A large proportion of the world’s nitrogen needs is derived from symbiotic nitrogen fixation (SNF), which contributes substantially to agricultural sustainability. The partnership between legumes and rhizobia result in the formation of specialised structures called root nodules. Within these nodules SNF is supported by the sucrose transported from the leaves to the nodules for respiration. The end products of SNF in soybean (Glycine max (L.) Merr.) root nodules, namely ureides, are transported to the upper parts of the plant to supply nitrogen. Symbiotic nitrogen fixation provides a vital advantage for the production of soybean compared with most grain crops in that soybean fixes the nitrogen required for its growth and for the production of the high-protein content in seed and oil. The process of SNF is dramatically affected by drought, salt, cold and heavy metal stresses. Since SNF is such an important yield-determining factor, a lack in understanding these complexes inevitably delays progress towards the genetic improvement of soybean genotypes and also complicates decisions with regard to the suitability of certain genotypes for the various soybean producing areas in South Africa. The largest soybean producing areas in South Africa are situated at high altitudes, with minimum daily temperatures which can be critically low and impeding the production of soybean. Soybean is chilling sensitive, with growth, development and yield being affected negatively at temperatures below 15°C. Dark chilling (low night temperature) stress has proved to be one of the most important restraints to soybean production in South Africa. Among the symptoms documented in dark chilling sensitive soybean genotypes are reduced growth rates, loss of photosynthetic capacity and pigment content, as well as premature leaf senescence and severely inhibited SNF. Existing knowledge about stress-induced nodule senescence is based on fragmented information in the literature obtained in numerous, and often diverse, legume species. The precise nature and sequence of events participating in nodule senescence has not yet been fully explained. The main objectives of this investigation were to characterise the natural senescence process in soybean nodules under optimal growth conditions and to characterise the alteration of the key processes of SNF in a chilling sensitive soybean genotype during dark chilling. Moreover, to establish whether recovery in nodule functionality following a long term dark chilling period occured, or whether nodule senescence was triggered, and if sensitive biochemical markers of premature nodule senescence could be identified. A known chilling sensitive soybean genotype, PAN809, was grown under controlled growth conditions in a glasshouse. To determine the baseline and change over time for key parameters involved in SNF, a study was conducted under optimal growing conditions over a period of 6 weeks commencing 4 weeks after sowing. The cluster of crown nodules were monitored weekly and analysis included nitrogenase activity, ureide content, respiration rate, leghemoglobin content, sucrose synthase (SS) activity and sucrose content. Further investigations focused on induced dark chilling effects on nodule function to determine the alterations in key parameters of SNF. Plants were subjected to dark chilling (6˚C) for 12 consecutive nights and kept at normal day temperatures (26˚C). The induced dark chilling was either only shoot (SC) exposure or whole plant chilling (WPC). These treatments were selected since, in some areas in South Africa cold nights result not only in shoot chilling (SC) but also in low soil temperatures causing direct chilling of both roots and shoots. To determine if premature nodule senescence was triggered, the recovery following 12 consecutive nights of chilling treatment was monitored for another 4 weeks. It was established that the phase of optimum nitrogenase activity under optimal growing conditions occurred during 4 to 6 weeks after sowing where after a gradual decline commenced. This decline was associated with a decline in nitrogenase protein content and an increase in ureide content. The stability of SS activity and nodule respiration showed that carbon-dependent metabolic processes were stable for a longer period than previously mentioned parameters. The negative correlation that was observed between nitrogenase activity and nodule ureide content pointed towards the possible presence of a feedback inhibition trigger on nitrogenase activity. A direct effect of dark chilling on nitrogenase activity and nodule respiration rate led to a decline in nodule ureide content that occurred without any limitations on the carbon flux of the nodules (i.e. stable sucrose synthase activity and nodule sucrose content). The effect on SC plants was much less evident but did indicate that currently unknown shoot-derived factors could be involved in the minor inhibition of SNF. It was concluded that the repressed rates of respiration might have led to increased O2 concentrations in the nodule, thereby inhibiting the nitrogenase protein and so the production of ureides. It was found that long term chilling severely disrupted nitrogenase activity and ureide synthesis in nodules. Full recovery in all treatments occurred after 2 weeks of suspension of dark chilling, however, this only occurred when control nodules already commenced senescence. This points toward reversible activation of the nitrogenase protein with no evidence in support of premature nodule senescence. An increase in intercellular air space area was induced by long term dark chilling in nodules, specifically by the direct chilling of nodules (WPC treatment). The delayed diminishment of intercellular air space area back to control levels following dark chilling may be an important factor involved in the recovery of nitrogenase activity because enlarged air spaces would have favoured gaseous diffusion, and hence deactivation of nitrogenase, in an elevated O2 environment (due to supressed nodule respiration rates). These findings revealed that dark chilling did not close the diffusion barrier, as in the case of drought and other stress factors, but instead opened it due to an increase in air space areas in all regions of the nodule. In conclusion, this study established that dark chilling did not initiate premature nodule senescence and that SNF demonstrated resilience, with full recovery possible following even an extended dark chilling period involving low soil temperatures. / Thesis(PhD (Botany))--North-West University, Potchefstroom Campus, 2013

