Samples were acquired using the Scheduled MRM™ Pro Algorithm in Analyst® Software. The information dependent acquisition (IDA) acquisition method consisted of a multiple reaction monitoring (MRM) survey scan coupled with an enhanced product ion scan for compound identity confirmation. Internal patterns were used as quality control of the process and to help determine retention times.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse-phase"],"chromatography_protocol":["Analysis was performed as previously described as previously described[1]. An XSELECT HSS XP (2.5 µm, 2.1 mm × 150 mm; Waters) with an UFLC XR HPLC (Shimadzu) was used for chromatographic separation. Mobile phase A was composed of 10 mM tributylamine (TBA), 10 mM acetic acid (pH 6.86), 5% methanol, and 2% 2-propanol; mobile phase B was 2-propanol. Oven temperature was 40°C. The optimized chromatographic gradient is described in Table 1. The autosampler temperature was 10°C and the injection volume was 10 µL with full loop injection.
McCloskey, D. and Ubhi, B.K., 2014. Quantitative and qualitative metabolomics for the investigation of intracellular metabolism. SCIEX Tech Note, pp.1-11.
Metabolites were determined from 15-20 mg of lyophilized cell biomass subjected to mechanical disruption in a Mini Bead Beater (Biospe Products) with 0.5 mm glass beads in the presence of 1 mL extraction buffer consisting of chloroform:methanol (3:7, v/v). Centrifugation at 5000 × g for 5 min at RT was then performed and these two steps were repeated until the pellets were found colorless. Supernatants were concentrated using speed-vacuum (Eppendorf concentrator Plus).
"],"organism":["Chlamydomonas reinhardtii"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS14720"],"author":["Inmaculada Couso. Spanish National Research Council. inmaculada.couso@ibvf.csic.es.","Rodrigo Bedera-García. Spanish National Research Council. rbedera@us.es."],"data_transformation_protocol":["Raw data was normalized using 10 µM paracetamol as an internal standard and the dry weight of each sample.
"],"study_factor":["HighLight","Genotype"],"submitter_email":["rbedera@us.es"],"sample_collection_protocol":["Four independent biological replicates were assessed for each strain and condition. The metabolite content determination was performed using cell pellets that firstly were lyophilized (Skadi-Europe TFD 8503), and stored at −20°C as described in Serrano-Perez et al. (2022) and Bedera-Garcia (2025).
Serrano-Perez, Emma, et al. 'Transcriptomic and metabolomic response to high light in the charophyte alga Klebsormidium nitens.' Frontiers in Plant Science 13 (2022): 855243.
Bedera-Garcia Rodrigo, et al. 'Inositol polyphosphates regulate resilient mechanisms in the green alga Chlamydomonas reinhardtii to adapt to extreme nutrient conditions.' Physiologia Plantarum 177.1 (2025): e70089.
"],"omics_type":["Metabolomics"],"study_design":["Metabolomics","targeted analysis","inositol polyphosphates","control","Targeted metabolomics","Whole Organism","AB SCIEX QTRAP 6500+","Mutant HL treatment","Photosynthesis regulation","Chlamydomonas reinhardtii","Mutant","Cell Signaling","Shimadzu Prominence UFLCXR","HL treatment"],"curator_keywords":["Metabolomics","targeted analysis","inositol polyphosphates","control","Targeted metabolomics","Whole Organism","AB SCIEX QTRAP 6500+","Mutant HL treatment","Chlamydomonas reinhardtii","Photosynthesis regulation","Mutant","Cell Signaling","Shimadzu Prominence UFLCXR","HL treatment"],"mass_spectrometry_protocol":["Analysis was performed as previously described as previously described[1]. A QTRAP® 6500+ System (SCIEX) operating in negative mode was used for targeted profiling using the Multiple Reaction Monitoring (MRM) approach. electrospray ionization parameters were optimized for 0.3mL/min flow rate, and are as follows: electrospray voltage of -4500 V, temperature of 500 °C, curtain gas of 40, CAD gas of 12, and gas 1 and 2 of 50 and 50 psi, respectively. Analyzer parameters were optimized for each compound using manual tuning. The instrument was mass calibrated with a mixture of polypropylene glycol (PPG) standards.
[1] McCloskey, D. and Ubhi, B.K., 2014. Quantitative and qualitative metabolomics for the investigation of intracellular metabolism. SCIEX Tech Note, pp.1-11.
