Inorganic Carbon Limitation Research Articles

Response to the CO2 concentrating mechanisms and transcriptional time series analysis of Ulva prolifera under inorganic carbon limitation

Ulva prolifera is a dominant species in green tides and has been affecting marine ecosystem for many years. Due to the low availability of CO2 in the environment, U. prolifera utilizes the CO2 concentrating mechanisms (CCMs) to increase intracellular inorganic carbon concentration. However, the transcriptional response mechanism and temporal changes of U. prolifera CCMs based on transcriptomics have not been thoroughly described. Therefore, we induced U. prolifera CCMs in a low CO2 environment to explore the dynamic regulation of CCMs expression under inadequate inorganic carbon supply. The results showed that inorganic carbon limitation increased the inorganic carbon affinity of U. prolifera, upregulating CCMs. The first 24 h of inorganic carbon environmental changes were the most active period for U. prolifera's expression regulation. U. prolifera gradually achieved a new steady state by regulating metabolic processes such as nucleic acids, energy, and ethylene-activated signaling pathways. In the carbon fixation system of U. prolifera, there are characteristics of both biophysical and biochemical CCMs. After 24 h of inorganic carbon limitation, the biophysical CCMs becomes more effective under conditions of inorganic carbon depletion. This study aids in exploring the CCMs of U. prolifera and their evolution in response to environmental changes.

Inorganic carbon limitation decreases ammonium removal and N2O production in the algae-nitrifying bacteria symbiosis system

Ammonium removal by a symbiosis system of algae (Chlorella vulgaris) and nitrifying bacteria was evaluated in a long-term photo-sequencing batch reactor under varying influent inorganic carbon (IC) concentrations (15, 10, 5 and 2.5 mmol L−1) and different nitrogen loading rate (NLR) conditions (270 and 540 mg-N L−1 d−1). The IC/N ratios provided were 2.33, 1.56, 0.78 and 0.39, respectively, for an influent NH4+-N concentration of 90 mg-N L−1 (6.43 mmol L−1). The results confirmed that both ammonium removal and N2O production were positively related with IC concentration. Satisfactory ammonium removal efficiencies (>98 %) and rates (29–34 mg-N gVSS−1 h−1) were achieved regardless of NLR levels under sufficient IC of 10 and 15 mmol L−1, while insufficient IC at 2.5 mmol L−1 led to the lowest ammonium removal rates of 0 mg-N gVSS−1 h−1. The ammonia oxidation process by ammonia oxidizing bacteria (AOB) played a predominant role over the algae assimilation process in ammonium removal. Long-time IC deficiency also resulted in the decrease in biomass and pigments of algae and nitrifying bacteria. IC limitation led to the decreasing N2O production, probably due to its negative effect on ammonia oxidation by AOB. The optimal IC concentration was determined to be 10 mmol L−1 (i.e., IC/N of 1.56, alkalinity of 500 mg CaCO3 L−1) in the algae-bacteria symbiosis reactor, corresponding to higher ammonia oxidation rate of ∼41 mg-N gVSS−1 h−1 and lower N2O emission factor of 0.13 %. This suggests regulating IC concentrations to achieve high ammonium removal and low carbon emission simultaneously in the algae-bacteria symbiosis wastewater treatment process.

