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Photosynthetic eukaryotes have metabolic pathways that occur in distinct subcellular compartments. However, because metabolites synthesized in one compartment, including fixed carbon compounds and reductant generated by photosynthetic electron flows, may be integral to processes in other compartments, the cells must efficiently move metabolites among the different compartments. This review examines the various photosynthetic electron flows used to generate ATP and fixed carbon and the trafficking of metabolites in the green alga Chlamydomomas reinhardtii; information on other algae and plants is provided to add depth and nuance to the discussion. We emphasized the trafficking of metabolites across the envelope membranes of the two energy powerhouse organelles of the cell, the chloroplast and mitochondrion, the nature and roles of the major mobile metabolites that move among these compartments, and the specific or presumed transporters involved in that trafficking. These transporters include sugar-phosphate (sugar-P)/inorganic phosphate (Pi) transporters and dicarboxylate transporters, although, in many cases, we know little about the substrate specificities of these transporters, how their activities are regulated/coordinated, compensatory responses among transporters when specific transporters are compromised, associations between transporters and other cellular proteins, and the possibilities for forming specific 'megacomplexes' involving interactions between enzymes of central metabolism with specific transport proteins. Finally, we discuss metabolite trafficking associated with specific biological processes that occur under various environmental conditions to help to maintain the cell's fitness. These processes include C4 metabolism in plants and the carbon concentrating mechanism, photorespiration, and fermentation metabolism in algae. [ABSTRACT FROM AUTHOR]

Plant physiologists set about comprehending the genesis of the C4 photosynthetic pathway after its discovery by Hatch and Slack. They discovered that a sophisticated combination of morphological and biochemical adaptations allowed the plant to concentrate CO2 around RuBisCO to achieve maximum efficiency. We categorize the evolutionary events leading to C4 photosynthesis, beginning with anoxygenic photosynthesis and the evolution of RuBisCO to the cooling of Earth by the Great Oxygenation Event that led to the oxygenic photosynthesis. The evolutionary descent of the C4 plants is a phenomenon that occurred around 30 million years ago. Due to industrialization and population growth, improved photosynthetic efficiency and carbon fixation of C4 plants could contest the current global scenario of rising CO2 concentration. C3 crops engineered with C4 traits, implemented on a large scale, could impact the climate globally. Here we discuss the various strategies used to introduce C4 traits in the C3 plants and the potential techniques to be considered for successful hybridization. [ABSTRACT FROM AUTHOR]

Hydrogen isotope ratios (δ²H) represent an important natural tracer of metabolic processes, but quantitative models of processes controlling H-fractionation in aquatic photosynthetic organisms are lacking. Here, we elucidate the underlying physiological controls of ²H/¹H fractionation in algal lipids by systematically manipulating temperature, light, and CO2(aq) in continuous cultures of the haptophyte Gephyrocapsa oceanica. We analyze the hydrogen isotope fractionation in alkenones (αalkenone), a class of acyl lipids specific to this species and other haptophyte algae. We find a strong decrease in the αalkenone with increasing CO2 (aq) and confirm αalkenone correlates with temperature and light. Based on the known biosynthesis pathways, we develop a cellular model of the δ²H of algal acyl lipids to evaluate processes contributing to these controls on fractionation. Simulations show that longer residence times of NADPH in the chloroplast favor a greater exchange of NADPH with ²H-richer intracellular water, increasing αalkenone. Higher chloroplast CO2(aq) and temperature shorten NADPH residence time by enhancing the carbon fixation and lipid synthesis rates. The inverse correlation of αalkenone to CO2(aq) in our cultures suggests that carbon concentrating mechanisms (CCM) do not achieve a constant saturation of CO2 at the Rubisco site, but rather that chloroplast CO2 varies with external CO2(aq). The pervasive inverse correlation of αalkenone with CO2(aq) in the modern and preindustrial ocean also suggests that natural populations may not attain a constant saturation of Rubisco with the CCM. Rather than reconstructing growth water, αalkenone may be a powerful tool to elucidate the carbon limitation of photosynthesis. [ABSTRACT FROM AUTHOR]

In reef-building corals (order Scleractinia) and giant clams (phylum Molluca), V-type H+-ATPase (VHA) in host cells is part of a carbon concentrating mechanism (CCM) that regulates photosynthetic rates of their symbiotic algae. Here, we show that VHA plays a similar role in the sea anemone Anemonia majano, a member of the order Actinaria and sister group to the Scleractinia, which in contrast to their colonial calcifying coral relatives is a solitary, soft-bodied taxa. Western blotting and immunofluorescence revealed that VHA was abundantly present in the host-derived symbiosome membrane surrounding the photosymbionts. Pharmacological inhibition of VHA activity in individual anemones resulted in an approximately 80% decrease of photosynthetic O2 production. These results extend the presence of a host-controlled VHA-dependent CCM to non-calcifying cnidarians of the order Actiniaria, suggesting it is widespread among photosymbiosis between aquatic invertebrates and Symbiodiniaceae algae.