A fundamental goal in ecology and evolutionary biology has been to understand how microevolutionary forces affect the origin and maintenance of mutualisms over ecological and evolutionary time scales. Mutualistic partners vary in the reciprocal benefits they provide, yet the role of partner variation on microevolutionary forces that impact the maintenance of mutualisms is unclear. Using the mutualism between legumes and nitrogen fixing symbionts, my dissertation investigated the ecological and evolutionary consequences of variation in partner quality. In the first experiment, I demonstrate how insect herbivory can change the costs and benefits of associating with exploiters, and that some degree of exploitation from non-beneficial rhizobia can reduce insect herbivory, thus removing the fitness advantage of associating purely with beneficial rhizobia. In the second study, I examine how rhizobia genotype modifies competition between hosts grown in kin and non-kin groups. I show that lower fitness in plant kin groups can simply be a by-product of genetic variance in plant size and non-linear relationships between plant size and fitness. I further show that the symbiotic community can change difference in fitness between kin and non-kin groups independent of these by-product effects. In my last chapter, I provide the first empirical evidence that an important mechanism for mutualism stability-- the ability for hosts to preferentially associate with beneficial rhizobia-- is genetically variable and can evolve in response to exploitation. I also show that host preference for beneficial rhizobia can be maintained in legume populations, even in the absence of exploitation. My dissertation provides insight into the potential evolutionary dynamics of stabilizing mechanisms by suggesting that the agents of selection that affect the level of host exploitation can come from biotic factors other than the exploiters themselves. My dissertation has also shown that inclusion of other ecological interactions, such as herbivory, can provide valuable perspective on fitness effects of symbionts on their hosts, and can even change our fundamental assumptions about the effects of exploitation on host fitness, which has formed the backbone of mutualism theory.

A fundamental goal in ecology and evolutionary biology has been to understand how microevolutionary forces affect the origin and maintenance of mutualisms over ecological and evolutionary time scales. Mutualistic partners vary in the reciprocal benefits they provide, yet the role of partner variation on microevolutionary forces that impact the maintenance of mutualisms is unclear. Using the mutualism between legumes and nitrogen fixing symbionts, my dissertation investigated the ecological and evolutionary consequences of variation in partner quality. In the first experiment, I demonstrate how insect herbivory can change the costs and benefits of associating with exploiters, and that some degree of exploitation from non-beneficial rhizobia can reduce insect herbivory, thus removing the fitness advantage of associating purely with beneficial rhizobia. In the second study, I examine how rhizobia genotype modifies competition between hosts grown in kin and non-kin groups. I show that lower fitness in plant kin groups can simply be a by-product of genetic variance in plant size and non-linear relationships between plant size and fitness. I further show that the symbiotic community can change difference in fitness between kin and non-kin groups independent of these by-product effects. In my last chapter, I provide the first empirical evidence that an important mechanism for mutualism stability-- the ability for hosts to preferentially associate with beneficial rhizobia-- is genetically variable and can evolve in response to exploitation. I also show that host preference for beneficial rhizobia can be maintained in legume populations, even in the absence of exploitation. My dissertation provides insight into the potential evolutionary dynamics of stabilizing mechanisms by suggesting that the agents of selection that affect the level of host exploitation can come from biotic factors other than the exploiters themselves. My dissertation has also shown that inclusion of other ecological interactions, such as herbivory, can provide valuable perspective on fitness effects of symbionts on their hosts, and can even change our fundamental assumptions about the effects of exploitation on host fitness, which has formed the backbone of mutualism theory.

