Short-term nutritional supplementation during management of pulmonary exacerbations in cystic fibosis: A controlled study, including effects of protein turnover

Effects of nutritional supplements on minimizing weight loss and abnormalities of protein turnover during pulmonary exacerbations in cystic fibrosis (CF) were studied by controlled trial. Patients received pulmonary therapy and either standard diet (n = 10) or adjunctive enteral supplements (n = 12). Initial protein turnover, measured by [15N]glycine kinetics, showed alterations of protein synthesis (P Syn) and catabolism (P Cat), which correlated with the degree of underweight, and negligible net protein deposition (P Dep). With treatment both groups had significant increases in mean body weight and forced expiratory volume in 1 s, expressed as percent predicted value for height (FEV1) by 3 wk, but a significant correlation between initial underweight and subsequent weight gain was observed only in supplemented patients. Mean P Syn and P Dep increased significantly (p < 0.001) only in the supplemented group. Pulmonary exacerbations in CF have important adverse effects on body-protein metabolism, similar to changes in protein-energy malnutrition and infection. These effects are reversed by short-term nutritional support. Strategic nutritional intervention should thus be considered in management, especially in malnourished patients.

Cannibalism contributes significantly to the diet of cultured sand crabs, Portunus pelagicus (L.): A dual stable isotope study

The significance of cannibalism in the diet of juvenile pond-cultured blue swimmer crabs (Portunus pelagicus (L.)) was investigated using dual stable isotope analysis of carbon and nitrogen. In a laboratory feeding experiment, δ15N demonstrated a constant trophic shift (Δδ15N ≈+ 1.6‰), and therefore seemed to be a reliable indicator for assessing trophic position for P. pelagicus. This agrees with previously reported trends. Difference in growth rate did not seem to influence δ15N values. In contrast, δ13C did not display consistent shifts between trophic levels (range of Δδ13C: + 1 to + 1.7‰). The results from the pond experiment showed that larger individuals had a more enriched δ15N than smaller individuals, which, when compared to the results from the laboratory experiment, indicates that larger individuals were at a higher trophic level. This is most likely due to cannibalism prevailing in the pond rather than a direct result of faster growth rate. Cannibalistic behaviour might further increase growth, resulting in the observed positive correlation between size and δ15N.

Metabolite mobilization in the stem galls of Parthenium hysterophorus induced by Epiblema strenuana inferred from the signatures of isotopic carbon and nitrogen and concentrations of total non-structural carbohydrates

Parthenium hysterophorus L. (Asteraceae) is a weed of national significance in Australia. Among the several arthropod agents introduced into Australia to control populations of P. hysterophorus biologically, Epiblema strenuana Walker (Lepidoptera: Tortricidae) is the most widespread and abundant agent. By intercepting the normal transport mechanisms of P. hysterophorus, the larvae of E. strenuana drain nutrients, other metabolic products, and energy, and place the host plant under intense metabolic stress. In this study, determinations of total non-structural carbohydrates (TNC) levels and carbon and nitrogen isotope ratios of fixed products in different parts of the plant tissue, including the gall, have been made to establish the function of gall as a sink for the nutrients. Values of δ13C and δ15N in galls were significantly different than those in proximal and distal stems, whereas the TNC levels were insignificant, when measured in the total population of P. hysterophorus, regardless of plant age. However, carbon, nitrogen, and TNC signatures presented significant results, when assayed in different developmental stages of P. hysterophorus. Carbon isotope ratios in galls were consistently more negative than those from the compared plant organs. Nitrogen isotope ratios in galls, on the contrary, were either similar to or less negative than the compared plant organs, especially within a single host-plant stage population (i.e., either rosette, preflowering, or flowering stage). TNC levels varied within compared plant populations. The stem distal to the gall functioned more efficiently as a nodal channel than the stem proximal to the gall, especially in the translocation of nitrogenous nutrients. Our findings indicate that the gall induced by E. strenuana functions as a sink for the assayed nutrients, although some variations have been observed in the patterns of nutrient mobilization. By creating a sink for the nutrients in the gall, E. strenuana is able to place the overall plant metabolism under stress, and this ability indicates E. strenuana has the necessary potential for use as a biological-control agent.