Due to the progressive climate change on our planet, scientists are interested in solving this issue since it threatens not only certain regions or countries but also the world’s ecosystems and economies. Therefore, minimizing carbon dioxide (CO2) emissions and reducing atmospheric levels are global priorities. Thus, it is necessary at this moment to develop an appropriate approach to reduce or stabilize CO2 levels in the atmosphere. However, CO2 capture projects are long-term, low-profitable, and high-risk environmental projects. Consequently, it is necessary to find an appropriate and sustainable CO2 capture approach that is efficient in reducing atmospheric CO2 levels while having a safe impact on the environment. Although carbon (C) is the key basic component used to produce biological compounds by photosynthetic organisms in terrestrial plants, the C pathway is a key factor affecting the capture of CO2 by photosynthetic organisms. Among photosynthetic organisms, Paulownia, a multipurpose tree, is popular around the world for its timber and its potential role in CO2 sequestration. Paulownia spp. belongs to the Paulowniaceae family and comprises a group of trees. These trees are primarily found in southeastern Asia, particularly in China, and have been intentionally grown for more than two millennia due to their ornamental, cultural, and medicinal value. The number of Paulownia species varies depending on taxonomic classification, ranging from 6 to 17. Among them, Paulownia tomentosa, Paulownia elongata, Paulownia fortunei, and Paulownia catalpifolia are the most widely recognized and favored species. The present review provides a comprehensive technical-economic scenario for the capture of one million tons of CO2 by Paulownia trees (as a terrestrial plant model, grown on 2,400 ha−1). P. tomentosa can be utilized in agroforestry systems to mitigate greenhouse gas (GHG) emissions within urban cities and emphasize the carbon storage potential of agroforestry. In conclusion, Paulownia trees as an environmental mass project showed great encouragement to investors and governments to expand these types of projects to achieve global climate goals by 2050.

Allometric relationships among the dimensions of leaves and their cells hold across diverse eudicotyledons, but have remained untested in the leaves of grasses. We hypothesised that geometric (proportional) allometries of cell sizes across tissues and of leaf dimensions would arise due to the coordination of cell development and that of cell functions such as water, nutrient and energy transport, and that cell sizes across tissues would be associated with light‐saturated photosynthetic rate. We tested predictions across 27 globally distributed C3 and C4 grass species grown in a common garden. We found positive relationships among average cell sizes within and across tissues, and of cell sizes with leaf dimensions. Grass leaf anatomical allometries were similar to those of eudicots, with exceptions consistent with the fewer cell layers and narrower form of grass leaves, and the specialised roles of epidermis and bundle sheath in storage and leaf movement. Across species, mean cell sizes in each tissue were associated with light‐saturated photosynthetic rate per leaf mass, supporting the functional coordination of cell sizes. These findings highlight the generality of evolutionary allometries within the grass lineage and their interlinkage with coordinated development and function. Summary statement: Allometries among leaf cell sizes and of cell sizes with leaf dimensions and photosynthetic rate hold across grass species, supporting their basis in developmental and functional coordination. [ABSTRACT FROM AUTHOR]

