Summary of Study ST001912

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 PR001206. The data can be accessed directly via it's Project DOI: 10.21228/M8TD6R 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.

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Study ID	ST001912
Study Title	Identification of effector metabolites using exometabolite profiling of diverse microalgae
Study Summary	We conducted an untargeted metabolomic analysis of non-polar exometabolites from four phylogenetically and ecologically diverse eukaryotic microalgal strains grown in the laboratory: freshwater Chlamydomonas reinhardtii, brackish Desmodesmus sp., marine Phaeodactylum tricornutum, and marine Microchloropsis salina. We analyzed both exometabolomes and cell pellet metabolomes to identify released metabolites based on relative enrichment in the exometabolomes.
Institute	Lawrence Berkeley National Laboratory
Last Name	Brisson
First Name	Vanessa
Address	7000 East Ave, Livermore, CA, 94550, USA
Email	brisson2@llnl.gov
Phone	5107177560
Submit Date	2021-08-23
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2021-09-10
Release Version	1

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Project:

Project ID:	PR001206
Project DOI:	doi: 10.21228/M8TD6R
Project Title:	LLNL Biofuels SFA
Project Summary:	The LLNL Biofuels SFA brings a systems biology approach to understanding interactions and resource allocation in bioenergy-relevant microbial communities.
Institute:	Lawrence Berkeley National Laboratory
Last Name:	Brisson
First Name:	Vanessa
Address:	7000 East Ave, Livermore, CA, 94550, USA
Email:	vlbrisson@lbl.gov
Phone:	510-717-7560
Funding Source:	DOE

Subject:

Subject ID:	SU001990
Subject Type:	Algae
Subject Species:	Chlamydomonas reinhardtii;Desmodesmus sp.;Phaeodactylum tricornutum;Microchloropsis salina
Taxonomy ID:	3055;91202;2950;2511165

Factors:

Subject type: Algae; Subject species: Chlamydomonas reinhardtii;Desmodesmus sp.;Phaeodactylum tricornutum;Microchloropsis salina (Factor headings shown in green)