Waste iron scraps promote anammox bacteria to resist inorganic carbon limitation

The anaerobic ammonium oxidation (anammox) process is adversely affected by the limitation of inorganic carbon (IC). In this research, a new technique was introduced to assist anammox biomass in counteracting the adverse effects of IC limitation by incorporating waste iron scraps (WIS), a cheap and easily accessible byproduct of lathe cutting. Results demonstrated that reducing the influent IC/TN ratio from 0.08–0.09 to 0.04 resulted in a 20 % decrease in the nitrogen removal rate (NRR) for the control reactor, with an average specific anammox activity (SAA) of 0.65 g N/g VSS/day. Nevertheless, the performance of the WIS-assisted anammox reactor remained robust despite the reduction in IC supply. In fact, the NRR and SAA of the WIS-assisted reactor exhibited substantial improvements, reaching approximately 1.86 kg/(m3·day) and 0.98 g N/g VSS/day, respectively. These values surpassed those achieved by the control reactor by approximately 39 % and 51 %, respectively. The microbial analysis confirmed that the WIS addition significantly stimulated the proliferation of anammox bacteria (dominated by Candidatus Kuenenia) under IC limitation. The anammox gene abundances in the WIS-assisted anammox reactor were 3–4 times higher than those in the control reactor. Functional genes prediction based on the KEGG database revealed that the addition of WIS significantly enhanced the relative abundances of genes associated with nitrogen metabolism, IC fixation, and central carbon metabolism. Together, the results suggested that WIS promoted carbon dioxide fixation of anammox species to resist IC limitation. This study provided a promising approach for effectively treating high ammonium-strength wastewater using anammox under IC limitation.

Metapangenomic investigation provides insight into niche differentiation of methanogenic populations from the subsurface serpentinizing environment, Samail Ophiolite, Oman.

Serpentinization reactions produce highly reduced waters that have hyperalkaline pH and that can have high concentrations of H2 and CH4. Putatively autotrophic methanogenic archaea have been identified in the subsurface waters of the Samail Ophiolite, Sultanate of Oman, though the strategies to overcome hyperalkaline pH and dissolved inorganic carbon limitation remain to be fully understood. Here, we recovered metagenome assembled genomes (MAGs) and applied a metapangenomic approach to three different Methanobacterium populations to assess habitat-specific functional gene distribution. A Type I population was identified in the fluids with neutral pH, while a Type II and "Mixed" population were identified in the most hyperalkaline fluids (pH 11.63). The core genome of all Methanobacterium populations highlighted potential DNA scavenging techniques to overcome phosphate or nitrogen limitation induced by environmental conditions. With particular emphasis on the Mixed and Type II population found in the most hyperalkaline fluids, the accessory genomes unique to each population reflected adaptation mechanisms suggesting lifestyles that minimize niche overlap. In addition to previously reported metabolic capability to utilize formate as an electron donor and generate intracellular CO2, the Type II population possessed genes relevant to defense against antimicrobials and assimilating potential osmoprotectants to provide cellular stability. The accessory genome of the Mixed population was enriched in genes for multiple glycosyltransferases suggesting reduced energetic costs by adhering to mineral surfaces or to other microorganisms, and fostering a non-motile lifestyle. These results highlight the niche differentiation of distinct Methanobacterium populations to circumvent the challenges of serpentinization impacted fluids through coexistence strategies, supporting our ability to understand controls on methanogenic lifestyles and adaptations within the serpentinizing subsurface fluids of the Samail Ophiolite.

The Influence of Symbiosis on the Proteome of the Exaiptasia Endosymbiont Breviolum minutum.

The cellular mechanisms responsible for the regulation of nutrient exchange, immune response, and symbiont population growth in the cnidarian-dinoflagellate symbiosis are poorly resolved. Here, we employed liquid chromatography-mass spectrometry to elucidate proteomic changes associated with symbiosis in Breviolum minutum, a native symbiont of the sea anemone Exaiptasia diaphana ('Aiptasia'). We manipulated nutrients available to the algae in culture and to the holobiont in hospite (i.e., in symbiosis) and then monitored the impacts of our treatments on host-endosymbiont interactions. Both the symbiotic and nutritional states had significant impacts on the B. minutum proteome. B. minutum in hospite showed an increased abundance of proteins involved in phosphoinositol metabolism (e.g., glycerophosphoinositol permease 1 and phosphatidylinositol phosphatase) relative to the free-living alga, potentially reflecting inter-partner signalling that promotes the stability of the symbiosis. Proteins potentially involved in concentrating and fixing inorganic carbon (e.g., carbonic anhydrase, V-type ATPase) and in the assimilation of nitrogen (e.g., glutamine synthase) were more abundant in free-living B. minutum than in hospite, possibly due to host-facilitated access to inorganic carbon and nitrogen limitation by the host when in hospite. Photosystem proteins increased in abundance at high nutrient levels irrespective of the symbiotic state, as did proteins involved in antioxidant defences (e.g., superoxide dismutase, glutathione s-transferase). Proteins involved in iron metabolism were also affected by the nutritional state, with an increased iron demand and uptake under low nutrient treatments. These results detail the changes in symbiont physiology in response to the host microenvironment and nutrient availability and indicate potential symbiont-driven mechanisms that regulate the cnidarian-dinoflagellate symbiosis.

