Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • Data Processing Description
• Parameters
• Instruments
• Deployments
• Project Information
• Program Information
• Funding

Metal quotas of two NE Pacific Line P diatom isolates and one Atlantic diatom isolate measured via ICP-MS after growth in zinc and cobalt media amendments.  Experiments with cultures collected from the GeoMICS expedition on the R/V Thomas G. Thompson (cruise TN280), along Line P in the NE Pacific, in May of 2012.

Location: Cultures collected from Northeast Pacific Line P Transect 48.8167 N 128.667 W

Metal quotas
Cellular metal quotas were measured by inductively coupled plasma mass spectrometry (ICP-MS). Biomass from replicate 25 mL matrix cultures of P. tricornutum CCMP632, P. delicatissima UNC1205, and Thalassiosira UNC1203 were pooled upon entering stationary phase and were centrifuged at 11,000 RPM (14,610 x g) for 40 minutes at 4°C. The cell pellet was resuspended in ~1 mL media and transferred to an acid-cleaned microcentrifuge tube. Cultures were centrifuged again for 30 min at 14,100 RPM (13,336 x g) at 4°C before the supernatant was discarded. The remaining cell pellet was acidified in 800 L of 5% nitric acid (Optima) containing 1 ppb indium for at least seven days. Solids were removed by centrifugation. No attempt was made to remove extracellular metals by washing cells with additional metal chelators in order to minimize processing blanks. Quota determinations therefore include contributions from both intracellular and extracellular pools. Process blank digestions containing acid but no cells were performed in parallel. Digests were diluted by a factor of 9 with 5% nitric acid 1 ppb indium solution before being analyzed in duplicate on a Thermo ICAP-Q plasma mass spectrometer calibrated to a multi-element standard curve (Spex Certiprep) over a range of 1 – 20 ppb. Samples were analyzed in KED mode after an 85s sample uptake window and element mass windows were scanned 3 times during measurements. The 1 ppb indium internal standard was used to correct for variation in sample delivery and plasma suppression between samples. Process blanks were subtracted from measured concentrations. Phosphorus concentrations were also measured by ICP-MS simultaneously and were calibrated to a standard curve ranging from 100 – 3,200 ppb using a 1 ppm certified P stock (Alfa Aesar Specpure). The seawater media base used for all growth experiments was similarly analyzed via ICP-MS using a 1:10 dilution of media base into 5% nitric acid 1 ppb indium and analyzed as above to determine background media concentrations of total Zn and Co (0.9 nmol L-1 and 0.1 nmol L-1, respectively).

Missing data identifiers in this dataset include:
* "nd" indicating no data
* Below Detection
* "contam" indicating the sample was contaminated

Isolation sources and locations
* Pseudonitzschia delicatissima UNC1205 and Thalassiosira UNC1203 were isolated from
station P8 of the Line P transect, 48.817N 128.666W
* Phaeodactylum tricornutum CCMP632 was ordered from Bigelow, the strains original
location of isolation was 54N 4W
* Thalassiosira pseudonana CCMP1335 was also from Bigelow, original location of
isolation was 40.756° N 72.82° W

BCO-DMO Data Manager Processing Notes:
* Combined three tables in sheet "Metal Quotas"  in originally submitted excel file "BCO-DMO LineP Metal Quotas.xlsx" and extracted to csv format.
* Column "culture" added to capture information in the title of each of the three original tables (P. delicatissima UNC1205, Thalassiosira UNC1203, P. tricornutum CCMP632)
* added a conventional header with dataset name, PI name, version date
* Missing data identifier in original data "NA" is displayed by default as "nd" in the bco-dmo data system.
* modified parameter names to conform with BCO-DMO naming conventions (only letters, numbers, and underscores)

NSF abstract:

Cobalt is important for many forms of marine life, yet it is one of the scarcest nutrients in the sea. Cobalt's oceanic abundance and distribution, along with other scarce nutrients, can influence the growth of microscopic plants (phytoplankton). This in turn can influence carbon cycles in the ocean and atmosphere. Therefore, knowledge of the controls on cobalt's abundance and chemical forms in seawater is a valuable component of our ability to understand the ocean's influence on global carbon cycling. Within phytoplankton and other marine microbes, metals such as cobalt, iron, nickel, and copper are used as critical components of enzymes responsible for key cellular reactions. Since these enzymes require metals to work, they are named metalloenzymes. Participating in a Pacific Ocean cruise from Alaska to Tahiti, this project will study the oceanic distributions of dissolved cobalt and the cellular content of a group of metalloenzymes known to influence biogeochemical cycles. The project will provide scientific impact by creating new knowledge about oceanic micronutrients in regions of economic interest with regard to fisheries and deep-sea mining. Measurement of proteins in the North Pacific will provide data of broad biological and chemical interest and will be made available through a new NSF-funded "EarthCube Ocean Protein Portal" data base. Educational impact will stem from participation of a graduate student and two young technicians, as well as the PI's development of a high school chemistry curriculum for use in two local high schools, thus allowing teachers to include real oceanic and environmental data at their first introduction to chemistry.

Cobalt has a complex biogeochemical cycle. Both its inorganic and organic forms are used by biology in the upper ocean and it is removed from solution by being scavenged in the intermediate and deep ocean. This scavenging removal results in cobalt having the smallest oceanic inventory of any biologically utilized element. Recent studies, however, have found that large dissolved cobalt plumes occur in major oxygen minimum zones due to a combination of less scavenging and additions from sedimentary and remineralization fluxes. The GP15 US GEOTRACES Pacific Meridional Transect (PMT) provides an opportunity to examine the influence of oxygen depletion on cobalt chemistry. Moreover, the study of the protein component of microbial communities using new proteomic techniques will provide evidence of how different major microorganisms respond to the chemical environment (e.g. through transporter production for specific nutrients and micronutrients) as well as the biochemical basis for metal requirements related to the use of specific metalloenzymes. Specifically, the PMT provides an opportunity to confirm that the Pacific oxygen minimum zones contain a large amount of cobalt and to test the hypotheses that simultaneous zinc scarcity could induce wide-scale biochemical substitution of cobalt for zinc in the North Pacific Ocean.

In support of obtaining deeper knowledge of major biogeochemically relevant proteins to inform a mechanistic understanding of global marine biogeochemical cycles.

A Gordon and Betty Moore Foundation Program.
​
Forging a new paradigm in marine microbial ecology:

Microbes in the ocean produce half of the oxygen on the planet and remove vast amounts of carbon dioxide, a greenhouse gas, from the atmosphere. Yet, we have known surprisingly little about these microscopic organisms. As we discover answers to some long-standing puzzles about the roles that marine microorganisms play in supporting the ocean’s food webs and driving global elemental cycles, we realized that we still need to learn much more about what these organisms do and how they do it—including how they evolved and contribute to our ocean's health and productivity.

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