Dataset: Diatom (Thalassiosira pseudonana) physiological data from experiments designed to study single-cell transcriptional profiling of nutrient acquisition heterogeneity in diatoms conducted in December of 2022

ValidatedRelease Date:2024-06-01Final no updates expectedDOI: 10.26008/1912/bco-dmo.918841.1Version 1 (2024-01-30)Dataset Type:experimental

Principal Investigator: Monica V. Orellana (University of Washington Friday Harbor Laboratories)

Co-Principal Investigator: Christopher Lausted (Institute for Systems Biology)

Scientist: Sui Huang (Institute for Systems Biology)

BCO-DMO Data Manager: Amber D. York (Woods Hole Oceanographic Institution)


Program: Ocean Carbon and Biogeochemistry (OCB)

Project: EAGER: Diatom Programmed Cell Death at Single-Cell Resolution (Diatom Death)


Abstract

This dataset includes physiological data for diatom Thalassiosira pseudonana grown during experiments conducted as part of a study of "Single-Cell transcriptional profiling of nutrient acquisition heterogeneity in diatoms." See "Related Datasets" section for T. pseudonana gene and cell information collected as part of the same study and experiments. Study description: Diatoms (Bacillariophyceae) are unicellular photosynthetic algae, accounting for about 40% of total marine primary prod...

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Cultures: Axenic Thalassiosira pseudonana (CCMP 1335, National Center for Marine Algae and Microbiota, Maine, USA; LSID urn:lsid:marinespecies.org:taxname:148934) batch cultures were grown in enriched artificial seawater (ESAW) medium modified with reduced levels of nitrate (170 μM) to characterize starvation. Before the experiment, the diatoms were acclimated to a constant 20 °C temperature and under a 12: 12 h dark: light diurnal cycle at 300 μmol photons·m−2·s−1 using cool fluorescent lights. The cultures were continuously equilibrated at ambient (420 ppm CO2 by bubbling mixed gasses (air and CO2 at 0.4 L/min) regulated using mass-flow controllers (GFC17, Aalborg) and monitored with a CO2 analyzer (model Q-S151; Qubit Systems) into a 1.5-L glass bioreactor system. The bioreactors were inoculated with 150,000 cells/ml of acclimated, axenic T. pseudonana and grown for 5 days on a 12: 12 h dark: light cycle. The pH was monitored spectrophotometrically (Dickson et al. 2007); the photochemical yield of photosystem II (variable fluorescence/maximum fluorescence Fv/Fm) was measured with an AquaPen AP100; and total dissolved nitrogen was measure using the Nitrite/Nitrate, Colorimetric Test Roche # 11746081001) kit after syringe-filtering (0.2μm) the samples. Samples were taken twice a day, in the middle of dark-time, and the middle of the light-time representing different growth conditions regulating the metabolism based on light and nitrogen on days one and two (Ashworth et al. 2013). On days three and four, samples were only taken during the light time for experiment b. This experiment was repeated to evaluate the recovery of the cells from starvation by amending NO3 (170 uM+/- 5uM) on day 5, after 70 hrs of starvation for T. pseudonana and sampled at T1: 0h, T2: 1h and T3: 3 hr, and T4: 6 hr after adding the supplemental nitrogen. Samples T1, T2, T3, were set on ice in the dark before analysis at when T4 was sampled.

Experiment Metadata (physiology, gene and cell information):

Experiment
Start_Date 12-4
Light:Dark 12:12
Lights_On 1:00 PM
Lights_(umols/m.s) 1000 umoles m-2s-1
CO2_ppm ~420
scRNAseq_Timepoints 18 hours, 30 hours, 42 hours, 54 hours, 68 hours, 80 hours, 92 hours
scRNAseq_Timepoint_Labels
Notes scRNA, Full ESAW nitrate limited (170uM). Starting innoculate = 150k/m

Related Datasets

IsRelatedTo

Dataset: T. pseudonana starve-recover experiments: Cell information
Relationship Description: Datasets were part of the same experiments conducted as part of a study of "Single-Cell transcriptional profiling of nutrient acquisition heterogeneity in diatoms."
Orellana, M. V., Lausted, C., Huang, S. (2024) Diatom (Thalassiosira pseudonana) cell information from experiments designed to study single-cell transcriptional profiling of nutrient acquisition heterogeneity in diatoms conducted in December of 2022. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2024-01-30 doi:10.26008/1912/bco-dmo.918860.1
IsRelatedTo

Dataset: T. pseudonana starve-recover experiments: Gene information
Relationship Description: Datasets were part of the same experiments conducted as part of a study of "Single-Cell transcriptional profiling of nutrient acquisition heterogeneity in diatoms."
Orellana, M. V., Lausted, C., Huang, S. (2024) Diatom (Thalassiosira pseudonana) gene information from experiments designed to study single-cell transcriptional profiling of nutrient acquisition heterogeneity in diatoms conducted in December of 2022. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2024-01-30 doi:10.26008/1912/bco-dmo.918852.1

Related Publications

Methods

Dickson, A.G.; Sabine, C.L. and Christian, J.R. (eds) (2007) Guide to best practices for ocean CO2 measurement. Sidney, British Columbia, North Pacific Marine Science Organization, 191pp. (PICES Special Publication 3; IOCCP Report 8). DOI: https://doi.org/10.25607/OBP-1342
Methods

Wolf, F. A., Angerer, P., & Theis, F. J. (2018). SCANPY: large-scale single-cell gene expression data analysis. Genome Biology, 19(1). https://doi.org/10.1186/s13059-017-1382-0