Background and Aims: To better understand C4 evolution in monocots, we characterized C3-C4 intermediate phenotypes in the grass genus Homolepis (subtribe Arthropogoninae)., Methods: Carbon isotope ratio (δ13C), leaf gas exchange, mesophyll (M) to bundle sheath (BS) tissue characteristics, organelle size and numbers in M and BS tissue, and tissue distribution of the P-subunit of glycine decarboxylase (GLDP) were determined for five Homolepis species and the C4 grass Mesosetum loliiforme from a phylogenetic sister clade. We generated a transcriptome-based phylogeny for Homolepis and Mesosetum species to interpret physiological and anatomical patterns in an evolutionary context, and to test for hybridization., Key Results: Homolepis contains two C3 (H. glutinosa, H. villaricensis), one weaker form of C2 termed sub-C2 (H. isocalycia), and two C2 species (H. longispicula, H. aturensis). Homolepis longispicula and H. aturensis express over 85% of leaf GDC in centripetal mitochondria within the BS, and have increased fractions of leaf chloroplasts, mitochondria and peroxisomes within the BS relative to H. glutinosa. Analysis of leaf gas exchange, cell ultrastructural, and transcript expression show M. loliiforme is a C4 plant of the NADP-malic enzyme subtype. Homolepis is comprised of two sister clades, one containing H. glutinosa and H. villaricensis and the second H. longispicula and H. aturensis. Homolepis isocalycia is of hybrid origin, with parents being H. aturensis and a common ancestor of the C3  Homolepis clade and H. longispicula., Conclusions: Photosynthetic activation of BS tissue in the sub-C2 and C2 species of Homolepis is similar to patterns observed in C3-C4 intermediate eudicots, indicating common evolutionary pathways from C3 to C4 photosynthesis in these disparate clades. Hybridization can diversify the C3-C4 intermediate character state and should be considered in reconstructing putative ancestral states using phylogenetic analyses., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Annals of Botany Company.)

Incorporating C4 photosynthetic traits into C3 crops is a rational approach for sustaining future demands for crop productivity. Using classical plant breeding, engineering this complex trait is unlikely to achieve its target. Therefore, it is critical and timely to implement novel biotechnological crop improvement strategies to accomplish this goal. However, a fundamental understanding of C3, C4, and C3–C4 intermediate metabolism is crucial for the targeted use of biotechnological tools. This review assesses recent progress towards engineering C4 photosynthetic traits in C3 crops. We also discuss lessons learned from successes and failures of recent genetic engineering attempts in C3 crops, highlighting the pros and cons of using rice as a model plant for short‐, medium‐ and long‐term goals of genetic engineering. This review provides an integrated approach towards engineering improved photosynthetic efficiency in C3 crops for sustaining food, fibre and fuel production around the globe. [ABSTRACT FROM AUTHOR]

Anthropogenic emissions of carbon dioxide (CO2) contribute to global warming. Limiting temperature rise requires negative emission techniques to retract the emitted CO2 from the atmosphere. Through photosynthesis, ecosystems naturally sequester and store carbon. Enhancing these processes forms the basis of biological sequestration strategies. Ecosystems are a sink of atmospheric CO2 and significantly impact the global carbon cycle. The fixed carbon is converted into biomass, a portion of which enters the soil carbon pool and can be sequestered for millennia. The formation of stable soil organic carbon (SOC) depends on land use, management practices, and the use of amendments. Employing best management practices and carbon boosting approaches such as conservation agriculture, agroforestry, biochar, afforestation, and restoration of wetlands can improve SOC stocks and create a positive soil carbon budget, especially in degraded ecosystems. . Carbon fixation by plants and microbes is fundamental to biological sequestration. Regulating the properties and expression of the enzymes involved and introducing novel pathways for carbon capture can enhance carbon fixation efficiency and positively affect yield. This review discusses biological carbon sequestration strategies highlighting the recent findings in the effects and potential of soil carbon boosting approaches in carbon mitigation and the prospects of genetic engineering in enhancing carbon fixation.

Autotrophic bacteria rely on various mechanisms to increase intracellular concentrations of inorganic forms of carbon (i.e., bicarbonate and CO2) in order to improve the efficiency with which they can be converted to organic forms. Transmembrane bicarbonate transporters and carboxysomes play key roles in accumulating bicarbonate and CO2, but other regulatory elements of carbon concentration mechanisms in bacteria are less understood. In this study, after analyzing the genomic regions around α-type carboxysome operons, we characterize a protein that is conserved across these operons but has not been previously studied. On the basis of a series of apo- and ligand-bound crystal structures and supporting biochemical data, we show that this protein, which we refer to as the carboxysome-associated PII protein (CPII), represents a new and distinct subfamily within the broad superfamily of previously studied PII regulatory proteins, which are generally involved in regulating nitrogen metabolism in bacteria. CPII undergoes dramatic conformational changes in response to ADP binding, and the affinity for nucleotide binding is strongly enhanced by the presence of bicarbonate. CPII therefore appears to be a unique type of PII protein that senses bicarbonate availability, consistent with its apparent genomic association with the carboxysome and its constituents.

