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MB Sample ID: SA151625

Local Sample ID:root-27
Subject ID:SU001725
Subject Type:Plant
Subject Species:Hordeum vulgare L.
Taxonomy ID:4513

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Subject:

Subject ID:SU001725
Subject Type:Plant
Subject Species:Hordeum vulgare L.
Taxonomy ID:4513

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
root-27SA151625FL017759Root_P deficiency-3Treatment

Collection:

Collection ID:CO001718
Collection Summary:Barley seedlings were allowed to grow up to two-leaf-stage in the normal hydroponic solution, and then exposed to various treatments after removing endosperm for eliminating its nutrient supply. The treatments included 20% PEG for drought stress, 150 mM NaCl for salt stress, 50 μM Al3+ at pH 4.5 for aluminum stress (Al stress, at the same time the nutrient solution with pH 4.5 and without Al addition used as control), 5 μM CdCl2 for cadmium stress (Cd stress), 0 mM N for N deficiency (replace KNO3, Ca(NO3).4H2O and (NH4)6Mo7O24 with 1mM CaCl2, 1mM KH2PO4 and 0.07 mM H2MoO4), 0 mM P for P deficiency (replace NH4H2PO4 with 1mM NH4Cl), 0 mM K for K deficiency (replace KNO3 with 1mM NaNO3). The hydroponic solution was renewed every four days. After twenty days of the treatments, the plants were harvested, and fresh and dry weight were recorded quickly. The dry tissues were used for mineral element concentration analysis with three biological replicates for each treatment. Meanwhile, fresh tissues were frozen in liquid nitrogen and stored at -80 ℃ for metabolite analysis with four biological replicates for each stress treatment.
Sample Type:Plant
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001738
Treatment Summary:The treatments included 20% PEG for drought stress, 150 mM NaCl for salt stress, 50 μM Al3+ at pH 4.5 for aluminum stress (Al stress, at the same time the nutrient solution with pH 4.5 and without Al addition used as control), 5 μM CdCl2 for cadmium stress (Cd stress), 0 mM N for N deficiency (replace KNO3, Ca(NO3).4H2O and (NH4)6Mo7O24 with 1mM CaCl2, 1mM KH2PO4 and 0.07 mM H2MoO4), 0 mM P for P deficiency (replace NH4H2PO4 with 1mM NH4Cl), 0 mM K for K deficiency (replace KNO3 with 1mM NaNO3).

Sample Preparation:

Sampleprep ID:SP001731
Sampleprep Summary:The frozen tissue was homogenized in liquid nitrogen and approximately 100 mg of powder sample was added into 2 ml vertical centrifuge tube. Then 1.4 ml of 100% methanol (pre-cooled at -20 °C) was added and vortexed thoroughly. Then the tubes were shaken for 10 min at 70 °C, followed by centrifugation at 11,000 g for 10 min. The supernatant was transferred into a new 10 ml sterile centrifuge tube, and 750 μl chloroform (−20 °C) and 1.5 ml deionized water (4 °C) were added. The mixture was vortexed and centrifuged at 2200 g for 15 min. The upper methanol/water phase was collected for metabolite analysis. A total of 150 μl collected solution was dried in a vacuum freeze dryer. The dried residue was derivatized in 40 μl of 15 mg ml-1 methoxylamine hydrochloride in pyridine for 120 min at 37 °C, followed by reaction with 70 μl of N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) reagent for another 30 min at 37 °C. After the reaction, the reacted solution was transferred to glass vial insert in 2 ml glass bottle (Agilent, USA) for GC-MS determination. Meanwhile, one blank derivatization reaction (with reagent) was also prepared as a control.

Combined analysis:

Analysis ID AN002694
Analysis type MS
Chromatography type GC
Chromatography system Agilent 7890A
Column Agilent DB5-MS (30m x 0.25mm, 0.25um)
MS Type API
MS instrument type GC-TOF
MS instrument name Agilent 7890A
Ion Mode UNSPECIFIED
Units pmoles/l

Chromatography:

Chromatography ID:CH001987
Instrument Name:Agilent 7890A
Column Name:Agilent DB5-MS (30m x 0.25mm, 0.25um)
Chromatography Type:GC

MS:

MS ID:MS002492
Analysis ID:AN002694
Instrument Name:Agilent 7890A
Instrument Type:GC-TOF
MS Type:API
MS Comments:The retention time (RT) and mass spectra of each compound were analyzed using AMDIS (Automated Mass Spectral Deconvolution and Identification System) software (http://chemdata.nist.gov/).
Ion Mode:UNSPECIFIED
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