Characterisation of the glnK-amtB operon and the involvement of AmtB in methylammonium uptake in Azorhizobium caulinodans

This work reports the characterisation of the Azorhizobium caulinodans amtB gene, the deduced protein sequence of which shares similarity to those of several ammonium transporters. amtB is located downstream from glnK, a glnB-like gene. It is cotranscribed with glnK from an NtrC- and σ⁵⁴-dependent promoter. glnK and amtB insertion mutant strains have been isolated. Methylammonium uptake was assayed in these strains and in other mutant strains in which the regulation of nitrogen metabolism is impaired. Our data suggest that the AmtB protein is an ammonium transporter, which is mainly regulated by NtrC in response to nitrogen availability. Received: 2 February 1998 / Accepted: 20 March 1998     

Plant and fungal responses to elevated atmospheric carbon dioxide in mycorrhizal seedlings of Pinus sylvestris

The effects of elevated CO₂ concentration upon the mycorrhizal relationships of Scots pine (Pinus sylvestris) seedlings were investigated. Plants were grown for 4 months with their shoots exposed to ambient (C_AMB=360 μl l⁻¹) or elevated (C_ELEV=700 μl l⁻¹) CO₂ environments while their root systems, either colonised by the mycorrhizal fungi Paxillus involutus or Suillus bovinus, or left in the non-mycorrhizal condition, were maintained in sealed dishes. In one series of these plants the effects of C_ELEV upon the extent of mycorrhizal development and upon their growth and nutrition were determined, while another series were transferred from the dishes after 1 month, to transparent observation chambers before being returned to the two CO₂ environments. In these chambers, the effects of C_ELEV upon development of the external mycelial systems of the two mycorrhizal fungi was determined by measuring the advance of the hyphal fronts of the mycorrhizal fungi across non-sterile peat from the colonised plants. The dry mass and number of mycorrhizal tips were significantly higher in C_ELEV than in the C_AMB condition in plants colonised by both fungi in the dishes. Yields of whole plants and of shoots were higher in the C_ELEV treatment whether or not they were grown in the mycorrhizal condition, but the greater yields were not associated in these sealed systems with enhanced nutrient gain. The dry mass of non-mycorrhizal plants was greater than that of those colonised by mycorrhizal fungi under elevated CO₂. This is thought to be attributable to the energetic cost of production of the larger mycorrhizal systems in this treatment. The extent of development of the mycorrhizal mycelial systems of both fungi was greatly increased in C_ELEV relative to that in C_AMB environments. It is hypothesised that increased allocation of carbon to mycorrhizal root systems and their associated mycelia would provide the potential for enhancement of nutrient acquisition in open systems of greater fertility.     

Rates of nitrogen mineralization across an elevation and vegetation gradient in the southern Appalachians

We measured nitrogen (N) transformation rates for six years to examine temporal variation across the vegetation and elevation gradient that exists within the Coweeta Hydrologic Laboratory. Net N mineralization and nitrification rates were measured using 28-day in situ closed core incubations. Incubations were conducted at various intervals, ranging from monthly during the growing season, to seasonally based on vegetation phenology. Vegetation types included oak-pine, cove hardwoods, low elevation mixed oak, high elevation mixed oak, and northern hardwoods. Elevations ranged from 782 to 1347 m. Nitrogen transformation rates varied with vegetation type. Mineralization rates were lowest in the oak-pine and mixed oak sites averaging <1.2 mg N kg soil-1 28 day-1. Rates in the cove hardwood site were greater than all other low elevation sites with an annual average of 3.8 mg N kg soil-1 28 day-1. Nitrogen mineralization was greatest in the northern hardwood site averaging 13 mg N kg soil-1 28 day-1. Nitrification rates were typically low on four sites with rates <0.5 mg N kg soil-1 28 day-1. However, the annual average nitrification rate of the northern hardwood site was 6 mg N kg soil-1 28 days-1. Strong seasonal trends in N mineralization were observed. Highest rates occurred in spring and summer with negligible activity in winter. Seasonal trends in nitrification were statistically significant only in the northern hardwood site. Nitrogen mineralization was significantly different among sites on the vegetation and elevation gradient. While N mineralization rates were greatest at the high elevation site, vegetation type appears to be the controlling factor.     

