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 or oxygenic phototrophs, generating local oxygen oases in the surface ocean below a predominantly anoxic atmosphere3-5.
The deposition of Banded Iron Formations (BIFs) from seawater throughout the Precambrian points to iron(II) (Fe(II)) as a major geochemical constituent of seawater, at least locally, during their deposition. Some of the largest BIFs are deep-water deposits, forming off the continental shelf and slope. The amount of Fe deposited is incompatible from a mass balance standpoint with predominantly continental (i.e., weathering) source. Therefore, much of the Fe must have been supplied from hydrothermal alteration of mafic or ultramafic seafloor crust6. Estimates of the rate of Fe deposited outboard of coastal environments are consistent with Fe(II) supplied to the surface ocean via upwelling7. In order for Fe to be transported in upwelling currents, must have been present in the reduced, mobile form — as Fe(II). The average oxidation state of Fe preserved in BIF is 2.4 8 and it is generally thought that BIF preserve Fe deposited as Fe(III), formed when upwelling Fe(II) was oxidized, possibly by oxygen. Therefore, exploring potential Fe(II) oxidation mechanisms along slope environments is important to understand how BIF formed. Moreover, refined geochemical characterization of marine sediments has identified that ferruginous conditions, where Fe(II) was present in an anoxic water column, were a persistent feature of the oceans throughout the Precambrian, and may not have been limited to just the time and place where BIF were deposited9. Therefore, for at least two billion years of Earth's history, redox interfaces between Fe(II) and O2 in the shallow oceans were likely commonplace.
Numerous studies utilize modern sites that are chemical and/or biological analogs of different features of the Precambrian ocean. A good example are ferruginous lakes where Fe(II) is stable and present in sunlit surface waters while photosynthetic activity (including by cyanobacteria) was detected10-13. The results of these studies provide insight into the geochemical and microbial characteristics of an oxic to anoxic/ferruginous chemocline. However these sites are generally physically stratified with little vertical mixing14, rather than the chemical interfaces occurring in an upwelling system, and are thought to support the most oxygen production in Precambrian time4.
A natural analogue to explore the development of a marine oxygen oasis beneath an anoxic atmosphere, and at an Fe(II)-rich upwelling system in sunlit surface water column is not available on the modern Earth. Therefore, a laboratory system that can simulate a ferruginous upwelling zone and also support the growth of cyanobacteria and photoferrotrophs is needed. The understanding and identification of microbial processes and their interaction with an upwelling aqueous medium that represents Precambrian seawater promotes the understanding and can complement the information gained from the rock record in order to fully understand the distinctive biogeochemical processes on ancient Earth.
Toward that end, a laboratory-scale column was designed in which Fe(II)-rich seawater medium (pH neutral) was pumped into the bottom of the column, and pumped out from the top. Illumination was provided at the top to create a 4 cm wide "photic zone" that supported the growth of cyanobacteria in the top 3 cm. Natural environments are generally stratified and stabilized by physicochemical gradients, like salinity or temperature. In order to stabilize the water column on a lab-scale, the column cylinder was packed with a porous glass bead matrix that helped to maintain the establishment of geochemical patterns that developed during the experiment. A continuous N2/CO2 gas flow was applied to flush the headspace of the column in order to maintain an anoxic atmosphere reflective of an ocean prior to the GOE15. After a constant flux of Fe(II) was established, cyanobacteria were inoculated throughout the column, and their growth was monitored by cell counts on samples removed through sampling ports. Oxygen was monitored in situ by placing oxygen-sensitive optode foils onto the inner wall of the column cylinder and measurements were made with an optical fiber from outside the column. Aqueous Fe speciation was quantified by removing samples from depth-resolved horizontal sampling ports and analyzed with the Ferrozine method. The abiotic control experiments and results demonstrate proof-of-concept — that a laboratory scale analog of the ancient water column, maintained in isolation from the atmosphere, is achievable. Cyanobacteria grew and produced oxygen, and the reactions between Fe(II) and oxygen were resolvable. Herein, the methodology for design, preparation, assembly, execution, and sampling of such a column are presented, along with results from an 84 hr run of the column while inoculated with the marine cyanobacterium Synechococcus sp. PCC 7002.
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…
The authors have nothing to disclose.
Mark Nordhoff assisted in the design and implementation of tubing connections. Ellen Struve helped to select and acquire equipment used.
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 |