Biological Nitrification Inhibition Research Articles

Biological nitrification inhibition activity in a soil-grown biparental population of the forage grass, Brachiaria humidicola

Utilization of biological nitrification inhibition (BNI) strategy can reduce nitrogen losses in agricultural systems. This study is aimed at characterizing BNI activity in a plant-soil system using a biparental hybrid population of Brachiaria humidicola (Bh), a forage grass with high BNI potential but of low nutritional quality. Soil nitrification rates and BNI potential in root-tissue were analyzed in a hybrid population (117), obtained from two contrasting Bh parents, namely CIAT 26146 and CIAT 16888, with low and high BNI activity, respectively. Observed BNI activity was validated by measuring archaeal (AOA) and bacterial (AOB) nitrifier abundance in the rhizosphere soil of parents and contrasting hybrids. Comparisons of the BNI potential of four forage grasses were conducted to evaluate the feasibility of using nitrification rates to measure BNI activity under field and pot grown conditions. High BNI activity was the phenotype most commonly observed in the hybrid population (72%). BNI activity showed a similar tendency for genotypes grown in pots and in the field. A reduction in AOA abundance was found for contrasting hybrids with low nitrification rates and high BNI potential. Bh hybrids with high levels of BNI activity were identified. Our results demonstrate that the microcosm incubation and the in vitro bioassay may be used as complementary methods for effectively assessing BNI activity. The full expression of BNI potential of Bh genotypes grown in the soil (i.e. low nitrification rates) requires up to one year to develop, after planting.

Further insights into underlying mechanisms for the release of biological nitrification inhibitors from sorghum roots

Sorghum roots release two categories of biological nitrification inhibitors (BNIs) – hydrophilic-BNIs and hydrophobic-BNIs. Earlier research indicated that rhizosphere pH and plasma membrane (PM) H+ATPase are functionally linked with the release of hydrophilic BNIs, but the underlying mechanisms are not fully elucidated. This study is designed to reveal further insights into the regulatory mechanisms of BNIs release in root systems, using three sorghum genetic stocks. Sorghum plants were grown in a hydroponic system with pH of nutrient solutions ranging from 3.0 9.0. Pharmacological agents [(fusicoccin and vanadate) and anion-channel blockers (−niflumic acid (NIF) and anthracene-9-carboxylate (A9C)] were applied to root exudate collection solutions; BNI activity was determined with luminescent Nitrosomonas europaea bioassay. Sorgoleone levels in root exudates and H+ excretion from roots were determined. Two-phase partitioning system is used to isolate root plasma membrane (PM) and H+ ATPase activity was determined. A decrease in rhizosphere pH improved the release of hydrophilic-BNIs from roots of all the three sorghum genotypes, but had no effect on the release of hydrophobic-BNIs. Hydrophobic-BNI activity and sorgoleone levels in root-DCM wash are positively correlated. Fusicoccin promoted H+extrusion and stimulated the release of hydrophilic-BNIs. Vanadate, in contrast, suppressed H+ extrusion and lowered the release of hydrophilic-BNIs. Anion-channel blockers did not inhibit the release of hydrophilic BNIs, but enhanced H+-extrusion and hydrophilic-BNIs release. Rhizosphere pH has a major influence on hydrophilic-BNIs release, but not on the release of hydrophobic-BNIs. The low rhizosphere pH stimulated PM-H+ ATPase activity; H+-extrusion is closely coupled with hydrophilic-BNIs release. Anion-channel blockers stimulated H+ extrusion and hydrophilic-BNIs release. Our results indicate that some unknown membrane transporters are operating the release of protonated BNIs, which may compensate for charge balance when transport of other anions is suppressed using anion-channel blockers. A new hypothesis is proposed for the release of hydrophilic-BNIs from sorghum roots.

