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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#3Ns b " (Lr#n), which can be transferred to wheat as T3BL.3Ns b S (denoted Lr#n-SA), where 3BS arm of chromosome 3B of wheat was replaced by 3Ns b S of L. racemosus We successfully introduced T3BL.3Ns b S 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.3Ns b S 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 N 2 O 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., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Nitrification results in poor nitrogen (N) recovery and negative environmental impacts in most agricultural systems. Some plant species release secondary metabolites from their roots that inhibit nitrification, a phenomenon known as biological nitrification inhibition (BNI). Here, we attempt to characterize BNI in sorghum (Sorghum bicolor). In solution culture, the effect of N nutrition and plant age was studied on BNI activity from roots. A bioluminescence assay using recombinant Nitrosomonas europaea was employed to determine the inhibitory effect of root exudates. One major active constituent was isolated by activity-guided HPLC fractionations. The structure was analysed using NMR and mass spectrometry. Properties and the 70% inhibitory concentration (IC(70)) of this compound were determined by in vitro assay. Sorghum had significant BNI capacity, releasing 20 allylthiourea units (ATU) g(-1) root DW d(-1). Release of BNI compounds increased with growth stage and concentration of supply. NH4+ -grown plants released several-fold higher BNI compounds than NO3- -grown plants. The active constituent was identified as methyl 3-(4-hydroxyphenyl) propionate. BNI compound release from roots is a physiologically active process, stimulated by the presence of NH4+. Methyl 3-(4-hydroxyphenyl) propionate is the first compound purified from the root exudates of any species; this is an important step towards better understanding BNI in sorghum.

Nitrification inhibitory activity was found in root tissue extracts of Brachiaria humidicola, a tropical pasture grass. Two active inhibitory compounds were isolated by activity-guided fractionation, using recombinant Nitrosomonas europaea containing luxAB genes derived from the bioluminescent marine gram-negative bacterium Vibrio harveyi. The compounds were identified as methyl-p-coumarate and methyl ferulate, respectively. Their nitrification inhibitory properties were confirmed in chemically synthesized preparations of each. The IC50 values of chemically synthesized preparations were 19.5 and 4.4 microM, respectively. The ethyl, propyl, and butyl esters of p-coumaric and ferulic acids inhibited nitrification, whereas the free acid forms did not show inhibitory activity.

Sorghum (Sorghum bicolor) roots are well-known for the release of Biological Nitrification Inhibitors (BNIs), such as sorgoleone (SGLN) and methyl 3–4-hydroxyphenyl propionate (MHPP). However, the deployment of sorghums with BNIs in agriculture practices has not been widely investigated and evaluated. In this study, sorghum and maize were intercropped in field and pot cultivation to investigate the Biological Nitrification Inhibition (BNI) effect of sorghum on the productivity of both crops and soil nitrification. Inorganic N content (NO 3 – and NH 4 +), N 2 O emission, and the abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in soil were analyzed. The results showed intercropping with sorghum have the potential to improve crop productivity, as indicated by the Land Equivalent Ratio greater than 1.0. Intercropping with sorghum reduced the transformation rate of NH 4 + to NO 3 – in soils and the emission of N 2 O, as compared to monoculture of maize. Additionally, the abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) under intercropping mode were reduced by 41% and 28% in field trials, and reduced by 59% and 23% in pot experiments, respectively. Incubation of the same soils with SGLN and/or MHPP showed that the combined treatment of both BNIs strongly inhibited nitrification and N 2 O emission, along with lower AOB and AOA abundances, as compared with those under the sole treatment of SGLN or MHPP. Our results indicated that the root exudates of sorghum play a significant role in inhibiting soil nitrification due to the overlapping or amplified effect of various BNIs, which contribute to the nitrogen nutrition of the neighbouring crops under intercropping mode. Therefore, intercropping with BNI crops provides an eco-friendly strategy to improve crop production with reduced N 2 O emission. • Intercropping mode of sorghum and maize reduced soil nitrification and N 2 O emission. • Intercropping mode decreased the abundance of AOA and AOB. • Effect of combined addition of SGLN and MHPP in soil was superior to sole addition. • Root exudates of sorghum were of significance in inhibiting soil nitrification. [ABSTRACT FROM AUTHOR]