Marine nitrogen (N) is an essential nutrient for all oceanic organisms. The cycling of N between biologically available and unavailable forms occurs through numerous reactions. Because of the vast number of reactions and chemical species involved, the N cycle is still not well understood. This dissertation focuses on understanding some of the reactions involved in the cycling of marine N, as well as improving techniques used to measure dissolved N2 gas. The largest loss term for global marine N is a reaction called denitrification. In this work, denitrification was measured in the sandy sediments of the Georgia continental shelf, an area where this reaction was thought to be unlikely because of the physical properties of the sediments. Nitrogen fixation, which is a reaction that produces biologically available N, was detected in Georgia estuarine sediments. N fixation was measured concurrently with denitrification in these sediments, resulting in a much smaller net loss of marine N than previously thought. Lastly, membrane inlet mass spectrometry (MIMS) is a technique that measures dissolved N2, the end product of denitrification and a reactant in N fixation reactions. This study suggests that N2 measurements by MIMS are influenced by O2 concentrations due to pressure differences inside of the ion source of the mass spectrometer. These findings seek to improve denitrification measurements using MIMS on samples with varying O2 concentrations. In conclusion, this dissertation suggests that the marine N cycle is more dynamic than has been suggested, due to the recognition of input and loss reactions in a wider range of marine and estuarine environments. However, improvements in the understanding of MIMS will help with direct measurements with reactions involved in the global marine N cycle.

This thesis focuses on the application of nanocrystalline transition metal oxide TiO2, WO3 and NiO thin films in new “green” building technologies. Specifically, their physicochemical properties in photocatalytic, self-cleaning and gas sensing applications are studied. There is an intimate connection between comfort issues, health, with connections to energy efficiency, leading to a need for intelligent building materials and green architecture. The importance of good indoor environment is augmented by the fact that modern man in developed countries spends some 90 % of his time inside buildings and vehicles. Poor air quality may lead to discomfort of the person inhabiting a building and in ultimately cause adverse health effects. Thin films of nanocrystalline TiO2 were prepared using reactive DC magnetron sputtering. Crystalline mesoporous films of WO3 and NiO were prepared using advanced gas deposition technique (AGD). The crystal structure, morphology, optical and chemical properties of the films were characterized by using grazing incidence X-ray diffraction (GIXRD), scanning electron microscopy (SEM), UV/Vis spectroscopy and X-ray photoelectron spectroscopy (XPS), respectively. The photocatalytic properties and adsorption of both organic and inorganic molecules on pure and functionalized films were probed by in situ Fourier transform infrared spectroscopy (FTIR). The gas sensing properties of sensors based on TiO2, WO3 and NiO were investigated by conductivity measurements and noise spectroscopy. It was found for the first time that NiO based thin film sensors can be used to detect H2S and NO2 at low temperatures – down to room temperature. Hybrid WO3 sensors functionalized with multiwalled carbon nanotubes (MWCNTs) were used to detect NO2, CO and NH3 gases. These hybrid gas sensors show improved recovery properties compared to unmodified WO3 sensors. TiO2 based gas sensors were able to detect low concentrations of H2S by noise spectroscopy provided that the sensors were irradiated by UV light. Furthermore we show that sulphur is photo-fixated in crystalline TiO2 films upon simultaneous SO2 gas exposure and UV irradiation. Studies of the kinetics and identity of the photo-fixated sulphur complexes show that these are formed by photo-induced reactions between oxygen and SO2 at oxygen surface vacancy sites in TiO2. The sulphur modified TiO2 films show interesting self-cleaning properties compared to the pure films. / Felaktigt tryckt som Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 739