Residual effect of BNI by Brachiaria humidicola pasture on nitrogen recovery and grain yield of subsequent maize

Background and AimsThe forage grass Brachiaria humidicola (Bh) has been shown to reduce soil microbial nitrification. However, it is not known if biological nitrification inhibition (BNI) also has an effect on nitrogen (N) cycling during cultivation of subsequent crops. Therefore, the objective of this study was to investigate the residual BNI effect of a converted long-term Bh pasture on subsequent maize (Zea mays L.) cropping, where a long-term maize monocrop field (M) served as control.MethodsFour levels of N fertilizer rates (0, 60, 120 and 240 kg N ha−1) and synthetic nitrification inhibitor (dicyandiamide) treatments allowed for comparison of BNI effects, while 15N labelled micro-plots were used to trace the fate of applied fertilizer N. Soil was incubated to investigate N dynamics.ResultsA significant maize yield increase after Bh was evident in the first year compared to the M treatment. The second cropping season showed an eased residual effect of the Bh pasture. Soil incubation studies suggested that nitrification was significantly lower in Bh soil but this BNI declined one year after pasture conversion. Plant N uptake was markedly greater under previous Bh compared with M. The N balance of the 15N micro-plots revealed that N was derived mainly (68–86%) from the mineralized soil organic N pool in Bh while plant fertilizer N recovery (18–24%) was not enhanced.ConclusionsApplied N was strongly immobilized due to long-term root turnover effects, while a significant residual BNI effect from Bh prevented re-mineralized N from nitrification resulting in improved maize performance. However, a significant residual Bh BNI effect was evident for less than one year only.

Using alternative forage species to reduce emissions of the greenhouse gas nitrous oxide from cattle urine deposited onto soil

Grazed pastures are a major contributor to emissions of the greenhouse gas nitrous oxide (N2O), and urine deposition from grazing animals is the main source of the emissions. Incorporating alternative forages into grazing systems could be an approach for reducing N2O emissions through mechanisms such as release of biological nitrification inhibitors from roots and increased root depth. Field plot and lysimeter (intact soil column) trials were conducted in a free draining Horotiu silt loam soil to test whether two alternative forage species, plantain (Plantago lanceolate L.) and lucerne (Medicago sativa L.), could reduce N2O emissions relative to traditional pasture species, white clover (Trifolium repens L.) and perennial ryegrass (Lolium perenne L.). The amounts of N2O emitted from the soil below each forage species, which all received the same cow urine at the same rates, was measured using an established static chamber method. Total N2O emissions from the plantain, lucerne and perennial ryegrass controls (without urine application) were generally very low, but emissions from the white clover control were significantly higher. When urine was applied in autumn or winter N2O emissions from plantain were lower compared with those from perennial ryegrass or white clover, but this difference was not found when urine was applied in summer. Lucerne had lower emissions in winter but not in other seasons. Incorporation of plantain into grazed pasture could be an approach to reduce N2O emissions. However, further work is required to understand the mechanisms for the reduced emissions and the effects of environmental conditions in different seasons.

Nitrogen transformations in modern agriculture and the role of biological nitrification inhibition.

The nitrogen (N)-use efficiency of agricultural plants is notoriously poor. Globally, about 50% of the N fertilizer applied to cropping systems is not absorbed by plants, but lost to the environment as ammonia (NH3), nitrate (NO3-), and nitrous oxide (N2O, a greenhouse gas with 300 times the heat-trapping capacity of carbon dioxide), raising agricultural production costs and contributing to pollution and climate change. These losses are driven by volatilization of NH3 and by a matrix of nitrification and denitrification reactions catalysed by soil microorganisms (chiefly bacteria and archaea). Here, we discuss mitigation of the harmful and wasteful process of agricultural N loss via biological nitrification inhibitors (BNIs) exuded by plant roots. We examine key recent discoveries in the emerging field of BNI research, focusing on BNI compounds and their specificity and transport, and discuss prospects for their role in improving agriculture while reducing its environmental impact.

