Biological Nitrification Inhibition Research Articles

Evidence of differences in nitrous oxide emissions and biological nitrification inhibition among Elymus grass species

Evidence of differences in nitrous oxide emissions and biological nitrification inhibition among Elymus grass species

Enlisting wild grass genes to combat nitrification in wheat farming: A nature-based solution.

Active nitrifiers and rapid nitrification are major contributing factors to nitrogen losses in global wheat production. Suppressing nitrifier activity is an effective strategy to limit N losses from agriculture. Production and release of nitrification inhibitors from plant roots is termed "biological nitrification inhibition" (BNI). Here, we report the discovery of a chromosome region that controls BNI production in "wheat grass" Leymus racemosus (Lam.) Tzvelev, located on the short arm of the "Lr#3Nsb" (Lr#n), which can be transferred to wheat as T3BL.3NsbS (denoted Lr#n-SA), where 3BS arm of chromosome 3B of wheat was replaced by 3NsbS of L. racemosus We successfully introduced T3BL.3NsbS into the wheat cultivar "Chinese Spring" (CS-Lr#n-SA, referred to as "BNI-CS"), which resulted in the doubling of its BNI capacity. T3BL.3NsbS from BNI-CS was then transferred to several elite high-yielding hexaploid wheat cultivars, leading to near doubling of BNI production in "BNI-MUNAL" and "BNI-ROELFS." Laboratory incubation studies with root-zone soil from field-grown BNI-MUNAL confirmed BNI trait expression, evident from suppression of soil nitrifier activity, reduced nitrification potential, and N2O emissions. Changes in N metabolism included reductions in both leaf nitrate, nitrate reductase activity, and enhanced glutamine synthetase activity, indicating a shift toward ammonium nutrition. Nitrogen uptake from soil organic matter mineralization improved under low N conditions. Biomass production, grain yields, and N uptake were significantly higher in BNI-MUNAL across N treatments. Grain protein levels and breadmaking attributes were not negatively impacted. Wide use of BNI functions in wheat breeding may combat nitrification in high N input-intensive farming but also can improve adaptation to low N input marginal areas.

Root exudation of contrasting drought-stressed pearl millet genotypes conveys varying biological nitrification inhibition (BNI) activity

Roots secrete a vast array of low molecular weight compounds into the soil broadly referred to as root exudates. It is a key mechanism by which plants and soil microbes interact in the rhizosphere. The effect of drought stress on the exudation process and composition is rarely studied, especially in cereal crops. This study focuses on comparative metabolic profiling of the exudates from sensitive and tolerant genotypes of pearl millet after a period of drought stress. We employed a combined platform of gas and liquid chromatography coupled to mass spectrometry to cover both primary and secondary metabolites. The results obtained demonstrate that both genotype and drought stress have a significant impact on the concentration and composition of root exudates. The complexity and function of these differential root exudates are discussed. To reveal the potential effect of root exudates on the soil microbial community after a period of drought stress, we also tested for biological nitrification inhibition (BNI) activity. The analysis revealed a genotype-dependent enhancement of BNI activity after a defined period of drought stress. In parallel, we observed a genotype-specific relation of elongated root growth and root exudation under drought stress. These data suggest that the drought stress-dependent change in root exudation can manipulate the microbial soil communities to adapt and survive under harsh conditions.