Diatoms play a fundamental role in marine food webs, and significantly contribute to global primary production and carbon sequestration into the deep ocean. In many offshore areas of the open ocean, iron (Fe) input is low, and its availability often limits phytoplankton biomass. Recently, gene sequences encoding ferritin, a nearly ubiquitous iron storage and detoxifying protein, have been identified in pennate diatoms such as Pseudo-nitzschia, but not in other Stramenopiles (which include centric diatoms, brown algae and some protist plant parasites) or Cryptophyte relatives. Members of this genus readily bloom upon addition of iron to Fe-limited waters, and are known to produce the neurotoxin domoic acid. Until now, the reason for the success of pennate diatoms in the open ocean was uncertain; however, expressing ferritin would allow pennate species to store Fe after a transient input, using it to dominate Fe stimulated algal blooms. Here, the ferritin gene was cloned from the coastal pennate diatom Pseudonitzschia multiseries, overexpressed in Escherichia coli, and purified using liquid chromatography. The ferritin protein sequence appears to encode a non-heme, ferritinlike di-iron carboxylate protein, while gel filtration chromatography and SDS-PAGE indicate that this ferritin is part of the 24 subunit maxi-ferritins. Spectroscopically monitoring the addition of Fe(II) to a buffered ferritin solution shows that the P. multiseries protein demonstrates ferroxidase activity, binding iron and storing it as Fe(III) in excess of 600 equivalents per protein shell. In keeping with the typical stoichiometry of the ferroxidase reaction, oxygen (O₂) is consumed in a 2 Fe:O₂ratio while hydrogen peroxide is produced concurrently. iii Diatoms evolved from secondary endosymbiosis involving eukaryotic red algae; however, a broad phylogenetic comparison suggests that P. multiseries ferritin was likely acquired via lateral gene transfer from cyanobacteria – not from its ancestral endosymbionts. Until recently, no other ferritins have been identified in diatoms, and the protein characterized here is unique in that it seems to be derived from a prokaryotic organism yet it occurs in a marine eukaryote. These findings have direct implications for the success of pennate diatoms in both Fe rich coastal waters and upon Fe addition in the open ocean.

Incldes bibliographical references / 1 v. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Agronomy, Waite Agricultural Research Institute, 1989