Effects of Epiphytic Biofilm Activity on the Photosynthetic Activity, pH and Inorganic Carbon Microenvironment of Seagrass Leaves (Zostera marina L.)

Epiphytic biofilms on seagrass leaves can lead to extreme microenvironmental conditions for the encapsulated leaf limiting both its photosynthesis and respiration. Yet, little is known about how the biological activity of the biofilm itself changes the seagrass phyllosphere microenvironment and dynamics. We used microsensors to measure O2 concentrations and pH gradients and calculate fluxes of O2, CO2 and bicarbonate (HCO3−) around seagrass leaves (Z. marina L.) covered with artificial, inactive biofilms and natural epiphytic biofilms. A sterilized seawater-agar matrix was used to make an artificial “inactive” biofilm on seagrass leaves with the same thickness as the natural leaf epiphytic biofilm, which impeded turbulent exchange of gases but did not have microbial activity. We compared the concentration profiles and fluxes of O2 and inorganic carbon of the “active” and “inactive” biofilm to investigate the effect of microbial activity and molecular diffusion in seagrass leaf biofilms. In light, the O2 flux of leaves with inactive biofilm was only 31% of the leaves with active biofilm, indicating that the photosynthesis of the microbial community in the biofilm makes up the majority of O2 production in the leaf microenvironment. During darkness, the O2 concentration profiles and O2 fluxes were almost identical in the “active” and “inactive” biofilms. The pH profiles showed the same trend with an increase in pH of ~1.0 in the “active” biofilms and ~0.3 pH units in the “inactive” biofilms in the light, and both showing a decrease of ~0.3 pH units in darkness compared to the bulk seawater. Our measurements thus demonstrate strong photosynthesis in the epiphyte layer driving phyllosphere basification and inorganic carbon limitation. The calculated CO2 concentration on the leaf surface decreased to 0.09 μmol L-1 in the epiphytic biofilm in the light compared to leaf surface CO2 concentrations of 13.8 μmol L-1 on bare seagrass leaves, and the CO2 influxes were only 3.0% and 5.4% of O2 effluxes for leaves with “active” and “inactive” biofilm, respectively. Calculations also showed that HCO3− influxes in light accounted for 91-97% of the total inorganic carbon influx to the seagrass leaf, although the HCO3− utilization via CO2 concentration mechanisms is energy-consuming. Besides increasing mass transfer impedance, leaf epiphytic biofilm activity thus strongly affects the seagrass leaf microenvironment in the light by inducing higher O2 concentration and pH, increasing CO2 limitation and reducing the leaf photosynthetic efficiency.

Effect of inorganic carbon limitation on the nitrogen removal performance of the single-stage reactor containing anammox and nitritation gel carriers

Herein, the effect of inorganic carbon (IC) limitation on the nitrogen removal performance of the single-stage reactor containing nitritation and anammox gel carriers was investigated. As a result of a continuous feeding test, the effluent ammonium concentration increased as the IC concentration decreased, indicating the deterioration of nitritation activity, not anammox. Furthermore, the sensitivity of IC to anammox and nitritation activity was investigated in anammox and nitritation reactors, respectively. Consequently, the relationship between the effluent IC concentration and nitritation rate was well described using the Michaelis-Menten equation. The apparent Km value of nitritation was calculated as 4.4mg-C L-1. In anammox reactor, it was calculated as 1.7mg-C L-1. These results revealed that the affinity of nitritation gel carriers to IC was lower than that of anammox, supporting that nitritation activity was easily deactivated by decrease in the IC concentration rather than anammox. Microbial community analysis revealed that Nitrosomonas europaea and Candidatus Jettenia asiatica were the dominant species of ammonium-oxidizing and anammox bacteria.