Rice husk biochar and crop residue amendment in subtropical cropping soils: Effect on biomass production, nitrogen use efficiency and greenhouse gas emissions

We investigated the effect of maize residues and rice husk biochar on biomass production, fertiliser nitrogen recovery (FNR) and nitrous oxide (N2O) emissions for three different subtropical cropping soils. Maize residues at two rates (0 and 10 t ha−1) combined with three rates (0, 15 and 30 t ha-1) of rice husk biochar were added to three soil types in a pot trial with maize plants. Soil N2O emissions were monitored with static chambers for 91 days. Isotopic 15N-labelled urea was applied to the treatments without added crop residues to measure the FNR. Crop residue incorporation significantly reduced N uptake in all treatments but did not affect overall FNR. Rice husk biochar amendment had no effect on plant growth and N uptake but significantly reduced N2O and carbon dioxide (CO2) emissions in two of the three soils. The incorporation of crop residues had a contrasting effect on soil N2O emissions depending on the mineral N status of the soil. The study shows that effects of crop residues depend on soil properties at the time of application. Adding crop residues with a high C/N ratio to soil can immobilise N in the soil profile and hence reduce N uptake and/or total biomass production. Crop residue incorporation can either stimulate or reduce N2O emissions depending on the mineral N content of the soil. Crop residues pyrolysed to biochar can potentially stabilise native soil C (negative priming) and reduce N2O emissions from cropping soils thus providing climate change mitigation potential beyond the biochar C storage in soils. Incorporation of crop residues as an approach to recycle organic materials and reduce synthetic N fertiliser use in agricultural production requires a thorough evaluation, both in terms of biomass production and greenhouse gas emissions.

On the Maximum Cloud Zone and the ITCZ over Indian, Longitudes during the Southwest Monsoon

An investigation is presented of the daily variation of the maximum cloud zone (MCZ) and the 7W mb trough in the Northern Hemisphere over the Indian longitudes 70–90°E during April–October for 1973–77. It is found that during June–September there are two favorable locations for a MCZ over these longitudes–on a majority of days the MCZ is present in the monsoon zone north of 15°N, and often a secondary MCZ occurs in the equatorial region (0–10°N). The monsoon MCZ gets established by northward movement of the MCZ occurring over the equatorial Indian ocean in April and May. The secondary MCZ appears intermittently, and is characterized by long spells of persistence only when the monsoon MCZ is absent. In each of the seasons studied, the MCZ temporarily disappeared from the mean summer monsoon location (15–28°N) about four weeks after it was established near the beginning of July. It is reestablished by the northward movement of the secondary MCZ, which becomes active during the absence of the monsoon MCZ, in a manner strikingly similar to that observed in the spring to summer transition. A break in monsoon conditions prevails just prior to the temporary disappearance of the monsoon MCZ. Thus we conclude that the monsoon MCZ cannot survive for longer than a month without reestablishment by the secondary MCZ. Possible underlying mechanisms are also discussed.

Tropical dryland agroforestry on clay soils: : Analysis of systems based on Acacia senegal in the Blue Nile region, Sudan

