Summary of Study ST003538

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 PR002177. The data can be accessed directly via it's Project DOI: 10.21228/M8723W This work is supported by NIH grant, U2C- DK119886.

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Study IDST003538
Study TitleMetabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3_2 genes enhancing soybean resistance to cyst nematodes
Study SummaryThe Gretchen Hagen 3 genes maintain endogenous hormone homeostasis by conjugating excess hormones with amino acids. Herein, we identified the members of the GH3 family in soybeans and analyzed their phylogeny, gene duplication, structure, domains, conserved motifs, cis-elements in promoter regions for stress responses, and functional characteristics. We found that GH3 genes are induced by pathogens in Group-II. Furthermore, 8 out of 16 Group-II genes responded to cyst nematode infection. Through functional analysis of eight GmGH3 genes via overexpression, they play a negative role in soybean resistance to cyst nematode. In addition, our metabolomic analysis showed that overexpression of Glyma.02G125600, Glyma.17G165500, and Glyma.13G284600 affected the content of salicylic acid and jasmonic acid. Our findings clarify the functional features of GH3 genes and reveal their involvement in plant hormone signaling pathways. This provides valuable insights into the complex molecular mechanisms underlying the interaction between soybeans and cyst nematode.
Institute
Shenyang Agricultural University
Last NameYang
First NameXiaowen
AddressCollege of Plant Protection
Email2020200130@stu.syau.edu.cn
Phone15702428682
Submit Date2024-09-27
Analysis Type DetailGC-MS
Release Date2024-11-15
Release Version1
Xiaowen Yang Xiaowen Yang
https://dx.doi.org/10.21228/M8723W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002177
Project DOI:doi: 10.21228/M8723W
Project Title:Metabolomic analysis of fluorescent hairy roots overexpressing the Gretchen Hagen 3 genes enhancing soybean resistance to cyst nematodes
Project Summary:Soybean root samples were collected, target metabolites were extracted, standard curves were established, methodology verification was performed, LC-MS/MS metabolites were detected, and data were analyzed.Metabolome analysis was conducted on FHRs overexpressing these genes to investigate how plant hormones play a role in soybean resistance to SCN. These results provide a new framework for understanding the interaction between soybean and SCN.
Institute:Shenyang Agricultural University
Last Name:Yang
First Name:Xiaowen
Address:College of Plant Protection
Email:2020200130@stu.syau.edu.cn
Phone:15702428682

Subject:

Subject ID:SU003667
Subject Type:Plant
Subject Species:Glycine max
Taxonomy ID:3847

Factors:

Subject type: Plant; Subject species: Glycine max (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Phenotype
SA387565CK-1Plant root control
SA387566CK-2Plant root control
SA387567CK-3Plant root control
SA387568CK-4Plant root control
SA387569CK-5Plant root control
SA387570CK-6Plant root control
SA387571GH3_2-1Plant root transgene
SA387572GH3_2-2Plant root transgene
SA387573GH3_2-3Plant root transgene
SA387574GH3_2-4Plant root transgene
SA387575GH3_2-5Plant root transgene
SA387576GH3_2-6Plant root transgene
Showing results 1 to 12 of 12

Collection:

Collection ID:CO003660
Collection Summary:Liquid nitrogen grinding samples, weighing a certain amount of samples into mass spectrometry water, vortex mixing, as diluted samples; Take a dilution sample of 100 μL and add 400 μL precipitator (acetonitrile: Water =1:1), vortex mixing, extraction at 4°C for 30 min, centrifugation at 12000 rpm and 4°C for 10 min, taking 300μL supernatant slowly through the extraction column, adding 500 μL eluent (30% acetonitrile), slowly through the extraction column, mixing the two times of flow solution, LC-MS analysis.
Sample Type:Plant roots

Treatment:

Treatment ID:TR003676
Treatment Summary:Take a dilution sample of 100μL and add 400 μL precipitator (acetonitrile: Water =1:1), vortex mixing, extraction at 4°C for 30 min, centrifugation at 12000 rpm and 4°C for 10 min, taking 300 μL supernatant slowly through the extraction column, adding 500 μL eluent (30% acetonitrile), slowly through the extraction column, mixing the two times of flow solution, LC-MS analysis.

Sample Preparation:

Sampleprep ID:SP003674
Sampleprep Summary:The concentration series of the standard solution were detected by LC-MS, and the ratio of the concentration of the standard product to the internal standard was taken as the horizontal coordinate and the ratio of the peak area of the standard product to the internal standard was taken as the vertical coordinate to investigate the linearity of the standard solution. QC samples were tested by inserting a number of needles at intervals in each batch of samples. The RSD of all QC samples was calculated with the concentration of the standard substance to evaluate stability. According to the established sample pretreatment and instrumental analysis methods, all samples were quantitatively analyzed.

Combined analysis:

Analysis ID AN005810
Analysis type MS
Chromatography type HILIC
Chromatography system AB Sciex QTRAP 6500+
Column Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
MS Type EI
MS instrument type QTRAP
MS instrument name AB Sciex QTRAP 6500+
Ion Mode POSITIVE
Units Peak area

Chromatography:

Chromatography ID:CH004413
Instrument Name:AB Sciex QTRAP 6500+
Column Name:Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
Column Temperature:35°C
Flow Gradient:The initial proportion of mobile phase B is set at 15%, which then linearly increases at a rate of 0.8% per minute until it reaches 45%, where it is held for 5 minutes to adequately elute moderately polar metabolites. Afterward, it increases at a rate of 2% per minute up to 90%, and finally remains at this composition for an additional 5 minutes to clean the chromatographic column and collect less polar metabolites.
Flow Rate:0.35 mL/min
Solvent A:100% Water; 5mM Ammonium Acetate (Adjusted to pH 4.5 with Acetic Acid)
Solvent B:100% Acetonitrile
Chromatography Type:HILIC

MS:

MS ID:MS005530
Analysis ID:AN005810
Instrument Name:AB Sciex QTRAP 6500+
Instrument Type:QTRAP
MS Type:EI
MS Comments:The concentration series of the standard solution were detected by LC-MS, and the ratio of the concentration of the standard product to the internal standard was taken as the horizontal coordinate and the ratio of the peak area of the standard product to the internal standard was taken as the vertical coordinate to investigate the linearity of the standard solution. QC samples were tested by inserting a number of needles at intervals in each batch of samples. The RSD of all QC samples was calculated with the concentration of the standard substance to evaluate stability. According to the established sample pretreatment and instrumental analysis methods, all samples were quantitatively analyzed.
Ion Mode:POSITIVE
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