Biological nitrification inhibition by rice root exudates in two different soils of Uruguay

ABSTRACT Rice root exudates can control nitrification by releasing biological nitrification inhibitors (BNIs), reducing nitrogen losses in agricultural soils. However, the inhibitory effect on nitrification and the abundance of ammonia oxidisers in different soil types remain unclear. Two temperate paddy soils with different organic matter contents were collected to investigate the impact on nitrification rates of two rice cultivar root exudates, El Paso 144 (O. sativa ssp. indica) and Tacuarí (O. sativa spp. japonica). Root exudates were extracted before the tillering growth stage, and their BNI potential was evaluated in a bioassay with luminescent Nitrosomonas europaea and in a 12-day soil microcosm incubation. While exudates from Tacuarí showed stronger BNI activity in the bioassay, its nitrification inhibition in both soils was similar to that of DCD. El Paso did not show BNI activity in Salto whose organic matter content was higher. The abundance of ammonia oxidisers was not affected by root exudates or DCD, but only ammonia-oxidising bacteria had a significant positive relationship with soil nitrate. Our results demonstrated that although the bioassay showed high BNI activity, its expression in soils varied depending on the rice cultivar and the type of soil, particularly with its organic matter content.

REVIEW ON KNOWLEDGE GAP IN Brachiaria GRASS RESEARCH AND UTILIZATION: ETHIOPIAN PERSPECTIVE

Still food security has not been attained fully in many tropical African countries including Ethiopia. However, the issue of food security achievement has been able to realize due to various setbacks among which low productivity of crops and livestock take the lion share. Among the various constraints the parasitic weed Striga, and Stemborer pests are responsible for lower crop yields in the region. Regarding livestock feed, shortage in terms of quantity and quality are the major impediment to the livestock sector. To achieve food security, increasing crop yield and livestock production is vital in Ethiopia and other tropical countries. Crop yields can be enhanced through the control of weeds using biological systems to increase food crop yield apart from chemical inputs. In case of livestock, full production and reproduction potential of animals can be met through fulfilling nutritional requirements of livestock. The major livestock feed resources in Ethiopia are natural pasture and crop residues. Both feed resources; however, are poor in nutritional value and they are listed as low maintenance feed category. Therefore, it is vital to intensify integrated crop- livestock production systems for sustainable economy and environment. Introducing forage grasses in the crop production system has been practiced in the tropics as push pull technology. In Ethiopia, Brachiaria grass is an emerging forage for integrated agricultural production that has been getting considerable recognition as an option to overcome the pests in crop production in the tropics due to its high adaptive and yielding as well as climate smart forages. In the country, Brachiaria is recently introduced by different organization in different agro-ecology of the country mainly as push-pull integrated agricultural system and considering its fodder potential for the livestock feed. Therefore, this review paper aimed to looking for the available research knowledge in Ethiopia and somewhere else in the glob for better utilization of Brachiaria grass in the integrated agricultural system. All available information regarding the research and utilization of Brachiaria grass were reviewed in the published papers. The review reveal that Brachiaria has many advantages over other grass species in terms of adaptation to drought and low fertility soils, ability to sequester carbon; increase nitrogen use efficiency through biological nitrification inhibition (BNI) and arrest greenhouse gas emissions. The knowledge has been established in quantifying the multiple contributions of Brachiaria grass inclusion as push pull technology in different parts of the world (South America, Kenya, Rwanda). Limited report showed that cut-and-carry system is the utilization practice of brachiaria grass grown the push pull integration. The potential of improved Brachiaria grass in Ethiopia to address the challenge of livestock feed scarcity and other environmental managements; however, remain unexploited/limited which calls researchers to work on. The review concluded that B. cultivars could have a significant contribution on both animal and cereal production in the tropics but limited research and utilization in Ethiopia.

Biological nitrification inhibition in maize\u2014isolation and identification of hydrophobic inhibitors from root exudates