Inorganic carbon limitation during nitrogen conversions in sponge-bed trickling filters for mainstream treatment of anaerobic effluent

Anaerobic sewage treatment is a proven technology in warm climate regions, and sponge-bed trickling filters (SBTFs) are an important post-treatment technology to remove residual organic carbon and nitrogen. Even though SBTFs can achieve a reasonably good effluent quality, further process optimization is hampered by a lack of mechanistic understanding of the factors influencing nitrogen removal, notably when it comes to mainstream anaerobically treated sewage. In this study, the factors that control the performance of SBTFs following anaerobic (i.e., UASB) reactors for sewage treatment were investigated. A demo-scale SBTF fed with anaerobically pre-treated sewage was monitored for 300 days, showing a median nitrification efficiency of 79% and a median total nitrogen removal efficiency of 26%. Heterotrophic denitrification was limited by the low organic carbon content of the anaerobic effluent. It was demonstrated that nitrification was impaired by a lack of inorganic carbon rather than by alkalinity limitation. To properly describe inorganic carbon limitation in models, bicarbonate was added as a state variable and sigmoidal kinetics were applied. The resulting model was able to capture the overall long-term experimental behaviour. There was no nitrite accumulation, which indicated that nitrite oxidizing bacteria were little or less affected by the inorganic carbon limitation. Overall, this study indicated the vital role of influent characteristics and operating conditions concerning nitrogen conversions in SBTFs treating anaerobic effluent, thus facilitating further process optimization.

Mixotrophic growth of the extremophile Galdieria sulphuraria reveals the flexibility of its carbon assimilation metabolism.

Summary Galdieria sulphuraria is a cosmopolitan microalga found in volcanic hot springs and calderas. It grows at low pH in photoautotrophic (use of light as a source of energy) or heterotrophic (respiration as a source of energy) conditions, using an unusually broad range of organic carbon sources. Previous data suggested that G. sulphuraria cannot grow mixotrophically (simultaneously exploiting light and organic carbon as energy sources), its photosynthetic machinery being repressed by organic carbon.Here, we show that G. sulphuraria SAG21.92 thrives in photoautotrophy, heterotrophy and mixotrophy. By comparing growth, biomass production, photosynthetic and respiratory performances in these three trophic modes, we show that addition of organic carbon to cultures (mixotrophy) relieves inorganic carbon limitation of photosynthesis thanks to increased CO2 supply through respiration. This synergistic effect is lost when inorganic carbon limitation is artificially overcome by saturating photosynthesis with added external CO2.Proteomic and metabolic profiling corroborates this conclusion suggesting that mixotrophy is an opportunistic mechanism to increase intracellular CO2 concentration under physiological conditions, boosting photosynthesis by enhancing the carboxylation activity of Ribulose‐1,5‐bisphosphate carboxylase‐oxygenase (Rubisco) and decreasing photorespiration.We discuss possible implications of these findings for the ecological success of Galdieria in extreme environments and for biotechnological applications.

Balancing Microalgae and Nitrifiers for Wastewater Treatment: Can Inorganic Carbon Limitation Cause an Environmental Threat?