Pikas (Ochotona princeps : Lagomorpha) as allogenic engineers in an alpine ecosystem

Pikas (Ochotona princeps: Lagomorpha) build caches of vegetation (“haypiles”), which serve as a food source during winter in alpine and subalpine habitats. Haypiles appear to degrade over time and form patches of nutrient-rich soils in barren talus and scree areas. We sampled soils underneath and next to haypiles, and plants growing on and near haypiles in an alpine cirque in northwestern Wyoming, USA, to determine the effects of pika food caches on N, C, and C/N ratios in soils and plants. We found that (1) haypile soils had significantly higher carbon and nitrogen levels and lower C/N ratios than both adjacent soils and soils in the general study area, (2) two of three plant species tested (Polemonium viscosum and Oxyria digyna) had significantly higher levels of tissue percent N when growing on haypile soils, and (3) total standing plant biomass at the study site increased with soil percent N suggesting that vegetation was nitrogen limited. Pikas may therefore function as allogenic ecosystem engineers by modulating nutrient availability to plants. Received: 5 July 1997 / Accepted: 30 November 1997     

Nitrogen relations of natural and disturbed plant communities in tropical Australia

Nitrogen relations of natural and disturbed tropical plant communities in northern Australia (Kakadu National Park) were studied. Plant and soil N characteristics suggested that differences in N source utilisation occur at community and species level. Leaf and xylem sap N concentrations of plants in different communities were correlated with the availability of inorganic soil N (NH⁺ ₄ and NO⁻ ₃). In general, rates of leaf NO⁻ ₃ assimilation were low. Even in communities with a higher N status, including deciduous monsoon forest, disturbed wetland, and a revegetated mine waste rock dump, levels of leaf nitrate reductase, xylem and leaf NO⁻ ₃ levels were considerably lower than those that have been reported for eutrophic communities. Although NO⁻ ₃ assimilation in escarpment and eucalypt woodlands, and wetland, was generally low, within these communities there was a suite of species that exhibited a greater capacity for NO⁻ ₃ assimilation. These “high- NO⁻ ₃ species” were mainly annuals, resprouting herbs or deciduous trees that had leaves with high N contents. Ficus, a high-NO⁻ ₃ species, was associated with soil exhibiting higher rates of net mineralisation and net nitrification. “Low-NO⁻ ₃ species” were evergreen perennials with low leaf N concentrations. A third group of plants, which assimilated NO⁻ ₃ (albeit at lower rates than the high-NO⁻ ₃ species), and had high-N leaves, were leguminous species. Acacia species, common in woodlands, had the highest leaf N contents of all woody species. Acacia species appeared to have the greatest potential to utilise the entire spectrum of available N sources. This versatility in N source utilisation may be important in relation to their high tissue N status and comparatively short life cycle. Differences in N utilisation are discussed in the context of species life strategies and mycorrhizal associations. Received: 5 July 1997 / Accepted: 13 July 1998     

Ferrous iron is transported across the peribacteroid membrane of soybean nodules

Symbiosomes and bacteroids isolated from soybean (Glycine max Merr.) nodules are able to take up ferrous iron. This uptake activity was completely abolished in the presence of ferrous-iron chelators. The kinetics of uptake were characterized by initially high rates of iron internalization, but no saturation was observed with increasing iron concentration. This process does not appear to involve the ferric reductase of the peribacteroid membrane. The transport of ferrous iron was inhibited by other transition metals, particularly copper. Ferrous iron was taken up by symbiosomes more efficiently than the ferric form. This indicates that the iron transport from the plant host cell to the microsymbiont in vivo may occur mainly as the ferrous form. Received: 11 February 1998 / Accepted: 29 May 1998     

Subcellular localization of glycoprotein epitopes during the development of lupin root nodules

Summary The monoclonal antibodies MAC236 and MAC265, raised against a soluble component of pea nodules, were used to elucidate the presence and subcellular localization of glycoprotein epitopes during the development of lupin (Lupinus albus L. cv. Multolupa) nodules, by means of immunocytochemistry and Western blot analysis. These antibodies recognize a single band of 95 kDa in pea, soybean and bean nodules, whilst two different bands of 240 and 135 kDa cross-react with MAC236 and MAC265 respectively in lupin nodules. This fact may indicate that the recognized epitopes can be present in different subcellular compartments and/or play different roles through the development of functional nodules. The results show that MAC265 is mainly associated with Bradyrhizobium infection and with the development of nodule primordium, in the first stages of nodulation. MAC265 is also detected when glycoprotein transport takes place across the cytoplasm and the cell wall, and also in the intercellular spaces of the middle cortex, attached to cell walls. The amount of MAC265 remains constant through nodule development. In contrast the amount of MAC236 increases with nodule age, parallel to the establishment of nitrogenase activity. This antibody is localized in cytoplasmic globules attached to the inner side of cell walls in the middle cortex, and mainly in the matrix filling the intercellular spaces of the middle and inner cortex. This main site of localization of MAC236 may indicate a role in the functioning of the oxygen diffusion barrier.     

