Summary of Study ST002917

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001813. The data can be accessed directly via it's Project DOI: 10.21228/M8C713 This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002917
Study TitleTransporter-mediated depletion of apoplastic proline directly contributes to plant pattern-triggered immunity against a bacterial pathogen
Study SummaryGC-MS analysis of apoplastic fluid extracted from arabidopsis plants treated with 100 nM flg22 or a mock treatment for 8 hours. Col-0 is wild type arabidopsis plants, QKO is a quadruple knockout mutant in the Col-0 background with knockouts in dde2-2, ein2-1, pad4-1, and sid2-2.
Institute
Oregon State University
DepartmentBotany and Plant Pathology
LaboratoryJeff C Anderson
Last NameRogan
First NameConner
AddressCordley Hall, 2701 SW Campus Way, Corvallis, OR 97331
Emailroganco@oregonstate.edu
Phone3146004945
Submit Date2023-08-28
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailGC-MS
Release Date2024-02-28
Release Version1
Conner Rogan Conner Rogan
https://dx.doi.org/10.21228/M8C713
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN004787
Analysis type MS
Chromatography type GC
Chromatography system Agilent 7890B
Column Agilent DB5-MS (30m x 0.25mm, 0.25um)
MS Type EI
MS instrument type Single quadrupole
MS instrument name Agilent 5977B
Ion Mode POSITIVE
Units Peak area

Chromatography:

Chromatography ID:CH003618
Chromatography Summary:For each sample, a 20 µL aliquot of AWF was lyophilized to dryness and resuspended in 10 µL of 30 mg/mL methoxyamine hydrochloride (Sigma-Aldrich) in pyridine (Sigma-Aldrich). The resuspended sample was incubated at 37°C and shaken at 1800 rpm for 90 minutes. After adding 20 µL of N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) with 1% trimethylchlorosilane (CovaChem), the sample was incubated at 37°C and shaken at 1800 rpm for 30 minutes. Each derivatized sample was injected with a 10:1 split into an Agilent 7890B GC system with a 30 m +10 m Duraguard x 0.25 mm x 0.25 μm DB-5MS+DG Agilent column. The oven temperature was kept at 60°C for 1 minute, then ramped to 300°C at a rate of 10°C/min and held at 300°C for 10 minutes. Analytes were detected with an Agilent 5977B MSD in EI mode scanning from 50 m/z to 600 m/z. Mass spectrum analysis, component identification and peak area quantification were performed with AMDIS (http://www.amdis.net/). To reduce erroneous identifications during automated feature identification, the FiehnLib library was used to create a custom library that included both identified compounds along with unidentified prominent peaks that were recorded as unknowns and designated by their respective retention times72. Statistics were performed with MetaboAnalyst73 and MetaboAnalystR74. An in house perl script was used to collate AMDIS-generated peak areas into an Excel spreadsheet. Peaks that were present in blank samples containing derivatization chemicals only were removed from the data set. Two to three technical replicates were analyzed for each sample and averaged to produce a single sample value. MetaboAnalyst settings were selected to replace missing values by 1/5 of the minimum positive value for each feature and normalize the peak areas of the features by the peak area of the internal ribitol standard in each respective sample.
Instrument Name:Agilent 7890B
Column Name:Agilent DB5-MS (30m x 0.25mm, 0.25um)
Column Temperature:60 ramped to 300
Flow Gradient:NA
Flow Rate:.52 mL/min helium gas
Injection Temperature:250
Internal Standard:ribitol
Solvent A:NA
Solvent B:NA
Chromatography Type:GC
Solvent C:NA
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