Summary of Study ST004459
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 PR002816. The data can be accessed directly via it's Project DOI: 10.21228/M8NZ73 This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST004459 |
| Study Title | Untargeted Metabolomics Reveals Tissue-Specific Metabolic Reprogramming and Adaptation Strategies in Astragalus membranaceus var. mongholicus Seedlings under Drought Stress |
| Study Type | Untargeted Metabolomics |
| Study Summary | This study investigated the metabolic responses of Astragalus seedlings to varying degrees of drought stress using non-targeted metabolomics. Seedlings were subjected to different drought treatments and analyzed via ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). A total of 1,926 metabolites were identified in positive ion mode and 1,178 in negative ion mode. Multivariate statistical analyses, including PCA and PLS-DA, revealed distinct metabolic profiles between control and drought-treated groups. Differential metabolites were identified in the SDR.vs.CKR comparison, 118 metabolites were significantly altered in positive mode (88 up-regulated, 30 down-regulated). |
| Institute | Yili Normal University |
| Department | College of Biological Science and Technology |
| Last Name | Kuerban |
| First Name | Tusong |
| Address | No. 448, Jiefang West Road (formerly known as Jiefang Road), Yining City |
| kurban910@163.com | |
| Phone | 0999-8996985 |
| Submit Date | 2025-11-30 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML |
| Analysis Type Detail | LC-MS |
| Release Date | 2026-01-06 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002816 |
| Project DOI: | doi: 10.21228/M8NZ73 |
| Project Title: | Deciphering the Metabolomic Landscape of Medicinal Plants in Response to Drought Stress |
| Project Type: | Untargeted Metabolomics |
| Project Summary: | Background: Astragalus membranaceus var. mongholicus is a high-value medicinal plant whose growth and medicinal compound accumulation are severely impacted by drought. Understanding its metabolic adaptation to water deficit is crucial for improving cultivation and quality. While physiological responses are documented, a comprehensive, untargeted metabolomic profile revealing the systemic reprogramming of metabolites across different tissues and drought intensities remains lacking. Objectives: This metabolomics study aimed to comprehensively characterize the drought-induced metabolic responses in A. mongholicus seedlings. Specifically, we sought to: 1) Profile the global metabolite changes in both aboveground and root tissues under light (LD), moderate (MD), and severe (SD) drought stress using liquid chromatography-mass spectrometry (LC-MS); 2) Identify and quantify differentially accumulated metabolites (DAMs) associated with each stress level; 3) Determine the key metabolic pathways perturbed by drought through KEGG enrichment analysis; and 4) Integrate metabolomic findings with physiological data to elucidate the functional role of metabolic changes in drought adaptation. Main Achievements: Untargeted LC-MS metabolomics identified a total of 921 DAMs (562 in positive ion mode, 359 in negative mode) in A. mongholicus seedlings under drought. The number and direction of metabolic changes exhibited clear tissue-specific and stress intensity-dependent patterns. Aboveground tissues responded primarily by down-regulating metabolites, with key up-regulations in the antioxidant metabolite glutathione and its related pathways (glutathione metabolism, cysteine and methionine metabolism) under LD. In roots, an active up-regulation of metabolites dominated, especially under SD. Pathway analysis revealed that roots reconfigured core amino acid metabolism (e.g., tryptophan, tyrosine, arginine, and proline metabolism) to potentially support energy supply and osmotic adjustment. The accumulation of osmolytes like proline and soluble sugars correlated with these metabolic pathway shifts. This study provides the first extensive metabolomic atlas of A. mongholicus under drought stress, uncovering a tissue-divergent strategy: aerial parts enhance antioxidant capacity, while roots rewire primary metabolism for resilience. These findings and the identified metabolic markers offer a valuable resource for understanding and improving drought tolerance in medicinal plants. |