Abstract Background Pyropia yezoensis (Rhodophyta) is widely cultivated in East Asia and plays important economic, ecological and research roles. Although inorganic carbon utilization of P. yezoensis has been investigated from a physiological aspect, the carbon concentration mechanism (CCM) of P. yezoensis remains unclear. To explore the CCM of P. yezoensis, especially during its different life stages, we tracked changes in the transcriptome, photosynthetic efficiency and in key enzyme activities under different inorganic carbon concentrations. Results Photosynthetic efficiency demonstrated that sporophytes were more sensitive to low carbon (LC) than gametophytes, with increased photosynthesis rate during both life stages under high carbon (HC) compared to normal carbon (NC) conditions. The amount of starch and number of plastoglobuli in cells corresponded with the growth reaction to different inorganic carbon (Ci) concentrations. We constructed 18 cDNA libraries from 18 samples (three biological replicates per Ci treatment at two life cycles stages) and sequenced these using the Illumina platform. De novo assembly generated 182,564 unigenes, including approximately 275 unigenes related to CCM. Most genes encoding internal carbonic anhydrase (CA) and bicarbonate transporters involved in the biophysical CCM pathway were induced under LC in comparison with NC, with transcript abundance of some PyCAs in gametophytes typically higher than that in sporophytes. We identified all key genes participating in the C4 pathway and showed that their RNA abundances changed with varying Ci conditions. High decarboxylating activity of PEPCKase and low PEPCase activity were observed in P. yezoensis. Activities of other key enzymes involved in the C4-like pathway were higher under HC than under the other two conditions. Pyruvate carboxylase (PYC) showed higher carboxylation activity than PEPC under these Ci conditions. Isocitrate lyase (ICL) showed high activity, but the activity of malate synthase (MS) was very low. Conclusion We elucidated the CCM of P. yezoensis from transcriptome and enzyme activity levels. All results indicated at least two types of CCM in P. yezoensis, one involving CA and an anion exchanger (transporter), and a second, C4-like pathway belonging to the PEPCK subtype. PYC may play the main carboxylation role in this C4-like pathway, which functions in both the sporophyte and gametophyte life cycles.

In reef-building corals (order Scleractinia) and giant clams (phylum Molluca), V-type H+-ATPase (VHA) in host cells is part of a carbon concentrating mechanism (CCM) that regulates photosynthetic rates of their symbiotic algae. Here, we show that VHA plays a similar role in the sea anemone Anemonia majano, a member of the order Actinaria and sister group to the Scleractinia, which in contrast to their colonial calcifying coral relatives is a solitary, soft-bodied taxa. Western blotting and immunofluorescence revealed that VHA was abundantly present in the host-derived symbiosome membrane surrounding the photosymbionts. Pharmacological inhibition of VHA activity in individual anemones resulted in an approximately 80% decrease of photosynthetic O2 production. These results extend the presence of a host-controlled VHA-dependent CCM to non-calcifying cnidarians of the order Actiniaria, suggesting it is widespread among photosymbiosis between aquatic invertebrates and Symbiodiniaceae algae.

In algae, it is well established that the pyrenoid, a component of the carbon-concentrating mechanism (CCM), is essential for efficient photosynthesis at low CO2. However, the signal that triggers the formation of the pyrenoid has remained elusive. Here, we show that, in Chlamydomonas reinhardtii, the pyrenoid is strongly induced by hyperoxia, even at high CO2 or bicarbonate levels. These results suggest that the pyrenoid can be induced by a common product of photosynthesis specific to low CO2 or hyperoxia. Consistent with this view, the photorespiratory by-product, H2O2, induced the pyrenoid, suggesting that it acts as a signal. Finally, we show evidence for linkages between genetic variations in hyperoxia tolerance, H2O2 signaling, and pyrenoid morphologies.

Ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (RuBisCO) is the carbon-fixing enzyme present in most photosynthetic organisms, converting CO2 into organic matter. Globally, photosynthetic efficiency in terrestrial plants has become increasingly challenged in recent decades due to a rapid increase in atmospheric CO2 and associated changes toward warmer and dryer environments. Well adapted for these new climatic conditions, the C4 photosynthetic pathway utilizes carbon concentrating mechanisms to increase CO2 concentrations surrounding RuBisCO, suppressing photorespiration from the oxygenase catalyzed reaction with O2. The energy efficiency of C3 photosynthesis, from which the C4 pathway evolved, is thought to rely critically on an uninterrupted supply of chloroplast CO2. Part of the homeostatic mechanism that maintains this constancy of supply involves the CO2 produced as a byproduct of photorespiration in a negative feedback loop. Analyzing the database of RuBisCO kinetic parameters, we suggest that in genera (Flaveria and Panicum) for which both C3 and C4 examples are available, the C4 pathway evolved only from C3 ancestors possessing much lower than the average carboxylase specificity relative to that of the oxygenase reaction (SC/O=SC/SO), and hence, the higher CO2 levels required for development of the photorespiratory CO2 pump (C2 photosynthesis) essential in the initial stages of C4 evolution, while in the later stage (final optimization phase in the Flaveria model) increased CO2 turnover may have occurred, which would have been supported by the higher CO2 levels. Otherwise, C4 RuBisCO kinetic traits remain little changed from the ancestral C3 species. At the opposite end of the spectrum, C3 plants (from Limonium) with higher than average SC/O, which may be associated with the ability of increased CO2, relative to O2, affinity to offset reduced photorespiration and chloroplast CO2 levels, can tolerate high stress environments. It is suggested that, instead of inherently constrained by its kinetic mechanism, RuBisCO possesses the extensive kinetic plasticity necessary for adaptation to changes in photorespiration that occur in the homeostatic regulation of CO2 supply under a broad range of abiotic environmental conditions.

Ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (RuBisCO) is the carbon-fixing enzyme present in most photosynthetic organisms, converting CO2 into organic matter. Globally, photosynthetic efficiency in terrestrial plants has become increasingly challenged in recent decades due to a rapid increase in atmospheric CO2 and associated changes toward warmer and dryer environments. Well adapted for these new climatic conditions, the C4 photosynthetic pathway utilizes carbon concentrating mechanisms to increase CO2 concentrations surrounding RuBisCO, suppressing photorespiration from the oxygenase catalyzed reaction with O2. The energy efficiency of C3 photosynthesis, from which the C4 pathway evolved, is thought to rely critically on an uninterrupted supply of chloroplast CO2. Part of the homeostatic mechanism that maintains this constancy of supply involves the CO2 produced as a byproduct of photorespiration in a negative feedback loop. Analyzing the database of RuBisCO kinetic parameters, we suggest that in genera (Flaveria and Panicum) for which both C3 and C4 examples are available, the C4 pathway evolved only from C3 ancestors possessing much lower than the average carboxylase specificity relative to that of the oxygenase reaction (S C/O= S C/ S O), and hence, the higher CO2 levels required for development of the photorespiratory CO2 pump (C2 photosynthesis) essential in the initial stages of C4 evolution, while in the later stage (final optimization phase in the Flaveria model) increased CO2 turnover may have occurred, which would have been supported by the higher CO2 levels. Otherwise, C4 RuBisCO kinetic traits remain little changed from the ancestral C3 species. At the opposite end of the spectrum, C3 plants (from Limonium) with higher than average S C/O, which may be associated with the ability of increased CO2, relative to O2, affinity to offset reduced photorespiration and chloroplast CO2 levels, can tolerate high stress environments. It is suggested that, instead of inherently constrained by its kinetic mechanism, RuBisCO possesses the extensive kinetic plasticity necessary for adaptation to changes in photorespiration that occur in the homeostatic regulation of CO2 supply under a broad range of abiotic environmental conditions. [ABSTRACT FROM AUTHOR]

Symbiotic dinoflagellate algae residing inside coral tissues supply the host with the majority of their energy requirements through the translocation of photosynthetically fixed carbon. The algae, in turn, rely on the host for the supply of inorganic carbon. Carbon must be concentrated as CO2 in order for photosynthesis to proceed, and here we show that the coral host plays an active role in this process. The host-derived symbiosome membrane surrounding the algae abundantly expresses vacuolar H(+)-ATPase (VHA), which acidifies the symbiosome space down to pH ∼ 4. Inhibition of VHA results in a significant decrease in average H(+) activity in the symbiosome of up to 75% and a significant reduction in O2 production rate, a measure of photosynthetic activity. These results suggest that host VHA is part of a previously unidentified carbon concentrating mechanism for algal photosynthesis and provide mechanistic evidence that coral host cells can actively modulate the physiology of their symbionts.