To control agronomic N losses and reduce environmental pollution, biological nitrification inhibition (BNI) is a promising strategy. BNI is an ecological phenomenon by which certain plants release bioactive compounds that can suppress nitrifying soil microbes. Herein, we report on two hydrophobic BNI compounds released from maize root exudation (1 and 2), together with two BNI compounds inside maize roots (3 and 4). On the basis of a bioassay-guided fractionation method using a recombinant nitrifying bacterium Nitrosomonas europaea, 2,7-dimethoxy-1,4-naphthoquinone (1, ED50 = 2 μM) was identified for the first time from dichloromethane (DCM) wash concentrate of maize root surface and named “zeanone.” The benzoxazinoid 2-hydroxy-4,7-dimethoxy-2H-1,4-benzoxazin-3(4H)-one (HDMBOA, 2, ED50 = 13 μM) was isolated from DCM extract of maize roots, and two analogs of compound 2, 2-hydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (HMBOA, 3, ED50 = 91 μM) and HDMBOA-β-glucoside (4, ED50 = 94 μM), were isolated from methanol extract of maize roots. Their chemical structures (1–4) were determined by extensive spectroscopic methods. The contributions of these four isolated BNI compounds (1–4) to the hydrophobic BNI activity in maize roots were 19%, 20%, 2%, and 4%, respectively. A possible biosynthetic pathway for zeanone (1) is proposed. These results provide insights into the strength of hydrophobic BNI activity released from maize root systems, the chemical identities of the isolated BNIs, and their relative contribution to the BNI activity from maize root systems.

Maize-Brachiaria intercropping: A strategy to supply recycled N to maize and reduce soil N2O emissions?

Nitrogen use in agriculture directly impacts food security, global warming, and environmental degradation. Forage grasses intercropped with maize produce feed for animals and or mulch for no-till systems. Forage grasses may exude nitrification inhibitors. It was hypothesized that brachiaria intercropping increases N recycling and maize grain yield and reduces nitrous oxide (N2O) emissions from soil under maize cropping. A field experiment was set up in December 2016 to test three cropping system (maize monocropped, maize intercropped with Brachiaria brizantha or with B. humidicola) and two N rates (0 or 150 kg ha-1). The grasses were sown with maize, but B. humidicola did not germinate well in the first year. B. brizantha developed slowly during the maize cycle because of shading but expanded after maize was harvested. The experiment was repeated in 2017/2018 when B. humidicola was replanted. N2O and carbon dioxide (CO2) emissions, maize grain yield and N content were measured during the two seasons. After the first maize harvest, the above- and below-ground biomass, C and N content of B. brizantha grown during fall-winter, and the biological nitrification inhibition potential of B. brizantha were evaluated. Maize yield responded to N fertilization (5.1 vs. 9.8 t ha-1) but not to brachiaria intercropping. B. brizantha recycled approximately 140 kg N ha-1 and left 12 t dry matter ha-1 for the second maize crop. However, the 2017/18 maize yields were not affected by the N recycled by B. brizantha, whereas N2O emissions were higher in the plots with brachiaria, suggesting that part of the recycled N was released too early after desiccation. Brachiarias showed no evidence of causing nitrification inhibition. The strategy of intercropping brachiarias did not increase maize yield, although it added C and recycled N in the system.

Biological nitrification inhibition and forage productivity of Megathyrsus maximus in Colombian dry tropics

Agronomic, nutritional, and environmental aspects are integrated to promote sustainable tropical grassland production. Biological nitrification inhibition (BNI) is a plant-based strategy to improve nitrogen use efficiency by grasses in which they suppress the pace of soil nitrification via exudation of inhibitory compounds. To evaluate the effect of BNI on the productive performance of Megathyrsus maximus under field conditions, we evaluated a collection of 27 germplasm accessions and commercial cultivars of the forage grass in the dry tropics of Colombia. We measured plant yield dry matter, nutrition quality parameters, and nitrification rates of soil at 22 months after pasture establishment. Our results highlighted germplasm accessions of superior agronomic performance (for dry matter production and nutrition quality) and high capacity to decrease nitrification. Although no relation was observed between agronomic aspects, nutritional aspects, and nitrification rates, we conclude that there is no agronomic or nutritional penalty on environmentally friendly grasses, and BNI could be adopted as a target trait in plant breeding programs toward the development of eco-efficient forages and contribute to the sustainable intensification of livestock systems.