| Institute: | Yili Normal University |
| Department: | College of Biological Science and Technology |
| Last Name: | Kuerban |
| First Name: | Tusong |
| Address: | No. 448, Jiefang West Road (formerly known as Jiefang Road), Yining City |
| Email: | kurban910@163.com |
| Phone: | 0999-8996985 |
Subject:
| Subject ID: | SU004626 |
| Subject Type: | Plant |
| Subject Species: | Astragalus membranaceus var. mongholicus |
| Taxonomy ID: | 139259 |
| Genotype Strain: | not specified |
| Age Or Age Range: | Seedlings, approximately 39 to 57 days post-sowing |
| Weight Or Weight Range: | Not measured in this study |
| Height Or Height Range: | Not measured in this study |
| Gender: | Not applicable |
| Species Group: | Fabaceae |
Factors:
Subject type: Plant; Subject species: Astragalus membranaceus var. mongholicus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Drought_Intensity |
|---|---|---|---|
| SA527257 | CKA_2 | Aboveground | Control |
| SA527258 | CKA_1 | Aboveground | Control |
| SA527259 | CKA_3 | Aboveground | Control |
| SA527260 | LDA_1 | Aboveground | Light Drought |
| SA527261 | LDA_2 | Aboveground | Light Drought |
| SA527262 | LDA_3 | Aboveground | Light Drought |
| SA527263 | MDA_1 | Aboveground | Moderate Drought |
| SA527264 | MDA_2 | Aboveground | Moderate Drought |
| SA527265 | MDA_3 | Aboveground | Moderate Drought |
| SA527266 | SDA_1 | Aboveground | Severe Drought |
| SA527267 | SDA_2 | Aboveground | Severe Drought |
| SA527268 | SDA_3 | Aboveground | Severe Drought |
| SA527269 | CKR_3 | Root | Control |
| SA527270 | CKR_1 | Root | Control |
| SA527271 | CKR_2 | Root | Control |
| SA527272 | LDR_1 | Root | Light Drought |
| SA527273 | LDR_2 | Root | Light Drought |
| SA527274 | LDR_3 | Root | Light Drought |
| SA527275 | MDR_1 | Root | Moderate Drought |
| SA527276 | MDR_2 | Root | Moderate Drought |
| SA527277 | MDR_3 | Root | Moderate Drought |
| SA527278 | SDR_1 | Root | Severe Drought |
| SA527279 | SDR_2 | Root | Severe Drought |
| SA527280 | SDR_3 | Root | Severe Drought |
| Showing results 1 to 24 of 24 |
Collection:
| Collection ID: | CO004619 |
| Collection Summary: | This study collected tissue samples from Astragalus membranaceus var. mongholicus seedlings under four levels of drought stress. Stress treatments included Control (CK), Light Drought (LD), Moderate Drought (MD), and Severe Drought (SD), corresponding to soil water content of 80-85%, 65-70%, 50-55%, and 35-40% of saturated soil water capacity, respectively. The stress period lasted 18 days. Both aboveground and root tissues were separately collected from each treatment group, with three biological replicates per group, resulting in a total of 24 samples. All samples were immediately frozen in liquid nitrogen upon collection and stored at -80°C for subsequent untargeted metabolomics analysis. |
| Sample Type: | Seedlings |
Treatment:
| Treatment ID: | TR004635 |
| Treatment Summary: | This study applied gradient drought stress treatments to Astragalus membranaceus var. mongholicus seedlings. Treatments consisted of four levels: Control (CK), Light Drought (LD), Moderate Drought (MD), and Severe Drought (SD), corresponding to soil water contents of 80-85%, 65-70%, 50-55%, and 35-40% of saturated soil water capacity, respectively. The stress treatment was initiated at the seedling stage and lasted for 18 days. Soil moisture was precisely maintained at the predetermined levels by daily replenishment of water loss using the weighing method in the evening. Upon termination of the treatment, both aboveground (shoot) and root tissues were separately collected from plants in each treatment group. Each group contained three independent biological replicates, resulting in a total of 24 tissue samples. All samples were immediately flash-frozen in liquid nitrogen upon collection and stored at -80°C for subsequent untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics profiling. |
Sample Preparation:
| Sampleprep ID: | SP004632 |
