Nitrogen acquisition in four co-existing species from an upland acidic grassland

The concentration of both nitrate and ammonium nitrogen was measured in soil taken from an upland acidic (pH 4.5) grassland habitat, containing four co-existing species, Deschampsia flexuosa (L.) Trin., Festuca ovina L., Juncus squarrosus L. and Nardus stricta L. Both nitrate and ammonium nitrogen were found to be present in the soil, in similarly small quantities. The effect of both sources of nitrogen on relative growth rate was studied, and an attempt was made to determine whether nitrate or ammonium nitrogen is the immediate source of nitrogen for these plants using assays of nitrate reductase (EC 1.6.6.2) and ammonium uptake. All four species showed larger growth rates on the same concentration of ammonium nitrogen compared to nitrate nitrogen. All species showed low activities of leaf nitrate reductase, even in plants grown on 18 mol nitrate m⁻³. Ammonium uptake activity appeared to be higher in species which showed the lowest nitrate reductase activity and least response to increasing nitrate concentration in the growth medium.     

The persistence of calcareous grassland species in the soil seed bank under developing and established scrub

The relationships between the composition of the soil seed bank, the field layer vegetation, and the scrub canopy were investigated along a 69 m transect, grading from incipient woodland, through scrub, into intensively rabbit-grazed calcareous grassland. The results are used to assess the persistence of species associated with open calcareous grassland in the seed bank under developing scrub. Scrub age, composition and density, changed along the transect from the woodland to open grassland. A total of 35 forb and grass species were found in the field layer. The pattern evident in the scrub layer was also reflected in the herbaceous vegetation. The field layer in the most closed portion of the transect, where the scrub was oldest, was dominated by shade-tolerant species normally associated with woodland habitats. The abundance of these species decreased along the transect as the scrub age declined, and the field layer became increasingly dominated by species typical of open grassland. A total of 47 species germinated from the seed bank. Few species were recorded in the seed bank along the entire length of the transect. Overall, the seed bank was dominated by Hypericum perforatum and Centaurium erythraea, which accounted for 38.2% and 28.6% of emerging seedlings respectively. As with a number of similar studies, the composition of the seed bank had a low correspondence with the composition of the field layer vegetation. The results also emphasise that the composition of the seed bank can be viewed as an ecological palimpsest, with germinable seed of species from each stage of the old-field succession occurring in the soil. The seed bank is an important component in the re-vegetation of an area after disturbance such as scrub removal. This study supports the findings of previous research in showing that relatively few characteristic calcareous grassland species form persistent seed banks. The soil seed bank would therefore appear to be of limited value in the restoration of such grassland following scrub removal.     

Nitrate reductase activity and growth response of forest species to ammonium and nitrate sources of nitrogen

Summary Three tree species,Eucalyptus regnans (F. Muell.),E. obliqua (L'Herit.),Pinus radiata (D. Don) were grown in sand culture with different proportions of nitrate and ammonium. Nitrate Reductase Activity (NRA) was induced in root tissue of all species and in leaf tissue of the eucalypts. An increasing proportion of nitrate resulted in increasing NRA in all species and hence NRA alone is no indication of N-preference. The highest NRA was found withE. regnans, a result which has also been obtained in the mature forest. The growth ofE. regnans was least with NH₄ ⁺ alone, whereas that ofE. obliqua was least with NO₃ ^– alone. The soils of matureE. regnans forest have a high potential for nitrification while those ofE. obliqua forest show little nitrification. Thus the preference for particular N sources shown by seedlings in culture is supported by related properties of mature forests. It is postulated however, that the inducibility of a high level of RNA in seedlings is more likely a result of a preference for NO₃ ^– than a cause.     

The ability of several high arctic plant species to utilize nitrate nitrogen under field conditions