Hygrophila polysperma is a heterophyllous amphibious plant. The growth of H. polysperma in submerged conditions is challenging due to the low CO2 environment, increased resistance to gas diffusion, and bicarbonate ion (HCO3–) being the dominant dissolved inorganic carbon source. The submerged leaves of H. polysperma have significantly higher rates of underwater photosynthesis compared with the terrestrial leaves. 4,4′-Diisothiocyanatostilbene-2,2′-disulfonate (DIDS), an anion exchanger protein inhibitor, and ethoxyzolamide (EZ), an inhibitor of internal carbonic anhydrase, repressed underwater photosynthesis by the submerged leaves. These results suggested that H. polysperma acclimates to the submerged condition by using HCO3– for photosynthesis. H. polysperma transports HCO3– into the leaf by a DIDS-sensitive HCO3– transporter and converted to CO2 by carbonic anhydrase. Additionally, proteome analysis revealed that submerged leaves accumulated fewer proteins associated with C4 photosynthesis compared with terrestrial leaves. This finding suggested that H. polysperma is capable of C4 and C3 photosynthesis in the terrestrial and submerged leaves, respectively. The ratio of phosphoenol pyruvate carboxylase to ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the submerged leaves was less than that in the terrestrial leaves. Upon anatomical observation, the terrestrial leaves exhibited a phenotype similar to the Kranz anatomy found among C4 plants; however, chloroplasts in the bundle sheath cells were not located adjacent to the vascular bundles, and the typical Kranz anatomy was absent in submerged leaves. These results suggest that H. polysperma performs proto-Kranz type photosynthesis in a terrestrial environment and shifts from a proto-Kranz type in terrestrial leaves to a HCO3– use photosynthesis in the submerged environments.

Hygrophila polysperma is a heterophyllous amphibious plant. The growth of H. polysperma in submerged conditions is challenging due to the low CO2 environment, increased resistance to gas diffusion, and bicarbonate ion (HCO3–) being the dominant dissolved inorganic carbon source. The submerged leaves of H. polysperma have significantly higher rates of underwater photosynthesis compared with the terrestrial leaves. 4,4′-Diisothiocyanatostilbene-2,2′-disulfonate (DIDS), an anion exchanger protein inhibitor, and ethoxyzolamide (EZ), an inhibitor of internal carbonic anhydrase, repressed underwater photosynthesis by the submerged leaves. These results suggested that H. polysperma acclimates to the submerged condition by using HCO3– for photosynthesis. H. polysperma transports HCO3– into the leaf by a DIDS-sensitive HCO3– transporter and converted to CO2 by carbonic anhydrase. Additionally, proteome analysis revealed that submerged leaves accumulated fewer proteins associated with C4 photosynthesis compared with terrestrial leaves. This finding suggested that H. polysperma is capable of C4 and C3 photosynthesis in the terrestrial and submerged leaves, respectively. The ratio of phosphoenol pyruvate carboxylase to ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the submerged leaves was less than that in the terrestrial leaves. Upon anatomical observation, the terrestrial leaves exhibited a phenotype similar to the Kranz anatomy found among C4 plants; however, chloroplasts in the bundle sheath cells were not located adjacent to the vascular bundles, and the typical Kranz anatomy was absent in submerged leaves. These results suggest that H. polysperma performs proto-Kranz type photosynthesis in a terrestrial environment and shifts from a proto-Kranz type in terrestrial leaves to a HCO3– use photosynthesis in the submerged environments. [ABSTRACT FROM AUTHOR]

Hydrogen isotope ratios (δ 2 H) represent an important natural tracer of metabolic processes, but quantitative models of processes controlling H-fractionation in aquatic photosynthetic organisms are lacking. Here, we elucidate the underlying physiological controls of 2 H/ 1 H fractionation in algal lipids by systematically manipulating temperature, light, and CO 2 (aq) in continuous cultures of the haptophyte Gephyrocapsa oceanica . We analyze the hydrogen isotope fractionation in alkenones (α alkenone ), a class of acyl lipids specific to this species and other haptophyte algae. We find a strong decrease in the α alkenone with increasing CO 2 (aq) and confirm α alkenone correlates with temperature and light. Based on the known biosynthesis pathways, we develop a cellular model of the δ 2 H of algal acyl lipids to evaluate processes contributing to these controls on fractionation. Simulations show that longer residence times of NADPH in the chloroplast favor a greater exchange of NADPH with 2 H-richer intracellular water, increasing α alkenone . Higher chloroplast CO 2 (aq) and temperature shorten NADPH residence time by enhancing the carbon fixation and lipid synthesis rates. The inverse correlation of α alkenone to CO 2 (aq) in our cultures suggests that carbon concentrating mechanisms (CCM) do not achieve a constant saturation of CO 2 at the Rubisco site, but rather that chloroplast CO 2 varies with external CO 2 (aq). The pervasive inverse correlation of α alkenone with CO 2 (aq) in the modern and preindustrial ocean also suggests that natural populations may not attain a constant saturation of Rubisco with the CCM. Rather than reconstructing growth water, α alkenone may be a powerful tool to elucidate the carbon limitation of photosynthesis., Competing Interests: Competing interests statement:The authors declare no competing interest.