| Sampleprep Summary: | Sample preparation for untargeted metabolomics analysis was performed as follows: Approximately 100 mg of plant tissue powder ground in liquid nitrogen was weighed and extracted with 500 μL of 80% methanol aqueous solution. The mixture was vortexed, incubated on ice for 5 min, and then centrifuged at 15,000 g and 4℃ for 20 min. The supernatant was collected and diluted with MS-grade water to a final methanol content of 35%, followed by another centrifugation under the same conditions (15,000 g, 4℃, 20 min). The resulting supernatant was subjected to LC-MS analysis. Chromatographic separation was carried out on a Hypersil Gold column (C18) maintained at 40℃, with a flow rate of 0.2 mL/min. Mobile phase A was 0.1% formic acid in water, and mobile phase B was methanol. Mass spectrometric detection was performed using an electrospray ionization (ESI) source in both positive and negative ion modes, with a scan range of m/z 100–1500. The spray voltage was set to 3.5 kV, sheath gas flow rate to 35 psi, auxiliary gas flow rate to 10 L/min, capillary temperature to 320°C, and auxiliary gas heater temperature to 350℃. |
Combined analysis:
| Analysis ID | AN007465 | AN007466 |
|---|---|---|
| Chromatography ID | CH005658 | CH005658 |
| MS ID | MS007161 | MS007162 |
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Fisher Vanquish UHPLC-Orbitrap Q Exactive HF | Thermo Fisher Vanquish UHPLC-Orbitrap Q Exactive HF |
| Column | Hypersil Gold column (100×2.1 mm, 1.9μm) | Hypersil Gold column (100×2.1 mm, 1.9μm) |
| MS Type | Other | Other |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH005658 |
| Chromatography Summary: | Untargeted metabolomics analysis was performed using reversed-phase liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Chromatographic separation was carried out on a Thermo Fisher Vanquish UHPLC system equipped with a Hypersil Gold C18 column (100 mm × 2.1 mm, 1.9 μm). The column temperature was maintained at 40°C with a flow rate of 0.2 mL/min. The mobile phase consisted of 0.1% formic acid in water (phase A) and methanol (phase B). A linear gradient elution program was employed as follows: 0-1.5 min, 2% B; 1.5-4.5 min, B phase increased linearly from 2% to 85%; 4.5-14.5 min, B phase increased linearly from 85% to 100%; 14.5-14.6 min, B phase rapidly decreased from 100% to 2%; followed by re-equilibration at 2% B until 17.5 min (total run time 17.5 min, note: based on a standard 12-min gradient plus equilibration time; actual run time may vary). All samples, including quality control (QC) samples, were analyzed under this single chromatographic condition. |
| Methods Filename: | Untargeted Metabolomics Experimental Methods-20241112 |
| Instrument Name: | Thermo Fisher Vanquish UHPLC-Orbitrap Q Exactive HF |
| Column Name: | Hypersil Gold column (100×2.1 mm, 1.9μm) |
| Column Temperature: | 40°C |
| Flow Gradient: | Linear gradient from 2% to 100% methanol (Solvent B) over 14.5 min at 0.2 mL/min, followed by a rapid reset to 2% B and re-equilibration. Total runtime was 17.5 min. |
| Flow Rate: | 0.2 mL/min |
| Solvent A: | 100% Water; 0.1% formic acid |
| Solvent B: | 100% Methanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS007161 |
| Analysis ID: | AN007465 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | Other |
| MS Comments: | Untargeted metabolomics in positive ESI mode. Key parameters: spray voltage 3.5 kV, capillary temp 320°C, sheath gas 35 psi, aux gas 10 L/min, S-lens RF 60, aux gas heater 350°C. Full scan m/z 100-1500. Data processed using XCMS for peak picking/alignment. Metabolites identified by matching to spectral libraries (mass tolerance 10 ppm). Features with CV >30% in QC samples were removed. |
| Ion Mode: | POSITIVE |
| MS ID: | MS007162 |
| Analysis ID: | AN007466 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | Other |
| MS Comments: | Untargeted metabolomics in negative ESI mode. Key parameters: spray voltage 3.5 kV, capillary temp 320°C, sheath gas 35 psi, aux gas 10 L/min, S-lens RF 60, aux gas heater 350°C. Full scan m/z 100-1500. Data processed using XCMS for peak picking/alignment. Metabolites identified by matching to spectral libraries (mass tolerance 10 ppm). Features with CV >30% in QC samples were removed. |
| Ion Mode: | NEGATIVE |