The first objective of this study is to assess the predictive capability of the ALBA (ALgae-BActeria) model for a pilot-scale (3.8 m2) high-rate algae-bacteria pond treating agricultural digestate. The model, previously calibrated and validated on a one-year data set from a demonstrative-scale raceway (56 m2), successfully predicted data from a six-month monitoring campaign with a different wastewater (urban wastewater) under different climatic conditions. Without changing any parameter value from the previous calibration, the model accurately predicted both online monitored variables (dissolved oxygen, pH, temperature) and off-line measurements (nitrogen compounds, algal biomass, total and volatile suspended solids, chemical oxygen demand). Supported by the universal character of the model, different scenarios under variable weather conditions were tested, to investigate the effect of key operating parameters (hydraulic retention time, pH regulation, kLa) on algae biomass productivity and nutrient removal efficiency. Surprisingly, despite pH regulation, a strong limitation for inorganic carbon was found to hinder the process efficiency and to generate conditions that are favorable for N2O emission. The standard operating parameters have a limited effect on this limitation, and alkalinity turns out to be the main driver of inorganic carbon availability. This investigation offers new insights in algae-bacteria processes and paves the way for the identification of optimal operational strategies.

Patterns of CO2 concentration and inorganic carbon limitation of phytoplankton biomass in agriculturally eutrophic lakes

Lake eutrophication is a pervasive problem globally, particularly serious in agricultural and densely populated areas. Whenever nutrients nitrogen and phosphorus do not limit phytoplankton growth directly, high growth rates will rapidly lead to biomass increases causing self-shading and light-limitation, and eventually CO2 depletion. The paradigm of phytoplankton limitation by nutrients and light is so pervasively established, that the lack of nutrient limitation is ordinarily interpreted as sufficient evidence for the condition of light limitation, without considering the possibility of limitation by inorganic carbon. Here, we firstly evaluated how frequently CO2 undersaturation occurs in a set of eutrophic lakes in the Pampa plains. Our results confirm that conditions of CO2 undersaturation develop much more frequently (yearly 34%, summer 44%) in these agriculturally impacted lakes than in deep, temperate lakes in forested watersheds. Secondly, we used Generalized Additive Models to fit trends in CO2 concentration considering three drivers: total incident irradiance, chlorophyll a concentration, and lake depth; in eight multi-year datasets from eutrophic lakes from Europe, North and South America, Asia and New Zealand. CO2 depletion was more often observed at high irradiance levels, and shallow water. CO2 depletion also occurred at high chlorophyll concentration. Finally, we identified occurrences of light- and carbon-limitation at the whole-lake scale. The different responses of chlorophyll a andCO2 allowed us to develop criteria for detecting conditions of CO2 limitation. For the first time, we provided whole-lake evidence of carbon limitation of phytoplankton biomass. CO2 increases and eutrophication represent two major and converging environmental problems that have additive and contrasting effects, promoting phytoplankton, and also leading to carbon depletion. Their interactions deserve further exploration and imaginative approaches to deal with their effects.

Dynamic Inventory of Intermediate Metabolites of Cyanobacteria in a Diurnal Cycle.

SummaryCyanobacteria are gaining importance both as hosts for photoautotrophic production of chemicals and as model systems for studies of diurnal lifestyle. The proteome and transcriptome of cyanobacteria have been closely examined under diurnal growth, whereas the downstream effects on the intermediary metabolism have not received sufficient attention. The present study focuses on identifying the cellular metabolites whose inventories undergo dramatic changes in a fast-growing cyanobacterium, Synechococcus elongatus PCC 11801. We identified and quantified 67 polar metabolites, whose inventory changes significantly during diurnal growth, with some metabolites changing by 100-fold. The Calvin-Benson-Bassham cycle intermediates peak at midday to support fast growth. The hitherto unexplored γ-glutamyl peptides act as reservoirs of amino acids. Interestingly, several storage molecules or their precursors accumulate during the dark phase, dispelling the notion that all biosynthetic activity takes place in the light phase. Our results will guide metabolic modeling and strain engineering of cyanobacteria.

A multi-parametric screening platform for photosynthetic trait characterization of microalgae and cyanobacteria under inorganic carbon limitation.

