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Summary

Abstract

Introduction

Protocol

Representative Results

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Acknowledgements

Materials

References

Environment

Laboratory Simulation of an Iron(II)-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

Published: July 24th, 2016

DOI:

10.3791/54251

1Department of Geosciences, University of Tuebingen, 2Department of Geological and Atmospheric Sciences, Iowa State University

We simulated a Precambrian ferruginous marine upwelling system in a lab-scale vertical flow-through column. The goal was to understand how geochemical profiles of O2 and Fe(II) evolve as cyanobacteria produce O2. The results show the establishment of a chemocline due to Fe(II) oxidation by photosynthetically produced O2.

A conventional concept for the deposition of some Precambrian Banded Iron Formations (BIF) proceeds on the assumption that ferrous iron [Fe(II)] upwelling from hydrothermal sources in the Precambrian ocean was oxidized by molecular oxygen [O2] produced by cyanobacteria. The oldest BIFs, deposited prior to the Great Oxidation Event (GOE) at about 2.4 billion years (Gy) ago, could have formed by direct oxidation of Fe(II) by anoxygenic photoferrotrophs under anoxic conditions. As a method for testing the geochemical and mineralogical patterns that develop under different biological scenarios, we designed a 40 cm long vertical flow-through column to simulate an anoxic Fe(II)-rich marine upwelling system representative of an ancient ocean on a lab scale. The cylinder was packed with a porous glass bead matrix to stabilize the geochemical gradients, and liquid samples for iron quantification could be taken throughout the water column. Dissolved oxygen was detected non-invasively via optodes from the outside. Results from biotic experiments that involved upwelling fluxes of Fe(II) from the bottom, a distinct light gradient from top, and cyanobacteria present in the water column, show clear evidence for the formation of Fe(III) mineral precipitates and development of a chemocline between Fe(II) and O2. This column allows us to test hypotheses for the formation of the BIFs by culturing cyanobacteria (and in the future photoferrotrophs) under simulated marine Precambrian conditions. Furthermore we hypothesize that our column concept allows for the simulation of various chemical and physical environments — including shallow marine or lacustrine sediments.

The Precambrian (4.6 to 0.541 Gy ago) atmosphere experienced a gradual build-up of photosynthetically produced oxygen (O2), perhaps punctuated by step changes at the so-called "Great Oxidation Event" (GOE) at approximately 2.4 Gy ago, and again in the Neoproterozoic (1 to 0.541 Gy ago) as atmospheric O2 approached modern levels1. Cyanobacteria are the evolutionary remnants of the first organisms capable of oxygenic photosynthesis2. Geochemical evidence and modeling studies support the role of shallow coastal environments in harboring active communities of cyanobacteria or organisms capable of oxygenic photosynthesis ....

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1. Preparation of Culturing Medium

Note: Information on the required equipment, chemicals and supplies for the preparation of the culture medium is listed in Table 1. Italic alphanumerical codes in brackets refer to the equipment itemized in Table 2 and shown in Figure 1.

  1. Prepare 5 L of Marine Phototroph (MP) medium (referred to hereafter as "medium") following the protocol of Wu et al.16. Adjust the pH .......

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Control experiment

Abiotic control experiments (10 days) demonstrated consistently low oxygen concentrations (O2 <0.15 mg/L) with no significant fluctuations in the Fe(II)-profile throughout the upwelling water column. The formation of precipitates (presumably Fe(III)(oxyhydr-)oxides) in the medium reservoir and the slight decrease in the overall Fe(II) concentration from 500 µM to.......

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Microbial communities in the Precambrian ocean were regulated by, or modified as a result of, their activity and the prevailing geochemical conditions. In interpreting the origins of BIF, researchers generally infer the presence or activity of microorganisms based on the sedimentology or geochemistry of BIF, e.g., Smith et al.23 and Johnson et al.24. The study of modern organisms in modern environments that have geochemical analogs to ancient environments is also a valuabl.......

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Mark Nordhoff assisted in the design and implementation of tubing connections. Ellen Struve helped to select and acquire equipment used.