The ability to utilize NO _inf3 ^sup– in seven high arctic plant species from Truelove Lowland, Devon Island, Canada was investigated, using an in vivo assay of maximum potential nitrate reductase (NR) activity and applications of ¹⁵N. Plant species were selected on the basis of being characteristic of nutrient-poor and nutrient-rich habitats. In all species leaves were the dominant site of NR activity. Root NR activity was negligible in all species except Saxifraga cernua. NO _inf3 ^sup– availability per se did not appear to limit NR activity of the species typically found on nutrient-poor sites (Dryas integrifolia, Saxifraga oppositifolia, and Salix arctica), or in Cerastium alpinum, as leaf NR activities remained low, even after NO _inf3 ^sup– addition. ¹⁵NO _inf3 ^sup– uptake was limited in D. integrifolia and Salix arctica. However, the lack of field induction of NR activity in C. alpinum and Saxifraga oppositifolia was not due to restricted nitrate uptake, as ¹⁵NO _inf3 ^sup– labelled NO _inf3 ^sup– entered the roots and shoots of both species. Leaf NR activity rates were low in three of the species typical of nutrient-rich habitats (O. digyna, P. radicatum and Saxifraga cernua), sampled from a site containing low soil NO _inf3 ^sup– . Additions of NO _inf3 ^sup– significantly increased leaf NR activity in these latter species, suggesting that potential NR activity was limited by the availability of NO _inf3 ^sup– . ¹⁵N labelled NO _inf3 ^sup– was taken up by O. digyna. P. radicatum and Saxifraga cernua. Although two species (D. integrifolia and Salix arctica) showed little utilization of NO _inf3 ^sup– , we concluded that five of the seven selected high arctic plant species (C. alpinum, O. digyna, P. radicatum, Saxifraga cernua and Saxifraga oppositifolia) do have the potential to utilize NO _inf3 ^sup– as a nitrogen source under field conditions, with the highest potential to utilize NO _inf3 ^sup– occurring in three of the species typically found on fertile habitats.     

Nitrate reductase activity and nitrogen compounds in xylem exudate of Juglans nigra seedlings: relation to nitrogen source and supply

Nitrogen (N) limits plant productivity and its uptake and assimilation may be regulated by N source, N availability, and nitrate reductase activity (NRA). Knowledge of how these factors interact to affect N uptake and assimilation processes in woody angiosperms is limited. We fertilized 1-year-old, half-sib black walnut (Juglans nigra L.) seedlings with ammonium (NH₄ ⁺) [as (NH₄)₂SO₄], nitrate (NO₃ ⁻) (as NaNO₃), or a mixed N source (NH₄NO₃) at 0, 800, or 1,600 mg N plant⁻¹ season⁻¹. Two months following final fertilization, growth, in vivo NRA, plant N status, and xylem exudate N composition were assessed. Specific leaf NRA was higher in NO₃ ⁻-fed and NH₄NO₃-fed plants compared to observed responses in NH₄ ⁺-fed seedlings. Regardless of N source, N addition increased the proportion of amino acids (AA) in xylem exudate, inferring greater NRA in roots, which suggests higher energy cost to plants. Root total NRA was 37% higher in NO₃ ⁻-fed than in NH₄ ⁺-fed plants. Exogenous NO₃ ⁻ was assimilated in roots or stored, so no difference was observed in NO₃ ⁻ levels transported in xylem. Black walnut seedling growth and physiology were generally favored by the mixed N source over NO₃ ⁻ or NH₄ ⁺ alone, suggesting NH₄NO₃ is required to maximize productivity in black walnut. Our findings indicate that black walnut seedling responses to N source and level contrast markedly with results noted for woody gymnosperms or herbaceous angiosperms.     

Plant, soil microbial and soil inorganic nitrogen responses to elevated CO2: a study in microcosms of Holcus lanatus

The impact of elevated atmospheric CO₂ concentrations on the nitrogen cycle was evaluated in a 2-month experiment in monospecific grassland microcosms (Holcus lanatus L.) grown on reconstituted grassland soil. The responses of the N pools in the plants, soil, and soil microbes were studied. The impact of high CO₂ on key stages of the N cycle, especially nitrification and denitrification processes, were also measured. Our study showed a strong plant response to high CO₂: total biomass increased by 76% (P < 0.001) and root length density increased by 77% (P = 0.010). However, total plant N was not significantly modified by high CO₂, because the percent N in the plant decreased by 40% (P < 0.001). We observed a large decrease in soil NO₃^– concentration under elevated CO₂ (–50%; P = 0.002). Soil ammonium concentrations were much less affected by CO₂ enrichment, and only in resin bags (–8%, P = 0.019). Soil nitrifying enzyme activity (NEA) had a tendency to increase (+17%; P = 0.061) and denitrifying enzyme activity (DEA) decreased (-12%; P = 0.013). We found evidence of increased microbial N sink (microbial N increased by 17%, P = 0.004). This and other studies suggest that rising CO₂ often reduces soil nitrate concentrations, which may lead to decreased nitrate leaching. Elevated CO₂ led to environmental conditions that were less favourable for denitrification in our study.     