Summary: C2 photosynthesis is a carbon concentrating mechanism that can increase net CO2 assimilation by capturing, concentrating and re‐assimilating CO2 released by photorespiration. Empirical and modelling studies indicate that C2 plants assimilate more carbon than C3 plants under high temperature, bright light, and low CO2 conditions. I argue that engineering C2 photosynthesis into C3 crops is a promising approach to improve photosynthetic performance under these – and temporally heterogeneous – environments, and review the modifications that may re‐create a C2 phenotype in C3 plants. Although a C2 engineering program would encounter many of the same challenges faced by C4 engineering programmes, the simpler leaf anatomical requirements make C2 engineering a feasible approach to improve crops in the medium term. [ABSTRACT FROM AUTHOR]

Membrane permeabilities to CO2 and HCO3− constrain the function of CO2 concentrating mechanisms that algae use to supply inorganic carbon for photosynthesis. In diatoms and green algae, plasma membranes are moderately to highly permeable to CO2 but effectively impermeable to HCO3−. Here, CO2 and HCO3− membrane permeabilities were measured using an 18O‐exchange technique on two species of haptophyte algae, Emiliania huxleyi and Calcidiscus leptoporus, which showed that the plasma membranes of these species are also highly permeable to CO2 (0.006–0.02 cm · s−1) but minimally permeable to HCO3−. Increased temperature and CO2 generally increased CO2 membrane permeabilities in both species, possibly due to changes in lipid composition or CO2 channel proteins. Changes in CO2 membrane permeabilities showed no association with the density of calcium carbonate coccoliths surrounding the cell, which could potentially impede passage of compounds. Haptophyte plasma‐membrane permeabilities to CO2 were somewhat lower than those of diatoms but generally higher than membrane permeabilities of green algae. One caveat of these measurements is that the model used to interpret 18O‐exchange data assumes that carbonic anhydrase, which catalyzes 18O‐exchange, is homogeneously distributed in the cell. The implications of this assumption were tested using a two‐compartment model with an inhomogeneous distribution of carbonic anhydrase to simulate 18O‐exchange data and then inferring plasma‐membrane CO2 permeabilities from the simulated data. This analysis showed that the inferred plasma‐membrane CO2 permeabilities are minimal estimates but should be quite accurate under most conditions. [ABSTRACT FROM AUTHOR]

Rapid changes on nutrient supply and CO2 concentration that occurred in the northern South China Sea (SCS) during the Early Oligocene, provides an ideal natural laboratory, allowing us to peer into the coccolithophores' physiology in the geological records. In this study, we established a new nannofossil assemblage index, termed as E* ratio, which is calculated by the relative abundance of eutrophic taxa and meso-oligotrophic taxa ( E ∗ = e e + c × 100 , where e is eutrophic taxa, and c is meso-oligotrophic taxa). Eutrophic taxa include small Reticulofenestra, Reticulofenestra lockeri group, Reticulofenestra bisecta group and Coccolithus pelagicus group, while meso-oligotrophic taxa include Cyclicargolithus spp. The E* ratio is well correlated with nutrient proxy during the Early Oligocene, while with different covarying patterns under the higher and lower CO2 condition. By comparing the assemblage changes to the published data, we suggest that coccolithophores may change the way they use carbon source and nutrient with the decline of CO2. Furthermore, this implies a possible initiation of the carbon concentrating mechanism. [ABSTRACT FROM AUTHOR]

