{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002148","ANALYSIS_ID":"AN003517","VERSION":"1","CREATED_ON":"March 21, 2022, 10:47 am"},

"PROJECT":{"PROJECT_TITLE":"Untargeted metabolomics of Quercus ilex leaves in Drought","PROJECT_TYPE":"LC-MS analysis","PROJECT_SUMMARY":"UPLC-MS analysis of samples from Quercus ilex leaves. The objective of the study is to characterize the leaf metabolome of Q. ilex in six-month-old seedlings and the changes that have taken place in response to drought stress by water withholding under high temperature and irradiance conditions.","INSTITUTE":"Universidad de Córdoba","DEPARTMENT":"Department Biochemistry and Molecular Biology","LABORATORY":"Agroforestry and Plant Biochemistry, Proteomics and Systems Biology","LAST_NAME":"López-Hidalgo","FIRST_NAME":"Cristina","ADDRESS":"Campus de Rabanales; Edificio C6, Planta Baja","EMAIL":"lopezhcristina@uniovi.es","PHONE":"626894948","FUNDING_SOURCE":"This work was supported by the University of Cordoba and financial support from the Spanish Ministry of Economy and Competitiveness (Project ENCINOMICS-2 PID2019-10908RB-100)","PUBLICATIONS":"Untargeted MS-based metabolomics analysis of the responses to drought stress in Quercus ilex leaf seedlings, and identification of putative compounds related to tolerance","CONTRIBUTORS":"Marta Tienda-Parrilla, Cristina López-Hidalgo, Víctor M. Guerrero-Sánchez, Álvaro Infantes-González, Rocío Valderrama, Jesús V. Jorrin-Novo, María-Dolores Rey"},

"STUDY":{"STUDY_TITLE":"Untargeted MS-based metabolomics analysis of the responses to drought stress in Quercus ilex leaf seedlings, and identification of putative compounds related to tolerance","STUDY_TYPE":"Untargeted MS-based metabolomics","STUDY_SUMMARY":"The effect and responses to drought stress have been analyzed in Quercus ilex seedlings by using a non-targeted metabolomic approach, implementing previous studies pub-lished in which other -omics platforms, transcriptomics, and proteomics, have been em-ployed. This work is aimed at characterizing the Q. ilex leaf metabolome, determining possible mechanisms and molecular markers of drought tolerance, and identifying puta-tive bioactive compounds. Six-month-old seedling leaves, subjected to drought stress im-posed by water withholding under high temperature and irradiance conditions, were col-lected when leaf fluorescence decreased by 20 % (day 17) and 45 % (day 24) relative to ir-rigated seedlings. A total of 3934 compounds were resolved, with 616 being variable, and 342 identified, belonging to five chemical families. Out of the identified compounds 33 were variable, mostly corresponding to amino acids, carboxylic acids, benzenoids, flavo-noids and isoprenoids. Epigallocatechin, ellagic acid, pulegone, indole-3-acrylic acid and dihydrozeatin-O-glucoside were up-accumulated under drought conditions at both sam-pling times. An integrated multi-omics analysis of phenolic compounds and related en-zymes was performed, revealing that some enzymes involved in the flavonoid pathways (chalcone synthase, anthocyanidin synthase and anthocyanidin reductase) were up-accumulated at day 24 in non-irrigated seedlings. Some putative markers of drought tol-erance to drought in Q. ilex are proposed for assisting breeding programs based on the se-lection of elite genotypes.","INSTITUTE":"new","DEPARTMENT":"Universidad de Córdoba","LABORATORY":"Agroforestry and Plant Biochemistry, Proteomics and Systems Biology","LAST_NAME":"López Hidalgo","FIRST_NAME":"Cristina","ADDRESS":"Campus de Rabanales; Edificio C6, Planta Baja","EMAIL":"lopezhcristina@uniovi.es","PHONE":"626894948"},

"SUBJECT":{"SUBJECT_TYPE":"Plant","SUBJECT_SPECIES":"Quercus ilex","TAXONOMY_ID":"58334"},
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"Subject ID":"-",
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"Subject ID":"-",
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],
"COLLECTION":{"COLLECTION_SUMMARY":"Out of the biological replicates, three asymptomatic (non-damaged) non-irrigated seedlings were randomly selected for metabolomics. All leaves were collected when the chlorophyll fluorescence dropped by 20 and 45 % in the non-irrigated seedlings compared to irrigated ones (at days 17 and 24, respectively) [7]. After collection, leaves from the three biological replicates per treatment and sampling time were washed with tap water, blot dried with filter paper, shock-frozen in liquid nitrogen, and stored at -80 °C until metabolite extraction.","SAMPLE_TYPE":"new","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"An experiment based on a completely randomized design was developed by using ten biological replicates per treatment [7]. Out of the biological replicates, three asymptomatic (non-damaged) non-irrigated seedlings were randomly selected for metabolomics. All leaves were collected when the chlorophyll fluorescence dropped by 20 and 45 % in the non-irrigated seedlings compared to irrigated ones (at days 17 and 24, respectively)","TREATMENT":"Drought","PLANT_PLOT_DESIGN":"Randomized design","PLANT_LIGHT_PERIOD":"28 W m-2 solar irradiance","PLANT_HUMIDITY":"41% humidity","PLANT_TEMP":"37 °C temperature","PLANT_GROWTH_STAGE":"Six-month-old seedlings","PLANT_METAB_QUENCH_METHOD":"Liquid N2","PLANT_STORAGE":"-80ºC"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Metabolites were extracted from leaves as described by Valledor et al., [41], with minor modifications. An extraction solution containing 600 µL of ice-cold methanol: chloroform: water (5:2:2) was added to 50 mg dry weight leaf tissue and vortexed for 10 sec. The mixture was sonicated (ultrasonic bath, 40 kHz for 10 min) and after centrifugation at 20,000 × g at 4°C for 4 mins, the supernatant was transferred to a new tube. Then, 200 µL of cold methanol: chloroform: water (5:2:2) was added to the pellets and the process was repeated once. After combining both supernatants, they were vacuum dried at 30°C (Speedvac, Eppendorf Vacuum Concentrator Plus/5301, Eppendorf, Leicestershire, UK). Dried extracts were reconstituted in methanol, centrifuged at 20,000×g for 10 min, and filtered through 0.22 μm PTPE membranes (Thermo Fisher Scientific, Courtaboeuf, France), and the filtrate was collected in 1.5 mL LC/MS certified sample vials."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000 RS","COLUMN_NAME":"Waters Acquity BEH C8 (100 x 2.1mm, 1.7um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive HF hybrid Orbitrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"The operating parameters, in positive ion mode, were sheath gas flow rate at 60 kV, auxiliary gas flow rate at 25 kV, sweep gas flow rate at 2 kV, spray voltage at 3 kV, capillary temperature at 320 °C, S-lens RF level at 50 kV, and auxiliary gas heater temperature at 400 °C. The Xcalibur v3.1 software was used for instrument control and data acquisition. Spectra data were acquired in a full scan (FS) mode at a resolution of 70,000 (full width half maximum, FWHM at m/z 200) for MS1, and a data dependent (dd-MS2/dd-SIM) manner for MS2, fragmenting the five most abundant precursor ions per MS1 scan (TopN, 5), acquiring MS/MS data between 200 and 2000 m/z at a resolution of 17.500.","MS_RESULTS_FILE":"ST002148_AN003517_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}