mb_sample_id	local_sample_id	Type
SA176935	79_E-FW-Cr-Inc_4	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176936	78_E-FW-Cr-Inc_3	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176937	76_E-FW-Cr-Inc_1	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176938	80_E-FW-Cr-Inc_5	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176939	77_E-FW-Cr-Inc_2	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176940	86_E-FW-DF-Inc_1	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176941	90_E-FW-DF-Inc_5	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176942	88_E-FW-DF-Inc_3	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176943	89_E-FW-DF-Inc_4	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176944	87_E-FW-DF-Inc_2	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176945	67_E-FW-TxCtrl-Inc_2	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176946	66_E-FW-TxCtrl-Inc_1	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176947	68_E-FW-TxCtrl-Inc_3	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176948	70_E-FW-TxCtrl-Inc_5	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176949	69_E-FW-TxCtrl-Inc_4	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176950	57_E-FW-TxCtrl-noInc_2	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176951	56_E-FW-TxCtrl-noInc_1	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176952	58_E-FW-TxCtrl-noInc_3	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176953	59_E-FW-TxCtrl-noInc_4	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176954	60_E-FW-TxCtrl-noInc_5	E - exometabolome \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176955	47_E-SW-DS-Inc_2	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA176956	49_E-SW-DS-Inc_4	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA176957	46_E-SW-DS-Inc_1	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA176958	50_E-SW-DS-Inc_5	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA176959	48_E-SW-DS-Inc_3	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA176960	38_E-SW-Ms-Inc_3	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA176961	39_E-SW-Ms-Inc_4	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA176962	40_E-SW-Ms-Inc_5	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA176963	36_E-SW-Ms-Inc_1	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA176964	37_E-SW-Ms-Inc_2	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA176965	27_E-SW-Pt-Inc_2	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA176966	28_E-SW-Pt-Inc_3	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA176967	26_E-SW-Pt-Inc_1	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA176968	30_E-SW-Pt-Inc_5	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA176969	29_E-SW-Pt-Inc_4	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA176970	16_E-SW-TxCtrl-Inc_1	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176971	18_E-SW-TxCtrl-Inc_3	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176972	19_E-SW-TxCtrl-Inc_4	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176973	17_E-SW-TxCtrl-Inc_2	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176974	20_E-SW-TxCtrl-Inc_5	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176975	8_E-SW-TxCtrl-noInc_3	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176976	9_E-SW-TxCtrl-noInc_4	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176977	7_E-SW-TxCtrl-noInc_2	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176978	10_E-SW-TxCtrl-noInc_5	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176979	6_E-SW-TxCtrl-noInc_1	E - exometabolome \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA177025	2_none-none-ExCtrl-noInc_2	none \| Medium:none \| Algal strain or abiotic control condition:ExCtrl - extraxtion control \| Incubation:noInc - not incubated
SA177026	4_none-none-ExCtrl-noInc_4	none \| Medium:none \| Algal strain or abiotic control condition:ExCtrl - extraxtion control \| Incubation:noInc - not incubated
SA177027	1_none-none-ExCtrl-noInc_1	none \| Medium:none \| Algal strain or abiotic control condition:ExCtrl - extraxtion control \| Incubation:noInc - not incubated
SA177028	5_none-none-ExCtrl-noInc_5	none \| Medium:none \| Algal strain or abiotic control condition:ExCtrl - extraxtion control \| Incubation:noInc - not incubated
SA177029	3_none-none-ExCtrl-noInc_3	none \| Medium:none \| Algal strain or abiotic control condition:ExCtrl - extraxtion control \| Incubation:noInc - not incubated
SA176980	81_P-FW-Cr-Inc_1	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176981	83_P-FW-Cr-Inc_3	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176982	85_P-FW-Cr-Inc_5	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176983	84_P-FW-Cr-Inc_4	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176984	82_P-FW-Cr-Inc_2	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:Cr - Chlamydomonas reinhardtii \| Incubation:Inc - incubated
SA176985	91_P-FW-DF-Inc_1	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176986	92_P-FW-DF-Inc_2	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176987	94_P-FW-DF-Inc_4	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176988	95_P-FW-DF-Inc_5	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176989	93_P-FW-DF-Inc_3	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:DF - Desmodesmus sp. \| Incubation:Inc - incubated
SA176990	71_P-FW-TxCtrl-Inc_1	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176991	95_P-FW-TxCtrl-Inc_5	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176992	72_P-FW-TxCtrl-Inc_2	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176993	74_P-FW-TxCtrl-Inc_4	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176994	73_P-FW-TxCtrl-Inc_3	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA176995	65_P-FW-TxCtrl-noInc_5	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176996	64_P-FW-TxCtrl-noInc_4	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176997	61_P-FW-TxCtrl-noInc_1	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176998	62_P-FW-TxCtrl-noInc_2	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA176999	63_P-FW-TxCtrl-noInc_3	P - pellet \| Medium:FW - freshwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA177000	53_P-SW-DS-Inc_3	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA177001	52_P-SW-DS-Inc_2	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA177002	51_P-SW-DS-Inc_1	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA177003	55_P-SW-DS-Inc_5	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA177004	54_P-SW-DS-Inc_4	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:DS - Desmodesmus sp. \| Incubation:Inc - incubated
SA177005	43_P-SW-Ms-Inc_3	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA177006	44_P-SW-Ms-Inc_4	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA177007	42_P-SW-Ms-Inc_2	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA177008	41_P-SW-Ms-Inc_1	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA177009	45_P-SW-Ms-Inc_5	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Ms - Microchloropsis salina \| Incubation:Inc - incubated
SA177010	31_P-SW-Pt-Inc_1	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA177011	34_P-SW-Pt-Inc_4	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA177012	32_P-SW-Pt-Inc_2	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA177013	33_P-SW-Pt-Inc_3	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA177014	35_P-SW-Pt-Inc_5	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:Pt - Phaeodactylum trocornutum \| Incubation:Inc - incubated
SA177015	24_P-SW-TxCtrl-Inc_4	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA177016	21_P-SW-TxCtrl-Inc_1	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA177017	22_P-SW-TxCtrl-Inc_2	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA177018	23_P-SW-TxCtrl-Inc_3	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA177019	55_P-SW-TxCtrl-Inc_5	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:Inc - incubated
SA177020	13_P-SW-TxCtrl-noInc_3	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA177021	15_P-SW-TxCtrl-noInc_5	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA177022	12_P-SW-TxCtrl-noInc_2	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA177023	14_P-SW-TxCtrl-noInc_4	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated
SA177024	11_P-SW-TxCtrl-noInc_1	P - pellet \| Medium:SW - saltwater \| Algal strain or abiotic control condition:TxCtrl - treatment control \| Incubation:noInc - not incubated

