MBOA in root exudates (and leachates) of winter/summer/mixed wheat cultivars
Ontology highlight
ABSTRACT:
Collection of irrigation water (leachates).
Hydroponic sampling of exudates to search for the BNI MBOA (and related compounds)
Aims:
- Quantify MBOA
- Search for other known BNIS
- Characterize exudate
LC-HRMS according to 10.1007/s00216-025-05818-y
INSTRUMENT(S): Orbitrap IQ-X
ORGANISM(S): Triticum Aestivum (ncbitaxon:4565)
SUBMITTER: 
christoph bueschl
PROVIDER: MSV000100410 | MassIVE | Thu Jan 08 06:46:00 GMT 2026
REPOSITORIES: MassIVE
Similar Datasets
Project description:Soil cultivation of spring wheat cultivar Remus. <br>
Hydroponic sampling of exudates to search for the BNI MBOA (and related compounds)<br>
Quantify MBOA<br>
Search for other known BNIS<br>
Characterize exudate<br>
<br>
LC-HRMS according to 10.1007/s00216-025-05818-y<br>
2026-01-08 | MSV000100411 | MassIVE
Project description:Sample prep + LC-MS measurement of barley exudates<br>
<br>
20 cultivars, 2 experimental groups (N-shock and control-schock), 4 replicates each<br>
<br>
Soil-Dipping-Hybrid setup<br>
Lyophilized & concentrated 30-fold (with lyophilization)<br>
Measured with Q Exactive HF, short method, 5uL injection volume<br>
For further details please refer to 10.1007/s00216-025-05818-y<br>
<br>
MS data processed with mzMine, features exported and formatted as inclusion lists, MSMS of pos/neg (separate) only of ammonium-schock samples
2026-01-08 | MSV000100408 | MassIVE
Project description:Goal :<br>
Detection of unknown degradation products of the nitrification inhibitors DMPP (SNI) and MBOA (BNI) in three different soils (Wieselburg, Marchfeld, and
Linz).<br>
<br>
List of samples:<br>
Same labeling scheme for all three soils (L, MF, WB)<br>
Sample NR | Timepoint | Treatment | weighed soil [g] | Which medium is it stored in<br>
1 | T0 (same day) | DMPP | 0.25 | HPLC-eluent<br>
5 | T0 (same day) | DMPP | 0.25 | HPLC-eluent<br>
6 | T0 (same day) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
10 | T0 (same day) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
11 | T0 (same day) | MBOA | 0.25 | Acetonirile<br>
15 | T0 (same day) | MBOA | 0.25 | Acetonirile<br>
16 | T0 (same day) | MBOA - stable isotope | 0.25 | Acetonirile<br>
20 | T0 (same day) | MBOA - stable isotope | 0.25 | Acetonirile<br>
21 | T0 (same day) | Control | 0.25 | HPLC-eluent<br>
25 | T0 (same day) | Control | 0.25 | Acetonirile<br>
26 | T1 (after 3 days) | DMPP | 0.25 | HPLC-eluent<br>
30 | T1 (after 3 days) | DMPP | 0.25 | HPLC-eluent<br>
31 | T1 (after 3 days) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
35 | T1 (after 3 days) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
36 | T1 (after 3 days) | MBOA | 0.25 | Acetonirile<br>
40 | T1 (after 3 days) | MBOA | 0.25 | Acetonirile<br>
41 | T1 (after 3 days) | MBOA - stable isotope | 0.25 | Acetonirile<br>
45 | T1 (after 3 days) | MBOA - stable isotope | 0.25 | Acetonirile<br>
46 | T1 (after 3 days) | Control | 0.25 | Acetonirile<br>
50 | T1 (after 3 days) | Control | 0.25 | HPLC-eluent<br>
51 | T2 (after 10 days) | DMPP | 0.25 | HPLC-eluent<br>
55 | T2 (after 10 days) | DMPP | 0.25 | HPLC-eluent<br>
56 | T2 (after 10 days) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
60 | T2 (after 10 days) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
61 | T2 (after 10 days) | MBOA | 0.25 | Acetonirile<br>
65 | T2 (after 10 days) | MBOA | 0.25 | Acetonirile<br>
66 | T2 (after 10 days) | MBOA - stable isotope | 0.25 | Acetonirile<br>
70 | T2 (after 10 days) | MBOA - stable isotope | 0.25 | Acetonirile<br>
71 | T2 (after 10 days) | Control | 0.25 | HPLC-eluent<br>
75 | T2 (after 10 days) | Control | 0.25 | Acetonirile<br>
