Nitrogenase Inhibition Limited Oxygenation of Earth's Proterozoic Atmosphere.

Cyanobacteria produced the oxygen that began to accumulate on Earth 2.5 billion years ago, at the dawn of the Proterozoic Eon. By 2.4 billion years ago, the Great Oxidation Event (GOE) marked the onset of an atmosphere containing oxygen. The oxygen content of the atmosphere then remained low for almost 2 billion years. Why? Nitrogenase, the sole nitrogen-fixing enzyme on Earth, controls the entry of molecular nitrogen into the biosphere. Nitrogenase is inhibited in air containing more than 2% oxygen: the concentration of oxygen in the Proterozoic atmosphere. We propose that oxygen inhibition of nitrogenase limited Proterozoic global primary production. Oxygen levels increased when upright terrestrial plants isolated nitrogen fixation in soil from photosynthetic oxygen production in shoots and leaves.

NO MECHANISTIC DEPENDENCE OF PHOTOSYNTHESIS ON CALCIFICATION IN THE COCCOLITHOPHORID EMILIANIA HUXLEYI (HAPTOPHYTA)(1).

There is still considerable uncertainty about the relationship between calcification and photosynthesis. It has been suggested that since calcification in coccolithophorids is an intracellular process that releases CO2 , it enhances photosynthesis in a manner analogous to a carbon-concentrating mechanism (CCM). The ubiquitous, bloom-forming, and numerically abundant coccolithophorid Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler was studied in nutrient-replete, pH and [CO2 ] controlled, continuous cultures (turbidostats) under a range of [Ca(2+) ] from 0 to 9 mM. We examined the long-term, fully acclimated photosynthesis-light responses and analyzed the crystalline structure of the coccoliths using SEM. The E. huxleyi cells completely lost their coccosphere when grown in 0 [Ca(2+) ], while thin, undercalcified and brittle coccoliths were evident at 1 mM [Ca(2+) ]. Coccoliths showed increasing levels of calcification with increasing [Ca(2+) ]. More robust coccoliths were noted, with no discernable differences in coccolith morphology when the cells were grown in either 5 or 9 mM (ambient seawater) [Ca(2+) ]. In contrast to calcification, photosynthesis was not affected by the [Ca(2+) ] in the media. Cells showed no correlation of their light-dependent O2 evolution with [Ca(2+) ], and in all [Ca(2+) ]-containing turbidostats, there were no significant differences in growth rate. The results show unequivocally that as a process, photosynthesis in E. huxleyi is mechanistically independent from calcification.

BICARBONATE STIMULATION OF CALCIFICATION AND PHOTOSYNTHESIS IN TWO HERMATYPIC CORALS(1).

A wide range of bicarbonate concentrations was used to monitor the kinetics of bicarbonate (HCO3 (-) ) use in both photosynthesis and calcification in two reef-building corals, Porites porites and Acropora sp. Experiments carried out close to the P. porites collection site in Barbados showed that additions of NaHCO3 to synthetic seawater proportionally increased the calcification rate of this coral until the concentration exceeded three times that of seawater (6 mM). Photosynthetic rates were also stimulated by HCO3 (-) addition, but these became saturated at a lower concentration (4 mM). Similar experiments on aquarium-acclimated colonies of Indo-Pacific Acropora sp. showed that calcification and photosynthesis in this coral were enhanced to an even greater extent than P. porites, with calcification continuing to increase above 8 mM HCO3 (-) , and photosynthesis saturating at 6 mM. Calcification rates of Acropora sp. were also monitored in the dark, and, although these were lower than in the light for a given HCO3 (-) concentration, they still increased dramatically with HCO3 (-) addition, showing that calcification in this coral is light stimulated but not light dependent.

Structural and physiological effects of calcium and magnesium in Emiliania huxleyi (Lohmann) Hay and Mohler.

In organisms which perform both photosynthesis and calcification, the fact that calcification proceeds faster in the light than in the dark has led to the long-established view that photosynthesis and calcification are closely coupled. It is now clear that calcification does not promote photosynthesis, but an enhancement of calcification by photosynthesis could still explain why calcification is faster in the light. To test this, the kinetics of the two processes were monitored over a wide range of calcium concentrations (0-50 mM) in the coccolithophore Emiliania huxleyi. The addition of 50 mM calcium strongly inhibited both processes, but when incubated in lower concentrations, rates of calcification increased up to 20 mM calcium whilst those of photosynthesis remained constant over the same range of calcium concentrations. So, rates of calcification are able to rise without a concomitant increase in photosynthetic rates. In addition, calcification rate and coccolith morphology responded similarly to changes in calcium concentrations; low calcification rates were associated with poor coccolith structure (undercalcification) and high calcification rates with perfectly formed coccoliths. Calcium concentration thus strongly influences calcification affecting both crystal structure and rate of calcite deposition. A similar structural analysis of coccoliths from cells grown in different magnesium concentrations showed that this ion is also essential for calcification, since strong signs of coccolith malformation and undercalcification were apparent at both low and high magnesium concentrations. In contrast with the calcium results, coccoliths were flawless only in the normal seawater concentration of 58 mM magnesium. We conclude that photosynthesis and calcification are not closely coupled and that calcification depends on a precise balance of both calcium and magnesium concentrations.

Modelling diatom growth in turbulent waters.

Algal models used as tools in the management of algal blooms may be inaccurate because representation of mixing processes is often oversimplified. A testable 3-D algal model for prediction of algal growth in turbulent surface waters was developed based on the Eulerian water quality model, HYDRO-3D. Out-door mesocosm experiments on the growth of the diatom Skeletonema costatum showed no evidence that diatom growth is significantly affected by light/dark fluctuations brought about by turbulent mixing, and no direct effects of turbulence on phytoplankton physiology were required in the algal model. The algal model was successfully calibrated and validated against mesocosm data and field data from Poplar Dock, London Docklands. Application of the model gave credible results for the hypothetical growth of S. costatum in Poplar Dock under a wide range of wind speeds and surface irradiances. However, differences between the results of a full 3-D simulation and a simplified 1-D representation of Poplar Dock were minimal, and no clear conclusions could be drawn on the superiority of 3-D models over 1-D models for simulation of complex flows in natural water bodies.

Acquisition and use of bicarbonate by Emiliania huxleyi.

• Bicarbonate acquisition mechanisms and the kinetics of dissolved inorganic carbon (DIC) use in photosynthesis and calcification were investigated in Emiliania huxleyi. • Photosynthesis was measured using O2 evolution and 14 C incorporation and calcification was measured with 14 C. Noncalcifying (coccolith-free) cells were produced from calcifying (coccolith-bearing) cells of the same strain of E. huxleyi, so that photosynthesis could be monitored independently from calcification. • Neither photosynthesis nor calcification was saturated at the ambient DIC concentration of seawater. In coccolith-bearing cells, both processes showed biphasic kinetics with DIC concentration, with a hiatus located at 1 mM. The same biphasic pattern and similar rates of photosynthesis were found in the coccolith-free cells. Inhibitor experiments showed that E. huxleyi acquires bicarbonate mainly by an anion exchange protein, but external carbonic anhydrase can be activated at low concentrations of DIC. • We conclude that the biphasic kinetics of photosynthesis and calcification are caused by the presence of two bicarbonate acquisition mechanisms and also, since calcification does not enhance photosynthesis in this coccolithophore, we question the current view that the two processes are tightly coupled.