing conditions nodules can become redundant and possibly parasitic.

Ce travail de thèse a pour but d'étudier la fixation d'azote marine dans les environnements riches mais déficitaires en azote (N), comparé au phosphore (P) dans un rapport N:P<16, grâce à l'utilisation d'approches complémentaire en culture in vitro et sur le terrain in situ. La première partie de ce travail a consisté à évaluer la réponse de la fixation d'azote de la cyanobactérie unicellulaire diazotrophe Crocosphaera watsonii face à des concentrations micromolaires en azote inorganique dissous (DIN) supposées inhiber l'activité de fixation d'azote : - suite à un apport sporadique, ou, - après une longue période d'acclimatation. Les résultats de ces études n'ont pas permis d'observer une inhibition des activités de fixation d'azote de cet organisme, laissant supposer que ce processus pourrait être actif dans une zone de l'Océan ayant ces même caractéristiques biogéochimiques : le Sud Est Tropical de l'Océan Pacifique (ETSP). En effet, cette zone est une des trois plus grandes zones de minimum d'oxygène (OMZ) de l'Océan et d'intenses processus de pertes de N (dénitrification et anammox) y ont lieu, résultant en un déficit de N par rapport au P. Des études présumaient que des processus inverses, de gains de N par la fixation d'azote, pourraient y être actifs mais aucune mesure à l'échelle du bassin n'y avait été faite car la fixation d'azote n'était supposée se produire que dans les environnements oligotrophes, comme les gyres subtropicaux. Dans le cadre d'un projet international, des missions océanographiques ont pu avoir lieu dans cette zone en Février 2010 pendant un évènement El Niño et en Mars-Avril 2011 pendant un évènement La Niña. / The objectif of these thesis was to study dinitrogen fixation in marine environments rich but deficient of nitrogen (N) compared to phosphorus (P) in a ratio N:P<16, by using complementary approaches in culture in vitro and in the field in situ. The first part of this work was to evaluate the response of nitrogen-fixing unicellular Cyanobacteria Crocosphaera watsonii faced with micromolar concentrations of dissolved inorganic nitrogen (DIN) supposed to inhibit nitrogen fixation activity : - after sporadic input, or, - after a long period of acclimatization. The results of these studies have failed to observe the inhibition of nitrogen fixation activities of this organism, suggesting that this process could be active in an area of the Ocean with these same biogeochemical characteristics : the Eastern Tropical South Pacific (ETSP). Indeed, this area is one of the three largest oxygen minimum zones (OMZ) of the Ocean, where intense processes of N losses (denitrification and anammox) took place, resulting in a deficit of N compared to P. Studies assumed that the inverse process, gain of N by nitrogen fixation, could be active in the ETSP but no measurements across the basin have been performed because nitrogen fixation was assumed to occur only in oligotrophic environments, such as the subtropical gyres. In the framework of an international project, cruises took place in this area in February 2010 during a El Niño event and in March-April 2011 during a La Niña event. Results of these two cruises have confirmed that nitrogen fixation was unexpectedly active with an intensity comparable to those reported in oligotrophic areas.

La fixation de N2, ou diazotrophie, est un processus biogéochimique majeur en raison de son apport en azote nouveau dans la couche de surface de l’océan. Cependant ses facteurs de contrôle restent mal connus. Le fer, dont les concentrations de surface sont faibles, est un élément potentiellement limitant de la fixation de N2 du fait du contenu en fer important de la nitrogénase. En raison de leur découverte récente, peu d’études ont été menées sur les cyanobactéries diazotrophes unicellulaires (UCYN) pouvant être responsables de ~50% de la fixation de N2 à l’échelle globale. Des expériences en culture ont permis de caractériser et quantifier pour la première fois la réponse d’une UCYN, Crocosphaera watsonii, face à la limitation en fer. En condition de limitation en fer, il a été observé une réduction de la croissance et des taux de fixation de N2 ainsi qu’une stratégie d’adaptation des cellules avec une diminution de leur volume. La stimulation de la croissance et de l’activité de C. watsonii cultivées en condition de limitation en fer suite à l’ajout d’une pluie saharienne artificielle a permis de mettre en évidence qu’une partie au moins du fer issu de poussières désertiques est biodisponible. En Atlantique subtropical Nord, où nous avons déterminé une forte contribution de la fixation de N2 à la production nouvelle, nous avons observé une limitation de la fixation de N2 et de la production primaire principalement par les phosphates et mis en évidence le rôle des métaux traces dans le contrôle de la fixation de N2. Un ajout de pluie saharienne a permis de stimuler systématiquement la fixation de N2 et la production primaire. / Despite the biogeochemical importance of N2 fixation, which represents the largest source of newly-Fixed nitrogen to the open ocean, some uncertainties remain about its controlling factors. Iron (Fe) is widely suspected as a key controlling factor due to the high Fe content of the nitrogenase complex and to its low concentration in oceanic surface seawaters. N2 fixation rates associated with unicellular N2 fixing cyanobacteria (UCYN) were estimated to be ~50% of the total N2 fixation at global scale, but as they have been recently discovered few studies have been conducted on these organisms. We performed culture experiments in order to quantify for the first time the response of an UCYN, Crocosphaera watsonii, to Fe limitation. Reduction of ambient Fe concentration led to significant decreases in growth rate and N2 fixation rates per cell and we observed an adaptive strategy to Fe limitation with a cell volume reduction. Then, the enhancement of growth and activity of C. watsonii under Fe limitation condition after artificial Saharan rain addition highlighted that at least a part of the Fe released by the dust is bioavailable. In subtropical North Atlantic, an important contribution of N2 fixation to new production was observed and we showed that primary production and N2 fixation were globally P-Limited. We revealed that trace metals play a key role in controlling N2 fixation in this area. Saharan rain addition stimulated N2 fixation, presumably by supplying these nutrients. All these results contribute to our knowledge of the control of oceanic N2 fixation and provide new insight about interactions between Fe, nitrogen and carbon biogeochemical cycles.