Microalgae and cyanobacteria are considered as important model organisms to investigate the biology of photosynthesis; moreover, they are valuable sources of biomolecules for several biotechnological applications. Understanding the species-specific traits of photosynthetic electron transport is extremely important, because it contributes to the regulation of ATP/NADPH ratio, which has direct/indirect links to carbon fixation and other metabolic pathways and thus overall growth and biomass production. In the present work, a cuvette-based setup is developed, in which a combination of measurements of dissolved oxygen, pH, chlorophyll fluorescence and NADPH kinetics can be performed without disturbing the physiological status of the sample. The suitability of the system is demonstrated using a model cyanobacterium Synechocystis sp. PCC6803, as well as biofuel-candidate microalgae species, such as Chlorella sorokiniana, Dunaliella salina and Nannochloropsis limnetica undergoing inorganic carbon (Ci) limitation. Inorganic carbon limitation, induced by photosynthetic Ci uptake under continuous illumination, caused a decrease in the effective quantum yield of PSII (Y(II)) and loss of oxygen-evolving capacity in all species investigated here; these effects were largely recovered by the addition of NaHCO3. Detailed analysis of the dark-light and light-dark transitions of NADPH production/uptake and changes in chlorophyll fluorescence kinetics revealed species- and condition-specific responses. These responses indicate that the impact of decreased Calvin-Benson cycle activity on photosynthetic electron transport pathways involving several sections of the electron transport chain (such as electron transfer via the QA-QB-plastoquinone pool, the redox state of the plastoquinone pool) can be analyzed with high sensitivity in a comparative manner. Therefore, the integrated system presented here can be applied for screening for specific traits in several significant species at different stages of inorganic carbon limitation, a condition that strongly impacts primary productivity.

Effect of inorganic carbon limitation on the conversion of organic carbon to total fatty acids by Monodus subterraneus

Traditional autotrophic microalgae exhibit low rates of organic carbon assimilation and conversion to useful compounds when switching to mixotrophic or heterotrophic growth. The goal of this study was to investigate the effect of inorganic carbon limitation on the efficiency of organic carbon (glycerol) assimilation and conversion to total fatty acids (TFAs) or the long-chain polyunsaturated fatty acid eicosapentaenoic acid (EPA). An oleaginous Monodus subterraneus was selected and six cultivation conditions were set, including Autotrophy-no aeration, Autotrophy-aeration, Mixotrophy-no aeration, Mixotrophy-no aeration & no Na2CO3, Mixotrophy-aeration, and Heterotrophy. The results showed M. subterraneus could utilize glycerol and grow under mixotrophic condition, while it was not occurred under heterotrophy. Superiority of mixotrophy to autotrophy on biomass productivity was more obvious under inorganic carbon limitation (no aeration or no Na2CO3) than inorganic carbon supply (aeration and existing Na2CO3 in the medium). CO2 limitation (no aeration) decreased content (of dry weight) and production (in medium) of TFAs, which was not evident in mixotrophy. CO2 limitation and inorganic carbon substrate stress largely improved the COD yield of TFAs and EPA under mixotrophic condition. TFA yield (%COD) under Mixotrophy-no aeration & no Na2CO3 was maximum (22.82%), and was almost two-fold higher than that under Mixotrophy-no aeration and nearly three-fold higher than that with Mixotrophy-aeration. EPA yield (% COD) under mixotrophy-no aeration & no Na2CO3 was maximum (6.58%). These results suggested that inorganic carbon limitation is a potentially useful method to enhance conversion of organic carbon to TFAs. Furthermore, the results suggest an application to obtain high value compounds (TFAs or EPA) combined with a high assimilation rate of waste glycerol from biodiesel and epichlorohydrin production by microalgae.