Source partitioning using N2O isotopomers and soil WFPS to establish dominant N2O production pathways from different pasture sward compositions.

Nitrous oxide (N2O) is a potent greenhouse gas (GHG) emitted from agricultural soils and is influenced by nitrogen (N) fertiliser management and weather and soil conditions. Source partitioning N2O emissions related to management practices and soil conditions could suggest effective mitigation strategies. Multispecies swards can maintain herbage yields at reduced N fertiliser rates compared to grass monocultures and may reduce N losses to the wider environment. A restricted-simplex centroid experiment was used to measure daily N2O fluxes and associated isotopomers from eight experimental plots (7.8 m2) post a urea-N fertiliser application (40 kg N ha-1). Experimental pastures consisted of differing proportions of grass, legume and forage herb represented by perennial ryegrass (Lolium perenne), white clover (Trifolium repens) and ribwort plantain (Plantago lanceolata), respectively. N2O isotopomers were measured using a cavity ring down spectroscopy (CRDS) instrument adapted with a small sample isotope module (SSIM) for the analysis of gas samples ≤20 mL. Site preference (SP = δ15Nα - δ15Nβ) and δ15Nbulk ((δ15Nα + δ15Nβ) / 2) values were used to attribute N2O production to nitrification, denitrification or a mixture of both nitrification and denitrification over a range of soil WFPS (%). Daily N2O fluxes ranged from 8.26 to 86.86 g N2O-N ha-1 d-1. Overall, 34.2% of daily N2O fluxes were attributed to nitrification, 29.0% to denitrification and 36.8% to a mixture of both. A significant diversity effect of white clover and ribwort plantain on predicted SP and δ15Nbulk indicated that the inclusion of ribwort plantain may decrease N2O emission through biological nitrification inhibition under drier soil conditions (31%-75% WFPS). Likewise, a sharp decline in predicted SP indicates that increased white clover content could increase N2O emissions associated with denitrification under elevated soil moisture conditions (43%-77% WFPS). Biological nitrification inhibition from ribwort plantain inclusion in grassland swards and management of N fertiliser source and application timing to match soil moisture conditions could be useful N2O mitigation strategies.

Relative efficacy and stability of biological and synthetic nitrification inhibitors in a highly nitrifying soil: Evidence of apparent nitrification inhibition by linoleic acid and linolenic acid

AbstractBiological nitrification inhibition is a plant‐mediated rhizosphere process where natural nitrification inhibitors can be produced and released by roots to suppress nitrifier activity in soil. Nitrification is one of the critical soil processes in the nitrogen (N) cycle, but unrestricted and rapid nitrification in agricultural systems can result in major losses of N from the plant–soil system (i.e., by NO3− leaching and gaseous N emissions). In this study, we explored the potential efficacy of biological nitrification inhibitors (linoleic acid [LA] and linolenic acid [LN]) and a proven efficient synthetic (dicyandiamide [DCD]) nitrification inhibitor on N dynamics, nitrous oxide (N2O) and carbon dioxide (CO2) emissions in a highly nitrifying soil. 14C‐labelled LA, LN and DCD mineralization was determined in a parallel experiment to explore the fate of inhibitors after application. We found that LA and LN had no effect on soil NH4+ concentrations, but significantly decreased NO3− concentrations. Soil that received DCD had lower NO3− and higher NH4+ concentrations than the control (soil without nitrification inhibitors). LA and LN increased the cumulative N2O and CO2 emissions when they were applied at high concentrations (635 or 1,270 mg kg−1 dry soil). LA and LN had a much greater mineralization rate than that of DCD: 47–56%, 37–61% and 2.7–5.5%, respectively, after 38 days incubation. We conclude that in contrast to the direct inhibition of nitrification caused by DCD, addition of LA and LN may cause apparent nitrification inhibition by promoting microbial immobilization of soil NH4+ and/or NO3−. Future studies on nitrification inhibitors need to clearly differentiate between the direct and indirect effects that result from addition of these compounds to soil.Highlights The efficacy and stability of nitrification inhibitors in a highly nitrifying soil were explored. This study supports efforts to mitigate N losses and improve nitrogen use efficiency of inputs. Addition of LA, LN and DCD can decrease NO3− concentration, but their modes of action may be different. The apparent effect of LA and LN on soil NO3− concentration could be indirect.