76 | T3 (after 14 days) | DMPP | 0.25 | HPLC-eluent<br>
80 | T3 (after 14 days) | DMPP | 0.25 | HPLC-eluent<br>
81 | T3 (after 14 days) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
85 | T3 (after 14 days) | DMPP - stable isotope | 0.25 | HPLC-eluent<br>
86 | T3 (after 14 days) | MBOA | 0.25 | Acetonirile<br>
90 | T3 (after 14 days) | MBOA | 0.25 | Acetonirile<br>
91 | T3 (after 14 days) | MBOA - stable isotope | 0.25 | Acetonirile<br>
95 | T3 (after 14 days) | MBOA - stable isotope | 0.25 | Acetonirile<br>
96 | T3 (after 14 days) | Control | 0.25 | Acetonirile<br>
100 | T3 (after 14 days) | Control | 0.25 | HPLC-eluent<br>
<br>
Concentrations:<br>
all smaples have 130ug of (NH4)2SO4<br>
DMPP samples have 1,3ug of DMPP per 1 g soil<br>
MBOA samples have 300ug of MBOA per 1 g soil<br>
for samples that are isotopically labeled the amount of Inhibitor was split between the unlabeled and labeled compound<br>
5ul of the respective solution was added to the sample.<br>
<br>
Measurements according to 10.1021/ac503290j, but with an Q Exactive HF instrument<br>
<br>
Results: <br>
D patterns unfortunately also always show up in the T0 controls --> so it's unlikely to be biological breakdown/degradation products<br>
<br>
2026-01-23 | MSV000100564 | MassIVE
Project description:Dilute + Shoot: Sample Preparation + Measurement<br>
500 uL of each samples were mixed with 500 uL MeOH + 0.2% FA<br>
Samples were vortexed<br>
Samples were centrifuged at 4000 rpm and 4C for 20 min<br>
Supernatents were transferred into a 2 mL HPLC glass vial with insert<br>
Measurement of 2 uL (or 20uL) with LC-HRMS (Q Exactive HF; Standard Metabolomics Method; FPS)<br>
for further details on the analytical method refer to 10.1007/s00216-025-05818-y<br>
2026-01-13 | MSV000100448 | MassIVE
Project description:Nitrosomonas europaea Nm50 BNI degradation experiment:<br>
- concentrate ~ 600 ml culture<br>
- resuspend to make motherculture - 270 ml (need 255 for experiments)<br>
- let acclimate back to 26C for ~20min- 1h without substrate<br>
- add 1 mM NH4Cl (final concentration) to mother culture bottle:<br>
- take T0 mother culture; take T1 mother culture (cca 20 min); T2 mother culture (40min); <br>
split to treatments after 60 min<br>
<br>
Treatments:<br>
prepare 150 ul Linolenic acid (LNL) mix (50% unlabeled; 50% 13C LNL) on ice; after using<br>
120 for inoculation of treatments; freeze the 30 ul mix @ -80<br>
prepare just medium with 0.5 mM NH4<br>
<br>
Biotic:<br>
LNL; 8 ul of (50% unlabeled; 50% 13C LNL mix) - 5 replicates; 17 ml culture each (5)<br>
PHA-LNL - 1 mM PHA+8 ul of (50% unlabeled; 50% 13C LNL mix) - 5 replicates; 17 ml<br>
culture each (5)<br>
control - nothing added - 5 replicates; 17 ml culture each (5)<br>
<br>
Abiotic:<br>
LNL - 8 ul of (50% unlabeled; 50% 13C LNL mix) - 5 replicates; 17 ml medium with 0.5 mM<br>
NH4+ (5)<br>
<br>
Sampling every 30-45 min; 5 timepoints<br>
<br>
A: LE + culture<br>
B: PHA-LE + culture<br>
C: culture only<br>
D: LE in media only<br>
<br>
LC-HRMS according to 10.1007/s00216-025-05818-y<br>
2026-01-08 | MSV000100409 | MassIVE
Project description:Nitrosomonas Europaea Nm50 culture<br>
<br>
- concentrated ~600 ml culture to 280 ml mother culture<br>
- let acclimate back to 26C for ~1h without substrate<br>
- add 1 mM NH4Cl (final concentration) to mother culture bottle:<br>
- T0 M1 - 15:30, <br>
T1 M1 - 16:30 (working, already nitrite visible),<br>
T2 - 18:00 (working, more nitrite clearly),<br>
T3 - 19:20 (still increasing, time to split)<br>
<br>
Split mother culture to 15 subcultures for treatment:<br>
LNH - linolenic 8 ul 98% (triplicates, 17 ml culture)<br>
LNL - linolenic 2 ul 98% (triplicates, 17 ml culture)<br>