Acacia senegal, the gum arabic producing tree, is the most important component in traditional dryland agroforestry systems in the Blue Nile region, Sudan. The aim of the present study was to provide new knowledge on the potential use of A. senegal in dryland agroforestry systems on clay soils, as well as information on tree/crop interaction, and on silvicultural and management tools, with consideration on system productivity, nutrient cycling and sustainability. Moreover, the aim was also to clarify the intra-specific variation in the performance of A. senegal and, specifically, the adaptation of trees of different origin to the clay soils of the Blue Nile region. In agroforestry systems established at the beginning of the study, tree and crop growth, water use, gum and crop yields, nutrient cycling and system performance were investigated for a period of four years (1999 to 2002). Trees were grown at 5 x 5 m and 10 x 10 m spacing alone or in mixture with sorghum or sesame; crops were also grown in sole culture. The symbiotic biological N2 fixation by A. senegal was estimated using the 15N natural abundance (δ15N) procedure in eight provenances collected from different environments and soil types of the gum arabic belt and grown in clay soil in the Blue Nile region. Balanites aegyptiaca (a non-legume) was used as a non-N-fixing reference tree species, so as to allow 15N-based estimates of the proportion of the nitrogen in trees derived from the atmosphere. In the planted acacia trees, measurements were made on shoot growth, water-use efficiency (as assessed by the δ13C method) and (starting from the third year) gum production. Carbon isotope ratios were obtained from the leaves and branch wood samples. The agroforestry system design caused no statistically significant variation in water use, but the variation was highly significant between years, and the highest water use occurred in the years with high rainfall. No statistically significant differences were found in sorghum or sesame yields when intercropping and sole crop systems were compared (yield averages were 1.54 and 1.54 ha-1 for sorghum and 0.36 and 0.42 t ha-1 for sesame in the intercropped and mono-crop plots, respectively). Thus, at an early stage of agroforestry system management, A. senegal had no detrimental effect on crop yield, but the pattern of resource capture by trees and crops may change as the system matures. Intercropping resulted in taller trees and larger basal and crown diameters as compared to the development of sole trees. It also resulted in a higher land equivalent ratio. When gum yields were analysed it was found that a significant positive relationship existed between the second gum picking and the total gum yield. The second gum picking seems to be a decisive factor in gum production and could be used as an indicator for the total gum yield in a particular year. In trees, the concentrations of N and P were higher in leaves and roots, whereas the levels of K were higher in stems, branches and roots. Soil organic matter, N, P and K contents were highest in the upper soil stratum. There was some indication that the P content slightly increased in the topsoil as the agroforestry plantations aged. At a stocking of 400 trees ha-1 (5 x 5 m spacing), A. senegal accumulated in the biomass a total of 18, 1.21, 7.8 and 972 kg ha-1of N, P, K and OC, respectively. Trees contributed ca. 217 and 1500 kg ha-1 of K and OC, respectively, to the top 25-cm of soil over the first four years of intercropping. Acacia provenances of clay plain origin showed considerable variation in seed weight. They also had the lowest average seed weight as compared to the sandy soil (western) provenances. At the experimental site in the clay soil region, the clay provenances were distinctly superior to the sand provenances in all traits studied but especially in basal diameter and crown width, thus reflecting their adaptation to the environment. Values of δ13C, indicating water use efficiency, were higher in the sand soil group as compared to the clay one, both in leaves and in branch wood. This suggests that the sand provenances (with an average value of -28.07 ) displayed conservative water use and high drought tolerance. Of the clay provenances, the local one (Bout) displayed a highly negative (-29.31 ) value, which indicates less conservative water use that resulted in high productivity at this particular clay-soil site. Water use thus appeared to correspond to the environmental conditions prevailing at the original locations for these provenances. Results suggest that A. senegal provenances from the clay part of the gum belt are adapted for a faster growth rate and higher biomass and gum productivity as compared to provenances from sand regions. A strong negative relationship was found between the per-tree gum yield and water use efficiency, as indicated by δ13C. The differences in water use and gum production were greater among provenance groups than within them, suggesting that selection among rather than within provenances would result in distinct genetic gain in gum yield. The relative δ15N values ( ) were higher in B. aegyptiaca than in the N2-fixing acacia provenances. The amount of Ndfa increased significantly with age in all provenances, indicating that A. senegal is a potentially efficient nitrogen fixer and has an important role in t agroforestry development. The total above-ground contribution of fixed N to foliage growth in 4-year-old A. senegal trees was highest in the Rahad sand-soil provenance (46.7 kg N ha-1) and lowest in the Mazmoom clay-soil provenance (28.7 kg N ha-1). This study represents the first use of the δ15N method for estimating the N input by A. senegal in the gum belt of Sudan. Key words: Acacia senegal, agroforestry, clay plain, δ13C, δ15N, gum arabic, nutrient cycling, Ndfa, Sorghum bicolor, Sesamum indicum

Nurmisäilörehun valkuaisen metaboliareittien mallintaminen lypsylehmän pötsissä dynaamisen mallin avulla