Showing results 1 to 95 of 95

Showing results 1 to 95 of 95

Collection:

Collection ID:	CO001983
Collection Summary:	Axenic algal were grown in glass 125 mL Erlenmeyer flasks that had previously been baked in a muffle furnace at 550°C for two hours to remove trace organics that could contaminate metabolomics analyses. Each flask contained 50 mL of the appropriate medium (defined freshwater or saltwater medium) and was capped with a foam stopper and covered with aluminum foil to avoid contamination during the experiment. Flasks were incubated with shaking at 90 rpm in a light incubator, with a 12-hour day/night light cycle. Daytime illumination was 3500 lux, and the incubator temperature was maintained at 22°C. Algal inocula were prepared by first growing algal stock cultures under experimental conditions (flasks, media, incubation) for one week to acclimate cultures to those conditions. Experimental flasks were inoculated by transferring 2 mL of inoculum culture to flasks with the appropriate medium. Cultures were harvested for metabolomic analysis from five replicate flasks for each alga after 8 days of incubation. Spent medium and cell pellets were separated by centrifugation. Half of the culture was transferred to a 50 mL falcon tube and centrifuged at 5000 g for eight minutes at 4°C. The supernatant was carefully transferred to a clean Falcon tube, and the remainder of the culture was added to the tube containing the pellet and the centrifugation procedure was repeated. After separation by centrifugation, the supernatant was filtered through a 0.45µm pore size syringe filter to remove residual algal cells. Filtered spent medium and cell pellets were frozen on dry ice and stored at -80°C until extraction.
Sample Type:	Algae
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR002002
Treatment Summary:	Treatments consisted of 4 different axenic algal strains growth separately for comparison: Chlamydomonas reinhardtii, Desmodesmus sp., Phaeodactylum tricornutum, and Microchloropsis salina. C. reinhartdii was grown in defined freshwater mediunm. Desmodesmus was grown separately in defined freshwater and saltwater medium. P. tricornutum and M. salina were grown separately in defined saltwater medium. Separate axenic controls were conducted for the defined freshwater and saltwater media. Metabolite extraction controls were also prepared using the same extraction protocol.

Treatment ID:

TR002002

Treatment Summary:

Treatments consisted of 4 different axenic algal strains growth separately for comparison: Chlamydomonas reinhardtii, Desmodesmus sp., Phaeodactylum tricornutum, and Microchloropsis salina. C. reinhartdii was grown in defined freshwater mediunm. Desmodesmus was grown separately in defined freshwater and saltwater medium. P. tricornutum and M. salina were grown separately in defined saltwater medium. Separate axenic controls were conducted for the defined freshwater and saltwater media. Metabolite extraction controls were also prepared using the same extraction protocol.