Biological nitrification inhibition by Brachiaria grasses mitigates soil nitrous oxide emissions from bovine urine patches

High nitrogen (N) concentration in bovine urine, which generally exceeds plant N uptake rates, results in the formation of hotspots of N loss when bovine urine is deposited on grazed pasture soils. High spatial variability in the distribution of urine patches in grazed pastures poses a major challenge to mitigate N losses. Some exudates from the roots of several tropical forage grasses were shown to inhibit the activity of soil nitrifiers; a process known as biological nitrification inhibition (BNI). We hypothesized that nitrate (NO3−) production and nitrous oxide (N2O) emissions from urine patches deposited on soils under forage grasses with high BNI capacity are lower than those with forage grasses with low BNI capacity. This hypothesis was tested using field plots of two tropical forage grass cultivars, Brachiaria humidicola cv. Tully (BT) and interspecific Brachiaria hybrid cv. Mulato (BM) which, correspondingly, have high and low BNI capacity. Nitrification rates and amoA gene copy numbers of ammonia oxidizing archaea (AOA) and bacteria (AOB) in soils under the two forage grasses were quantified before and after urine and water (control) application, as well, an additional experiment was conducted to quantify denitrification potential. Moreover, soil N2O emissions from simulated urine (0.123 kg N m−2) and water patches were monitored over a 29-day period. Results showed a greater suppression of nitrification, denitrification and AOA abundance in soils under BT than those under BM. Positive relationships (p < 0.05) existed between AOA and AOB abundance and NO3− contents in soils under BM. Bovine urine resulted in higher cumulative N2O fluxes from soils under BM (80 mg N2O-N m−2) compared to those under BT (32 mg N2O-N m−2). Consequently, N2O emission factors were higher for soils under BM (0.07%) than under BT (0.00002%). We conclude that tropical forage grasses with high BNI capacity play a key role in mitigating N2O emissions from bovine urine patches in archaea-dominated soils. This suggests that wide-spread adoption of tropical forage grasses with high BNI capacity may have a great potential to tighten N cycling in grazed pastures.

Potential Role of Fungal Endophytes in Biological Nitrification Inhibition in Brachiaria Grass Species

Brachiaria species have the ability to suppress nitrification in soil by releasing an inhibitory compound called ‘brachialactone’ from its roots; a process termed biological nitrification inhibition (BNI). This study tested the hypothesis that endophytic association with Brachiaria grass improves BNI activity of root tissues and reduces nitrification in Brachiaria-cultivated soil. Four cultivars of Brachiaria [i.e., B. decumbens (Basilisk), B. humidicola (Tully), B. brizantha (Marandu)], and one hybrid (Cayman) were evaluated for their BNI potentials under greenhouse and field conditions. In each experiment, plants were grown with (E+) and without (E-) endophyte inoculation, and harvested after eight months of growth. Root tissues and rhizosphere soil were taken from 0-30 cm depth and analyzed for BNI activity and nitrification, using bioluminescence assays and soil incubation, respectively. In the greenhouse experiment, endophyte association reduced BNI activity of root tissues in at least two cultivars (Basilisk and Marandu; by 13% and 6%, respectively); and this corresponded with 9% and 10% higher rates of nitrification (for Basilisk and Marandu, respectively) in soils grown with endophyte-infected plants than in the control. Under field conditions, endophyte association increased rates of nitrification in Marandu and Cayman by a similar magnitude of 12%, compared with endophyte-free control. In both experiments, Tully and Basilisk were essentially the most outstanding candidates for low-nitrifying forage systems, as shown by their high BNI activity and/or low rates of nitrification. The study also showed that cultivating soils with Brachiaria grasses could offer more agronomic and environmental benefits due to low N loss through nitrification than leaving the soils bare. However, further research to identify endophyte species that could suppress soil nitrifying microbes may enhance BNI process in Brachiaria.

