Summary of Study ST003035
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 PR001888. The data can be accessed directly via it's Project DOI: 10.21228/M8P728 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST003035 |
| Study Title | Central Transcriptional Regulator Controls Growth and Carbon Storage under High Light Stress in Photosynthetic Microalgae Model Strains |
| Study Type | Algae |
| Study Summary | Carbon capture efficiency and biochemical storage are some of the primary drivers of photosynthetic productivity and by extension crop yield. To elucidate the mechanisms governing yield phenotypes and carbon allocation regulatory elements, we selected two microalgae strains as simplified models of photosynthetic crops. The Picochlorum celeri TG2 isolate is one of the fastest growing algae and in this work is juxtaposed to a closely related, slower growing, isolate, TG1, of the same species with less than 2% genomic divergence. Through the application of a comprehensive systems biology light-stress response study, we observed a stark difference in carbon assimilation and storage rates, with the slower growing isolate accumulating almost three times the amount of starch compared to the fast-growing isolate. We characterized the carbon storage rates and allocation dynamics, with metabolic bottlenecks, and transport rates of intermediates underlying the variations in growth and composition in high light using instationary 13C-fluxomics experiments. High light stress analysis of transcriptomic dynamics during acclimation of the strains from low to high light identified a widespread response with up to 73% the annotated gene set significantly differentially expressed after only 1 hour. Broad transcriptional regulatory control was inferred by a rapid depletion of a global diel-responsive transcription factor closely related to a circadian-regulator in plants, as the single most distinct transcription factor. Transferring this factor to the slower variant increased yield, specific growth rate, and carbohydrate accumulation of the selected engineered strain, providing further evidence for a coordinating regulatory mechanism for this complex phenotype. |
| Institute | National Renewable Energy Lab |
| Department | Biosciences |
| Laboratory | Laurens Lab |
| Last Name | Laurens |
| First Name | Lieve |
| Address | 15013 Denver West Paekway, Golden, CO 80401 |
| lieve.laurens@nrel.gov | |
| Phone | +1 720-273-6534 |
| Submit Date | 2023-12-18 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-26 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN004976 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Vanquish |
| Column | Waters ACQUITY Premier BEH Amide (150 x 2.1mm, 1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE |
| Units | Raw Area Count |
Chromatography:
| Chromatography ID: | CH003757 |
| Chromatography Summary: | The metabolites were separated using hydrophilic interaction chromatography (HILIC) using a BEH Amide column (1.7 µm, 2.1 mm X 150 mm, ACQUITY Premier BEH Amide, Waters Corporation) via a gradient method. Solvent A comprised of 20 mM ammonium acetate and 15 mM ammonium hydroxide in 97% 18.2 mΩ-cm water and 3% acetonitrile whereas solvent B comprised of 20 mM ammonium acetate and 15 mM ammonium hydroxide in 95% acetonitrile and 5% 18.2 mΩ-cm water. All the chemicals used were LC-MS grade. The chromatography method used a constant flow rate of 0.2 mL/min and a linear gradient to enable separation of a wide range of metabolites followed by a column regeneration step as follows: 90% solvent B for 1 min followed by a linear gradient down to 75% solvent B for 23 min, a linear gradient down to 45% solvent B in 2 min, a linear gradient down to 25% B in 4 min, followed by a step to the starting composition of 90% B and hold for 6 min for column regeneration for a total run time of 36 min. The column temperature was maintained at 25°C and injection volume was set to 20 µL. Metabolite extracts were diluted such that the final injection solvent was 75% acetonitrile. Data was collected using a Thermo Scientific Q-Exactive mass spectrometer in negative ion mode. The scan ranged from 75 to 1000 m/z with a resolution of 140,000. AGC target was set to 3e6 while maximum IT was 200 ms. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY Premier BEH Amide (150 x 2.1mm, 1.7um) |
| Column Temperature: | 25 |
| Flow Gradient: | 90% solvent B for 1 min followed by a linear gradient down to 75% solvent B for 23 min, a linear gradient down to 45% solvent B in 2 min, a linear gradient down to 25% B in 4 min, followed by a step to the starting composition of 90% B and hold for 6 min for column regeneration for a total run time of 36 min |
| Flow Rate: | 0.2 mL/min |
| Internal Standard: | 1-13C Leucine |
| Solvent A: | 20 mM ammonium acetate and 15 mM ammonium hydroxide in 97% 18.2 mΩ-cm water and 3% acetonitrile |
| Solvent B: | 20 mM ammonium acetate and 15 mM ammonium hydroxide in 95% acetonitrile and 5% 18.2 mΩ-cm water |
| Randomization Order: | Samples were randomized |
| Chromatography Type: | HILIC |