Sample Preparation:

Sampleprep ID:	SP001996
Sampleprep Summary:	In order to disrupt cells for metabolite extraction, cell pellets were resuspended in 1 mL sterile water, frozen at -80°C, and lyophilized. Lyophilized pellets were disrupted using a sterilized steel ball and vortexing three times for five seconds. Disrupted pellets were then resuspended in 50 mL sterile water and filtered and frozen in the same manner as spent medium. Thus, the spent medium and cell pellet samples corresponded to the same original sample volume. Samples for LC-MS/MS metabolomics analysis were extracted by solid phase extraction using Waters Sep-Pak C18 3 mL cartridges. Because chlorophyll and biomass content were similar across samples, samples were normalized to the same volume (30 mL). Cartridges were first preconditioned with 3 mL methanol, and the rinsed with 6 mL ultrapure water. Metabolites were extracted by passing 30 mL of sample through the cartridge. Columns with extracted metabolites were rinsed with 6 mL ultrapure water. Metabolites were eluted with 1 mL methanol into a 1.5 mL polypropylene tube and the dried in a vacuum centrifuge with a refrigerated vapor trap. Dried metabolite extracts resuspended in 150 µL methanol with matrix control internal standards, vortexed for 5 to 10 seconds, and sonicated in a water bath sonicator with ice for ten minutes. Samples were centrifuged at 10,000 g for five minutes at 4°C to pellet insoluble salts and proteins. Supernatant was transferred to a 0.2 µm pore size centrifuge filter device, and centrifuged at 5,000 g for five minutes at 4°C. Filtrate was transferred to an autosampler vial for LC-MS/MS analysis.

Sampleprep ID:

SP001996

Sampleprep Summary:

In order to disrupt cells for metabolite extraction, cell pellets were resuspended in 1 mL sterile water, frozen at -80°C, and lyophilized. Lyophilized pellets were disrupted using a sterilized steel ball and vortexing three times for five seconds. Disrupted pellets were then resuspended in 50 mL sterile water and filtered and frozen in the same manner as spent medium. Thus, the spent medium and cell pellet samples corresponded to the same original sample volume. Samples for LC-MS/MS metabolomics analysis were extracted by solid phase extraction using Waters Sep-Pak C18 3 mL cartridges. Because chlorophyll and biomass content were similar across samples, samples were normalized to the same volume (30 mL). Cartridges were first preconditioned with 3 mL methanol, and the rinsed with 6 mL ultrapure water. Metabolites were extracted by passing 30 mL of sample through the cartridge. Columns with extracted metabolites were rinsed with 6 mL ultrapure water. Metabolites were eluted with 1 mL methanol into a 1.5 mL polypropylene tube and the dried in a vacuum centrifuge with a refrigerated vapor trap. Dried metabolite extracts resuspended in 150 µL methanol with matrix control internal standards, vortexed for 5 to 10 seconds, and sonicated in a water bath sonicator with ice for ten minutes. Samples were centrifuged at 10,000 g for five minutes at 4°C to pellet insoluble salts and proteins. Supernatant was transferred to a 0.2 µm pore size centrifuge filter device, and centrifuged at 5,000 g for five minutes at 4°C. Filtrate was transferred to an autosampler vial for LC-MS/MS analysis.

Combined analysis:

Analysis ID	AN003109	AN003110
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Agilent 1290	Agilent 1290
Column	Agilent Sorbax Eclipse Plus C18 RRHD (50 x 2.1 mm, 1.8 um)	Agilent Sorbax Eclipse Plus C18 RRHD (50 x 2.1 mm, 1.8 um)
MS Type	ESI	ESI
MS instrument type	Single quadrupole	Single quadrupole
MS instrument name	Thermo Q Exactive Orbitrap	Thermo Q Exactive Orbitrap
Ion Mode	NEGATIVE	POSITIVE
Units	peak height	peak height

Chromatography:

Chromatography ID:	CH002296
Instrument Name:	Agilent 1290
Column Name:	Agilent Sorbax Eclipse Plus C18 RRHD (50 x 2.1 mm, 1.8 um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS002891
Analysis ID:	AN003109
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Single quadrupole
MS Type:	ESI
MS Comments:	Feature detection conducted using MZmine 2
Ion Mode:	NEGATIVE

MS ID:	MS002892
Analysis ID:	AN003110
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Single quadrupole
MS Type:	ESI
MS Comments:	Feature detection conducted using MZmine 2
Ion Mode:	POSITIVE