#METABOLOMICS WORKBENCH Lsiviya_20250926_081112 DATATRACK_ID:6494 STUDY_ID:ST004271 ANALYSIS_ID:AN007109 PROJECT_ID:PR002697 VERSION 1 CREATED_ON October 9, 2025, 10:27 pm #PROJECT PR:PROJECT_TITLE Metabolomic profiles of sweet potato (Ipomoea batatas) cultivars PR:PROJECT_SUMMARY In the current syudy, four distinct sweet potato cultivars—Atacama, Jane, PR:PROJECT_SUMMARY Blesbok, and Bellevue—grown in South Africa were compared biochemically using PR:PROJECT_SUMMARY metabolomics methods. UHPLC-qTOF MS, an untargeted metabolomics technique, was PR:PROJECT_SUMMARY used to evaluate the metabolite profiles of the four cultivars. PR:INSTITUTE Nelson Mandela University PR:LAST_NAME Rauwane PR:FIRST_NAME Molemi PR:ADDRESS University Way, Summerstrand, Gqeberha, 6019 PR:EMAIL molemi.rauwane@mandela.ac.za PR:PHONE +27415044554 PR:FUNDING_SOURCE Water Research Commission (Project No. C2023/2024-01262) #STUDY ST:STUDY_TITLE Metabolomic profiles of sweet potato (Ipomoea batatas) cultivars: Insights into ST:STUDY_TITLE biochemical differences ST:STUDY_SUMMARY Sweet potato (Ipomoea batatas L.) is regarded as a crucial crop for enhancing ST:STUDY_SUMMARY food security. The crop offers several opportunities to enhance agricultural ST:STUDY_SUMMARY productivity, particularly in developing nations, for small-scale farmers. ST:STUDY_SUMMARY Greater emphasis has been made on sweet potato studies due to the crop’s ST:STUDY_SUMMARY nutritional value, including metabolites which are important in the plants’ ST:STUDY_SUMMARY defence system. Metabolomics has become a useful tool for improving our ST:STUDY_SUMMARY understanding of primary and secondary metabolism in plants. Hence, the current ST:STUDY_SUMMARY study aimed at using metabolomics tools to determine biochemical differences ST:STUDY_SUMMARY between four different sweet potato cultivars (Atacama, Jane, Blesbok and ST:STUDY_SUMMARY Bellevue) grown in South Africa. An untargeted metabolomics approach ,UHPLC-qTOF ST:STUDY_SUMMARY MS, was used to assess the metabolite profiles of the four cultivars. ST:STUDY_SUMMARY Biochemical analysis identified metabolites belonging to various classes, with ST:STUDY_SUMMARY flavonoids being the most significantly enriched class. Key metabolites included ST:STUDY_SUMMARY kaempferol 7,4’-dimethyl ether 3-O-sulfate, 7-hydroxy-3-methylflavone, ST:STUDY_SUMMARY myricetin 3,3’-digalactoside, 7β,16α-diacetoxywithanolide D and ST:STUDY_SUMMARY myristoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine. The results from this ST:STUDY_SUMMARY study also indicate differential expression of these metabolites which could be ST:STUDY_SUMMARY due to cultivar specific pathways that may be influenced by genetics, ST:STUDY_SUMMARY environmental factors and stress tolerance introduced through plant breeding . ST:STUDY_SUMMARY This study provides insights into the metabolic differences of sweet potato ST:STUDY_SUMMARY cultivars, which offers potential biomarkers contributing towards crop ST:STUDY_SUMMARY improvement. ST:INSTITUTE Nelson Mandela University ST:LAST_NAME Rauwane ST:FIRST_NAME Molemi ST:ADDRESS University Way, Summerstrand, Gqeberha, 6019 ST:EMAIL molemi.rauwane@mandela.ac.za ST:PHONE +27415044554 #SUBJECT SU:SUBJECT_TYPE Plant SU:SUBJECT_SPECIES Ipomoea batatas SU:TAXONOMY_ID 4120 SU:GENDER Not applicable #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS S1 Figlan_22042024_S1 Cultivar:Atacama | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S1.mzML SUBJECT_SAMPLE_FACTORS S2 Figlan_22042024_S2 Cultivar:Atacama | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S2.mzML SUBJECT_SAMPLE_FACTORS S3 Figlan_22042024_S3 Cultivar:Atacama | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S3.mzML SUBJECT_SAMPLE_FACTORS S4 Figlan_22042024_S4 Cultivar:Atacama | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S4.mzML SUBJECT_SAMPLE_FACTORS S5 Figlan_22042024_S5 Cultivar:Atacama | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S5.mzML SUBJECT_SAMPLE_FACTORS S6 Figlan_22042024_S6 Cultivar:Atacama | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S6.mzML SUBJECT_SAMPLE_FACTORS S7 Figlan_22042024_S7 Cultivar:Atacama | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S7.mzML SUBJECT_SAMPLE_FACTORS S8 Figlan_22042024_S8 Cultivar:Atacama | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S8.mzML