Obwohl Böden des Kronendachs (canopy soils) deutlich zur oberiridischen labilen Biomasse beitragen können, werden sie oft in Studien über Nährstoffkreisläufe übersehen. In Wäldern mit einem großen Vorkommen an Böden im Kronendach, wie beispielsweise jene in tropischen Bergregionen, könnte dies zu einem unvollständigen Verständnis der Gesamt-Nährstoffprozesse des Waldes beitragen. Böden im Kronendach sind Ansammlungen organischen Materials, welche gewöhnlich auf Zweigen von Bäumen tropischer Wälder zu finden sind. Sie bestehen in erster Linie aus zersetztem epiphytischen Material aber umfassen auch herunterfallendes Laub, Staub, wirbellose Tiere, Pilze und Mikroorganismen. Es gibt nur eine Handvoll Studien, die Stickstoff (N) Kreisläufe und/oder Treibhausgas (THG) Flüsse in Böden des Kronendachs untersucht haben und keine hat versucht die tatsächlichen Feldraten zu bestimmen oder herauszufinden, wie sich diese Böden – welche besonders sensibel gegenüber atmosphärischen Prozessen sind – mit Nährstoffdeposition ändern könnten. Diese Dissertation stellt die Ergebnisse einer Forschungsstudie dar, welche N-Umsatzraten und THG Flüsse von Böden des Kronendachs quantifiziert und untersucht, wie diese Raten durch zunehmende Mengen an N und Phosphor (P) im Boden verändert werden. In Gebieten mit atmosphärischer N- und P-Deposition, erhalten Böden des Kronendaches sowohl direkte als auch indirekte Nährstoffeinträge auf Grund von angereichertem Bestandsniederschlag und Pflanzenstreu. Es wurden folgende Umsatzraten in Böden des Kronendachs tropische Bergwälder entlang eines Höhengradienten (1000 m , 2000 m , 3000 m) gemessen: (1) asymbiotische biologische N2-Fixierung, (2) Netto- und Brutto-N-Transformation, und (3) Kohlendioxid (CO2), Methan (CH4) und Lachgas (N2O) Flüsse. Zudem wurden indirekte Auswirkungen von N-und P-Gaben, die auf dem Waldboden ausgebracht wurden, untersucht. Umsatzraten der N2-Fixierung, des N Kreislaufes und von THG Flüssen, welche in Böden des Kronendachs gemessen wurden, wurden mit denen vom Waldboden verglichen (entweder als Teil dieser Arbeit oder in parallelen Studien von zwei anderen Mitgliedern unserer Arbeitsgruppe), um die Aktivität von Böden des Kronendachs in den Kontext des gesamten Waldes zu stellen. N2-Fixierung wurde mit der Acetylenreduktionsmethode, Netto-N-Umsatzraten wurden mittels in situ Inkubationen (buried bag method) und Brutto-N-Umsatzraten wurden mit der 15N-Verdünnungsmethode (15N pool dilution technique) bestimmt. Gasflüsse wurden sowohl unter Verwendung statischer Kammern gemessen, deren Sockel permanent im Boden angebracht waren, als auch unter Verwendung regelmäßig entfernter intakter Bodenproben, die zur Gasmessung in luftdichten Einweckgläsern inkubiert wurden. Messungen der N2-Fixierung und des N Kreislaufes erfolgten während der Regen- und Trockenzeit im Feld unter Verwendung intakter Bodenproben. THG Messungen wurden fünf Mal während des Zeitraumes von einem Jahr durchgeführt. Der Waldboden unserer Standorte war 4 Jahre lang zweimal im Jahr mit moderaten Mengen an N ( 50 kg N ha-1 Jahr-1) und P (10 kg P ha-1 Jahr-1) gedüngt worden und umfasste folgende Behandlungen: Kontrolle, N-, P- und N+P-Zugaben. Das Kronendach trug 7-13 % zur gesamten Boden N2-Fixierung (Kronendach + Waldboden) bei, welche zwischen 0,8 und 1,5 kg N ha-1 Jahr-1 lag. N2-Fixierungsraten