Hedgerows of the fodder tree legume Leucaena leucocephala (Lam.) de Wit ssp. glabrata (Rose Zárate) (leucaena) planted with companion grass (leucaena-grass pasture) form a productive, profitable and sustainable tropical pasture in northern Australia. Leucaena is renowned for its longevity (>30 years) and productivity under regular grazing, and this is a key factor in its profitability. To-date graziers and researchers have not been concerned about the sustainability of commercial leucaena-grass pastures, which are rarely fertilized. However, nutrient depletion could be expected after many years of nutrient removal under heavy grazing without replenishment, even on soils of moderate initial fertility. This study investigated the long-term productivity of leucaena-grass pastures in relation to nutrient depletion in Queensland. Experimental trials were conducted at 3 research stations and 6 commercial cattle properties. Prior to conducting field trials, a postal survey of leucaena growers ascertained perceived changes in leucaena, grass and livestock productivity over time. Physical and management factors influencing long-term pasture productivity were also explored. Graziers reported that leucaena productivity had declined in 58% of aging pastures. Lower livestock productivity was associated with declining leucaena growth, even though grass growth remained vigorous. Leucaena growth decline was more frequent on soil types of marginal initial fertility. Maintenance fertilizer was not applied to most (98%) leucaena pastures surveyed despite significant amounts of nutrient removal, particularly phosphorus (P) and sulphur (S), occurring over prolonged periods of moderate to high grazing pressure. It was predicted that under current management practices large areas of commercial leucaena pasture will be affected by soil nutrient depletion over the next 10 years. The effect of age of leucaena plants on pasture productivity was investigated in pastures aged from 8 to 38 years. Leucaena growth, expressed as rainfall use efficiency (RUE), declined with age (from 4.0 to 1.9 kg total dry matter (DM)/ha/mm), as did leaf nitrogen (N), P and S concentrations. Leucaena productivity decline was attributed to P and S deficiency restricting growth and symbiotic dinitrogen (N2) fixation. Composition of interrow grass changed from native grass dominance before leucaena establishment to green panic (Panicum maximum var. trichoglume) dominance in the aging leucaena pastures, particularly adjacent to leucaena hedgerows. This was attributed to increased soil Navailability. Leucaena and grass roots were concentrated in the topsoil; however, leucaena roots did extend beyond 1 m depth while grass roots did not. Changes in topsoil organic carbon (OC) and total nitrogen (TN) resulting from the planting of leucaena hedgerows into native grass pastures and previously cropped soils were studied. Topsoil OC and TN contents increased significantly under leucaena pasture (OC from 81-290 kg/ha/year and TN from 12-24 kg/ha/yr). Since TN and OC showed similar trends, there was no significant effect on carbon:N ratios. Leucaena contributed to soil OC both directly via plant part decomposition, and indirectly, via enhanced grass growth in the inter-row. Lower topsoil OC accumulation rates (81 kg/ha/yr) were observed in the older leucaena-grass pastures related to the decline in yield and vigour of leucaena in these aging pastures. The amount of carbon dioxide equivalent (CO2-e) accumulated in soil OC in productive leucaena-grass pasture was estimated to be higher than the amount of CO2-e emitted in methane from beef production from these pastures, thus positively impacting on their greenhouse gas balance. Leucaena responses to P and/or S applications were evaluated in a 30 year-old leucaenagrass pasture. Leucaena RUE and symbiotic N2 fixation were restricted by S deficiency. Sulfur concentration in leaf tissue and high N:S ratio were useful indicators of S deficiency. Although leucaena growth and its nutritional status were little affected by P application, symbiotic N2 fixation did respond significantly to P application. Leucaena and grass responses to fertiliser applications were further evaluated at a variety of soil types and environments on 8 sites in Queensland. Increased leucaena RUE (from 3.1 to 4.6 kg total DM/ha/mm) and enhanced nutritional status at most sites showed that leucaena plants were restricted by P and/or S deficiency. The major factors contributing to the P and S deficiencies were: a) inherent low soil P and/or S fertility, b) nutrient removal by cropping and grazing, c) shallow soils, d) acid soils, and e) grass competition for available water and nutrients. Inter-row cultivation (with or without fertiliser) had little effect on leucaena growth but significantly increased grass RUE (from 4.7 to 7.0 kg total DM/ha/mm) at some sites probably due to enhanced mineralization of N. Leaf P and S concentrations were not reliable indicators of deficiencies of these nutrients, possibly due to inadequate leaf sampling conditions. The effects of ambient temperature, water stress and phenological development of plant on nutrient concentrations in leucaena leaf was investigated to determine whether leaf tissueanalysis can reliably predict nutrient deficiencies. The youngest fully expanded leaf (YFEL) was established as the most appropriate leaf tissue for predicting nutritional status of leucaena plants since the YFEL: (a) was an easily identifiable tissue in which nutrient shifts were at a minimum; (b) provided information for readily mobile (N, P and potassium) and variably mobile (S, copper and zinc) nutrients, thus simplifying leaf collection; and (c) facilitated comparison of data from leaves of similar physiological age in different growing conditions and sites. Nutrient concentrations in YFEL were significantly influenced by water stress and phenological stage of plant development (particularly flowering and pod filling) through the mechanism of rate of leaf appearance. Chronological age of the YFEL, an indicator of leaf appearance, varied from 12 to >70 days depending upon plant phenological stage, being >140 days under prolonged water stress. It was found that nutrient concentrations in leucaena YFEL can only be interpreted against critical concentrations if plants are actively growing (October-April) in a vegetative stage and YFEL are <20 days old. This will occur if there is no water stress for ≥28 days prior to sampling. A close correlation existed between chronological age of YFEL and leaf calcium (Ca) concentration. Calcium concentration could be used to assess the age of YFEL and thereby determine the suitability of tissue samples for nutrient analysis and interpretation. Leaves with Ca concentrations >0.7% DM should be discarded as they are likely to be too old (>20 days). The research program has identified that leucaena established on non-alluvial soils need to be provided with regular maintenance P and S fertiliser to promote symbiotic N2 fixation and to maintain high RUE. At present, many leucaena pastures are likely to be suffering undiagnosed nutrient deficiencies that will be limiting pasture and animal productivity and enterprise profitability. Youngest fully expanded leaf analysis can be used as a predictive tool to diagnose nutrient deficiencies provided the recommended protocol is followed. Further investigation is required to: a) assess the duration of responses to applied fertiliser to determine frequency of application; b) investigate the rate of maintenance fertiliser P and S that has to be applied to maintain leucaena symbiotic N2 fixation and RUE at a desired level to benefit both forage quality and quantity, and soil fertility; c) study methods of fertiliser placement for adequate and timely supply of nutrients, particularly P, to leucaena roots; and d) confirm the use of Ca concentration in YFEL as a predictor of optimum leaf age for the range of soils and areas where leucaena is grown.

Thesis (M.P.H.)--University of Texas Health Science Center at Houston, School of Public Health, 2008. / Source: Masters Abstracts International, Volume: 46-06, page: 3262. Adviser: John R. Herbold. Includes bibliographical references.

Thesis (M.S.)--Medical College of Ohio, 2005. / "In partial fulfillment of the requirements for the degree of Master of Science in Biomedical Sciences." Major advisor: Nabil Ebraheim. Includes abstract. Document formatted into pages: iii, 57 p. Title from title page of PDF document. Bibliography: pages 35-42,49-56.