Controls on inorganic N species transformations and potential leaching in freely drained sub-soils of heavily N-impacted acid grassland

In N-polluted, minimally managed soils, recent research has shown that ammonium-N, as well as nitrate-N, may move down through soil profiles. There is a need, therefore, to reassess what controls N species transformations below the rooting depth in such soils. The changes with depth down to 1 m in extractable ammonium-N and nitrate-N concentrations have been studied for two heavily N-impacted acid grassland soils near York, UK, for fresh soils and for soils incubated after either deionised water (d.w.) or ammonium-N spiking. Although in close proximity, the profiles showed marked differences in their relationships of ammonium- and nitrate-N to soil pH, C%, N% and C:N ratio. One profile was slightly more acidic at the surface, which seems to have changed the distribution of organic matter throughout the soil profile. Trends in ammonium production with depth were clearer when ammonium-N concentrations were recalculated on the basis of ammonium-N per unit mass of soil organic C. This then allowed a consistent trend with soil pH to be discerned for both profiles. Ammonium-N spiking showed that ammonium substrate availability was limiting potential net nitrification rate at 20–60 cm depth for both profiles. Potential mineral-N production was considerable at depth, which would facilitate transport of N to surface waters and/or groundwaters.     

Nitrification and nitrogen accumulation in the early stages of primary succession on Mt. Fuji

The relationship between nitrification potential and nitrogen accumulation was studied in an early successional sere on Mt. Fuji. Soil organic nitrogen accumulated with the invasion ofPolygonum cuspidatum and successively withMiscanthus oligostachyus and other species. Laboratory incubation experiments showed a higher nitrification potential at theM. oligostachyus state. The numbers of nitrifying bacteria increased with the progress of succession. No significant difference in nitrate reductase activity was found between pioneer and succeeding species. The soil solution at theM. oligostachyus stage contained a lower level of nitrate than rainwater, while that of the bare ground and theP. cuspidatum stage contained a higher nitrate level than rainwater. It was concluded that the high nitrate levels in the soil solution of the bare ground and theP. cuspidatum stage were due to lower nitrate-absorbing activity, leading to loss of nitrogen with precipitation, while the lower nitrate levels at theM. oligostachyus stage when higher nitrification activity occurred were due to higher nitrate-absorbing activity, preventing net loss of nitrogen from the ecosystem.     

Nitrogen acquisition from inorganic and organic sources by boreal forest plants in the field

A wide range of recent studies have indicated that organic nitrogen may be of great importance to plant nitrogen (N) nutrition. Most of these studies have, however, been conducted in laboratory settings, excluding important factors for actual plant uptake, such as competition, mycorrhizal associations and soil interactions. In order to accurately evaluate the importance of different N compounds to plant N nutrition, field studies are crucial. In this study, we investigated short- as well as long-term plant nitrogen uptake by Deschampsia flexuosa, Picea abies and Vaccinium myrtillus from ¹⁵NO₃^–, ¹⁵NH₄⁺ and (U-¹³C, ¹⁵N) arginine, glycine or peptides. Root N uptake was analysed after 6 h and 64 days following injections. Our results show that all three species, irrespective of their type of associated mycorrhiza (arbuscular, ecto- or ericoid, respectively) rapidly acquired similar amounts of N from the entire range of added N sources. After 64 days, P. abies and V. myrtillus had acquired similar amounts of N from all N sources, while for D. flexuosa, the uptake from all N sources except ammonium was significantly lower than that from nitrate. Furthermore, soil analyses indicate that glycine was rapidly decarboxylated after injections, while other organic compounds exhibited slower turnover. In all, these results suggest that a wide range of N compounds may be of importance for the N nutrition of these boreal forest plants, and that the type of mycorrhiza may be of great importance for N scavenging, but less important to the N uptake capacity of plants.     

Interactions between plant and insect diversity in the restoration of lowland calcareous grasslands in southern Britain

Abstract. The lowland calcareous grasslands of northwestern Europe are highly prized by ecologists and conservationists as a result of the diversity of their plant and invertebrate communities. Large areas of such grasslands have been lost this century as a result of changes in agricultural land use. Recent changes in agricultural policies, in particular the introduction of agri-environmental incentive schemes, have resulted in an increasing area being managed for the restoration of these communities. This paper reviews the management techniques employed in the restoration of lowland calcareous grasslands and the factors that govern their success. Constraints on the enhancement of the plant diversity of restoration sites include high soil fertility and the presence of undesirable species in the soil seed bank. However, it is thought that the primary constraint is the availability of propagules from which new populations can be established. Similarly, the dispersal mode and ability of insect species is likely to be the major factor limiting the enhancement of insect diversity. Evaluation of the success of restoration management usually involves monitoring changes in the plant community. However, as a result of their short life-cycles and sensitivity to small-scale environmental conditions, insects may respond more rapidly to changes resulting from restoration management and therefore provide better indicators of success. With the exception of a few high-profile butterfly species, the use of insects as indicator taxa has largely been neglected in terrestrial systems. This paper illustrates their potential use with reference to lowland calcareous grasslands in southern Britain.     