Influence of biological nitrification inhibition by forest tree species on soil denitrifiers and N2O emissions

Some forest tree species are able to carry out a process known as biological nitrification inhibition, BNI, i.e. they inhibit nitrifiers through the production of specific compounds. We tested the hypothesis that, by restricting N supply to NO2−- and N2O-reducers, BNI would decrease potential N2O production and consumption and in situ N2O emissions. as compared to soils under trees without BNI capacity. Soils were collected from long-term monocultures (>43 ys) of three tree species without BNI capacity (Fagus sylvatica, Pinus nigra and Pseudotsuga menziesii) and two tree species with BNI capacity (Abies nordmanniana and Picea abies). The level of limitation of denitrification by NO3− was high for species with BNI capacity and low for species without BNI capacity, and was correlated with potential nitrification rates and the abundances of genes specifically harboured by ammonia oxidizing archaea and Nitrobacter. However, potential denitrification and actual N2O emissions did not reflect the tree BNI status, and denitrification limitation by soil carbon was higher than limitation by N under three tree species. Structural equation modelling revealed that the ratio between the gene copy abundances of nitrite-reducers and N2O-reducers was the microbial variable that best explained N2O emissions, along with soil pH and moisture. In addition, the NO3− concentration in the soil solution at 60 cm depth increased with the potential nitrification-to-denitrification ratio, suggesting a higher risk of NO3− leaching under some tree species like Douglas fir.

Revisiting plant biological nitrification inhibition efficiency using multiple archaeal and bacterial ammonia-oxidising cultures

Nitrification is a major process within the nitrogen (N) cycle leading to global losses of N, including fertiliser N, from natural and agricultural systems and producing significant nitrous oxide emissions. One strategy for the mitigation of these losses involves nitrification inhibition by plant-derived biological nitrification inhibitors (BNIs). Cultivation-based studies of BNIs, including screening for new compounds, have predominantly investigated inhibition of a single ammonia-oxidising bacterium (AOB), Nitrosomonas europaea, even though ammonia oxidation in soil is usually dominated by ammonia-oxidising archaea (AOA), especially in acidic soils, and AOB Nitrosospira sp., rather than Nitrosomonas, in fertilised soils. This study aimed to assess the sensitivity of ammonia oxidation by a range of AOA and AOB pure cultures to BNIs produced by plant roots (methyl 3-(4-hydroxyphenyl) propionate, sakuranetin and 1,9-decanediol) and shoots (linoleic acid, linolenic acid and methyl linoleate). AOA were generally more sensitive to BNIs than AOB, and sensitivity was greater to BNIs produced by shoots than those produced by roots. Sensitivity also varied within AOA and AOB cultures and between different BNIs. In general, N. europaea was not a good indicator of BNI inhibition, and findings therefore highlight the limitations of use of a single bioassay strain and suggest the use of a broader range of strains that are more representative of natural soil communities.

Rhizosphere pH and cation-anion balance determine the exudation of nitrification inhibitor 3-epi-brachialactone suggesting release via secondary transport.