Achieving nitratation repression in an SBR at mainstream conditions through inorganic carbon limitation

This study investigated the effects of the influent inorganic carbon (IC) availability on nitrifying microbial activity of a sequencing batch reactor (SBR) operated for partial-nitritation (PN) at mainstream conditions (temperature of 15 °C and low influent N concentration). By operating the reactor with influent ammonium to inorganic carbon centration ratio (NH4+-N/IC) of 0.40 ± 0.01 mg N·mg C−1, which results in an excess of IC for full nitritation, the oxidized ammonium fully nitrified, and nitrite accumulation was not reached. Limiting influent IC (influent NH4+-N/IC of 0.73 ± 0.03 mg N·mg C−1) favored nitrite-oxidizing bacteria (NOB) suppression while nitritation was maintained, leading to nitrite accumulation of over 80%. Low bulk liquid IC reduced pH values and this one also favored the achievement of higher free nitrous acid (FNA) levels, but it did not stop nitrite oxidation to nitrate. PN control was also favored by keeping low bulk oxygen concentration during all the aeration phase (below 0.5 mg O2·L−1). Results obtained by batch assays showed that the ratio between the ammonium (rNH4+batch) and nitrite oxidation rates (rNO2−batch) dropped from 0.92 (without IC limitation) to 0.04 (with IC limitation). The results obtained highlight the critical IC role in the microbial interactions of the partial-nitritation process and present new insights in the direction of obtaining NH4+/NO2− for mainstream nitrite short-cut nitrogen removal process.

Growth of Cyanobacteria Is Constrained by the Abundance of Light and Carbon Assimilation Proteins

Cyanobacteria must balance separate demands for energy generation, carbon assimilation, and biomass synthesis. We used shotgun proteomics to investigate proteome allocation strategies in the model cyanobacterium Synechocystis sp. PCC 6803 as it adapted to light and inorganic carbon (Ci) limitation. When partitioning the proteome into seven functional sectors, we find that sector sizes change linearly with growth rate. The sector encompassing ribosomes is significantly smaller than in E.coli, which may explain the lower maximum growth rate in Synechocystis. Limitation of light dramatically affects multiple proteome sectors, whereas the effect of Ci limitation is weak. Carbon assimilation proteins respond more strongly to changes in light intensity than to Ci. Acoarse-grained cell economy model generally explains proteome trends. However, deviations from model predictions suggest that the large proteome sectors for carbon and light assimilation are not optimally utilized under some growth conditions and mayconstrain the proteome space available to ribosomes.

Effect of carbon limitation on photosynthetic electron transport in Nannochloropsis oculata

This study describes the impacts of inorganic carbon limitation on the photosynthetic efficiency and operation of photosynthetic electron transport pathways in the biofuel-candidate microalga Nannochloropsis oculata. Using a combination of highly-controlled cultivation setup (photobioreactor), variable chlorophyll a fluorescence and transient spectroscopy methods (electrochromic shift (ECS) and P700 redox kinetics), we showed that net photosynthesis and effective quantum yield of Photosystem II (PSII) decreased in N. oculata under carbon limitation. This was accompanied by a transient increase in total proton motive force and energy-dependent non-photochemical quenching as well as slightly elevated respiration. On the other hand, under carbon limitation the rapid increase in proton motive force (PMF, estimated from the total ECS signal) was also accompanied by reduced conductivity of ATP synthase to protons (estimated from the rate of ECS decay in dark after actinic illumination). This indicates that the slow operation of ATP synthase results in the transient build-up of PMF, which leads to the activation of fast energy dissipation mechanisms such as energy-dependent non-photochemical quenching. N. oculata also increased content of lipids under carbon limitation, which compensated for reduced NAPDH consumption during decreased CO2 fixation. The integrated knowledge of the underlying energetic regulation of photosynthetic processes attained with a combination of biophysical methods may be used to identify photo-physiological signatures of the onset of carbon limitation in microalgal cultivation systems, as well as to potentially identify microalgal strains that can better acclimate to carbon limitation.