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Name Company Catalog Number Comments
Widdel flask (5 L) Ochs 110015 labor-ochs.de
Glass bottles (5 L) Rotilabo Y682.1 carlroth.com
Glass pipettes (5 mL) 51714 labor-ochs.de
0.22 µm Steritop filter unit (0.22 µm Polyethersulfone membrane) Millipore X337.1 carlroth.com
Aluminum foil
Sterile Luer Lock glass syringe, filled with cotton C681.1 carlroth.com
Luer Lock stainless steel needles (150 mm, 1.0 mm ID) 201015 labor-ochs.de
NaCl Sigma 433209 sigmaaldrich.com
MgSO4 Sigma 208094 sigmaaldrich.com
CaCl2 Sigma C4901 sigmaaldrich.com
NH4Cl Sigma A9434 sigmaaldrich.com
KH2PO4 Sigma P5655 sigmaaldrich.com
KBr Sigma P3691 sigmaaldrich.com
KCl Sigma P9541 sigmaaldrich.com
Glass cylinder Y310.1 carlroth.com
Glass wool 7377.2 carlroth.com
Glass beads (ø 0.55 - 0.7 mm) 11079105 biospec.com
Butyl rubber stopper (ø 1.2 cm) 271024 labor-ochs.de
Petri Dish, glass (ø 8.0 cm) T939.1 carlroth.com
Polymers glue OTTOSEAL S68 adchem.de
Optical oxygen sensor foil (for oxygen analysis, see below) – on request – presens.de
Rubber tubing (35 mm, 7 mm ID) 770350 labor-ochs.de
Luer Lock tube connector (3.0 mm, luer lock male = LLM) P343.1 carlroth.com
Luer Lock tube connector (3.0 mm, luer lock female = LLF) P335.1 carlroth.com
Rubber tubing (25 mm, 0.72 mm ID) 2600185 newageindustries.com
Rubber tubing (50 mm, 7 mm ID) 770350 labor-ochs.de
Luer Lock stainless steel needle (150 mm, 1.0 mm ID) 201015 labor-ochs.de
Luer Lock glass syringe (10 mL) C680.1 carlroth.com
Loose cotton 
Butyl rubber stopper (ø 1.75 cm) 271050 labor-ochs.de
Stainless steel needle (40 mm, 1.0 mm ID) Sterican 4665120 bbraun.de
Luer Lock stainless steel needle (150 mm, 1.5 mm ID) 201520 labor-ochs.de
position: Luer Lock female connector part at C.7
Polymers glue OTTOSEAL S68 adchem.de
Stainless steel needle (120 mm, 0.7 mm ID) Sterican 4665643 bbraun.de
Rubber tubing (40 mm, 0.74 mm ID) 2600185 newageindustries.com
Heat shrink tubing (35 mm, 3 mm ID shrunk) 541458 - 62 conrad.de
Tube clamp STHC-C-500-4 tekproducts.com
Luer Lock tube connector (1.0 mm, LLF) P334.1 carlroth.com
Luer Lock plastic cap (LLM) CT69.1 carlroth.com
Glass bottle (5 L) Rotilabo Y682.1 carlroth.com
Butyl rubber stopper (for GL45) 444704 labor-ochs.de
Stainless steel capillary (300 mm, 0.74 mm ID) 56736 sigmaaldrich.com
Stainless steel capillary (50 mm, 0.74 mm ID) 56737 sigmaaldrich.com
Shrink tubing (35 mm, 3 mm ID shrunk) 541458 - 62 conrad.de
Rubber tubing (100 mm, 0.74 mm ID) 2600185 newageindustries.com
Luer Lock tube connector (1.0 mm, LLF) P334.1 carlroth.com
Luer Lock glass syringe (10 mL) C680.1 carlroth.com
Loose cotton 
Butyl rubber stopper (ø 1.75 cm) 271050 labor-ochs.de
Stainless Steel needle (40 mm, 0.8 mm ID) Sterican 4657519 bbraun.de
Luer Lock glass syringe (5 mL) C679.1 carlroth.com
Butyl rubber stopper (ø 1.75 mm) 271050 labor-ochs.de
Stainless steel needle (40 mm, 0.8 mm ID) Sterican 4657519 bbraun.de
Rubber tubing (40 mm, 0.74 mm ID) 2600185 newageindustries.com
Glass bottle (2 L) Rotilabo X716.1 carlroth.com
Butyl rubber stopper (for GL45) 444704 labor-ochs.de
Stainless steel capillary (50 mm, 0.74 mm ID) 56736 sigmaaldrich.com
Rubber tubing (30 mm x 0.74 mm ID) 2600185 newageindustries.com
Rubber tubing (100 mm x 0.74 mm ID) 2600185 newageindustries.com
Luer Lock tube connector (1.0 mm, LLF) P334.1 carlroth.com
Luer Lock 3-way connector (LLF, 2x LLM) 6134 cadenceinc.com
Light source Samsung SI-P8V151DB1US samsung.com
Peristalic pump Ismatec EW-78017-35 coleparmer.com
Pumping tubing (0.89 mm ID) EW-97628-26 coleparmer.com
Stainless steel capillary (200 mm, 0.74 mm ID) 56736 sigmaaldrich.com
Stainless steel capillary (400 mm, 0.74 mm ID) 56737 sigmaaldrich.com
Supel-Inert Foil (Tedlar - PFC) gas pack (10 L) 30240-U sigmaaldrich.com
Rubber tube (30 mm, 6 mm ID) 770300 labor-ochs.de
Luer Lock tube connector (3.0 mm, LLM) P343.1 carlroth.com
Luer Lock tube connector (3.0 mm, LLF) P335.1 carlroth.com
Gas-tight syringe (20 mL) C681.1 carlroth.com
Bunsen burner
Fiber optic oxygen meter for oxygen quantification Presens TR-FB-10-01 presens.de
Vacuum pump
Silicone glue for oxygen optodes Presens PS1 presens.de

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