Biological nitrification inhibition (BNI) of Brachiaria humidicola has been attributed to nitrification-inhibiting fusicoccanes, most prominently 3-epi-brachialactone. However, its release mechanism from B. humidicola roots remains elusive. Two hydroponic experiments were performed to investigate the role of rhizosphere pH and nutritional N form in regulating 3-epi-brachialactone release by B. humidicola and verify the underlying release pathway. Low rhizosphere pH and NH4 + nutrition promoted 3-epi-brachialactone exudation. However, the substitution of NH4 + by K+ revealed that the NH4 + effect was not founded in a direct physiological response to the N form but was related to the cation-anion balance during nutrient uptake. Release of 3-epi-brachialactone correlated with the transmembrane proton gradient ΔpH and NH4 + uptake (R2 = 0.92 for high ~6.8 and R2 = 0.84 for low ~4.2 trap solution pH). This corroborated the release of 3-epi-brachialactone through secondary transport, with the proton motive force (ΔP) defining transport rates across the plasma membrane. It was concluded that 3-epi-brachialactone release cannot be conceptualized as a regulated response to soil pH or NH4 + availability, but merely as the result of associated changes in ΔP.

Biological nitrification inhibition by sorghum root exudates impacts ammonia-oxidizing bacteria but not ammonia-oxidizing archaea

Sorghum has a great capacity to release biological nitrification inhibitors (BNIs), but the inhibitory effect on nitrification and ammonia oxidizer populations under planted soil conditions is unclear. A pot experiment with three nitrogen (N) application rates (0, 50, and 200 mg N kg−1) was set up to detect the influence of sorghum growth on soil nitrification and investigate the function of blocking the activity of ammonia oxidizers. A 15N-labeled experiment was also conducted to detect the N form absorbed by sorghum. Sorghum root exudates were collected at 30 days after transplanting to hydroponic culture and added into cultured soil to determine the shifts in the populations of nitrifiers. The 15N labeling experiment showed that the uptake rate by sorghum of ammonium N fertilizer was 24% and that of nitrate N fertilizer was 9%, indicating that sorghum was an ammonium using plant. Compared with unplanted soil, sorghum planting had a significant inhibitory effect on the nitrification process even at the high-N fertilizer rates. Autotrophic nitrification was the prevailing process, and sorghum root exudation inhibited this process as much as dicyandiamide (DCD, 10 mg kg−1). Root exudates had a significant inhibitory effect on ammonia-oxidizing bacteria (AOB) but had no effect on ammonia-oxidizing archaea (AOA).

Sorgoleone production enhances mycorrhizal association and reduces soil nitrification in sorghum

The control of nitrogen (N) flows via nitrates leaching and nitrous oxide emission is crucial for a safe environment. Some plants have shown abilities to suppress soil nitrification by releasing biological inhibitors, such as sorgoleone in sorghum. We hypothesized that the N uptake capacity of plants partly influences soil nitrification, and so do the intensity of the arbuscular mycorrhizal fungi (AMF)-sorghum symbiosis. Therefore, this study clarified the relationships between sorghum biological nitrification inhibition levels and AMF and fungi abundances in rhizosphere soils and sorghum roots by qPCR. The varieties 296B, IS32234, and IS20205, which release low (L-sorg), medium (M-sorg), and high (H-sorg) sorgoleone, respectively, were grown in φ 12 cm × 1 m pipes. Including a control without plant, they received 0 and 120 kg N ha−1 as ammonium sulfate solution. Samples were collected from 0 to 10, 10–30, and 30–80 cm soil layers. AMF and fungi populations at 31 days were significantly higher in H-sorg followed by M-sorg soils. The application of 120 kg N ha−1 (70 days) substantially increased AMF’s abundance in the 0–10 cm horizon. This result reveals that N-rich environments favor AMF expression. The AMF increase was more marked in the H-sorg and M-sorg soils, indicating that the levels of sorgoleone released may also influence the AMF symbiosis. Although H-sorg harbored more endophytic fungi, we did not observe any difference in root AMF colonization. The higher sorgoleone varieties showed higher plant biomass and N amounts, reflecting better N absorption and assimilation. The study reveals that sorgoleone, which confers sorghum its nitrification inhibition ability, also strengthens the sorghum-AMF symbiosis, N uptake and reduces the ammonia pool subjected to nitrification, and possibly the loss of nitrogen.

