Grazing experiment 7: Cellular carbohydrate data for low-high pCO2 acclimated Rhodomonas sp. cultures, 2011-2016 (E Hux Response to pCO2 project)

Data Type: experimental
Version Date: 2016-12-14

» Planktonic interactions in a changing ocean: Biological responses of Emiliania huxleyi to elevated pCO2 and their effects on microzooplankton (E Hux Response to pCO2)
Olson, Brady M.Western Washington University (WWU)Principal Investigator
Love, BrookeWestern Washington University (WWU)Co-Principal Investigator
Strom, SuzanneWestern Washington University (WWU)Co-Principal Investigator
Still, Kelly AnnWestern Washington University (WWU)Student
Copley, NancyWoods Hole Oceanographic Institution (WHOI BCO-DMO)BCO-DMO Data Manager

Dataset Description

Related Reference:
Still, Kelly Ann, Microzooplankton grazing, growth and gross growth efficiency are affected by pCO2 induced changes in phytoplankton biology. (Masters Thesis) Western Washington University.

Acquisition Description

The phytoplankton Rhodomonas sp. CCMP 755 was grown semi-continuously in atmosphere controlled chambers at three different CO2 treatment concentrations; Ambient (400ppmv), Moderate (750ppmv), and High (1000ppmv).  Cultures were diluted daily starting day 4 with pre-equilibrated media containing f/50 nutrients.  Some of the culture removed was used to evaluate chemical parameters.  Samples for carbohydrate mass were taken by gravity filtering 30 ml of culture through muffled GF/F filters. Filters were wrapped in aluminum foil and stored at -80°C until analysis.  Prior to analysis, filters were extracted in 1 ml of 95% H2SO4 and 1 ml of nanopure water in a sonication bath for 30 minutes and then 20 hours at room temperature.  After extraction, samples were centrifuged at 10,000 rpm for 6 minutes.  1.6 ml of the extract was combined with 4 ml of concentrated H2SO4 and 0.8 ml of 10% phenol and allowed to react for 30 minutes.  Samples were analyzed colorimetrically using a Spec20D+ spectrophotometer at 485 nm and compared with standards made from fructose.

Processing Description

Picograms per ml for carbohydrate were calculated based on standard curves using fructose and were then normalized to per cell based on cell counts for the sample day.

BCO-DMO Processing Notes:
- added conventional header with dataset name, PI name, version date
- modified parameter names to conform with BCO-DMO naming conventions
- nd (no data) was entered into all blank cells

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samplesample ID that names the treatment replicate and the day of semi-continuous culture or standard concentration unitless
nm485485 nm wavelength value unitless?
fructose_1_6mlcarbohydrate content in fructose picograms (pg) per 1.6 ml extract
fructose_mlcarbohydrate content per milliliter
cell_mlcell concentration in the culture cells/milliliter
vol_filt_mlvolume of culture filtered for sample milliliters
cells_on_filtertotal cells on the filter cells
cells_per_ml_extractcells per ml of extract milliliters
fructose_ug_cellcarbohydrate as fructose micrograms/cell
fructose_pg_cellcarbohydrate as fructose picograms/cell

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Dataset-specific Instrument Name
Spec20D+ spectrophotometer
Generic Instrument Name
Generic Instrument Description
An instrument used to measure the relative absorption of electromagnetic radiation of different wavelengths in the near infra-red, visible and ultraviolet wavebands by samples.

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Start Date
End Date
laboratory experiments

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Project Information

Planktonic interactions in a changing ocean: Biological responses of Emiliania huxleyi to elevated pCO2 and their effects on microzooplankton (E Hux Response to pCO2)

Description from NSF award abstract:
The calcifying Haptophyte Emiliania huxleyi appears to be acutely sensitive to the rising concentration of ocean pCO2. Documented responses by E. huxleyi to elevated pCO2 include modifications to their calcification rate and cell size, malformation of coccoliths, elevated growth rates, increased organic carbon production, lowering of PIC:POC ratios, and elevated production of the active climate gas DMS. Changes in these parameters are mechanisms known to elicit alterations in grazing behavior by microzooplankton, the oceans dominant grazer functional group. The investigators hypothesize that modifications to the physiology and biochemistry of calcifying and non-calcifying Haptophyte Emiliania huxleyi in response to elevated pCO2 will precipitate alterations in microzooplankton grazing dynamics. To test this hypothesis, they will conduct controlled laboratory experiments where several strains of E. huxleyi are grown at several CO2 concentrations. After careful characterization of the biochemical and physiological responses of the E. huxleyi strains to elevated pCO2, they will provide these strains as food to several ecologically-important microzooplankton and document grazing dynamics. E. huxleyi is an ideal organism for the study of phytoplankton and microzooplankton responses to rising anthropogenic CO2, the effects of which in the marine environment are called ocean acidification; E. huxleyi is biogeochemically important, is well studied, numerous strains are in culture that exhibit variation in the parameters described above, and they are readily fed upon by ecologically important microzooplankton.

The implications of changes in microzooplankton grazing for carbon cycling, specifically CaCO3 export, DMS production, nutrient regeneration in surface waters, and carbon transfer between trophic levels are profound, as this grazing, to a large degree, regulates all these processes. E. huxleyi is a model prey organism because it is one of the most biogeochemically influential global phytoplankton. It forms massive seasonal blooms, contributes significantly to marine inorganic and organic carbon cycles, is a large producer of the climatically active gas DMS, and is a source of organic matter for trophic levels both above and below itself. The planned controlled study will increase our knowledge of the mechanisms that drive patterns of change between trophic levels, thus providing a wider array of tools necessary to understand the complex nature of ocean acidification field studies, where competing variables can confound precise interpretation.

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Funding SourceAward
NSF Division of Ocean Sciences (NSF OCE)

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