Marine heatwaves and decreased light availability interact to erode the ecophysiological performance of habitat-forming kelp species.

Coastal marine ecosystems are threatened by a range of anthropogenic stressors, operating at global, local, and temporal scales. We investigated the impact of marine heatwaves (MHWs) combined with decreased light availability over two seasons on the ecophysiological responses of three kelp species (Laminaria digitata, L. hyperborea, and L. ochroleuca). These species function as important habitat-forming foundation organisms in the northeast Atlantic and have distinct but overlapping latitudinal distributions and thermal niches. Under low-light conditions, summertime MHWs induced significant declines in biomass, blade surface area, and Fv/Fm values (a measure of photosynthetic efficiency) in the cool-water kelps L. digitata and L. hyperborea, albeit to varying degrees. Under high-light conditions, all species were largely resistant to simulated MHW activity. In springtime, MHWs had minimal impacts and in some cases promoted kelp performance, while reduced light availability resulted in lower growth rates. While some species were negatively affected by summer MHWs under low-light conditions (particularly L. digitata), they were generally resilient to MHWs under high-light conditions. As such, maintaining good environmental quality and water clarity may increase resilience of populations to summertime MHWs. Our study informs predictions of how habitat-forming foundation kelp species will be affected by interacting, concurrent stressors, typical of compound events that are intensifying under anthropogenic climate change.

Shading by giant kelp canopy can restrict the invasiveness of Undaria pinnatifida (Laminariales, Phaeophyceae).

The spread of non-indigenous and invasive seaweeds has increased worldwide, and their potential effects on native seaweeds have raised concern. Undaria pinnatifida is considered among the most prolific non-indigenous species. This species has expanded rapidly in the Northeast Pacific, overlapping with native communities such as the iconic giant kelp forests (Macrocystis pyrifera). Canopy shading by giant kelp has been argued to be a limiting factor for the presence of U. pinnatifida in the understory, thus its invasiveness capacity. However, its physiological plasticity under light limitation remains unclear. In this work, we compared the physiology and growth of juvenile U. pinnatifida and M. pyrifera sporophytes transplanted to the understory of a giant kelp forest, to juveniles growing outside of the forest. Extreme low light availability compared to that outside (~0.2 and ~4.4 mol photon ⋅ m-2 ⋅ d-1 , respectively) likely caused a "metabolic energy crisis" in U. pinnatifida, thus restricting its photoacclimation plasticity and nitrogen acquisition, ultimately reducing its growth. Despite M. pyrifera juveniles showing photoacclimatory responses (e.g., increases in photosynthetic efficiency and lower compensation irradiance, Ec ), their physiological/vegetative status deteriorated similarly to U. pinnatifida, which explains the low recruitment inside the forest. Generally, our results revealed the ecophysiological basis behind the limited growth and survival of juvenile U. pinnatifida sporophytes in the understory.

Limits to photosynthesis: seasonal shifts in supply and demand for CO2 in Scots pine.

Boreal forests undergo a strong seasonal photosynthetic cycle; however, the underlying processes remain incompletely characterized. Here, we present a novel analysis of the seasonal diffusional and biochemical limits to photosynthesis (Anet ) relative to temperature and light limitations in high-latitude mature Pinus sylvestris, including a high-resolution analysis of the seasonality of mesophyll conductance (gm ) and its effect on the estimation of carboxylation capacity ( VCmax ). We used a custom-built gas-exchange system coupled to a carbon isotope analyser to obtain continuous measurements for the estimation of the relevant shoot gas-exchange parameters and quantified the biochemical and diffusional controls alongside the environmental controls over Anet . The seasonality of Anet was strongly dependent on VCmax and the diffusional limitations. Stomatal limitation was low in spring and autumn but increased to 31% in June. By contrast, mesophyll limitation was nearly constant (19%). We found that VCmax limited Anet in the spring, whereas daily temperatures and the gradual reduction of light availability limited Anet in the autumn, despite relatively high VCmax . We describe for the first time the role of mesophyll conductance in connection with seasonal trends in net photosynthesis of P. sylvestris, revealing a strong coordination between gm and Anet , but not between gm and stomatal conductance.