Impacts of pasture species and ruminant urine on N2O emissions and nitrogen transforming microbial communities in soil mesocosms

ABSTRACT Multispecies grasslands can alter nitrogen (N) cycling processes. A mesocosm experiment was performed comparing bare soil, soil planted with perennial ryegrass (Lolium perenne), white clover (Trifolium repens), plantain (Plantago lanceolata), and a mixed sward with all three species. We aimed to assess the plant effects on: ruminant urine-induced dry matter yields, plant N uptake, soil N2O emissions, and genes encoding nitrifier and denitrifier N transformations. After 35 days urine application increased both dry matter in the plantain (11%) and mixed (19%) swards, and N uptake in the ryegrass (63%), plantain (120%) and mixed (103%) swards. Urine increased cumulative N2O emissions, and while there was a trend for greater N uptake to cause lower emissions this was not significant. Urine addition elevated inorganic-N concentrations with few differences between plant treatments. Ammonia oxidising bacteria increased under urine but plantain did not inhibit this response. On days 3–10, increases in nirK and nosZI were indicative of enhanced nitrifier-denitrification and N2O scavenging by denitrifiers, respectively. The elevation of nosZI under urine-affected soil warrants further investigation in relation to potential pasture species effects. Consistent with previous in situ studies this study showed no evidence of plantain affecting soil inorganic-N concentrations via biological nitrification inhibition.

Urochloa Grasses Swap Nitrogen Source When Grown in Association with Legumes in Tropical Pastures

The degradation of tropical pastures sown with introduced grasses (e.g., Urochloa spp.) has dramatic environmental and economic consequences in Latin America. Nitrogen (N) limitation to plant growth contributes to pasture degradation. The introduction of legumes in association with grasses has been proposed as a strategy to improve N supply via symbiotic N2 fixation, but the fixed N input and N benefits for associated grasses have hardly been determined in farmers’ pastures. We have carried out on-farm research in ten paired plots of grass-alone (GA) vs. grass-legume (GL) pastures. Measurements included soil properties, pasture productivity, and sources of plant N uptake using 15N isotope natural abundance methods. The integration of legumes increased pasture biomass production by about 74%, while N uptake was improved by two-fold. The legumes derived about 80% of their N via symbiotic N2 fixation. The isotopic signature of N of grasses in GA vs. GL pastures suggested that sources of grass N are affected by sward composition. Low values of δ15N found in some grasses in GA pastures indicate that they depend, to some extent, on N from non-symbiotic N2 fixation, while δ15N signatures of grasses in GL pastures pointed to N transfer to grass from the associated legume. The role of different soil–plant processes such as biological nitrification inhibition (BNI), non-symbiotic N2 fixation by GA pastures and legume–N transfer to grasses in GL pastures need to be further studied to provide a more comprehensive understanding of N sources supporting the growth of grasses in tropical pastures.

Gross nitrogen transformations in tropical pasture soils as affected by Urochloa genotypes differing in biological nitrification inhibition (BNI) capacity