LEH - linoleic 8 ul 98% (triplicates, 17 ml culture)<br>
LEL - linoleic 2 ul 98% (triplicates, 17 ml culture)<br>
C - control, nothing added (just 17 ml, culture)<br>
<br>
shake up really well after addition:<br>
T1 19:40 (500 ul samples for MS)<br>
T2 21:30 (500 ul samples for MS)<br>
T3 23:00 (500 ul samples for MS)<br>
T4 1:30 - 1:50 (500 ul samples for MS)<br>
<br>
Harvest at T4 by filtration 15 ml from each, freeze @ -80C<br>
<br>
LC-HRMS measurments according to 10.1007/s00216-025-05818-y<br>
2026-01-05 | MSV000100368 | MassIVE
Project description:Goal :<br>
Finding of bio-degradation products of several native SNIs and BNIs in Trichoderma reesei QM6a liquid minimal medium (12C and 13C) cultures.<br>
<br>
Minimal medium<br>
Amount for 500 mL medium | Actual | amount Substance<br>
1.7500 g | 1.75115 g | (NH4)2SO4<br>
2.5000 g | 2.51394 g | KH2PO4<br>
0.6200 g | 0.63039 g | MgSO4 . 7 H2O<br>
0.3125 g | 0.20447 g | NaCl<br>
10 mL | 2 x 5 mL | 50X Trace element solution<br>
ad 490 mL | 490 mL | autoclaved ELGA water<br>
plus either 0.25g D-Glucose-13C or 0.25g D-Glucose-12C<br>
<br>
Tested BNIs/SNIs (1.50mg each in D2O)<br>
3,4-Dimethyl-1H-pyrazole phosphate (DMPP) [SOS 896]<br>
Methyl p-coumerate (MHPA) [no SOS]<br>
2-Amino-4-chloro-6-methylpyrimidine (ACMP) [no SOS]<br>
Coixol (MBOA) [no SOS]<br>
Karanjin<br>
Sakuranetin<br>
Syringic acid (SA) [SOS 899]<br>
3,4-Dihydroxybenzaldehyde (DHBA) [SOS 900]<br>
(R)-(+)-Limonene [SOS 277]<br>
<br>
Trichoderma reesei QM6a spores on two MEX plates,<br>
incubated with QM6a on 2024-08-22 (four days).<br>
Transfer to BOKU IFA-Tulln.<br>
Continued incubation at 28C, 90% relative humidity in constant darkness.<br>
<br>
Inoculation and pre-cultivation:<br>
Weighted glucose and filled up to respective volume with prepared minimal medium without carbon source<br>
Sterile filtered prepare medium (at Lamina)<br>
Inoculated medium with 15 uL spore solution per well (12C glucose) or 7 uL spore solution per well (U-13C6 glucose) respectively<br>
Incubation at 28C, 70 % relative humidity in constant darkness in Memmert HPP260 Incubator<br>
<br>
Treatment of cultures with SNI and BNI solutions and addition of more glucose<br>
<br>
Sampling after 7 days<br>
<br>
Measurements according to 10.1021/ac503290j, but with an Q Exactive HF instrument<br>
<br>
2026-01-23 | MSV000100562 | MassIVE
Project description:Root exudates contain specialised metabolites that affect the plant’s root microbiome. How host-specific microbes cope with these bioactive compounds, and how this ability shapes root microbiomes, remains largely unknown. We investigated how maize root bacteria metabolise benzoxazinoids, the main specialised metabolites of maize. Diverse and abundant bacteria metabolised the major compound in the maize rhizosphere MBOA and formed AMPO. AMPO forming bacteria are enriched in the rhizosphere of benzoxazinoid-producing maize and can use MBOA as carbon source. We identified a novel gene cluster associated with AMPO formation in microbacteria. The first gene in this cluster, bxdA encodes a lactonase that converts MBOA to AMPO in vitro. A deletion mutant of the homologous bxdA genes in the genus Sphingobium, does not form AMPO nor is it able to use MBOA as a carbon source. BxdA was identified in different genera of maize root bacteria. Here we show that plant-specialised metabolites select for metabolisation-competent root bacteria. BxdA represents a novel benzoxazinoid metabolisation gene whose carriers successfully colonize the maize rhizosphere and thereby shape the plant’s chemical environmental footprint
2024-04-05 | GSE263275 | GEO
Project description:Measurement of DMPP, MBOA, Methyl p-coumerate (MHPA) and Limonene in soil extracts.<br>