Phytoplankton growth rates in the Amundsen Sea (Antarctica) during summer: The role of light.

In the Amundsen Sea, significant global warming accelerates ice melt, and is consequently altering many ocean properties such as sea ice concentration, surface freshening, water column stratification, and underwater light properties. To examine the influence of light, which is one of the fundamental factors for phytoplankton growth, incubation experiments and field surveys were performed during the austral summer of 2016. In the incubation experiments, phytoplankton abundance and carbon biomass significantly increased with increasing light levels, probably indicating light limitation. Growth rates of the small pennates (mean 0.42 d-1) increased most rapidly with an increase in light, followed by those of Phaeocystis antarctica (0.31 d-1), and the large diatoms (0.16 d-1). A short-term study during the field survey showed that phytoplankton distribution in the surface layer was likely controlled by different responses to light and the sinking rate of each species. These results suggest that the approach adopted by previous studies of explaining phytoplankton ecology as a characteristic of two major taxa, namely diatoms and P. antarctica, in the coastal Antarctic waters might cause errors owing to oversimplification and misunderstanding, since diatoms comprise several species that have different ecophysiological characteristics.

Temporal dependence of chlorophyll a-nutrient relationships in Lake Taihu: Drivers and management implications.

Eutrophication and its associated algal blooms are principal environmental challenges confronting lakes worldwide. The empirical relationships between nutrient (total nitrogen, TN; total phosphorus, TP) and chlorophyll a (Chla) level are widely used as a theoretical basis for lake eutrophication management. Here, seasonal environmental variables and Chla from 2005 to 2020 in Chinese shallow eutrophic Lake Taihu were examined and Chla-nutrient equations in the entire period and annually from 2005 to 2020 were explored using 95% quantile regression model. The results showed robust linear relationships of logChla-logTN and logChla-logTP in the vast majority of cases. Based on Chla-nutrient equations in the entire study period, 0.69 mg/L TN and 52 µg/L TP are recommended as nutrient threshold in Lake Taihu. Furthermore, the results revealed increasing Chla sensitivity to nutrient for each study month (i.e. February, May, August, and November) from 2005 to 2020, whose drivers included increase in water temperature and water level, decrease in wind speed, mass ratio of nitrogen to phosphorus, and grazing effect. It is noteworthy that atmospheric stilling is likely to be the key climatic factor promoting annual peak Chla in Lake Taihu. For one, the deviations of the sub-index of Trophic State Index indicated that light is a critical limiting factor of summer Chla in Lake Taihu. For another, calmer water mainly due to atmospheric stilling decreased near 40% non-algal turbidity and a substantially increased buoyant cyanobacteria during the study period, improving phytoplankton "light niche". Thus, increasing algal sensitivity to nutrient occurred until the additional algal-turbidity induce further light limitations or the exhaustion of TN (or TP) cause nutrient limitation. Given atmospheric stilling is a global phenomenon, this study would affect future algal bloom mitigation efforts in shallow lakes as temperature is always the focus in the recent literatures on global climate change.

Coastal darkening substantially limits the contribution of kelp to coastal carbon cycles.

Macroalgal-dominated habitats are rapidly gaining recognition as important contributors to marine carbon cycles and sequestration. Despite this recognition, relatively little is known about the production and fate of carbon originating from these highly productive ecosystems, or how anthropogenic- and climate-related stressors affect the role of macroalgae in marine carbon cycles. Here, we examine the impact of increasing turbidity on carbon storage, fixation and loss in southern hemisphere kelp forests. We quantified net primary production (NPP) and biomass accumulation (BA), and estimated carbon release via detritus and dissolved organic carbon (DOC) across a large-scale turbidity gradient. We show that increased turbidity, resulting in a 63% reduction in light, can result in a 95% reduction in kelp productivity. When averaged annually, estimates of NPP and BA per plant at high-light sites were nearly six and two times greater than those at low-light sites, respectively. Furthermore, the quantity of carbon fixed annually by kelp forests was up to 4.7 times greater than that stored as average annual standing stock. At low-light sites, the majority of C goes directly into tissue growth and is subsequently eroded. In contrast, excess production at high-light sites accounts for up to 39% of the total carbon fixed and is likely released as DOC. Turbidity is expected to increase in response to climate change and our results suggest this will have significant impacts on the capacity of kelp forests to contribute to carbon sequestration pathways. In addition to demonstrating that turbidity significantly reduces the quantity of carbon fixed by kelp forests, and subsequently released as detritus, our results highlight the negative impacts of turbidity on a large source of previously unaccounted for carbon.

