Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • BCO-DMO Processing Description
• Parameters
• Instruments
• Deployments
• Project Information
• Funding

Methods on EN667:
One liter (L) samples were collected via CTD rosette for particulate lipid, carbohydrate, and organic carbon samples. Samples were filtered onto 47-millimeter (mm) 0.2-micrometer (um) Durapore filters (lipids, carbs; Millipore) and 25 mm 0.7 um GF/F filters (POC; Whatman), flash frozen in liquid nitrogen, and stored in LN2 headspace until extraction.

Carbohydrate quantiifcation was performed according to a "standard addition" protocol as follows: particulate carbohydrates were filtered onto 47 mm 0.2 um pore-size Durapore filters, flash frozen at sea, and returned to lab. In the lab, filter was cut in half, one half was "spiked" with a known-concentration monosaccharide standard mix, and both halves were then analyzed separately using the following extraction and analysis procedure: sample filter halves were extracted in 1.8 milliliter (mL) 0.4 molar (M) hydrochloric acid for 20 hours at 100 degrees Celsius (C), dried down at 60 degrees C in a vacuum centrifuge, resuspended in milliQ water, and analyzed on a Thermo Dionex ICS 6000 anion exchange chromatography tandem pulsed amperometric detector. The instrumental ionization response factor was calculated separately for each sample to account for matrix effects, and was calculated using the difference in instrument response peak area between the non-"spiked" and "spiked" filter halves for each analyte peak (i.e. via "standard addition" protocol). The final concentration of the non-"spiked" filter half (i.e. half of the filtered sample volume) was then calculated using that response factor. Samples were then blank corrected using the average of three 2-L filtered seawater blanks collected from the surface of the Sargasso Sea in September 2023. Both raw and blank-corrected per-liter carbohydrate (galactose and glucose) values are reported in molarity (M).

Methods on SR2320:
One liter samples were collected via CTD rosette for particulate lipid, carbohydrate, and organic carbon samples. Samples were filtered onto 47 mm 0.2 um Durapore filters (lipids, carbs; Millipore) and 25 mm 0.7 um GF/F filters (POC; Whatman), flash frozen in liquid nitrogen, and stored in LN2 headspace until extraction.

Carbohydrate quantiifcation was performed according to a "standard addition" protocol as follows: particulate carbohydrates were filtered onto 47 mm 0.2 um pore-size Durapore filters, flash frozen at sea, and returned to lab. In the lab, filter was cut in half, one half was "spiked" with a known-concentration monosaccharide standard mix, and both halves were then analyzed separately using the following extraction and analysis procedure: sample filter halves were extracted in 1.8 mL 0.4 M hydrochloric acid for 20 hours at 100 degrees C, dried down at 60 degrees C in a vacuum centrifuge, resuspended in milliQ water, and analyzed on a Thermo Dionex ICS 6000 anion exchange chromatography tandem pulsed amperometric detector. The instrumental ionization response factor was calculated separately for each sample to account for matrix effects, and was calculated using the difference in instrument response peak area between the non-"spiked" and "spiked" filter halves for each analyte peak (i.e. via "standard addition" protocol). The final concentration of the non-"spiked" filter half (i.e. half of the filtered sample volume) was then calculated using that response factor. Samples were then blank corrected using the average of three 2-L filtered seawater blanks collected from the surface of the Sargasso Sea in September 2023. Both raw and blank-corrected per-liter particulate carbohydrate (galactose and glucose) values are reported in molarity (M).

Methods on AE2320:
Two liter samples were collected via CTD rosette for particulate lipid, carbohydrate, and organic carbon samples. Samples were filtered onto 47 mm 0.2 um Durapore filters (lipids, carbs; Millipore) and 25 mm 0.7 um GF/F filters (POC; Whatman), flash frozen in liquid nitrogen, and stored in LN2 headspace until extraction.