Nitrogen nutrition of Salvinia natans: Effects of inorganic nitrogen form on growth,morphology, nitrate reductase activity and uptake kinetics of ammonium and nitrate

In this study we assessed the growth, morphological responses, and N uptake kinetics of Salvinia natans when supplied with nitrogen as NO₃⁻, NH₄⁺, or both at equimolar concentrations (500 μM). Plants supplied with only NO₃⁻ had lower growth rates (0.17 ± 0.01 g g⁻¹ d⁻¹), shorter roots, smaller leaves with less chlorophyll than plants supplied with NH₄⁺ alone or in combination with NO₃⁻ (RGR = 0.28 ± 0.01 g g⁻¹ d⁻¹). Ammonium was the preferred form of N taken up. The maximal rate of NH₄⁺ uptake (V_max) was 6–14 times higher than the maximal uptake rate of NO₃⁻ and the minimum concentration for uptake (C_min) was lower for NH₄⁺ than for NO₃⁻. Plants supplied with NO₃⁻ had elevated nitrate reductase activity (NRA) particularly in the roots showing that NO₃⁻ was primarily reduced in the roots, but NRA levels were generally low (<4 μmol NO₂⁻ g⁻¹ DW h⁻¹). Under natural growth conditions NH₄⁺ is probably the main N source for S. natans, but plants probably also exploit NO₃⁻ when NH₄⁺ concentrations are low. This is suggested based on the observation that the plants maintain high NRA in the roots at relatively high NH₄⁺ levels in the water, even though the uptake capacity for NO₃⁻ is reduced under these conditions.     

Nitrogen retention efficiency and nitrogen losses of a managed and phytodiverse temperate grassland

Maintaining nitrogen retention efficiency (NRE) is crucial in minimizing N losses when intensifying management of temperate grasslands. Our aim was to evaluate how grassland management practices and sward compositions affect NRE (1 − N losses/soil available N), defined as the efficiency with which soil available N is retained in an ecosystem. A three-factorial grassland management experiment was established with two fertilization treatments (without and combined N, phosphorus and potassium fertilization), two mowing frequencies (cut once and thrice per year) and three sward compositions (control, monocot- and dicot-enhanced swards). We measured N losses as leaching and nitrous oxide emissions, and soil available N as gross N mineralization rates. Fertilization increased N losses due to increased nitrification and decreased microbial N immobilization, and consequently decreased NRE. Intensive mowing partly dampened high N losses following fertilization. Sward compositions influenced NRE but not N losses: control swards that developed for decades under extensive management had the highest NRE, whereas monocot-enhanced sward had the lowest NRE. NRE was highly correlated with microbial NH₄⁺ immobilization and microbial biomass and only marginally correlated with plant N uptake, underlining the importance of microbial N retention in the soil-plant system. Microbial N retention is reflected in NRE but not in indices commonly used to reflect plant response. NRE was able to capture the effects of sward composition and fertilization whereas N losses were only sensitive to fertilization; thus, NRE is a better index when evaluating environmental sustainability of sward compositions and management practices of grasslands.     

Nitrogen transformations in microenvironments of river beds and riparian zones

The soil of flooded riparian zones, the rhizosphere of riparian plants, biofilms at solid surfaces in the river, and the surface layer of sediments all constitute important environments for the oxidative or reductive transformations of inorganic nitrogen compounds. The exact microzonation and coupling of the processes have recently been studied intensively with ¹⁵N enrichment methods and microsensors for NH₄⁺, NO₂⁻, NO₃⁻, and N₂O. Microsensor analyses of gradients in sediments and biofilms have shown that nitrate production takes place in an aerobic surface zone that has a maximum thickness of a few millimeters in most shallow-water sediments and may be as thin as 100 μm in biofilms from very eutrophic environments. In the anoxic zone, denitrification is also concentrated in a zone of maximum a few millimeters, and typically half of the nitrate produced by nitrification is denitrified while the other half escapes to the water. The supply of nitrate from above is primarily controlled by the oxic layer acting as a diffusion barrier, and therefore denitrification is generally a linear function of the nitrate concentration in the water. The overlying water is thus a much more important source of nitrate for denitrification if the concentration is high. The rate and location of denitrification are also affected by bioturbating animals, benthic microphytes, plants, and bacteria performing dissimilatory nitrate reduction to ammonium (DNRA).     