Biological nitrification inhibition by weeds: wild radish, brome grass, wild oats and annual ryegrass decrease nitrification rates in their rhizospheres

This study investigated the ability of several plant species commonly occurring as weeds in Australian cropping systems to produce root exudates that inhibit nitrification via biological nitrification inhibition (BNI). Seedlings of wild radish (Raphanus raphanistrum), great brome grass (Bromus diandrus), wild oats (Avena fatua), annual ryegrass (Lolium rigidum) and Brachiaria humidicola (BNI-positive control) were grown in hydroponics, and the impact of their root exudates on NO3– production by Nitrosomonas europaea was measured in a pure-culture assay. A pot study (soil-based assay) was then conducted to confirm the ability of the weeds to inhibit nitrification in whole soils. All of the tested weeds slowed NO3– production by N. europaea in the pure-culture assay and significantly inhibited potential nitrification rates in soil-based assays. Root exudates produced by wild radish were the most inhibitory, slowing NO3– production by the pure culture of N. europaea by 53 ± 6.1% and completely inhibiting nitrification in the soil-based assay. The other weed species all had BNI capacities comparable to that of B. humidicola and significantly higher than that previously reported for wheat cv. Janz. This study demonstrates that several commonly occurring weed species have BNI capacity. By altering the N cycle, and retaining NH4+ in the soils in which they grow, these weeds may gain a competitive advantage over species (including crops) that prefer NO3–. Increasing our understanding of how weeds compete with crops for N may open avenues for novel weed-management strategies.

A colourimetric microplate assay for simple, high throughput assessment of synthetic and biological nitrification inhibitors

A simple, rapid, colourimetric method for screening biological nitrification inhibitors in plants is presented. Our approach combines the use of the Griess assay to track the rate of nitrite (NO2 −) production by pure cultures of ammonia oxidising bacteria in the presence and absence of nitrification inhibitors with a simple method for collecting root exudates from plants. NO2 − formation was tracked colourimetically on a microplate reader over 9 h of incubation. The advantage of this method is that it provides a simple, high throughput means of measuring biological nitrification inhibition in root exudates, using wild-type bacterial cultures. NO2 − formation rates and inhibition levels measured using the high through-put method were highly correlated with those measured by tracking NO2 − formation using a segmented flow analyser. The method was able to quantify inhibition of Nitrosomonas europaea by the synthetic nitrification inhibitors allythiourea (AT), dicyandiamide (DCD) and 3,4,-dimethylpyrazole phosphate (DMPP) with IC50 values similar to those reported in the literature. The method detected biological nitrification inhibition (BNI) in root exudates from Brachiaria humidicola and the lack of BNI in root exudates from wheat cv. Janz with minimal alteration of the exudates prior to testing. The results also showed that the more common soil ammonia oxidising bacterium (AOB), Nitrosospira multiformis, was much less sensitive to AT and DCD than N. europaea but had similar sensitivity to DMPP. This method provides a potentially useful way of screening large numbers of root exudate samples allowing for phenotyping of the BNI trait in crop and pasture populations which will be required for the trait to be introduced into commercial varieties.

Production of Biomass Crops Using Biowastes on Low-Fertility Soil: 2. Effect of Biowastes on Nitrogen Transformation Processes.

Increasing production of biowastes, particularly biosolids (sewage sludge), requires sustainable management strategies for their disposal. Biosolids can contain high concentrations of nutrients; hence, land application can have positive effects on plant growth and soil fertility, especially when applied to degraded soils. However, high rates of biosolids application may result in excessive nitrogen (N) leaching, which can be mitigated by blending biosolids with other biowastes, such as sawdust. We aimed to determine the effects of biosolids and sawdust on growth and N uptake by sorghum, rapeseed, and ryegrass as well as N losses via leaching. Plants were grown in a greenhouse over a 5-mo period in a low-fertility soil amended with biosolids (1250 kg N ha), biosolids-sawdust (0.5:1), or urea (200 kg N ha). Urea application increased biomass production of sorghum and ryegrass but proved insufficient for rapeseed on low-fertility soil. Biosolids application increased plant N concentrations in ryegrass and rapeseed and increased N uptake into the seeds of sorghum, increasing seed quality. Biosolids application did result in lower N leaching compared with urea, irrespective of plant species, and N leaching was unaffected by mixing the biosolids with sawdust. There was an indication of biological nitrification inhibition in the rhizosphere of sorghum. Rapeseed had similar growth and N uptake into biomass in biosolids and biosolids-sawdust treatments and hence was the most promising species with regard to recycling fresh sawdust in combination with high rates of biosolids on low-fertility soil.

