This study provides with original data sets on the physiology of the unicellular diazotrophic cyanobacterium Crocosphaera watsonii WH8501, maintained in continuous culture in conditions of obligate diazotrophy. Cultures were exposed to a 12:12 light-dark regime, representative of what they experience in nature and where growth is expected to be balanced. Nitrogen and carbon metabolism were monitored at high frequency and their dynamics was compared with the cell cycle. Results reveal a daily cycle in the physiological and biochemical parameters, tightly constrained by the timely decoupled processes of N(2) fixation and carbon acquisition. The cell division rate increased concomitantly to carbon accumulation and peaked 6 h into the light. The carbon content reached a maximum at the end of the light phase. N(2) fixation occurred mostly during the dark period and peaked between 9 and 10 h into the night, while DNA synthesis, reflected by DNA fluorescence, increased until the end of the night. Consequently, cells in G1- and S-phases present a marked decrease in their C:N ratio. Nitrogen acquisition through N(2) fixation exceeded 1.3- to 3-fold the nitrogen requirements for growth, suggesting that important amounts of nitrogen are excreted even under conditions supposed to favour balanced, carbon and nitrogen acquisitions.

Light:dark (12:12 h) quantification of cellular carbohydrate fluxes was conducted with the unicellular, N 2 -fixing cyanobacterium Crocosphaera watsonii WH8501 grown in continuous cultures under conditions of diazotrophy. As commonly observed in photoautotrophs, C. watsonii accumulates photosynthetic products as carbohydrates, which thereafter serve as an internal energy reserve for energy-demanding processes. Our goal was to further discriminate between cellular pools with different sizes to observe their respective dynamics. Total cellular carbohydrate as well as the cellular carbohydrate pools showed a diel cycle of accumulation and decrease. Total cellular carbohydrate contents represented 45 and 32% of the cellular carbon content at the end of the light and dark period, respectively. The observed decrease in cellular carbohydrate was not only due to carbohydrate catabolism but also to their release in the environment as extracellular polymeric substances (EPS), including soluble EPS and transparent exopolymeric particles (TEP). The occurrence of such excretion during the exponential growth of C. watsonii appears as part of the metabolic regulations that participate in maintaining a daily carbon:nitrogen balanced growth. About 10% of the total carbon cell content is excreted daily as soluble EPS. TEP, which are formed from dissolved precursors (including some soluble EPS) released by C. watsonii, increased during the dark period, when they represented up to 3.6% of total carbon cell content. Such recurrent release of carbohydrate by populations of C. watsonii is expected to stimulate the microbial loop in warm, oligotrophic oceans.

In the present study depth profiles of light, oxygen, pH and photosynthetic performance in an artificial biofilm of the green alga Halochlorella rubescens in a porous substrate photobioreactor (PSBR) were recorded with microsensors. Biofilms were exposed to different light intensities (50-1,000 μmol photons m(-2) s(-1) ) and CO2 levels (0.04-5% v/v in air). The distribution of photosynthetically active radiation showed almost identical trends for different surface irradiances, namely: a relatively fast drop to a depth of about 250 µm, (to 5% of the incident), followed by a slower decrease. Light penetrated into the biofilm deeper than the Lambert-Beer Law predicted, which may be attributed to forward scattering of light, thus improving the overall light availability. Oxygen concentration profiles showed maxima at a depth between 50 and 150 μm, depending on the incident light intensity. A very fast gas exchange was observed at the biofilm surface. The highest oxygen concentration of 3.2 mM was measured with 1,000 μmol photons m(-2) s(-1) and 5% supplementary CO2. Photosynthetic productivity increased with light intensity and/or CO2 concentration and was always highest at the biofilm surface; the stimulating effect of elevated CO2 concentration in the gas phase on photosynthesis was enhanced by higher light intensities. The dissolved inorganic carbon concentration profiles suggest that the availability of the dissolved free CO2 has the strongest impact on photosynthetic productivity. The results suggest that dark respiration could explain previously observed decrease in growth rate over cultivation time in this type of PSBR. Our results represent a basis for understanding the complex dynamics of environmental variables and metabolic processes in artificial phototrophic biofilms exposed to a gas phase and can be used to improve the design and operational parameters of PSBRs.

We analysed the effect of photoperiod length (PPL) (16:8 and 8:16 h of light-dark regime, named long and short PPL, respectively) on the temporal orchestration of the two antagonistic, carbon and nitrogen acquisitions in the unicellular, diazotrophic cyanobacterium Crocosphaera watsonii strain WH8501 growing diazotrophically. Carbon and nitrogen metabolism were monitored at high frequency, and their patterns were compared with the cell cycle progression. The oxygen-sensitive N2 fixation process occurred mainly during the dark period, where photosynthesis cannot take place, inducing a light-dark cycle of cellular C : N ratio. Examination of circadian patterns in the cell cycle revealed that cell division occurred during the midlight period, (8 h and 4 h into the light in the long and short PPL conditions, respectively), thus timely separated from the energy-intensive diazotrophic process. Results consistently show a nearly 5 h time lag between the end of cell division and the onset of N2 fixation. Shorter PPLs affected DNA compaction of C. watsonii cells and also led to a decrease in the cell division rate. Therefore, PPL paces the growth of C. watsonii: a long PPL enhances cell division while a short PPL favours somatic growth (biomass production) with higher carbon and nitrogen cell contents.

No abstract

We propose a dynamical model depicting nitrogen (N 2) fixation (diazotrophy) in a unicellular cyanobacterium, Crocosphaera watsonii, grown under light limitation and obligate diazotrophy. In this model, intracellular carbon and nitrogen are both divided into a functional pool and a storage pool. An internal pool that explicitly describes the nitrogenase enzyme is also added. The model is successfully validated with continuous culture experiments of C. watsonii under three light regimes, indicating that proposed mechanisms accurately reproduce the growth dynamics of this organism under various light environments. Then, a series of model simulations is run for a range of light regimes with different photoperiods and daily light doses. Results reveal how nitrogen and carbon are coupled, through the diel cycle, with nitrogenase dynamics, whose activity is constrained by the light regime. We finally identify optimal productivity conditions.