Root production is determined by radiation flux in a temperate grassland community

Accurate knowledge of the response of root turnover to a changing climate is needed to predict growth and produce carbon cycle models. A soil warming system and shading were used to vary soil temperature and received radiation independently in a temperate grassland dominated by Holcus lanatus L. Minirhizotrons allowed root growth and turnover to be examined non‐destructively. In two short‐term (8 week) experiments, root responses to temperature were seasonally distinct. Root number increased when heating was applied during spring, but root death increased during autumnal heating. An experiment lasting 12 months demonstrated that any positive response to temperature was short‐lived and that over a full growing season, soil warming led to a reduction in root number and mass due to increased root death during autumn and winter. Root respiration was also insensitive to soil temperature over much of the year. In contrast, root growth was strongly affected by incident radiation. Root biomass, length, birth rate, number and turnover were all reduced by shading. Photosynthesis in H. lanatus exhibited some acclimation to shading, but assimilation rates at growth irradiance were still lower in shaded plants. The negative effects of shading and soil warming on roots were additive. Comparison of root data with environmental measurements demonstrated a number of positive relationships with photosynthetically active radiation, but not with soil temperature. This was true both across the entire data set and within a shade treatment. These results demonstrate that root growth is unlikely to be directly affected by increased soil temperatures as a result of global warming, at least in temperate areas, and that predictions of net primary productivity should not be based on a positive root growth response to temperature.     

A field study of the effects of elevated CO2 on plant biomass and community structure in a calcareous grassland

The effects of elevated CO₂ on plant biomass and community structure have been studied for four seasons in a calcareous grassland in northwest Switzerland. This highly diverse, semi-natural plant community is dominated by the perennial grass Bromus erectus and is mown twice a year to maintain species composition. Plots of 1.3 m² were exposed to ambient or elevated CO₂ concentrations (n = 8) using a novel CO₂ exposure technique, screen-aided CO₂ control (SACC) starting in March 1994. In the 1st year of treatment, the annual harvested biomass (sum of aboveground biomass from mowings in June and October) was not significantly affected by elevated CO₂. However, biomass increased significantly at elevated CO₂ in the 2nd (+20%, P = 0.05), 3rd (+21%, P = 0.02) and 4th years (+29%, P = 0.02). There were no detectable differences in root biomass in the top 8 cm of soil between CO₂ treatments on eight out of nine sampling dates. There were significant differences in CO₂ responsiveness between functional groups (legumes, non-leguminous forbs, graminoids) in the 2nd (P = 0.07) and 3rd (P < 0.001) years of the study. The order of CO₂ responsiveness among functional groups changed substantially from the 2nd to the 3rd year; for example, non-leguminous forbs had the smallest relative response in the 2nd year and the largest in the 3rd year. By the 3rd year of CO₂ exposure, large species-specific differences in CO₂ response had developed. For five important species or genera the order of responsiveness was Lotus corniculatus (+271%), Carex flacca (+249%), Bromus erectus (+33%), Sanguisorba minor (no significant CO₂ effect), and six Trifolium species (a negative response that was not significant). The positive CO₂ responses in Bromus and Carex were most closely related to increases in tiller number. Species richness was not affected by CO₂ treatment, but species evenness increased under elevated CO₂ (modified Hill ratio; P = 0.03) in June of the 3rd year, resulting in a marginally significant increase in species diversity (Simpson's index; P = 0.09). This and other experiments with calcareous grassland plants show that elevated atmospheric CO₂ concentrations can substantially alter the structure of calcareous grassland communities and may increase plant community biomass. Received: 12 July 1997 / Accepted: 14 September 1998     

Long-term changes in vegetation and soil chemistry in a calcareous and sandy semi-natural grassland

Calcicole plant species are vulnerable to acidification and fertilization, caused by deposition and changes in land use, since they are adapted to nutrient-poor calcareous conditions. In this study we used vegetation data (vascular plants, bryophytes and lichens) from 1964 and 1985 and stored soil samples from 1966 to investigate long-term soil chemistry and vegetation changes in a semi-natural, sandy calcareous grassland in southern Sweden. In the re-investigation in 2008 we found that increased decalcification due to acidification could not be verified. The plant community had changed from stress-tolerant calcareous grassland towards a community promoted by higher nutrient availability. Furthermore, the cover of species indicating calcareous conditions had decreased. A decline in the cover of species adapted to alkaline, phosphorus-poor conditions may be due to increased nutrient availability, but there were also indications that the vegetation had changed due to overgrowth by woody plants. This long-term impoverishment of the plant community highlights the need for appropriate management of calcareous grasslands, in order to limit the nutrients available in the soil and prevent overgrowth by shrubs and trees.