System-level analysis of metabolic trade-offs during anaerobic photoheterotrophic growth in Rhodopseudomonas palustris.

BACKGROUND: Living organisms need to allocate their limited resources in a manner that optimizes their overall fitness by simultaneously achieving several different biological objectives. Examination of these biological trade-offs can provide invaluable information regarding the biophysical and biochemical bases behind observed cellular phenotypes. A quantitative knowledge of a cell system's critical objectives is also needed for engineering of cellular metabolism, where there is interest in mitigating the fitness costs that may result from human manipulation. RESULTS: To study metabolism in photoheterotrophs, we developed and validated a genome-scale model of metabolism in Rhodopseudomonas palustris, a metabolically versatile gram-negative purple non-sulfur bacterium capable of growing phototrophically on various carbon sources, including inorganic carbon and aromatic compounds. To quantitatively assess trade-offs among a set of important biological objectives during different metabolic growth modes, we used our new model to conduct an 8-dimensional multi-objective flux analysis of metabolism in R. palustris. Our results revealed that phototrophic metabolism in R. palustris is light-limited under anaerobic conditions, regardless of the available carbon source. Under photoheterotrophic conditions, R. palustris prioritizes the optimization of carbon efficiency, followed by ATP production and biomass production rate, in a Pareto-optimal manner. To achieve maximum carbon fixation, cells appear to divert limited energy resources away from growth and toward CO2 fixation, even in the presence of excess reduced carbon. We also found that to achieve the theoretical maximum rate of biomass production, anaerobic metabolism requires import of additional compounds (such as protons) to serve as electron acceptors. Finally, we found that production of hydrogen gas, of potential interest as a candidate biofuel, lowers the cellular growth rates under all circumstances. CONCLUSIONS: Photoheterotrophic metabolism of R. palustris is primarily regulated by the amount of light it can absorb and not the availability of carbon. However, despite carbon's secondary role as a regulating factor, R. palustris' metabolism strives for maximum carbon efficiency, even when this increased efficiency leads to slightly lower growth rates.

A leaf-level biochemical model simulating the introduction of C2 and C4 photosynthesis in C3 rice: gains, losses and metabolite fluxes.

This work aims at developing an adequate theoretical basis for comparing assimilation of the ancestral C3 pathway with CO2 concentrating mechanisms (CCM) that have evolved to reduce photorespiratory yield losses. We present a novel model for C3 , C2 , C2  + C4 and C4 photosynthesis simulating assimilatory metabolism, energetics and metabolite traffic at the leaf level. It integrates a mechanistic description of light reactions to simulate ATP and NADPH production, and a variable engagement of cyclic electron flow. The analytical solutions are compact and thus suitable for larger scale simulations. Inputs were derived with a comprehensive gas-exchange experiment. We show trade-offs in the operation of C4 that are in line with ecophysiological data. C4 has the potential to increase assimilation over C3 at high temperatures and light intensities, but this benefit is reversed under low temperatures and light. We apply the model to simulate the introduction of progressively complex levels of CCM into C3 rice, which feeds > 3.5 billion people. Increasing assimilation will require considerable modifications such as expressing the NAD(P)H Dehydrogenase-like complex and upregulating cyclic electron flow, enlarging the bundle sheath, and expressing suitable transporters to allow adequate metabolite traffic. The simpler C2 rice may be a desirable alternative.

High carbon storage in carbon-limited trees.