<br>
Nitrification inhibitors extraction protocols<br>
BNIs extraction with acetonitrile:<br>
<br>
Solutions:<br>
Acetonitrile HPLC grade <br>
<br>
Protocol:<br>
1. Weight 2g of soil and add NH4+ solution and the BNIs as powder (MHPA, MBOA) or liquid (limonene)<br>
2. Add 4mL acetonitrile<br>
3. Shake the mixture vigorously by hand before being putting the centrifuge tubes horizontally on an orbital shaker for 30 min (200 rpm)<br>
4. Centrifuge at 10000 rpm for 10mins <br>
5. Filter supernatant through a 0.45um<br>
6. Store -20C<br>
<br>
DMPP extraction:<br>
Solutions:<br>
HPLC-eluent (212 ml methanol + 788 ml of a 1 mM H3PO4 and 5 mM KH2PO4 buffer <br>
2 M KCl <br>
1 M NaOH<br>
di-ethyl-ether<br>
Protocol:<br>
1. Weigh 2 g of field moist soil into 50 mL tapered polypropylene centrifuge tube and add NH4 with DMPP solutions (100uL for Marchfeld, 30uL for Wieselburg)<br>
2. Add 2 mL HPLC-eluent <br>
3. Add 8 mL 2 M KCl <br>
4. Shake the mixture vigorously by hand before being putting the centrifuge tubes horizontally on an orbital shaker for 60 min (170 rpm)<br>
5. Centrifuge the extracts at 10000 rpm for 5 min. Decant and transfer to fresh 50mL centrifuge tube<br>
6. Add 400uL NaOH + 8mL di-ethylether <br>
7. Shake 10mins and then centrifuge at 10000rpm for 10mins <br>
8. Transfer 8mL of upper di-ethylether in a 50mL falcon tube containing 2mL HPLC eluent<br>
9. Evaporate di-ehtylether overnight <br>
10. Transfer about 2mL into an eppendrof <br>
11. Store -20C<br>
<br>
Measurements according to 10.1021/ac503290j, but with an Q Exactive HF instrument<br>
<br>
2026-01-23 | MSV000100563 | MassIVE
Project description:Nitrosomonas europaea Nm50 BNI degradation experiment - MBOA and D-DMPP<br>
- concentrate ~ 600 ml culture<br>
- resuspend to make mother-culture - 50 ml in 100 ml shot bottle. Feed 1 mM NH4Cl (final concentration) to mother culture bottle to keep active till 25th and setup<br>
- Keep at 28C overnight<br>
<br>
After two days<br>
- Wash remaining 25 ml of mother culture (2x 12.5 in one spin column 50 ml tube: use "spent/already used 2nd tube as Tara with 12.5 ml water, during centrifugation); wash 3x with medium (check if nitrite is gone, resuspend in 170 ml medium - need 160 for setup + mother culture samples)<br>
- let acclimate for ~20min - 40 min without substrate (lunch)<br>
- prepare 84 ml medium with 0.5 mM ammonium for abiotic controls<br>
- add 1 mM ammonium to mother culture, sample M0, M1 (20 min), M2 (40 min)<br>
<br>
D-DMPP mix (790% D, 30% notz labeled) dissolved in MQ to 0.05 M (50 mM). [17.95 mg D-DMPP + 5.3 mg DMPP in 2.4 mL MQ]<br>
Split mother culture into the following incubations, in small plastic vials I will give you:<br>
A: 5 x 8 ml culture; mother stock 42; no DMPP (just 40 ul medium)<br>
B: 5 X 8 ml culture mother stock 42 + 1 uM DMPP final concentration (2.1 ul emulsion)<br>
C: 5 X 8 ml culture mother stock 42 + 2.5 uM DMPP final concentration (21 ul emulsion)<br>
D: 5 x 8 ml culture mother stock 42 + 10 uM DMPP final concentration (210 ul emulsion)<br>
ABIOTICS:<br>
E: 5x 8 ml medium (with 0.5 ammonium); mother stock 42 + 1 uM DMPP final concentration (2.1 ul emulsion)<br>
F: 5x 8 ml medium (with 0.5 ammonium) mother stock 42 + 10 uM DMPPfinal concentration (210 ul emulsion)<br>
Take T0<br>
Sample every 20-30-60 min; 0.5 ml for N assay and 0.5 ml for analytics (for E and F only T0, T2, T4)<br>
<br>
Sample preparation d-DMPP samples:<br>
1.Centrifuge samples in Eppendorf tubes at 30,000 g for 10 minutes at 4C<br>
2.Transfer 300 uL supernatant into HPLC vials with 300/400 uL inserts<br>
<br>
Measurements according to 10.1021/ac503290j, but with an Q Exactive HF instrument<br>
<br>
2026-01-23 | MSV000100561 | MassIVE