Biological nitrification inhibition by rice root exudates and its relationship with nitrogen‐use efficiency

Microbial nitrification in soils is a major contributor to nitrogen (N) loss in agricultural systems. Some plants can secrete organic substances that act as biological nitrification inhibitors (BNIs), and a small number of BNIs have been identified and characterized. However, virtually no research has focused on the important food crop, rice (Oryza sativa). Here, 19 rice varieties were explored for BNI potential on the key nitrifying bacterium Nitrosomonas europaea. Exudates from both indica and japonica genotypes were found to possess strong BNI potential. Older seedlings had higher BNI abilities than younger ones; Zhongjiu25 (ZJ25) and Wuyunjing7 (WYJ7) were the most effective genotypes among indica and japonica varieties, respectively. A new nitrification inhibitor, 1,9-decanediol, was identified, shown to block the ammonia monooxygenase (AMO) pathway of ammonia oxidation and to possess an 80% effective dose (ED80 ) of 90ngμl-1 . Plant N-use efficiency (NUE) was determined using a 15 N-labeling method. Correlation analyses indicated that both BNI abilities and 1,9-decanediol amounts of root exudates were positively correlated with plant ammonium-use efficiency and ammonium preference. These findings provide important new insights into the plant-bacterial interactions involved in the soil N cycle, and improve our understanding of the BNI capacity of rice in the context ofNUE.

Identification of several wheat landraces with biological nitrification inhibition capacity

Background and aims Nitrification is the first step in several pathways that lead to losses of nitrogen from agricultural systems. Biological nitrification inhibition (BNI) refers to the ability of some plant species to release chemicals from their roots that inhibit microbial ammonia oxidation thereby decreasing nitrification rates. BNI has been found in the wheat relative Leymus racemosus but not in Triticum aestivum. The aim of this work was to assess a number of landraces of Triticum aestivum for BNI ability.

Development of a Biochar-Plant-Extract-Based Nitrification Inhibitor and Its Application in Field Conditions

The global use of nitrogen (N) fertilizer has increased 10-fold in the last fifty years, resulting in increased N losses via nitrate leaching to groundwater bodies or from gaseous emissions to the atmosphere. One of the biggest problems farmers face in agricultural production systems is the loss of N. In this context, novel biological nitrification inhibitors (BNI) using biochar (BC) as a renewable matrix to increase N use efficiency, by reducing nitrification rates, have been evaluated. The chemical and morphological characteristics of BC were analyzed and BC-BNI complexes were formulated using plant extracts from pine (Pinus radiata), eucalyptus (Eucalyptus globulus) and peumo (Cryptocarya alba). In field experiments, fertilizer and treatments, based on crude plant extracts and BC-BNI complexes, were applied and the effect on nitrification was periodically monitored, and at the laboratory level, a phytotoxicity assay was performed. The biochar-peumo (BCPe) complex showed the highest nitrification inhibition (66%) on day 60 after application compared with the crude plant extract, suggesting that BCPe complex protects the BNI against biotic or abiotic factors, and therefore BC-BNI complexes could increase the persistence of biological nitrification inhibitors. None of the biochar complexes had toxic effect on radish plants.

Transcriptional response of plasma membrane H+-ATPase genes to ammonium nutrition and its functional link to the release of biological nitrification inhibitors from sorghum roots

Aims Sorghum (Sorghum bicolor) roots release biological nitrification inhibitors (BNIs) to suppress soil nitrification. Presence of NH4+ in the rhizosphere stimulates BNIs release and it is hypothesized to be functionally associated with plasma membrane (PM) H+-ATPase activity. However, whether the H+-ATPase is regulated at the transcriptional level, and if so, which isoforms of the H+-ATPases are involved in BNIs release are not known. Also, it is not clear whether the stimulation on BNIs release from roots is due to NH4+ uptake or its assimilation, which are addressed in this study.