Several tropical grasses, particularly Urochloa humidicola (Rendle) Schweick.) (Syn. Brachiaria humidicola), have been associated with low soil nitrate content and reduced nitrogen (N) losses from pasture systems. Previous studies have detected that root exudates of certain Urochloa genotypes can inhibit ammonium oxidation in in vitro bioassay with Nitrosomonas and reduce net nitrification in soils, a phenomenon termed biological nitrification (BNI). However, net nitrification rates reflect the result of several N transformation processes that co-occur and together determine changes in NO3− concentrations in soils. Thus, to better understand the mechanisms underlying BNI, gross rates of nitrification, ammonification and inorganic N immobilization need to be assessed. In this study we used, for the first time, stable isotopes techniques to disentangle the different gross N transformation processes that determine net nitrification rates in soil of Urochloa genotypes differing in BNI. Intact soil cores were collected from two experimental sites in Colombia under different Urochloa genotypes, classified as low-BNI and high-BNI based on net nitrification rates determined in previous studies. Soil cores were labeled with (15NH4)2SO4 and K15NO3 to quantify gross N ammonification and nitrification, respectively and calculate the immobilization rates. Our results do not confirm lower gross nitrification rates under the high-BNI genotypes, despite reduced potential net nitrification rates and lower abundance of ammonia oxidising organisms, hence not supporting the widely accepted mechanism of BNI. Instead, the low potential nitrification rates could be explained by increased inorganic N immobilization under high-BNI genotypes, both in NH4+-N and NO3−-N form. The lack of differences might be explained by a strong N-limitation in our experiment, unlike previous BNI studies when large amounts of fertilizers were applied. Our results suggest that a combination of different mechanisms, particularly stimulation of N immobilization may be responsible for the BNI capacity observed as low NO3− soil content and reduced N losses.

Nitrifying Microbes in the Rhizosphere of Perennial Grasses are Modified by Biological Nitrification Inhibition.

Soil nitrification (microbial oxidation of ammonium to nitrate) can lead to nitrogen leaching and environmental pollution. A number of plant species are able to suppress soil nitrifiers by exuding inhibitors from roots, a process called biological nitrification inhibition (BNI). However, the BNI activity of perennial grasses in the nutrient-poor soils of Australia and the effects of BNI activity on nitrifying microbes in the rhizosphere microbiome have not been well studied. Here we evaluated the BNI capacity of bermudagrass (Cynodon dactylon L.), St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze), saltwater couch (Sporobolus virginicus), seashore paspalum (Paspalum vaginatum Swartz.), and kikuyu grass (Pennisetum clandestinum) compared with the known positive control, koronivia grass (Brachiaria humidicola). The microbial communities were analysed by sequencing 16S rRNA genes. St. Augustinegrass and bermudagrass showed high BNI activity, about 80 to 90% of koronivia grass. All the three grasses with stronger BNI capacities suppressed the populations of Nitrospira in the rhizosphere, a bacteria genus with a nitrite-oxidizing function, but not all of the potential ammonia-oxidizing archaea. The rhizosphere of saltwater couch and seashore paspalum exerted a weak recruitment effect on the soil microbiome. Our results demonstrate that BNI activity of perennial grasses played a vital role in modulating nitrification-associated microbial populations.

Ammonia-oxidizing bacteria dynamics affected by plantain under synthetic cattle urine patches

Plantain has been suggested as a nitrous oxide (N2O) and nitrate (NO3-) leaching mitigation option as it may induce biological nitrification inhibition (BNI) via plantain root exudation, which affects the activity of ammonia-oxidizing bacteria. This preliminary study compared the abundance of the ammonia monooxygenase gene (amoA) in soils containing either plantain and white clover, or ryegrass and white clover. Plants were sown in pots and grown in a greenhouse. Two months after sowing, synthetic cattle urine was applied to half the pots, and rhizosphere and bulk soil samples were collected 30 and 90 days after urine application. The abundance of the amoA gene was measured using real time quantitative PCR. The abundance of amoA genes in rhizosphere soil around ryegrass plants and in bulk soil under ryegrass/white clover were higher in pots treated with urine than the no-urine controls. AmoA gene abundance in bulk soil under plantain/white clover was higher in pots treated with urine (P<0.05) but not in rhizosphere soil around plantain plants (P>0.05) compared with the control. Furthermore, amoA gene copy numbers in the rhizosphere soil around plantain plants were lower than for ryegrass plants (P<0.05). However, there was no difference in the abundance of amoA genes in bulk soil of either combination of plant species evaluated (P>0.05). The results suggest that, in the time frame of our experiment, plantain could induce a BNI effect in the rhizosphere soil but not in the bulk soil.