Geochemistry and microbial community composition across a range of acid mine drainage impact and implications for the Neoarchean‐Paleoproterozoic transition

AbstractStreams impacted by acid mine drainage (AMD, also known as acid rock drainage) represent local environmental and ecological disasters; however, they may also present an opportunity to study microbial communities in environments analogous to past conditions. Neoarchean continents had streams and rivers replete with detrital pyrites. Following the emergence of oxygenic photosynthesis, Cyanobacteria colonized streams and rivers on continental surfaces. The combination of labile detrital pyrite grains and locally produced O2 generated by Cyanobacteria produced ideal conditions for pyrite oxidation similar to that found at modern AMD‐impacted sites. To explore the connection of modern sites to ancient conditions, we sampled sites that exhibited a range of AMD‐impact (e.g., pH from 2.1 to 7.9 [Fe2+] up to 5.2 mmol/L [SO42−] from 0.3 to 52.4 mmol/L) and found (i) nearly all analytes correlated to sulfate concentration; (ii) all sites exhibited the predominance of a single taxon most closely related to Ferrovum myxofaciens, an Fe‐oxidixing betaproteoabacterium capable of carbon and nitrogen fixation, and (iii) signs of potential inorganic carbon limitation and nitrogen cycling. From these findings and building on the work of others, we present a conceptual model of continental surfaces during the Neoarchean and Paleoproterozoic linking local O2 production to pyrite oxidation on continental surfaces to sulfate production and delivery to nearshore environments. The delivery of sulfate drives sulfate reduction and euxinia—favoring anoxygenic photosynthesis over cyanobacterial O2 generation in near‐continent/shelf marine environments.

Influence of substrate and pH on the diversity of the aeroterrestrial alga Klebsormidium (Klebsormidiales, Streptophyta): a potentially important factor for sympatric speciation

:Our knowledge of the processes involved in speciation of microalgae remains highly limited. In the present study, we investigated a potential role of ecological speciation processes in diversification of the filamentous green alga Klebsormidium. We examined 12 strains representing four different genotypes. The strains were collected from sandstone and limestone rocks and were cultivated at five different pH levels ranging from pH 4 to pH 8. We determined the responses of the 12 strains to the experimental pH conditions by (1) measuring the effective quantum yield of photosystem II, and (2) determining the growth rates after cultivation at different pH levels. Strong differences were found between the results obtained by these two methods. Direct counting of cells revealed a strong ecological differentiation of strains of Klebsormidium isolated from different substrate types. Strains isolated from limestone showed the highest growth rates at higher pH levels; whereas, the strains isolated from sandstone exhibited two distinct growth responses with optima at pH 5 and 6, respectively. In contrast, the effective quantum yield of photosystem II was always down-regulated at lower pH values, probably due to dissolved inorganic carbon limitation. In general, we determined distinct ecophysiological differentiation among distantly and closely related lineages, thereby corroborating our hypothesis that the sympatric speciation of terrestrial algae is driven by ecological divergence. We clearly showed that pH is a critical ecological factor that influences the diversity of autotrophic protists in terrestrial habitats.

NdhV subunit regulates the activity of type-1 NAD(P)H dehydrogenase under high light conditions in cyanobacterium Synechocystis sp. PCC 6803.

The cyanobacterial NAD(P)H dehydrogenase (NDH-1) complexes play crucial roles in variety of bioenergetic reactions. However, the regulative mechanism of NDH-1 under stressed conditions is still unclear. In this study, we detected that the NDH-1 activity is partially impaired, but the accumulation of NDH-1 complexes was little affected in the NdhV deleted mutant (ΔndhV) at low light in cyanobacterium Synechocystis sp. PCC 6803. ΔndhV grew normally at low light but slowly at high light under inorganic carbon limitation conditions (low pH or low CO2), meanwhile the activity of CO2 uptake was evidently lowered than wild type even at pH 8.0. The accumulation of NdhV in thylakoids strictly relies on the presence of the hydrophilic subcomplex of NDH-1. Furthermore, NdhV was co-located with hydrophilic subunits of NDH-1 loosely associated with the NDH-1L, NDH-1MS′ and NDH-1M complexes. The level of the NdhV was significantly increased at high light and deletion of NdhV suppressed the up-regulation of NDH-1 activity, causing the lowered the photosynthetic oxygen evolution at pH 6.5 and high light. These data indicate that NdhV is an intrinsic subunit of hydrophilic subcomplex of NDH-1, required for efficient operation of cyclic electron transport around photosystem I and CO2 uptake at high lights.