"],"metabolite_name":["Sucrose","Citrate","Glucose","UDP-Glucose","ADP","Fructose1,6P","Sedoheptulose7P","3Phosphoglycerate","Histidine","Phosphoenolpyruvate","Tyrosine","AMP","Threonine","Succinate","Methionine","Glutamate","cis-Aconitate","Malate","Isocitrate","Sucrose6P","Glyceraldehyde3P","alpha-ketoglutarate","Serine","DihydroxyacetoneP","Cystine","Paracetamol","NADH","L-citrulline","6Phosphogluconic","Arginine","Ac-CoA","NADP","Ribulose1,5P","Glucose6P","ADP-Glucose","Shikimate","GSSG","Alanine","Phenylalanine","Fructose6P","Pyruvate","Tryptophan","Aspartate","Ribose5P","Erythrose4P","NAD","Fumarate","Valine","Asparagine","ATP","Glucosylglycerol","Glutamine"],"additional_accession":[]},"is_claimable":false,"name":"Inositol Pyrophosphates Mediate Chloroplast Lipid Remodeling and Nuclear Gene Repression during High-Light Acclimation in Chlamydomonas reinhardtii","description":"Microalgae are photosynthetic organisms capable of autotrophic growth. Their applicability in multiple industrial fields has been largely studied, thanks to their ability to fixate CO2 into high added value organic products like fatty acids and carotenoids. However, our under-standing of the cellular signaling networks that control carbon flux and acclimation to envi-ronmental stress remains incomplete. In this study, we used the Chlamydomonas reinhard-tii mutant strain vip1-1, which carries a loss-of-function mutation in the hexakisphosphate kinase responsible for the synthesis of inositol pyrophosphates InsP7 and InsP8 (PP-InsPs), to investigate the role of these molecules during high-light acclimation. Our results indicate that PP-InsPs participate in the regulation of carbon storage in the form of starch and their deficiency increases TAGs levels in the algal cells. They also impact chloroplast-specific lipid remodeling by modifying membrane composition and fluidity through fatty acid desatu-rations and glycerolipid composition. In addition, our findings suggest that PP-InsPs are in-volved in chloroplast-nucleus communication, where they coordinate transcriptional repres-sion of photosynthesis associated nuclear genes (PhANGs), fatty acid desaturases and lipid synthases, contributing to cellular acclimation to high light. We also found that PP-InsPs modulating effect extended to protein synthesis and accumulation of Calvin-Benson-Bassham cycle intermediates. Therefore, we propose that PP-InsPs function as integratory molecules that balance carbon allocation between storage and structural pools, in response to environmental cues such as high light. These data uncover a novel function of PP-InsPs in high light acclimation and potentially in chloroplast-nucleus communication, providing new insights that may help engineering more resilient and efficient strains.","dates":{"publication":"2026-06-09","submission":"2026-06-08"},"accession":"MTBLS14720","cross_references":{"MetaboLights":["MTBLC16467","MTBLC18050","MTBLC32650","MTBLC16977","MTBLC17115","MTBLC16857","MTBLC15971","MTBLC17992","MTBLC17376","MTBLC16414","MTBLC16643","MTBLC17895","MTBLC17295","MTBLC17053","MTBLC16015","MTBLC16828","MTBLC17138","MTBLC15846","MTBLC57723","MTBLC57579","MTBLC60880","MTBLC18066","MTBLC17794","MTBLC18021","MTBLC16908","MTBLC18009","MTBLC456216","MTBLC16905","MTBLC30616","MTBLC15351","MTBLC15954","MTBLC15741","MTBLC18012","MTBLC30915","MTBLC57498","MTBLC91033","MTBLC17858","MTBLC16349","MTBLC46195","MTBLC48153","MTBLC16108","MTBLC17797","MTBLC48928","MTBLC17234","MTBLC15721","MTBLC6650","MTBLC32816","MTBLC16947","MTBLC30887","MTBLC32805","MTBLC16119","MTBLC16710"],"ChEBI":["CHEBI:16467","CHEBI:18050","CHEBI:32650","CHEBI:16977","CHEBI:17115","CHEBI:16857","CHEBI:15971","CHEBI:17992","CHEBI:17376","CHEBI:16414","CHEBI:16643","CHEBI:17895","CHEBI:17295","CHEBI:17053","CHEBI:16015","CHEBI:16828","CHEBI:17138","CHEBI:15846","CHEBI:57723","CHEBI:57579","CHEBI:60880","CHEBI:18066","CHEBI:17794","CHEBI:18021","CHEBI:16908","CHEBI:18009","CHEBI:456216","CHEBI:16905","CHEBI:30616","CHEBI:15351","CHEBI:15954","CHEBI:15741","CHEBI:18012","CHEBI:30915","CHEBI:57498","CHEBI:91033","CHEBI:17858","CHEBI:16349","CHEBI:46195","CHEBI:48153","CHEBI:16108","CHEBI:17797","CHEBI:48928","CHEBI:17234","CHEBI:15721","CHEBI:6650","CHEBI:32816","CHEBI:16947","CHEBI:30887","CHEBI:32805","CHEBI:16119","CHEBI:16710"]}}