M. L. van Beusichem

Role of nitrifier denitrification in the production of nitrous oxide

Abstract Nitrifier denitrification is the pathway of nitrification in which ammonia (NH 3 ) is oxidized to nitrite (NO 2 − ) followed by the reduction of NO 2 − to nitric oxide (NO), nitrous oxide (N 2 O) and molecular nitrogen (N 2 ). The transformations are carried out by autotrophic nitrifiers. Thus, nitrifier denitrification differs from coupled nitrification–denitrification, where denitrifiers reduce NO 2 − or nitrate (NO 3 − ) that was produced by nitrifiers. Nitrifier denitrification contributes to the development of the greenhouse gas N 2 O and also causes losses of fertilizer nitrogen in agricultural soils. In this review article, present knowledge about nitrifier denitrification is summarized in order to give an exact definition, to spread awareness of its pathway and controlling factors and to identify areas of research needed to improve global N 2 O budgets. Due to experimental difficulties and a lack of awareness of nitrifier denitrification, not much is known about this mechanism of N 2 O production. The few measurements carried out so far attribute up to 30% of the total N 2 O production to nitrifier denitrification. Low oxygen conditions coupled with low organic carbon contents of soils favour this pathway as might low pH. As nitrifier denitrification can lead to substantial N 2 O emissions, there is a need to quantify this pathway in different soils under different conditions. New insights attained through quantification experiments should be used in the improvement of computer models to define sets of conditions that show where and when nitrifier denitrification is a significant source of N 2 O. This may subsequently render the development of guidelines for low-emission farming practices necessary.

Uptake and distribution of cadmium in maize inbred lines

Genotypic variation in uptake and distribution of cadmium (Cd) was studied in 19 inbred lines of maize (Zea mays L.). The inbred lines were grown for 27 days on an in situ Cd-contaminated sandy soil or for 20 days on nutrient solution culture with 10 µg Cd L-1. The Cd concentrations in the shoots showed large genotypic variation, ranging from 0.9 to 9.9 µg g-1 dry wt. for the Cd-contaminated soil and from 2.5 to 56.9 µg g-1 dry wt. for the nutrient solution culture. The inbred lines showed a similar ranking for the Cd concentrations in the shoots for both growth media (r2=0.89). Two main groups of inbreds were distinguished: a group with low shoot, but high root Cd concentrations (shoot: 7.4±5.3 µg g-1 dry wt.; root: 206.0±71.2 µg g-1 dry wt.; ‘shoot Cd excluder’) and a group with similar shoot and root Cd concentrations (shoot: 54.2±3.4 µg g-1 dry wt.; root: 75.6±11.2 µg g-1 dry wt.; ‘non-shoot Cd excluder’). The classification of the maize inbred lines and the near equal whole-plant Cd uptake between the two groups demonstrates that internal distribution rather than uptake is causing the genotypic differences in shoot Cd concentration of maize inbred lines. Zinc (Zn), a micronutrient chemically related to Cd, showed an almost similar distribution pattern for all maize inbred lines. The discrepancy in the internal distribution between Cd and Zn emphasizes the specificity of the Cd distribution in maize inbred lines.

Effect of nitrogen supply rate on disease resistance in tomato depends on the pathogen.

The aim of this study was to investigate the effect of tissue nitrogen concentration, as a consequence of nitrogen supply rate, on the susceptibility of tomato plants to three pathogens. We varied tissue N concentration by supplying N at different rates by adding nitrate in different, exponentially increasing amounts to the nutrient solution on which the tomato plants were grown. Separate experiments were carried out to test susceptibility of tomato plants to the bacterial speck-causing Pseudomonas syringae pv tomato, to the wilt agent Fusarium oxysporum f.sp. lycopersici and to tomato powdery mildew caused by Oidium lycopersicum. The effect of tissue N concentration appeared to be highly pathogen-dependent: there was no effect on susceptibility to F. oxysporum, but susceptibility to P. syringae and O. lycopersicum increased significantly with increasing N concentration. We have previously demonstrated the opposite for susceptibility to Botrytis cinerea: decreasing susceptibility with increasing N concentration. The apparent contradictory effects are discussed in relation to the effect of N supply on both the nutritional value of the plant tissue to the pathogen and on the concentration of resistance-related compounds. We conclude that the effect of changing both characteristics on disease susceptibility is highly pathogen-specific and is probably dependent on differences in resource requirements of the pathogen or the sensitivity of the pathogen to plant resistance reactions or on both these factors.

Phytochelatins as biomarkers for heavy metal stress in maize (Zea mays L.) and wheat (Triticum aestivum L.): combined effects of copper and cadmium.