veränderten sich nur geringfüging mit der Höhenstufe, waren aber in der Trockenzeit deutlich höher als in der Regenzeit. N2-Fixierung im Waldboden wurde in N-Parzellen im Vergleich zu Kontroll- und P-Parzellen gehemmt, währen sie in Böden des Kronendachs in P-Parzellen im Vergleich zu Kontrollparzellen stimuliert wurde. Böden des Kronendachs trugen bis zu 23% zur gesamten mineralischen N-Produktion (Kronendach + Waldboden) bei; Brutto-N-Mineralisierung in Böden des Kronendachs lag zwischen 1,2 und 2,0 mg N kg-1 d-1. In Kontrollparzellen nahmen Brutto-Umsatzraten von Ammonium (NH4+) mit zunehmender Höhe ab, wohingegen Brutto-Umsatzraten von Nitrat (NO3-) keinen klaren Trend mit der Höhenstufe aufwiesen, aber signifikant durch die Saison beeinflusst wurden. Effekte durch Nährstoff-Zugabe unterschieden sich je nach Höhenstufe, aber kombinierte N+P-Zugabe erhöhte in der Regel auf allen Höhenstufen die N-Umsatzraten. CO2 Emissionsraten von Böden des Kronendachs berechnet auf der Basis der Fläche von Gaskammern (10,5 bis 109,5 mg CO2-C m-2 h-1) waren ähnlich denen vom Waldboden ähnlich und nahmen mit zunehmender Höhenstufe ab. Emissionen vom Kronendach, berechnet auf der Basis der Waldfläche (0,15 bis 0,51 Mg CO2-C m-2 h-1), machten jedoch nur 5-11% der gesamten Boden-CO2 Emissionen (Kronendach + Waldboden) aus. CH4 Flüsse (-0,07 bis 0,02 kg CH4-C ha-1 Jahr-1) und N2O Flüsse (0,00 bis 0,01 kg N2O-N ha-1 Jahr-1) von Böden des Kronendachs machten weniger als 5% der Gesamtflüsse von Böden aus. P-Zugabe reduzierte CH4 Emissionen in allen Höhenstufen, so dass Böden des Kronendachs als leichte CH4 Senken agierten (-10,8 bis -2,94 μg CH4-C m-2 h-1). Nur in 2000 m wurden Böden des Kronendachs unter N Zugabe zu leichten N2O Quellen (2,43 ± 3,72 μg N2O-N m-2 h-1), wohingegen P Zugabe die CO2 emissionen um ungefähr 50% reduzierte. Die Ergebnisse zeigen, dass Böden des Kronendachs eine aktive Mikrobengemeinschaft besitzen, welche wertvolle Dienstleistungen hinsichtlich von Nährstoffkreisläufen für das Ökosystem des Kronendachs erbringt. Zusätzlich, war der Nährstoffkreislauf der Böden des Kronendachs in unseren Wäldern eindeutig an die Nährstoffverfügbarkeit des Waldbodens gekoppelt, was im Gegensatz zu Theorien steht, die besagen dass Böden des Kronendachs vom Nährstoffkreislauf der Waldböden entkoppelt seien. Wir haben festgestellt, dass Böden des Kronendachs in höheren Lagen eher einen wesentlichen Anteil des gesamten Wald-Nährstoffkreislaufes ausmachen; dies sollte in Studien berücksichtigt werden, die sich mit Nährstoffkreisläufen solcher Gegenden beschäftigen. Langfristige atmosphärische N- und P-Deposition verfügt über das Potenzial, die Dynamik von Nährstoffflüssen im Kronendach erheblich zu verändern. N-Deposition könnte die N2-Fixierung hemmen, wobei “hotspots“ weiterhin in Bereichen mit großen Mengen an P vorkommen. Interne N-Kreisläufe in Böden des Kronendachs werden wahrscheinlich durch N -und P-Deposition stimuliert werden, aber chronischen Nährstoffzugabe könnte auch zu erhöhten mineralischen N-Verlusten aus dem Bodensystem des Kronendachs führen. THG-relevante Prozesse in Böden des Kronendachs werden wahrscheinlich auch auf N- und P-Deposition reagieren, aber mit Ausnahme von CO2-Emissionen ist es unwahrscheinlich, dass Gasflüsse von Böden des Kronendachs wesentlich zum gesamten THG-Budget des Waldes beitragen.