SUBJECT_SAMPLE_FACTORS S25 Figlan_22042024_S25 Cultivar:Jane | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S25.mzML SUBJECT_SAMPLE_FACTORS S26 Figlan_22042024_S26 Cultivar:Jane | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S26.mzML SUBJECT_SAMPLE_FACTORS S27 Figlan_22042024_S27 Cultivar:Jane | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S27.mzML SUBJECT_SAMPLE_FACTORS S28 Figlan_22042024_S28 Cultivar:Jane | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S28.mzML SUBJECT_SAMPLE_FACTORS S29 Figlan_22042024_S29 Cultivar:Jane | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S29.mzML SUBJECT_SAMPLE_FACTORS S30 Figlan_22042024_S30 Cultivar:Jane | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S30.mzML SUBJECT_SAMPLE_FACTORS S31 Figlan_22042024_S31 Cultivar:Jane | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S31.mzML SUBJECT_SAMPLE_FACTORS S32 Figlan_22042024_S32 Cultivar:Jane | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S32.mzML SUBJECT_SAMPLE_FACTORS S49 Figlan_22042024_S49 Cultivar:Blesbok | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S49.mzML SUBJECT_SAMPLE_FACTORS S50 Figlan_22042024_S50 Cultivar:Blesbok | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S50.mzML SUBJECT_SAMPLE_FACTORS S51 Figlan_22042024_S51 Cultivar:Blesbok | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S51.mzML SUBJECT_SAMPLE_FACTORS S52 Figlan_22042024_S52 Cultivar:Blesbok | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S52.mzML SUBJECT_SAMPLE_FACTORS S53 Figlan_22042024_S53 Cultivar:Blesbok | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S53.mzML SUBJECT_SAMPLE_FACTORS S54 Figlan_22042024_S54 Cultivar:Blesbok | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S54.mzML SUBJECT_SAMPLE_FACTORS S55 Figlan_22042024_S55 Cultivar:Blesbok | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S55.mzML SUBJECT_SAMPLE_FACTORS S56 Figlan_22042024_S56 Cultivar:Blesbok | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S56.mzML SUBJECT_SAMPLE_FACTORS S73 Figlan_22042024_S73 Cultivar:Bellevue | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S73.mzML SUBJECT_SAMPLE_FACTORS S74 Figlan_22042024_S74 Cultivar:Bellevue | Sample source:leaves Biological Replication number=1; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S74.mzML SUBJECT_SAMPLE_FACTORS S75 Figlan_22042024_S75 Cultivar:Bellevue | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S75.mzML SUBJECT_SAMPLE_FACTORS S76 Figlan_22042024_S76 Cultivar:Bellevue | Sample source:leaves Biological Replication number=2; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S76.mzML SUBJECT_SAMPLE_FACTORS S77 Figlan_22042024_S77 Cultivar:Bellevue | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S77.mzML SUBJECT_SAMPLE_FACTORS S78 Figlan_22042024_S78 Cultivar:Bellevue | Sample source:leaves Biological Replication number=3; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S78.mzML SUBJECT_SAMPLE_FACTORS S79 Figlan_22042024_S79 Cultivar:Bellevue | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S79.mzML SUBJECT_SAMPLE_FACTORS S80 Figlan_22042024_S80 Cultivar:Bellevue | Sample source:leaves Biological Replication number=4; RAW_FILE_NAME(Raw file name)=Figlan_22042024_S80.mzML #COLLECTION CO:COLLECTION_SUMMARY Sweet potato trials were carried out at the Agricultural Research Council, CO:COLLECTION_SUMMARY Roodeplaat- Vegetable, Industrial and Medicinal Plants Research Institute CO:COLLECTION_SUMMARY (ARC-VIMP), Pretoria, South Africa (25 ̊59” S, 28 ̊35” E; 1189 m CO:COLLECTION_SUMMARY altitude). Experimental trials were conducted during February 2024 to April CO:COLLECTION_SUMMARY 2024. Cuttings were obtained from greenhouse propagation at the ARC-VIMP, CO:COLLECTION_SUMMARY whereby the cultivars Atacama, IIAM-Jane, Bellevue and Blesbok were pre-selected CO:COLLECTION_SUMMARY for the study. Each cultivar was cut approximately 30 cm long and planted three CO:COLLECTION_SUMMARY nodes below the soil surface. Plants were assembled in four replicates of a CO:COLLECTION_SUMMARY randomized complete block design (RCBD). Plants were watered once a week to CO:COLLECTION_SUMMARY field capacity and fertilized with multifeed and LAN (28) when required. CO:COLLECTION_SUMMARY Harvested leaf material were pulverised using liquid nitrogen and 200 mg of the CO:COLLECTION_SUMMARY powdered material was