Heavy metal contaminated soils often show increased levels of more than one metal, e.g. copper (Cu), cadmium (Cd), zinc (Zn), lead (Pb) or nickel (Ni). In case such soils are used for crop production, prediction of yield reduction or quality decline due to heavy metals in the soil is inadequate when based only on chemical soil analysis. The use of biomarkers such as phytochelatins (PC), non-protein thiols specifically induced in plants upon exposure to heavy metals, may be an additional tool or diagnostic criterion in heavy metal research and in practice. In the present work, Cu and Cd uptake and induction of PC synthesis are studied with hydroponically grown maize and wheat plants exposed to mixtures of the two metals. We observed a close positive relationship between the concentrations of Cd and PC in the plant shoot material. A decreased shoot concentration of Cd after addition of Cu, due to metal competition at common root absorption sites, coincided with lower shoot PC levels. Also differences in metal uptake and xylary metal transport among the two plant species were reflected in corresponding differences in PC concentration. The observed direct relationship between shoot PC concentration and the degree of metal-induced growth inhibition makes the use of PC promising for the purpose tested for.

Effects of soil moisture content and temperature on methane uptake by grasslands on sandy soils

Aerobic grasslands may consume significant amounts of atmospheric methane (CH4). We aimed (i) to assess the spatial and temporal variability of net CH4 fluxes from grasslands on aerobic sandy soils, and (ii) to explain the variability in net CH4 fluxes by differences in soil moisture content and temperature. Net CH4 fluxes were measured with vented closed flux chambers at two sites with low N input on sandy soils in the Netherlands: (i) Wolfheze, a heather grassland, and (ii) Bovenbuurtse Weilanden, a grassland which is mown twice a year. Spatial variability of net CH4 fluxes was analysed using geostatistics. In incubation experiments, the effects of soil moisture content and temperature on CH4 uptake capacity were assessed. Temporal variability of net CH4 fluxes at Wolfheze was related to differences in soil temperature (r2 of 0.57) and soil moisture content (r2 of 0.73). Atmospheric CH4 uptake was highest at high soil temperatures and intermediate soil moisture contents. Spatial variability of net CH4 fluxes was high, both at Wolfheze and at Bovenbuurtse Weilanden. Incubation experiments showed that, at soil moisture contents lower than 5% (w/w), CH4 uptake was completely inhibited, probably due to physiological water stress of methanotrophs. At soil moisture contents higher than 50% (w/w), CH4 uptake was greatly reduced, probably due to the slow down of diffusive CH4 and O2 transport in the soil, which may have resulted in reduced CH4 oxidation and possibly some CH4 production. Optimum soil moisture contents for CH4 uptake were in the range of 20 – 35% (w/w), as prevailing in the field. The sensitivity of CH4 uptake to soil moisture content may result in short-term variability of net atmospheric CH4 uptake in response to precipitation and evapotranspiration, as well as in long-term variability due to changing precipitation patterns as a result of climate change.

Nitrogen availability and susceptibility of tomato leaves to Botrytis cinerea

The aim of this study was to investigate the effect of nitrogen availability on susceptibility of tomato leaves to the fungal pathogen Botrytis cinerea. Plants with varying nitrogen availability were grown by adding N daily in exponentially increasing amounts to a nutrient solution at different rates. Leaves of plants grown at low nitrogen availability had a high leaf C/N ratio (21 g g-1) and were about 2.5 times more susceptible to primary lesion formation by B. cinerea compared to plant grown at high nitrogen availability, which had a low leaf C/N ratio (11 g g-1). Leaf C/N ratio accounted for 95% of variation in susceptibility. This relationship between C/N ratio and susceptibility persisted when plants were grown with exponential P addition and optimal N supply, and was thus independent of plant growth rate or related factors. We could not explain the effect of nitrogen availability by variation in the most obvious N-based resistance compound α-tomatine because more susceptible leaves with a high C/N ratio contained more α-tomatine. These leaves also contained more soluble carbohydrates. The level of soluble carbohydrates correlated positively with susceptibility, independent of the growth method. We therefore suggest that the effect of N availability on susceptibility must be explained by variation in levels of soluble carbohydrates and speculate about the role of these carbohydrates in the infection process.