Tämän tutkimuksen tarkoituksena oli määrittää ammoniumtypen, säilörehun liukoisen ja säilörehun liukenemattoman valkuaisen metaboloitumista pötsissä sekä kuvata ammmoniumtypen sekä säilörehun liukoisen ja liukenemattoman fraktion virtauskinetiikkaa alempaan ruoansulatuskanavaan. Koska typen metaboliaa märehtijän elimistössä on tutkittu paljon ja siitä on vakiintuneita käsityksiä, tässä tutkimuksessa keskityttiin metaboliareittien suuruuden arviointiin sekä typen virtausnopeuden estimointiin WinSAAM -ohjelmistolla rakennetun dynaamisen mallin avulla. Koe suoritettiin ajanjaksolla 2.3.-26.4.2002 Maa- ja elintarviketalouden tutkimuskeskuksessa (MTT) Jokioisilla, kotieläintuotannon tutkimusosaston koe-eläintallilla. Koemallina oli 4 x 4 latinalainen neliö, jossa oli neljä eläintä, neljä jaksoa ja neljä koekäsittelyä. Koe-eläiminä olivat neljä usean kerran poikinutta ayrshire-lypsylehmää. Lehmien poikimisesta oli kulunut keskimäärin 68 pv. (± 27 pv). Lehmät saivat 10 kg/pv väkirehuseosta ja hapolla (AIV 2000, 5,4 l/tn) säilöttyä esikuivattua säilörehua vapaasti siten, että jätettä jäi 5-10 %. Typellisten yhdisteiden metaboliareittejä mallinnettaessa havaintoaineistona käytettiin pötsin ammoniumtypen, bakteeritypen, alkueläintypen, liukenemattoman typen ja valkuaisen C-fraktion 15N-rikastumisen perusteella laskettua taustarikastumisen ylittävää poolikokoa. Havaintoaineiston perusteella rakennettiin ammoniumtypen (Malli 1), säilörehun liukoisen ei-ammoniumtypen (SNAN) (Malli 2) ja säilörehun liukenemattoman typen (Malli 3) metaboliareittejä kuvaavat mallit. Malli 1 estimoi ammoniumtypen imeytymisen, ulosvirtauksen ja synteesin bakteeritypeksi erinomaisesti. Alkueläintypen metabolian parametriarvoissa oli enemmän hajontaa kuin ammoniumtypen tai bakteeritypen metabolian parametriarvoissa. Mallin 1 mukaan 38 % ammoniumtypestä imeytyi suoraan pötsin seinämän läpi, 35 % syntetisoitui bakteeritypeksi ja 9 % alkueläintypeksi. Loppuosa (18 %) ammoniumtypestä virtasi nestefaasin mukana satakertaan. Malli 2 estimoi säilörehun SNAN:n metaboitumisen ammonium- ja bakteeritypeksi erinomaisesti. Myös ammoniumtypen metabolia ja bakteeritypen ulosvirtauksen estimointi onnistui erinomaisesti. Malli 2 ei antanut kovin luotettavaa estimaattia alkueläintypen metaboliasta. Mallin 2 mukaan 29 % säilörehun SNAN:stä imeytyi ammoniumtyppenä pötsin seinämän läpi, 10 % virtasi ulos pötsistä ammoniumtypen muodossa, 42 % syntetisoitui bakteeritypeksi ja 7 % alkueläintypeksi. Lisäksi 13 % SNAN:stä ohitti pötsin hajoamatta. Malli 3 estimoi typen metaboliareiteille erittäin tarkat parametriarvot. Tässä mallissa myös alkueläinten metabolialle saatiin luotettavat parametriarvot. Mallin 3 mukaan 57 % säilörehun liukenemattomasta typestä ohitti pötsin hajoamatta, 24 % syntetisoitui bakteeritypeksi ja 18 % alkueläintypeksi. Nurmisäilörehuun perustuvalla dieetillä hyvin suuri osuus satakertaan virtaavasta mikrobivalkuaisesta on peräisin ei-ammoniumtyppi -lähteistä, eli liukenemattomasta rehuvalkuaisesta, aminohapoista ja peptideistä. Koska lyhyt- ja pitkäketjuisten peptidien osuus SNAN fraktiosta on suuri ja koska jopa 13 % säilörehun SNAN fraktiosta voi ohittaa pötsin hajoamatta, on liukoisten typpifraktioiden ja varsinkin peptidien rooli pötsin typpimetaboliassa hyvin merkittävä.

