URLhttps://www.bco-dmo.org/dataset/816576
Download URLhttps://www.bco-dmo.org/dataset/816576/data/download
Media Type text/tab-separated-values
Created June 23, 2020
Modified June 30, 2020
State Final no updates expected

Acquisition Description

Location: North Atlantic, western flank of the mid-Atlantic Ridge 22.75589 N 46.08125 W

Methodology:

Prior to the extraction, we freeze-dried, ground and sieved sediment samples to less than 125 μm (Ruttenberg 1992). For a given sample, we weighed four sample replicates (2 g) and placed each in 250 mL HDPE bottles. Sodium dithionite (F.W. 147.12 g/mol; 7.4 g) was added to each sample split, followed by 200 mL of citrate-bicarbonate solution (pH 7.6). This step produces effervescence, so the solution should be added slowly to the sample. We shook samples for 8 h and then centrifuged them at 3,700 rpm for 15 min. We filtered the supernatants with a 0.4 μm polycarbonate filter. We took 20 mL aliquots from the filtrate for each sample split for MRP and total P analyses, and kept them refrigerated until analysis within 24 h. We added 200 mL of ultrapure water to the solid residue for each sample split as a wash step after the above reductive step, shook samples for 2 h, and then centrifuged them at 3,700 rpm for 15 min. We filtered the supernatants with 0.4 μm polycarbonate filters and set aside 20 mL of filtrate from each sample split for MRP and total P analyses. We then extracted the solid sample residues in 200 mL of sodium acetate buffer (pH 4.0) for 6 h. At the end of this extraction step, we centrifuged the bottles at 3,700 rpm for 15 min, filtered the supernatants with 0.4 μm polycarbonate filters and took a 20 mL aliquot of filtrate from each sample split for MRP and total P analyses. We added 200 mL of ultrapure water to the solid residue for each sample split as a wash step, shook samples for 2 h, and then centrifuged them at 3,700 rpm for 15 min.  We filtered the supernatants with 0.4 μm polycarbonate filters and set aside 20 mL of filtrate from each sample split for MRP and total P analyses. We repeated the water rinse step, and collected aliquots for MRP and total P analyses as in the previous steps. The concentrations of  TP were determined as described below.

Solid sediment sample residues following the pretreatment described above were transferred to two 50 mL centrifuge tubes (2 sample replicates combined per tube). We added 20 mL of 0.25 M NaOH + 0.05 M Na2EDTA solution to each tube, vortexed until all sediment was resuspended and then shook samples for 6 h at room temperature (Cade-Menun et al. 2005). We used a solid to solution ratio of 1:5 for this step to minimize the amount of freeze-dried material that will need to be dissolved for the 31P NMR experiments. Large amounts of salts from the NaOH-EDTA concentrated in NMR samples lead to higher viscosity and increase line broadening on NMR spectra (Cade-Menun and Liu 2014). We chose an extraction time of 6 h to improve total P recovery while limiting the degradation of natural P compounds in the sample. At the end of the extraction, samples were centrifuged at 3,700 rpm for 15 min and supernatants decanted into 50 mL centrifuge tubes. We collected a 500 μL aliquot from each sample, which we diluted with 4.5 mL of ultrapure water. These were refrigerated until analysis for total P content on the ICP-OES. The sample residues and supernatants were frozen on a slant to maximize the exposed surface area during the lyophilization step; this was done immediately after the removal of the 500 μL aliquot. Once completely frozen, the uncapped tubes containing supernatants and residues were freeze-dried over the course of 48 h. Each tube was covered with parafilm with small holes from a tack to minimize contamination. Freeze-dried supernatants from identical sample splits were combined and dissolved in 500 μL each of ultrapure water, D2O, NaOH-EDTA and 10 M NaOH prior to 31P NMR analysis.  The D2O is required as signal lock in the spectrometer (Cade-Menun and Liu 2014). Sample pH was maintained at a pH > 12 to optimize peak separation (Cade-Menun 2005; Cade-Menun and Liu 2014). Sample pH was assessed with a glass electrode, and verified with pH paper to account for the alkaline error caused by the high salt content of our samples (Covington 1985).

Freeze-dried sample residues were ashed in crucibles at 550oC for 2 h and then extracted in 25 mL of 0.5 M sulfuric acid for 16 h (Olsen and Sommers 1982; Cade-Menun and Lavkulich 1997). We centrifuged samples at 3,700 rpm for 15 min, filtered supernatants with 0.4 μm polycarbonate filters, and measured P content on an ICP-OES. 