Methane emissions from wet grasslands on peat soil in a nature preserve

The area of wet grasslands on peat soil in the Netherlands is slowly increasing at the expense of drained, agriculturally used grasslands. This study aimed (i) to assess the contribution of wet grasslands on peat soil to methane (CH4) emissions, and (ii) to explain differences among sites and between years in order to improve our understanding of controlling factors. For these purposes, a field study was conducted in the period 1994–1996 in the nature preserve “Nieuwkoopse Plassen”, which is a former peat mining and agricultural area. Net CH4 emissions were measured weekly to monthly with vented closed flux chambers at three representative sites, and at ditches near these sites. Three-years average of CH4 emissions was 7.9 g CH4 m−2 yr−1 for Drie Berken Zudde, 13.3 for Koole, and 20.4 for Brampjesgat. Ditches near the sites emitted 4.2–22.5 g CH4 m−2 yr−1. The time-course of CH4 emissions for all experimental sites and years was fit with a multiple linear regression model with ground water level and soil temperature as independent variables. Lowering or raising the ground water level by 5 cm could decrease or increase CH4 emissions by 30–50%. Therefore, ground water level management of these grasslands should be done with care.

Nitrous oxide production, its source and distribution in urine patches on grassland on peat soil

Urine patches are considered to be important sites for nitrous oxide (N2O) production through nitrification and denitrification due to their high concentration of nitrogen (N). The aim of the present study was to determine the microbial source and size of production of N2O in different zones of a urine patch on grassland on peat soil. Artificial urine was applied in elongated patches of 4.5 m. Four lateral zones were distinguished and sampled for four weeks using an intact soil core incubation method. Incubation of soil cores took place without any additions to the headspace to determine total N2O production, with acetylene addition to determine total denitrification (N2O+N2), and with methyl fluoride to determine the N2O produced through denitrification.Nitrous oxide production was largest in the centre and decreased towards the edge of the patch. Maximum N2O production was about 50 mg N m−2 d−1 and maximum denitrification activity was 70 mg N m−2 d−1. Nitrification was the main N2O producing process. Nitrous oxide production through denitrification was only of significance when denitrification activity was high. Total N loss through nitrification and denitrification over 31 days was 4.1 g N per patch which was 2.2% of the total applied urine-N.

Determinants of spatial variability of methane emissions from wet grasslands on peat soil

Methane (CH4) emissions from soils, representing the consequence of CH4 production, CH4 consumption and CH4 transport, are poorly characterised and show a large spatial variability. This study aimed to assess the determinants of field-scale spatial variability of CH4 emissions from wet grasslands on peat soil. Mean CH4 emission rates of a three-year experiment at 18 plots distributed over three sites in the nature preserve “Nieuwkoopse Plassen” on peat soil in the Netherlands were related to CH4 production and CH4 consumption capacities of soil layers, and to soil and vegetation characteristics. Spatial variability of CH4 emissions and possible determining factors was high. Annual CH4 emissions ranged from 3 to 37 g CH4 m-2 yr-1. Coefficients of variation (CV) of CH4 emissions were on average 37% among sites and 83% within sites. Most important determinants of spatial variability were CH4 production capacity (average: 211 ng CH4 g-1 dry soil h-1; CV: 131%) and aboveground biomass of sedges (Carex spp.) (average: 0.45 g dm-2; CV: 127%) (P < 0.01). Sedges may affect CH4 emissions by stimulating CH4 transport from anaerobic layers to the surface via their vascular system and/or by serving as substrate for methanogens. For extrapolation of CH4 emissions to larger areas, best results will be obtained by using factors that are easy to determine, like vegetation.

Nitrogen loss from grassland on peat soils through nitrous oxide production

Nitrous oxide (N2O) in soils is produced through nitrification and denitrification. The N2O produced is considered as a nitrogen (N) loss because it will most likely escape from the soil to the atmosphere as N2O or N2. Aim of the study was to quantify N2O production in grassland on peat soils in relation to N input and to determine the relative contribution of nitrification and denitrification to N2O production. Measurements were carried out on a weekly basis in 2 grasslands on peat soil (Peat I and Peat II) for 2 years (1993 and 1994) using intact soil core incubations. In additional experiments distinction between N2O from nitrification and denitrification was made by use of the gaseous nitrification inhibitor methyl fluoride (CH3F).Nitrous oxide production over the 2 year period was on average 34 kg N ha-1 yr-1 for mown treatments that received no N fertiliser and 44 kg N ha-1 yr-1 for mown and N fertilised treatments. Grazing by dairy cattle on Peat I caused additional N2O production to reach 81 kg N ha-1 yr-1. The sub soil (20–40 cm) contributed 25 to 40% of the total N2O production in the 0–40 cm layer. The N2O production:denitrification ratio was on average about 1 in the top soil and 2 in the sub soil indicating that N2O production through nitrification was important. Experiments showed that when ratios were larger than l, nitrification was the major source of N2O. In conclusion, N2O production is a significant N loss mechanism in grassland on peat soil with nitrification as an important N2O producing process.