How weed control and fertilisation influence tree physiological processes and growth at early establishment in an exotic F1 hybrid pine plantation of subtropical Australia

Purpose This study investigated how nitrogen (N) nutrition and key physiological processes varied under changed water and nitrogen competition resulting from different weed control and fertilisation treatments in a 2-year-old F1 hybrid (Pinus elliottii Engelm var. elliottii × P. caribaea var. hondurensis Barr. ex Golf.) plantation on a grey podzolic soil type, in Southeast Queensland. Materials and methods The study integrated a range of measures including growth variables (diameter at ground level (DGL), diameter at breast height (DBH) and height (H)), foliar variables (including foliar N concentration, foliar δ13C and δ15N) and physiological variables (including photosynthesis (An), stomatal conductance (gs), transpiration (E), intrinsic water use efficiency (WUEi) (A/gs) and xylem pressure potential (ΨXPP)) to better understand the mechanisms influencing growth under different weed control and fertilisation treatments. Five levels of weed control were applied: standard (routine), luxury, intermediate, mechanical and nil weed control, all with routine fertilisation plus an additional treatment, routine weed control and luxury fertilisation. Relative weed cover was assessed at 0.8, 1.1 and 1.6 years after plantation establishment to monitor the effectiveness of weed control treatments. Soil investigation included soil ammonium (NH4 +-N), nitrate (NO3 −-N), potentially mineralizable N (PMN), gravimetric soil moisture content (MC), hot water extractable organic carbon (HWETC), hot water extractable total N (HWETN), total C, total N, stable C isotope composition (δ13C), stable N isotope composition (δ15N), total P and extractable K. Results and discussion There were significant relationships between foliar N concentrations and relative weed cover and between tree growth and foliar N concentration or foliar δ15N, but initial site preparation practices also increased soil N transformations in the planting rows reducing the observable effects of weed control on foliar δ15N. A positive relationship between foliar N concentration and foliar δ13C or photosynthesis indicated that increased N availability to trees positively influenced non-stomatal limitations to photosynthesis. However, trees with increased foliar N concentrations and photosynthesis were negatively related to xylem pressure potential in the afternoons which enhanced stomatal limitations to photosynthesis and WUEi. Conclusions Luxury and intermediate weed control and luxury fertilisation positively influenced growth at early establishment by reducing the competition for water and N resources. This influenced fundamental key physiological processes such as the relationships between foliar N concentration, A n, E, gs and ΨXPP. Results also confirmed that time from cultivation is an important factor influencing the effectiveness of using foliar δ15N as an indicator of soil N transformations.

Effects of weed control and fertilization at early establishment on tree nitrogen and water use in an exotic F1 hybrid pine of subtropical Australia

Purpose We investigated the effects of weed control and fertilization at early establishment on foliar stable carbon (δ13C) and nitrogen (N) isotope (δ15N) compositions, foliar N concentration, tree growth and biomass, relative weed cover and other physiological traits in a 2-year old F1 hybrid (Pinus elliottii var. elliottii (Engelm) × Pinus caribaea var. hondurensis (Barr. ex Golf.)) plantation grown on a yellow earth in southeast Queensland of subtropical Australia. Materials and methods Treatments included routine weed control, luxury weed control, intermediate weed control, mechanical weed control, nil weed control, and routine and luxury fertilization in a randomised complete block design. Initial soil nutrition and soil fertility parameters included (hot water extractable organic carbon (C) and total nitrogen (N), total C and N, C/N ratio, labile N pools (nitrate (NO3 −) and ammonium (NH4 +)), extractable potassium (K+)), soil δ15N and δ13C. Relative weed cover, foliar N concentrations, tree growth rate and physiological parameters including photosynthesis, stomatal conductance, photosynthetic nitrogen use efficiency, foliar δ15N and foliar δ13C were also measured at early establishment. Results and discussion Foliar N concentration at 1.25 years was significantly different amongst the weed control treatments and was negatively correlated to the relative weed cover at 1.1 years. Foliar N concentration was also positively correlated to foliar δ15N and foliar δ13C, tree height, height growth rates and tree biomass. Foliar δ15N was negatively correlated to the relative weed cover at 0.8 and 1.1 years. The physiological measurements indicated that luxury fertilization and increasing weed competition on these soils decreased leaf xylem pressure potential (Ψxpp) when compared to the other treatments. Conclusions These results indicate how increasing N resources and weed competition have implications for tree N and water use at establishment in F1 hybrid plantations of southeast Queensland, Australia. These results suggest the desirability of weed control, in the inter-planting row, in the first year to maximise site N and water resources available for seedling growth. It also showed the need to avoid over-fertilisation, which interfered with the balance between available N and water on these soils.