The response of soil CO2 flux to changes in atmospheric CO2, nitrogen supply and plant diversity

We measured soil CO₂ flux over 19 sampling periods that spanned two growing seasons in a grassland Free Air Carbon dioxide Enrichment (FACE) experiment that factorially manipulated three major anthropogenic global changes: atmospheric carbon dioxide (CO₂) concentration, nitrogen (N) supply, and plant species richness. On average, over two growing seasons, elevated atmospheric CO₂ and N fertilization increased soil CO₂ flux by 0.57 µmol m^?2 s^?1 (13% increase) and 0.37 µmol m^?2 s^?1 (8% increase) above average control soil CO₂ flux, respectively. Decreases in planted diversity from 16 to 9, 4 and 1 species decreased soil CO₂ flux by 0.23, 0.41 and 1.09 µmol m^?2 s^?1 (5%, 8% and 21% decreases), respectively. There were no statistically significant pairwise interactions among the three treatments. During 19 sampling periods that spanned two growing seasons, elevated atmospheric CO₂ increased soil CO₂ flux most when soil moisture was low and soils were warm. Effects on soil CO₂ flux due to fertilization with N and decreases in diversity were greatest at the times of the year when soils were warm, although there were no significant correlations between these effects and soil moisture. Of the treatments, only the N and diversity treatments were correlated over time; neither were correlated with the CO₂ effect. Models of soil CO₂ flux will need to incorporate ecosystem CO₂ and N availability, as well as ecosystem plant diversity, and incorporate different environmental factors when determining the magnitude of the CO₂, N and diversity effects on soil CO₂ flux.     

An economic assessment of algal turf scrubber technology for treatment of dairy manure effluent

Controlling the input of nitrogen (N) and phosphorus (P) from dairies and other livestock operations into the surrounding air- and water-sheds poses both technical and economic challenges to the agricultural community. The purpose of this paper is to assess the economics of algal turf scrubber treatment technology at the farm-scale for a hypothetical 1000-cow dairy. Costs were developed for farms with and without anaerobic pretreatment. The majority of capital costs were due to land preparation, installation of liner material, and engineering fees. The majority of operational costs were due to energy requirements for biomass drying, pumping water, and repayment of capital investment. On farms using anaerobic pretreatment, waste heat from burning of biogas could be used to offset the energy requirements of biomass drying. In addition, biogas combustion exhaust gas could then be recycled back to the algal system to supply dissolved inorganic carbon for optimal algal production and pH control. Under the best case (algal system coupled with anaerobic digestion pretreatment), the yearly operational costs per cow, per kg N, per kg P, and per kg of dried biomass were $454, $6.20, $31.10, and $0.70, respectively. Without anaerobic digestion pretreatment, the yearly operational costs were 36% higher, amounting to $631 per cow, $8.70 per kg N, $43.20 per kg P, and $0.97 per kg of dried biomass. For perspective, a recent survey of 36 Maryland dairy farms found long-term annual profits of about $500 per cow. As no market currently exists for manure grown algal biomass, our cost analysis does not include any value of the biomass generated during manure treatment. In addition, there are a variety of potential uses for the algal biomass from manure treatment that could defray treatment costs. Future opportunities for dairies to participate in nutrient trading approaches to watershed nutrient management may also become important.     

Reduced community diversity in semi-natural meadows in southern Sweden, 1965–1990

Two data sets, one from the 1960s and one from 1990, from continuously managed semi-natural meadows (semi-natural grasslands used for hay-cutting) in Småland, southern Sweden, were analysed to describe the vegetation and in an attempt to characterise changes that have occurred in the vegetation. Based on a classification of the data set, nine plant communities were recognised. The main vegetational differences, as revealed by an ordination, were due to variation in soil moisture, which ranged from wet to dry.During the investigation period, the amount of hay meadow area decreased, particularly the area of wet-moist meadows. In addition, the total variation in the vegetation diminished, and three plant communities more or less disappeared. The turnover index of species in the data set was 36%, and most of the species that were lost had initially been uncommon. Lost species included rare taxa, such as Gentianella campestris and Linum catharticum, while the new species tended to be common in the region. Annual and biennial species accounted for a greater proportion of the lost taxa than did perennial species. The mean cover of species per plot and the relative abundance of graminoids per plot increased.The recorded changes in the vegetation may be related to the increased loads of nitrogen pollutants as well as to a decrease in management intensity. The diminished area is related to changes in land-use. The meadow sites in Småland have been part of a landscape rich in grasslands, but today they have a more or less relict status. They differ from grasslands found elsewhere in Sweden and Scandinavia. In Sweden, the majority (68%) of endangered vascular plants belong to the agricultural landscape. To ensure the survival of individual species it is important to preserve all types of meadows, not only a few selected ones, since no two meadows are alike.