The concentrations of nonstructural carbohydrates (NSCs) in plant tissues are commonly used as an indicator of total plant carbon (C) supply; but some evidence suggests the possibility for high NSC concentrations during periods of C limitation. Despite this uncertainty, NSC dynamics have not been investigated experimentally under long-term C limitation. We exposed saplings of 10 temperate tree species differing in shade tolerance to 6% of ambient sunlight for 3 yr to induce C limitation, and also defoliated one species, Carpinus betulus, in the third season. Growth and NSC concentrations were monitored to determine C allocation. Shade strongly reduced growth, but after an initial two-fold decrease, NSC concentrations of shaded saplings recovered to the level of unshaded saplings by the third season. NSC concentrations were generally more depleted under shade after leaf flush, and following herbivore attacks. Only under shade did artificial defoliation lead to mortality and depleted NSC concentrations in surviving individuals. We conclude that, irrespective of shade tolerance, C storage is maintained under prolonged shading, and thus high NSC concentrations can occur during C limitation. Yet, our results also suggest that decreased NSC concentrations are indicative of C limitation, and that additional leaf loss can lead to lethal C shortage in deep shade.

Light-optimized growth of cyanobacterial cultures: Growth phases and productivity of biomass and secreted molecules in light-limited batch growth.

Cyanobacteria are photosynthetic microorganisms whose metabolism can be modified through genetic engineering for production of a wide variety of molecules directly from CO2, light, and nutrients. Diverse molecules have been produced in small quantities by engineered cyanobacteria to demonstrate the feasibility of photosynthetic biorefineries. Consequently, there is interest in engineering these microorganisms to increase titer and productivity to meet industrial metrics. Unfortunately, differing experimental conditions and cultivation techniques confound comparisons of strains and metabolic engineering strategies. In this work, we discuss the factors governing photoautotrophic growth and demonstrate nutritionally replete conditions in which a model cyanobacterium can be grown to stationary phase with light as the sole limiting substrate. We introduce a mathematical framework for understanding the dynamics of growth and product secretion in light-limited cyanobacterial cultures. Using this framework, we demonstrate how cyanobacterial growth in differing experimental systems can be easily scaled by the volumetric photon delivery rate using the model organisms Synechococcus sp. strain PCC7002 and Synechococcus elongatus strain UTEX2973. We use this framework to predict scaled up growth and product secretion in 1L photobioreactors of two strains of Synechococcus PCC7002 engineered for production of l-lactate or L-lysine. The analytical framework developed in this work serves as a guide for future metabolic engineering studies of cyanobacteria to allow better comparison of experiments performed in different experimental systems and to further investigate the dynamics of growth and product secretion.

Theory of turbid microalgae cultures.

Microalgae can be cultivated in closed or open photobioreactors (PBR). In these systems, light rapidly decreases as it passes through the culture due to the turbidity of the medium. Thus, microalgae experiment different light intensities depending on their position in the medium. In this paper, we study theoretically how the growth rate of microalgae is affected by different factors; incident light intensity, form of the PBR, microalgae population density, turbidity of non-microalgae components, and light path-length of the reactor. We show that for different types of PBR the average growth rate is completely determined by the incident light intensity and the optical depth. In the case of vertical cylindrical PBRs illuminated from above (e.g. race-way or panel-type reactors), we described (and we prove under general assumptions) in details the dependence of the AGR on the aforementioned factors. Finally, we discuss some implications of our analysis; the occurrence of the Allee effect, if light ostensibly limits or inhibits the growth rate in outdoor cultures, and how the geometry of the PBR affects microalgae growth rate and productivity.

Dissolved organic carbon and unimodal variation in sexual signal coloration in mosquitofish: a role for light limitation?

Natural selection plays an important role in the evolution of sexual communication systems. Here, we assess the effect of two well-known selection agents, transmission environment and predation, on interpopulation variation in sexual signals. Our model system is a series of 21 populations of Bahamian mosquitofish subjected to independent variation in optical conditions and predation risk. We show that optically diverse environments, caused by locally variable dissolved organic carbon concentrations, rather than spatial variation in predation, drove divergence in fin coloration (fin redness). We found a unimodal pattern of phenotypic variation along the optical gradient indicating a threshold-type response of visual signals to broad variation in optical conditions. We discuss evolutionary and ecological mechanisms that may drive such a pattern as well as the implications of non-monotonic clines for evolutionary differentiation.