Aspects and Applications of Pulse Sequence Design for Solution-State Nuclear Magnetic Resonance Spectroscopy

NMR spectroscopy enables the study of biomolecules from peptides and carbohydrates to proteins at atomic resolution. The technique uniquely allows for structure determination of molecules in solution-state. It also gives insights into dynamics and intermolecular interactions important for determining biological function. Detailed molecular information is entangled in the nuclear spin states. The information can be extracted by pulse sequences designed to measure the desired molecular parameters. Advancement of pulse sequence methodology therefore plays a key role in the development of biomolecular NMR spectroscopy. A range of novel pulse sequences for solution-state NMR spectroscopy are presented in this thesis. The pulse sequences are described in relation to the molecular information they provide. The pulse sequence experiments represent several advances in NMR spectroscopy with particular emphasis on applications for proteins. Some of the novel methods are focusing on methyl-containing amino acids which are pivotal for structure determination. Methyl-specific assignment schemes are introduced for increasing the size range of 13C,15N labeled proteins amenable to structure determination without resolving to more elaborate labeling schemes. Furthermore, cost-effective means are presented for monitoring amide and methyl correlations simultaneously. Residual dipolar couplings can be applied for structure refinement as well as for studying dynamics. Accurate methods for measuring residual dipolar couplings in small proteins are devised along with special techniques applicable when proteins require high pH or high temperature solvent conditions. Finally, a new technique is demonstrated to diminish strong-coupling induced artifacts in HMBC, a routine experiment for establishing long-range correlations in unlabeled molecules. The presented experiments facilitate structural studies of biomolecules by NMR spectroscopy.

The use of metabolomics to monitor simultaneous changes in metabolic variables following supramaximal low volume high intensity exercise

High Intensity Exercise (HIE) stimulates greater physiological remodeling when compared to workload matched low-moderate intensity exercise. This study utilized an untargeted metabolomics approach to examine the metabolic perturbations that occur following two workload matched supramaximal low volume HIE trials. In a randomized order, 7 untrained males completed two exercise protocols separated by one week; 1) HIE150%: 30 x 20s cycling at 150% VO2peak, 40s passive rest; 2) HIE300%: 30 x 10s cycling at 300% VO2peak, 50 s passive rest. Total exercise duration was 30 minutes for both trials. Blood samples were taken at rest, during and immediately following exercise and at 60 minutes post exercise. Gas chromatography-mass spectrometry (GC-MS) analysis of plasma identified 43 known metabolites of which 3 demonstrated significant fold changes (HIE300% compared to the HIE150% value) during exercise, 14 post exercise and 23 at the end of the recovery period. Significant changes in plasma metabolites relating to lipid metabolism [fatty acids: dodecanoate (p=0.042), hexadecanoate (p=0.001), octadecanoate (p=0.001)], total cholesterol (p=0.001), and glycolysis [lactate (p=0.018)] were observed following exercise and during the recovery period. The HIE300% protocol elicited greater metabolic changes relating to lipid metabolism and glycolysis when compared to HIE150% protocol. These changes were more pronounced throughout the recovery period rather than during the exercise bout itself. Data from the current study demonstrate the use of metabolomics to monitor intensity-dependent changes in multiple metabolic pathways following exercise. The small sample size indicates a need for further studies in a larger sample cohort to validate these findings.

Short-term plant-decomposer feedbacks in grassland plants

Plant species differ in their effects on ecosystem productivity and it is recognised that these effects are partly due to plant species-specific influences on soil processes. Until recently, however, not much attention was given to the potential role played by soil biota in these species-specific effects. While soil decomposers are responsible for governing the availability of nutrients for plant production, they simultaneously depend on the amount of carbon provided by plants. Litter and rhizodeposition constitute the two basal resources that plants provide to soil decomposer food webs. While it has been shown that both of these can have effects on soil decomposer communities that differ among plant species, the putative significance of these effects for plant nitrogen (N) acquisition is currently understudied. My PhD work aimed at clarifying whether the species-specific influences of three temperate grassland plants on the soil microfood-web, through rhizodeposition and litter, can feed back to plant N uptake. The methods and approach used (15N labelling of plant litter in microcosm experiments) revealed to be an effective combination of tools in studying these feedbacks. Plant effects on soil organisms were shown to differ significantly between plant species and the effects could be followed across several trophic levels. The labelling of litter further permitted the evaluation of plant acquisition of N derived from soil organic matter. The results show that the structure of the soil microfood-web can have a significant role in plant N acquisition when the structure is experimentally manipulated, such as when comparing systems consisting of microbes to those consisting of microbes and their grazers. However, despite this, the results indicate that differences in N uptake from soil organic matter between different plant species are not related to the effects these species exert on the structure of the soil microfood-web. Rather, these differences in N uptake seem to be determined by other species-specific traits of live plants and their litter. My results thus indicate that different resources provided by different plant species may not induce species-specific decomposer feedbacks on plant N uptake from soil organic matter. This further suggests that the species-specific plant effects on soil decomposer communities may not, at least in the short term, have significant consequences on plant production.