Carbohydrate quantiifcation was performed according to a "standard addition" protocol as follows: particulate carbohydrates were filtered onto 47 mm 0.2 um pore-size Durapore filters, flash frozen at sea, and returned to lab. In the lab, filter was cut in half, one half was "spiked" with a known-concentration monosaccharide standard mix, and both halves were then analyzed separately using the following extraction and analysis procedure: sample filter halves were extracted in 1.8 mL 0.4 M hydrochloric acid for 20 hours at 100 degrees C, dried down at 60 degrees C in a vacuum centrifuge, resuspended in milliQ water, and analyzed on a Thermo Dionex ICS 6000 anion exchange chromatography tandem pulsed amperometric detector. The instrumental ionization response factor was calculated separately for each sample to account for matrix effects, and was calculated using the difference in instrument response peak area between the non-"spiked" and "spiked" filter halves for each analyte peak (i.e. via "standard addition" protocol). The final concentration of the non-"spiked" filter half (i.e. half of the filtered sample volume) was then calculated using that response factor. Samples were then blank corrected using the average of three 2-L filtered seawater blanks collected from the surface of the Sargasso Sea in September 2023. Both raw and blank-corrected per-liter particulate carbohydrate (galactose and glucose) values are reported in molarity (M).

- Imported the three original CSV files (SR2310_RIPPLE_2_GalGlu_data.csv, EN667_RIPPLE_1_GalGlu_data.csv, AE2320_RIPPLE_3_GalGlu_data.csv) into the BCO-DMO system.
- Treated "NA" as a missing value (missing values are empty/blank in the final CSV file).
- Concatenated the three files into a single file, adding a column for Cruise_Name.
- Converted the original "Time_UTC" column (format %m/%d/%Y %H:%M, UTC) into ISO 8601 format and renamed it to "ISO_DateTime_UTC".
- Renamed fields to comply with BCO-DMO naming conventions.
- Converted Lat from degrees-decimal_minutes to decimal degrees, applied only to rows where Cruise_ID == EN667.
- Converted Long from degrees-decimal_minutes to decimal degrees, applied only to rows where Cruise_ID == EN667.
- Rounded Lat and Long to maximum 8 decimal places (only values exceeding that precision affected).
- Saved the final file as "997842_v1_carbohydrates_ripple_cruises.csv".

The Production and Fate of Fats in the Upper Ocean Phytoplankton, microscopic photosynthetic organisms in the ocean, produce a type of fat called triaclyglycerols (TAGs). A recent discovery in the North Pacific Ocean showed that a significant percentage of primary production from phytoplankton is devoted to producing TAGs. This suggests that TAGs may serve as a source of energy for phytoplankton at night, when they are unable to generate energy from photosynthesis. Phytoplankton are the base of the food web in the ocean. Therefore, it is important to understand how they create and store energy. This project will investigate the role of fats in the ocean. Specifically, this research will look at: 1) factors that affect the production and use of TAGs by phytoplankton and 2) how TAGs contribute to the global carbon cycle. This project will support the training and education of graduate students. This project will also provide resources for mentoring high school students, support summer research experiences for high school and undergraduate students, and offer field trips for 7th grade students.

Triacylglycerols (TAGs) are one of the most abundant classes of lipids in the ocean. A new discovery suggests that TAGs are also a very dynamic class of biochemicals. A recent study in the surface waters of the North Pacific subtropical gyre (NPSG) showed that TAGs doubled in concentration between sunrise and sunset daily, accounting for 16 to 42% of net primary production by eukaryotic nanophytoplankton (Becker et al., 2018). These results show that TAGs are an vital component of the physiology of eukaryotic phytoplankton and that TAGs contribute significantly to the carbon cycle of the NPSG. Based on estimates from this study, daytime production of TAGs in the subtropical gyres accounted for 4 to 6 percent of total global primary production. Outside of subtropical gyres, the production rates of TAGs are entirely unknown, particularly in regions where primary production rates are higher and eukaryotic phytoplankton are more dominant. Thus, the contribution of TAGs to the global ocean carbon cycle is almost certainly underestimated. There are major outstanding questions about TAGs. What environmental factors affect rates of net TAG production? What fraction of net TAG production is exported in sinking particles? Do TAGs play a role in the food web of the euphotic zone? How much of the TAGs produced during the day do phytoplankton themselves consume at night? These questions will be answered using state-of-the-art lipidomics, in situ observations, isotope-tracing techniques, incubations, and on-deck experiments. This study will provide significant advances in our understanding of TAG metabolism in phytoplankton, elucidate the roles that TAGs play in the marine carbon cycle, constrain their global importance by studying TAGs in multiple disparate environments, and set the groundwork for future research on these fascinating and vital molecules.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.