The effects of pH and pCO2 on photosynthesis and respiration in the diatom Thalassiosira weissflogii.

The response of marine phytoplankton to the ongoing increase in atmospheric pCO2 reflects the consequences of both increased CO2 concentration and decreased pH in surface seawater. In the model diatom Thalassiosira weissflogii, we explored the effects of varying pCO2 and pH, independently and in concert, on photosynthesis and respiration by incubating samples in water enriched in H218O. In long-term experiments (~6-h) at saturating light intensity, we observed no effects of pH or pCO2 on growth rate, photosynthesis or respiration. This absence of a measurable response reflects the very small change in energy used by the carbon concentrating mechanism (CCM) compared to the energy used in carbon fixation. In short-term experiments (~3 min), we also observed no effects of pCO2 or pH, even under limiting light intensity. We surmise that in T. weissflogii, it is the photosynthetic production of NADPH and ATP, rather than the CO2-saturation of Rubisco that controls the rate of photosynthesis at low irradiance. In short-term experiments, we observed a slightly higher respiration rate at low pH at the onset of the dark period, possibly reflecting the energy used for exporting H+ and maintaining pH homeostasis. Based on what is known of the biochemistry of marine phytoplankton, our results are likely generalizable to other diatoms and a number of other eukaryotic species. The direct effects of ocean acidification on growth, photosynthesis and respiration in these organisms should be small over the range of atmospheric pCO2 predicted for the twenty-first century.

Brownification increases winter mortality in fish.

In northern climates, winter is a bottleneck for many organisms. Low light and resource availability constrains individual foraging rates, potentially leading to starvation and increased mortality. Increasing input of humic substances to aquatic ecosystems causes brownification of water and hence a further decrease of light availability, which may lead to further decreased foraging rates and starvation mortality during winter. To test this hypothesis, we measured the effects of experimentally increased humic water input on consumption and survival of young-of-the-year three-spined stickleback (Gasterosteus aculeatus) over winter in large outdoor enclosures. Population densities were estimated in autumn, and the following spring and food availability and consumption were monitored over winter. As hypothesized, mortality was higher under humic (76%) as compared to ambient conditions (64%). In addition, body condition and ingested prey biomass were lower under humic conditions, even though resource availability was not lower under humic conditions. Light conditions were significantly poorer under humic conditions. This suggests that increased mortality and decreased body condition and ingested prey biomass were not due to decreased resource availability but due to decreased search efficiency in this visual feeding consumer. Increased future brownification of aquatic systems may, therefore, negatively affect both recruitment and densities of fish.

Ectomycorrhizal fungi reduce the light compensation point and promote carbon fixation of Pinus thunbergii seedlings to adapt to shade environments.

We examined the effects of three ectomycorrhizal (ECM) symbionts on the growth and photosynthesis capacity of Japanese black pine (Pinus thunbergii) seedlings and estimated physiological and photosynthetic parameters such as the light compensation point (LCP), biomass, and phosphorus (Pi) concentration of P. thunbergii seedlings. Through this investigation, we documented a new role of ectomycorrhizal (ECM) fungi: enhancement of the survival and competitiveness of P. thunbergii seedlings under low-light condition by reducing the LCP of seedlings. At a CO2 concentration of 400 ppm, the LCP of seedlings with ECM inoculations was 40-70 µmol photons m-2 s-1, significantly lower than that of non-mycorrhizal (NM) seedlings (200 µmol photons m-2 s-1). In addition, photosynthetic carbon fixation (Pn) increased with light intensity and CO2 level, and the Pn of ECM seedlings was significantly higher than that of NM seedlings; Pisolithus sp. (Pt)- and Laccaria amethystea (La)-mycorrhizal seedlings had significantly lower Pn than Cenococcum geophilum (Cg)-mycorrhizal seedlings. However, La-mycorrhizal seedlings exhibited the highest fresh weight, relative water content (RWC), and the lowest LCP in the mycorrhizal group. Concomitantly, ECM seedlings showed significantly increased chlorophyll content of needles and higher Pi concentrations compared to NM seedlings. Overall, ECM symbionts promoted growth and photosynthesis while reducing the LCP of P. thunbergii seedlings. These findings indicate that ECM fungi can enhance the survival and competitiveness of host seedlings under low light.