Total P concentrations in sediment extracts were measured using inductively coupled plasma optical emission spectroscopy (ICP-OES). Standards were prepared with the same solutions as those used for the extraction procedure in order to minimize matrix effects on P measurements. Sediment extracts and standards (0 μM, 3.2 μM, 32 μM and 320 μM) were diluted to lower salt content to prevent salt buildup on the nebulizer (1:20 dilution for step 1, 1:10 for steps 2 – 4). Concentration data from both wavelengths (213 nm and 214 nm) were averaged to obtain extract concentrations for each sample. The detection limit for P on this instrument for both wavelengths is 0.4 μM. The MRP concentrations were measured on a QuikChem 8000 automated ion analyzer. Standards were prepared with the same solutions used for the extraction step to minimize matrix effects on P measurements. Sediment extracts and standards (0 – 30 μM PO4) were diluted ten-fold to prevent matrix interference with color development. The detection limit for P on this instrument is 0.2 μM. We derived MUP concentrations by subtracting MRP from total P concentrations.

Processing Description

Data were processed in Excel.

BCO-DMO Data Manager Processing Notes:
* Data from originally submitted Excel file Data_TP.xlsx exported as csv.  Sheets for steps 1-4 combined into one data table.  
* added a conventional header with dataset name, PI name, version date
* modified parameter names to conform with BCO-DMO naming conventions
* blank values in this dataset are displayed as “nd” for “no data.” nd is the default missing data identifier in the BCO-DMO system.
* Date format changed to ISO 8601 format yyyy-mm-dd

Instruments

Also referred to as an Inductively coupled plasma atomic emission spectroscope (ICP-AES). These instruments pass nebulised samples into an inductively-coupled gas plasma (8-10000 K) where they are atomised and excited. The de-excitation optical emissions at characteristic wavelengths are spectroscopically analysed. It is often used in the detection of trace metals.

Parameters

Extract [sample_descrip]
Details
Extract
Extract solution
text description of sample collected
Details
Step
Step in the sequential extraction scheme (1-4)
text description of sample collected
Dilution [sample_descrip]
Details
Dilution
Sample dilution
text description of sample collected
Sample_ID [sample]
Details
Sample_ID
Sample ID, unique sample identifier

unique sample identification or number; any combination of alpha numeric characters; precise definition is file dependent

Analyte_Name [sample_descrip]
Details
Analyte_Name
Element analyzed
text description of sample collected
Int_Corr [unknown]
Details
Int_Corr
Intensity (corrected)
association with a community-wide standard parameter is not yet defined
RSD_Corr_Int [unknown]
Details
RSD_Corr_Int
Relative standard deviation (RSD) of corrected intensity
association with a community-wide standard parameter is not yet defined
Conc_Calib [P]
Details
Conc_Calib
Calibrated concentration of total phosphorous

P (Phosphorus). May be reported in parts per million, nanomoles per liter, or other units. Refer to dataset metadata for units.

Date [date]
Details
Date

Date the samples were analyzed in ISO 8601 format yyyy-mm-dd

date; generally reported in GMT as YYYYMMDD (year; month; day); also as MMDD (month; day); EqPac dates are local Hawaii time. ISO_Date format is YYYY-MM-DD (http://www.iso.org/iso/home/standards/iso8601.htm)

Dataset Maintainers

NameAffiliationContact
Adina PaytanUniversity of California-Santa Cruz (UC Santa Cruz)
Delphine DefforeyUniversity of California-Santa Cruz (UC Santa Cruz)
Amber YorkUniversity of California-Santa Cruz (UC Santa Cruz)
Amber YorkUniversity of California-Santa Cruz (UC Santa Cruz)
Amber YorkWoods Hole Oceanographic Institution (WHOI BCO-DMO)

BCO-DMO Project Info

Project Title Potential phosphorus uptake mechanisms of the deep sedimentary biosphere
Acronym Deep sea sediments
URLhttps://www.bco-dmo.org/project/664073
Created November 7, 2016
Modified November 7, 2016
Project Description

The goal of this project is to explore potential microbial P uptake mechanisms in marine sediments beneath the North Atlantic Gyre and their effects on the relative distribution of organic P compounds as a function of burial depth and changing redox conditions. We use a combination of metagenomic analyses and solution 31P nuclear magnetic resonance spectroscopy (31P NMR) to investigate (1) the presence of microbial functional genes pertaining to P uptake and metabolism and (2) the possible P substrates for the deep biosphere in these oligotrophic sediments.

NSF C-DEBI Award #156246 to Dr. Adina Paytan

NSF C-DEBI Award #157598 to Dr. Delphine Defforey 

Data Project Maintainers
NameAffiliationRole
Adina PaytanUniversity of California-Santa Cruz (UC Santa Cruz)Principal Investigator
Benjamin J. TullyTexas A&M University (TAMU)Co-Principal Investigator
Jason B. SylvanUniversity of Southern California (USC)Co-Principal Investigator
Delphine DefforeyUniversity of California-Santa Cruz (UC Santa Cruz)Co-Principal Investigator
Barbara J. Cade-MenunAgriculture and Agri-Food Canada (AGR GC)Co-Principal Investigator
Brandi Kiel ReeseTexas A&M, Corpus Christi (TAMU-CC)Co-Principal Investigator
Laura A. ZinkeUniversity of Southern California (USC)Co-Principal Investigator
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