Denitrification losses from an intensively managed sub-tropical pasture: Impact of soil moisture on the partitioning of N2 and N2O emissions

Intensively managed pastures in subtropical Australia under dairy production are nitrogen (N) loaded agro-ecosystems, with an increased pool of N available for denitrification. The magnitude of denitrification losses and N2:N2O partitioning in these agro-ecosystems is largely unknown, representing a major uncertainty when estimating total N loss and replacement. This study investigated the influence of different soil moisture contents on N2 and N2O emissions from a subtropical dairy pasture in Queensland, Australia. Intact soil cores were incubated over 15 days at 80% and 100% water-filled pore space (WFPS), after the application of 15N labelled nitrate, equivalent to 50 kg N ha−1. This setup enabled the direct quantification of N2 and N2O emissions following fertilisation using the 15N gas flux method. The main product of denitrification in both treatments was N2. N2 emissions exceeded N2O emissions by a factor of 8 ± 1 at 80% WFPS and a factor of 17 ± 2 at 100% WFPS. The total amount of N-N2 lost over the incubation period was 21.27 kg ± 2.10 N2-N ha−1 at 80% WFPS and 25.26 kg ± 2.79 kg ha−1 at 100% WFPS respectively. N2 emissions remained high at 100% WFPS, while related N2O emissions decreased. At 80% WFPS, N2 emissions increased constantly over time while N2O fluxes declined. Consequently, N2/(N2 + N2O) product ratios increased over the incubation period in both treatments. N2/(N2 + N2O) product ratios responded significantly to soil moisture, confirming WFPS as a key driver of denitrification. The substantial amount of fertiliser lost as N2 reveals the agronomic significance of denitrification as a major pathway of N loss for sub-tropical pastures at high WFPS and may explain the low fertiliser N use efficiency observed for these agro-ecosystems.

X-ray crystallographic and NMR studies of pantothenate synthetase provide insights into the mechanism of homotropic inhibition by pantoate

The structural basis for the homotropic inhibition of pantothenate synthetase by the substrate pantoate was investigated by X-ray crystallography and high-resolution NMR spectroscopic methods. The tertiary structure of the dimeric N-terminal domain of Escherichia coli pantothenate synthetase, determined by X-ray crystallography to a resolution of 1.7 Å, showed a second molecule of pantoate bound in the ATP-binding pocket. Pantoate binding to the ATP-binding site induced large changes in structure, mainly for backbone and side chain atoms of residues in the ATP binding HXGH(34–37) motif. Sequence-specific NMR resonance assignments and solution secondary structure of the dimeric N-terminal domain, obtained using samples enriched in 2H, 13C, and 15N, indicated that the secondary structural elements were conserved in solution. Nitrogen-15 edited two-dimensional solution NMR chemical shift mapping experiments revealed that pantoate, at 10 mm, bound at these two independent sites. The solution NMR studies unambiguously demonstrated that ATP stoichiometrically displaced pantoate from the ATP-binding site. All NMR and X-ray studies were conducted at substrate concentrations used for enzymatic characterization of pantothenate synthetase from different sources [Jonczyk R & Genschel U (2006) J Biol Chem 281, 37435–37446]. As pantoate binding to its canonical site is structurally conserved, these results demonstrate that the observed homotropic effects of pantoate on pantothenate biosynthesis are caused by competitive binding of this substrate to the ATP-binding site. The results presented here have implications for the design and development of potential antibacterial and herbicidal agents.