Patterns of shading tolerance determined from experimental light reduction studies of seagrasses.

A review and analysis of the experimental literature on seagrass shading evaluated the relationships among experimental light reduction, experimental duration, additional modifying factors and common meadow-scale seagrass response metrics to determine whether there were consistent statistical relationships. Modifying factors included study latitude, field site depth, season of experiment initiation, rhizome connectivity (severed, intact), experiment type (field, mesocosm), and seagrass life history strategy. Highly significant, best fit linear regression models were found for both biomass and shoot density reduction that included light reduction, duration and other modifying variables, although unexplained variation in the data were high. Duration of light limitation affected extent of response for both metrics, and unexplained variance was greatly reduced by analysis of data from durations >60d for shoot density and for >60d <120d for biomass. Life history strategy was also a significant factor in three of four regression models. While the slopes of the responses were relatively similar for biomass and shoot density, unexplained variation was generally greater for shoot density than biomass in models for data pooled across species. There were highly significant, best fit regression models found for both biomass and shoot density for both genus and species level analyses, with the extent and duration of light reduction the most important model factors. Season of experiment, rhizome status, latitude, and experiment type all were also included in multiple models. Biomass regression models again tended to have lesser unexplained variation than shoot density models. Life history was invariant within genus and species, and separate analyses for data divided among Colonizing, Opportunistic, and Persistent strategies found relatively similar, best fit regression models among strategies. However, the mean percent reduction of both biomass and shoot density was generally lower for the Persistent strategy than for the other two life histories, suggesting a greater buffering capacity against effects of light reduction for such species. Overall, biomass based models explained more of the variance in seagrass response to light reduction than shoot density, and may be the preferred response variable for meadow-scale impact assessments. The relationships observed may inform management decisions by helping define the scope of expected responses of seagrasses in general to the range of factors that may reduce light availability to seagrasses.

Mechanisms that increase the growth efficiency of diatoms in low light.

Photoacclimation was studied in Thalassiosira pseudonana to help understand mechanisms underlying the success of diatoms in low-light environments, such as coastal and deep mixing ecosystems. Light harvesting and other cell characteristics were combined with oxygen and carbon production measurements to assess the water-splitting reaction at PSII ([Formula: see text]) and intermediate steps leading to net carbon production (NPPC). These measurements revealed that T. pseudonana is remarkably efficient at converting harvested light energy into biomass, with at least 57 % of [Formula: see text] retained as NPPC across all light-limited growth rates examined. Evidence for upregulation of ATP generation pathways that circumvent carbon fixation indicated that high growth efficiency at low light levels was at least partly due to increases in the efficiency of ATP production. Growth rate-dependent demands for ATP and NADPH were reflected in carbon composition and in unexpected shifts in the light-limited slope (α) of photosynthesis-irradiance relationships generated from chlorophyll-specific (14)C-uptake. Overall, these results suggest that pathway gating of carbon and energy flow depends on light availability and is a key factor promoting the efficiency of diatom growth at low light intensities.

Assessment of the Effects of Light Availability on Growth and Competition Between Strains of Planktothrix agardhii and Microcystis aeruginosa.