transferred into a 2 ml centrifuge tube. CO:COLLECTION_PROTOCOL_FILENAME Sweetpotato_protocol.pdf CO:SAMPLE_TYPE Leaves CO:COLLECTION_LOCATION Agricultural Research Council- Roodeplaat Vegetable and Ornamental Plants CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY No treatment TR:PLANT_GROWTH_LOCATION Agricultural Research Council- Vegetable, Industrial and Medicinal Plant, TR:PLANT_GROWTH_LOCATION Roodeplaat, South Africa TR:PLANT_PLOT_DESIGN 242 m2 TR:PLANT_ESTAB_DATE 5 February 2024 TR:PLANT_HARVEST_DATE 3 April 2024 TR:PLANT_HARVEST_METHOD Young leaves were harvested in 50ml tubes and snapped frozen in liquid nitrogen TR:PLANT_STORAGE -80C #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Harvested leaf material were pulverised using liquid nitrogen and 200 mg of the SP:SAMPLEPREP_SUMMARY powdered material was transferred into a 2 ml centrifuge tube. Sample extraction SP:SAMPLEPREP_SUMMARY process was carried using a modified methodology outlined by Maserumule et SP:SAMPLEPREP_SUMMARY al.,2023. A1.5 ml of 80% ice-cold methanol (HPLC grade, Minema Chemicals) was SP:SAMPLEPREP_SUMMARY added to each sample, and the mixture was vortexed for 30 seconds. Samples were SP:SAMPLEPREP_SUMMARY sonicated for 2 hours in 4˚C cold water, and then centrifuged for 5 minutes at SP:SAMPLEPREP_SUMMARY 2850 ref per minute (rpm) at 4°C. The supernatant was then transferred to a 2 SP:SAMPLEPREP_SUMMARY ml tube and stored at 4°C. Filtration of extracted samples was performed using SP:SAMPLEPREP_SUMMARY 0.22 µm nylon filters into glass vials containing 500 µl inserts (Alwsci SP:SAMPLEPREP_SUMMARY Techologies, 6x31 mm). Maserumule M, Rauwane M, Madala NE, Ncube E, Figlan S. SP:SAMPLEPREP_SUMMARY 2023 Defence-related metabolic changes in wheat (Triticum aestivum L.) seedlings SP:SAMPLEPREP_SUMMARY in response to infection by Puccinia graminis f. sp. tritici. Front Plant Sci SP:SAMPLEPREP_SUMMARY 14. (doi:10.3389/fpls.2023.1166813) SP:SAMPLEPREP_PROTOCOL_FILENAME Sweetpotato_protocol.pdf SP:PROCESSING_STORAGE_CONDITIONS Described in summary SP:EXTRACT_STORAGE 4℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu LCMS-9030 CH:COLUMN_NAME Shim pack Velox C18 (100 x 2.1 mm, 2.7 um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% methanol; 0.1% formic acid CH:FLOW_GRADIENT 2 minutes 10% B equilibration; 10–60% B induced throughout 3–5 minutes. From CH:FLOW_GRADIENT 5-8 minutes, the conditions were altered from 60–90% B, and from 8–11 CH:FLOW_GRADIENT minutes, the gradient was maintained at 90%. At one minute (11-12 minutes), the CH:FLOW_GRADIENT gradient was returned to initial conditions of 90-60% at 1 minute, which was CH:FLOW_GRADIENT followed by a 1-minute column equilibration period CH:FLOW_RATE 0.45 ml/min CH:COLUMN_TEMPERATURE 55 CH:METHODS_FILENAME Sweetpotato_protocol.pdf #ANALYSIS AN:ANALYSIS_TYPE MS AN:OPERATOR_NAME Prof Ntakadzeni Madala AN:ANALYSIS_PROTOCOL_FILE Sweetpotato_protocol.pdf #MS MS:INSTRUMENT_NAME Shimadzu LCMS-9030 MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS The raw data acquired from LC-qTOF-MS/MS were converted into mzML files on Lab MS:MS_COMMENTS solutions (Shim-pack UFLC SHIMADZU CBM20A). The data was then pre-processed MS:MS_COMMENTS using XCMS online (https://xcmsonline.scripps.edu/). For pre-processing, MS:MS_COMMENTS UPLC/UHD Q-TOF negative mode parameter was used, with adjustments to the MS:MS_COMMENTS parameter. Feature detection centWave method was performed with a maximum MS:MS_COMMENTS threshold (ppm) set to 5, a signal/noise threshold set to 7, prefilter peaks MS:MS_COMMENTS were set to 3, prefilter intensity was set to 700 and noise filter was set to MS:MS_COMMENTS 15. Using the obiwarp method, retention time was corrected with a profstep set MS:MS_COMMENTS to 0.5 m/z. Alignment was performed using minimum fraction (minfrac) and a width MS:MS_COMMENTS of overlapping m/z for peak density (mzmid) set to 0.025 m/z. Kruskal-Wallis MS:MS_COMMENTS non-parametric statistical test methods was applied to determine the difference MS:MS_COMMENTS between all four cultivars. Post-hoc analysis was performed with fold change MS:MS_COMMENTS threshold set to 1.5 and p-value threshold set to 0.05, followed by data MS:MS_COMMENTS normalization set to median fold change. MS:MS_RESULTS_FILE ST004271_AN007109_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END