Acidification in Rhizospheric Soil of Field-Grown Sorghum Decreases Nitrification Activity

To date, most studies on biological nitrification inhibition (BNI) in sorghum have been performed with plants grown in hydroponic systems. However, the current study was conducted to determine whether or not sorghum inhibits nitrification in fields of Alfisols, and clarify the mechanism that results in inhibition of soil nitrification in the field. Nitrification activity in the rhizosphere of sorghum (Sorghum bicolor (L.) Moench) i.e. soil attached to its roots within a few millimeters was measured and compared with those in adjacent bulk soil. Sweet sorghum (6 varieties) and grain sorghum (3 varieties) were cultivated in 4 Alfisol fields in a semi-arid tropical region of India during the 2010 or 2011 rainy seasons. Soil samples were collected three times during the growing season. Nitrification activity in the rhizospheric soil was significantly lower than that in the bulk soil during 8 out of 12 samplings while the pH (H2O, 1:2) of the rhizospheric soil was significantly lower than that of the bulk soil in 10 out of 12 samplings. Acidification of the soil by sulfuric acid decreased the nitrification activity to a comparable extent, as emerged in the rhizospheric soils. Our results indicate that acidification of soil around roots would be one of the main causes of nitrification inhibition by sorghum in the field.

A 2-yr field assessment of the effects of chemical and biological nitrification inhibitors on nitrous oxide emissions and nitrogen use efficiency in an intensively managed vegetable cropping system

The application of nitrification inhibitors (NIs) is effective in suppressing nitrification and N2O emissions while promoting crop yields in many agroecosystems. However, the inhibitory effects of different NIs for vegetable production under soil and environmental conditions in China are not fully understood. To evaluate the effects of chemical and biological NIs on N2O emissions and the nitrogen use efficiency (NUE), a 2-yr field experiment with four treatments (regular urea (Urea), urea+dicyandiamide (DCD), urea+nitrapyrin (CP) and urea+biological nitrification inhibitor (BNI)) performed in triplicate was carried out in an intensive vegetable field using the static chamber and gas chromatography method. The results showed that the CP and BNI treatments shifted the main form of soil inorganic nitrogen (N) from nitrate (NO3−), which was the case for the Urea and DCD treatments, to ammonium (NH4+). The variations in soil temperature, moisture and NO3− content regulated the seasonal fluctuations of N2O emissions. Moreover, the DCD treatment did not significantly affect N2O or agronomic NUE relative to the Urea treatment, while CP and BNI significantly decreased annual N2O emissions by 16.5% and 18.1% and improved NUE by 12.6% and 6.7%, respectively. Thus, a markedly lower global warming potential (GWP) and greenhouse gas intensity (GHGI) was observed in the CP and BNI treatments relative to the Urea and DCD treatments. The results demonstrated that the NIs played important roles in enhancing yields and reducing N2O emissions from the vegetable ecosystem and that the CP and BNI treatments are suitable for marketing in China.

Use of the nitrification inhibitor dicyandiamide (DCD) does not mitigate N2O emission from bovine urine patches under Oxisol in Northwest Brazil

Animal production systems are important sources of greenhouse gases (GHGs), especially methane (CH4) and nitrous oxide (N2O). GHG emissions from urine patches have been extensively studied in temperate climates, with few studies under tropical conditions. Here we examined the driving factors of N2O and CH4 emission from urine patches in the tropics, as well as the role of the nitrification inhibitor DCD (dicyandiamide) in mitigating emissions. We hypothesized that the high temperature and periodical rainfall can increase GHG emissions from urine patches through accelerating mineralization of urine-N. We measured CH4 and N2O emissions from beef cattle urine (360 kg N ha−1) in Rondonia state (Brazil, tropical climate), during two different seasons (winter and summer), with and without the application of DCD (10 kg ha−1). No effects of DCD on cumulative N2O emissions were detected in summer, but DCD retarded the main emission peak. During winter DCD increased N2O emissions from 10.8 to 39.2 mg N–N2O m−2 (p ≤ 0.05). Emission factors averaged 0.4 % for summer and 0.1 % for winter, which is significantly lower than the IPCC default value of 1 %. The climate, associated with soil (acidic pH, WFPS and low N content) and plant properties (biological nitrification inhibition) resulted in a low emission factor. We concluded that the IPCC default emission factor for tropical systems may be reduced, and that the application of DCD is not recommended in such systems.