In this study, we tested the hypothesis that Planktothrix agardhii strains isolated from a tropical water body were better competitors for light than Microcystis aeruginosa strains. These cyanobacteria are common in eutrophic systems, where light is one of the main drivers of phytoplankton, and Planktothrix is considered more shade-adapted and Microcystis more high-light tolerant. First, the effect of light intensities on growth was studied in batch cultures. Next, the minimum requirement of light (I*) and the effect of light limitation on the outcome of competition was investigated in chemostats. All strains showed similar growth at 10 µmol photons m(-2) s(-1), demonstrating the ability of the two species to grow in low light. The optimum light intensity was lower for P. agardhii, but at the highest light intensity, Microcystis strains reached higher biovolume, confirming that P. agardhii has higher sensitivity to high light. Nonetheless, P. agardhii grew in light intensities considered high (500 µmol photons m(-2) s(-1)) for this species. M. aeruginosa showed a higher carrying capacity in light-limited condition, but I* was similar between all the strains. Under light competition, Microcystis strains displaced P. agardhii and dominated. In two cases, there was competitive exclusion and in the other two P. agardhii managed to remain in the system with a low biovolume (≈15%). Our findings not only show that strains of P. agardhii can grow under higher light intensities than generally assumed but also that strains of M. aeruginosa are better competitors for light than supposed. These results help to understand the co-occurrence of these species in tropical environments and the dominance of M. aeruginosa even in low-light conditions.

Hydrocarbon production in high density Botryococcus braunii race B continuous culture.

Continuous cultures of Botryococcus braunii race B were maintained at photosynthetic cell densities as high as 20 g dry weight per liter for up to 3 months. Growth associated triterpene hydrocarbon accumulation was nearly constant at 22.5% of dry weight for a range of growth rates maintained by daily replacement of 5-15% of the respective cultures. The ability to achieve high cell concentrations and oil levels of roughly 5 g triterpene oil/L resulted from a combination of high light (∼ 1/4 full sun for 15 h/day) and replenishing stoichiometrically balanced growth medium. Due to light-limited growth conditions, cell concentration dropped nearly linearly with increased dilution rate. This reduction in cell number resulted in increased productivity per cell at higher dilution rates and was accompanied by a dramatic increase in algae colony size from 0.09 to 0.343 mm at high dilution rate. This change in colony size resulted in an equally dramatic change in optical density (OD) per gram dry weight, which precluded use of simple correlations of OD and cell concentration. A trickle-film photobioreactor was also demonstrated as a scalable approach to achieving these ultra-high cell concentrations. Additional media analysis revealed a steady increase in photobioreactor conductivity suggesting an accumulation of ions may be the reason for rapid culture crash and washout observed at all dilution rates after several months of continuous operation. The volumetric productivity of 22.5 mg oil/L/photo-h reported here is more than an order of magnitude higher than previous reports for B. braunii race B, reflecting the high cell densities used in this work and substantiating a higher metabolic rate for B. braunii race B than previously surmised from its relatively long doubling times.

Mechanistic modeling of sulfur-deprived photosynthesis and hydrogen production in suspensions of Chlamydomonas reinhardtii.

The ability of unicellular green algal species such as Chlamydomonas reinhardtii to produce hydrogen gas via iron-hydrogenase is well known. However, the oxygen-sensitive hydrogenase is closely linked to the photosynthetic chain in such a way that hydrogen and oxygen production need to be separated temporally for sustained photo-production. Under illumination, sulfur-deprivation has been shown to accommodate the production of hydrogen gas by partially-deactivating O2 evolution activity, leading to anaerobiosis in a sealed culture. As these facets are coupled, and the system complex, mathematical approaches potentially are of significant value since they may reveal improved or even optimal schemes for maximizing hydrogen production. Here, a mechanistic model of the system is constructed from consideration of the essential pathways and processes. The role of sulfur in photosynthesis (via PSII) and the storage and catabolism of endogenous substrate, and thus growth and decay of culture density, are explicitly modeled in order to describe and explore the complex interactions that lead to H2 production during sulfur-deprivation. As far as possible, functional forms and parameter values are determined or estimated from experimental data. The model is compared with published experimental studies and, encouragingly, qualitative agreement for trends in hydrogen yield and initiation time are found. It is then employed to probe optimal external sulfur and illumination conditions for hydrogen production, which are found to differ depending on whether a maximum yield of gas or initial production rate is required. The model constitutes a powerful theoretical tool for investigating novel sulfur cycling regimes that may ultimately be used to improve the commercial viability of hydrogen gas production from microorganisms.