Nitrification Inhibition Potential of Brachiaria humidicola

An incubation experiment was conducted to determine the nitrification inhibition potential of Brachiaria humidicola (B. humidicola) and their effect on nitrification process. The pots soil was mixed 2 mg of nitrogen through ammonium sulphate. Seven treatments were evaluated viz. control, four root extracts of B. humidicola called as biological nitrification inhibitors (BNIs) (i.e., 70 % ethyl alcohol, 40 % ethyl alcohol, phosphate buffer solution and 2 M KCl salt solution extracts) and two standard chemical inhibitors i.e. dicyandiamide and neem oil coating. The amount of NH4 +-N was reduced 20.66–11.91 μg g−1 soil and NO3 −-N increased 28.89–31.18 μg g−1 soil from 14th to 22nd day time interval. Percent nitrification inhibition was more in BNIs (70 and 40 % alcohol extract) treated soils compared to plant based and synthetic nitrification inhibitors. The nitrification inhibition by B. humidicola also varied it was maximum (64.71 %) observed at 14th day over 22nd day (49.63 %).

Biological nitrification inhibition in sorghum: the role of sorgoleone production

Nitrification and denitrification are the two most important processes that contribute to greenhouse gas emission and inefficient use of nitrogen. Suppressing soil nitrification through the release of nitrification inhibitors from roots is a plant function, and termed “Biological Nitrification Inhibition (BNI)”. We report here the role and contribution of sorgoleone release to sorghum-BNI function. Three sorghum genotypes (Hybridsorgo, IS41245 and GDLP 34-5-5-3) were evaluated for their capacity to release sorgoleone, which has BNI-activity, in hydroponic and soil culture. Sorgoleone released was measured using HPLC; BNI-activity was determined using a luminescent recombinant Nitrosomonas europaea assay. Sorgoleone production and BNI-activity release by roots are closely associated (1 μg of sorgoleone is equivalent to 1 ATU activity in assay). Purified sorgoleone inhibited Nitrosomonas activity and suppressed soil nitrification. Sorghum genotypes release varying quantity of sorgoleone; GDLP 34-5-5-3 and Hybridsorgo showed higher capacity for both sorgoleone release and BNI-activity than did IS41245. In soil culture, GDLP 34-5-5-3 released higher quantity of sorgoleone into the rhizosphere, which had higher BNI-activity, and suppressed soil nitrification to a greater extent than did by IS41245. These results demonstrate genetic differences for sorgoleone release and its functional link with BNI-capacity; there is potential for genetic improvement of sorghum BNI-capacity and deployment of this in low-nitrifying sorghum production systems.

Biological nitrification inhibition (BNI) in Brachiaria pastures: A novel strategy to improve eco-efficiency of crop-livestock systems and to mitigate climate change

Up to 70% of the nitrogen (N) fertilizers applied to agricultural systems is lost due to nitrification and denitrification. Nitrification is a microbiological process that generates nitrate (NO3 ) and promotes the loss of N fertilizers by leaching and denitrification. Nitrification and denitrification are the only known biological processes that generate nitrous oxide (N2O), a powerful greenhouse gas contributing to global warming. There is an urgent need to suppress nitrification processes in soil to improve N recovery and N use efficiency (NUE) of agricultural systems and to mitigate climate change (Subbarao et al. 2012). Certain Brachiaria grasses (B. humidicola) can suppress soil nitrification by releasing biological nitrification inhibitors (BNIs) from roots, thereby reducing N2O emissions. This phenomenon, termed biological nitrification inhibition (BNI), has been the subject of recent research to characterize and validate the concept under field conditions (Subbarao et al. 2009). Advances on 3 aspects of BNI research are reported here: (1) gene quantification of soil nitrifying microorganisms to determine BNI activity in B. humidicola; (2) screening of B. humidicola breeding materials to identify hybrids with contrasting levels of BNI; and (3) quantification of the BNI residual effect from B. humidicola on N recovery and agronomic NUE of a subsequent maize crop.