Background: Pyropia yezoensis (Rhodophyta) is widely cultivated in East Asia and plays important economic, ecological and research roles. Although inorganic carbon utilization of P. yezoensis has been investigated from a physiological aspect, the carbon concentration mechanism (CCM) of P. yezoensis remains unclear. To explore the CCM of P. yezoensis, especially during its different life stages, we tracked changes in the transcriptome, photosynthetic efficiency and in key enzyme activities under different inorganic carbon concentrations. Results: Photosynthetic efficiency demonstrated that sporophytes were more sensitive to low carbon (LC) than gametophytes, with increased photosynthesis rate during both life stages under high carbon (HC) compared to normal carbon (NC) conditions. The amount of starch and number of plastoglobuli in cells corresponded with the growth reaction to different inorganic carbon (Ci) concentrations. We constructed 18 cDNA libraries from 18 samples (three biological replicates per Ci treatment at two life cycles stages) and sequenced these using the Illumina platform. De novo assembly generated 182,564 unigenes, including approximately 275 unigenes related to CCM. Most genes encoding internal carbonic anhydrase (CA) and bicarbonate transporters involved in the biophysical CCM pathway were induced under LC in comparison with NC, with transcript abundance of some PyCAs in gametophytes typically higher than that in sporophytes. We identified all key genes participating in the C4 pathway and showed that their RNA abundances changed with varying Ci conditions. High decarboxylating activity of PEPCKase and low PEPCase activity were observed in P. yezoensis. Activities of other key enzymes involved in the C4-like pathway were higher under HC than under the other two conditions. Pyruvate carboxylase (PYC) showed higher carboxylation activity than PEPC under these Ci conditions. Isocitrate lyase (ICL) showed high activity, but the activity of malate synthase (MS) was very low. Conclusion: We elucidated the CCM of P. yezoensis from transcriptome and enzyme activity levels. All results indicated at least two types of CCM in P. yezoensis, one involving CA and an anion exchanger (transporter), and a second, C4-like pathway belonging to the PEPCK subtype. PYC may play the main carboxylation role in this C4-like pathway, which functions in both the sporophyte and gametophyte life cycles. [ABSTRACT FROM AUTHOR]

Highlights • Karenia brevis is not affected by changes in environmentally relevant CO 2 concentrations. • Karenia brevis maintains an efficient and regulated CO 2 concentrating mechanism (CCM). • The CCM can sustain dense blooms Karenia brevis. • Rising CO 2 can potentially elevate the negative socioeconomic effects of Karenia brevis. Abstract Karenia brevis is a marine dinoflagellate commonly found in the Gulf of Mexico and important both ecologically and economically due to its production of the neurotoxin brevetoxin, which can cause respiratory illness in humans and widespread death of marine animals. K. brevis strains have previously shown to be sensitive to changes in CO 2 , both in terms of growth as well as toxin production. Our study aimed to understand this sensitivity by measuring underlying mechanisms, such as photosynthesis, carbon acquisition, and photophysiology. K. brevis (CCFWC-126) did not show a significant response in growth, cellular composition of carbon and nitrogen, nor in photosynthetic rates between p CO 2 concentrations of 150, 400, or 780 µatm. However, a strong response in its acquisition of inorganic carbon was found. Half saturation values for CO 2 increased from 1.5 to 3.3 µM, inorganic carbon preference switched from HCO 3 − to CO 2 (14–56% CO 2 usage), and external carbonic anhydrase activity was downregulated by 23% when comparing low and high p CO 2. We conclude that K. brevis must employ an efficient and regulated CO 2 concentrating mechanism (CCM) to maintain constant carbon fixation and growth across p CO 2 levels. No statistically significant correlation between CO 2 and brevetoxin content was found, yet a positive trend with enhanced p CO 2 was detected. This study is the first explaining how this socioeconomically important species is able to efficiently supply inorganic carbon for photosynthesis, which can potentially prolong bloom situations. This study also highlights that elevated CO 2 concentrations, as projected for a future ocean, can affect underlying physiological processes of K. brevis , some of which could lead to increases in cellular brevetoxin production and therefore increased impacts on coastal ecosystems and economies. [ABSTRACT FROM AUTHOR]

Heterosigma akashiwo is a raphidophyte known for forming ichthyotoxic blooms. In order to predict the potential impacts of rising CO 2 on H. akashiwo it is necessary to understand the factors influencing growth rates over a range of CO 2 concentrations. Here we examined the physiology and gene expression response of H. akashiwo to concentrations from 200 to 1000 ppm CO 2. Growth rate data were combined from this and previous studies and fit with a CO 2 limitation-inhibition model that revealed an apparent growth optimum around 600–800 ppm CO 2. Physiological changes included a significant increase in C:N ratio at ∼800 ppm CO 2 and a significant decrease in hydrogen peroxide concentration at ∼1000 ppm. Whole transcriptome sequencing of H. akashiwo revealed sharp distinctions in metabolic pathway gene expression between ∼600 and ∼800 ppm CO 2. Hierarchical clustering by co-expression identified groups of genes with significant correlations to CO 2 and growth rate. Genes with significant differential expression with CO 2 included carbon concentrating mechanism genes such as beta-carbonic anhydrases and a bicarbonate transporter, which may underpin shifts in physiology. Genes involved in cell motility were significantly changed by both elevated CO 2 and growth rate, suggesting that future ocean conditions could modify swimming behavior in this species. [ABSTRACT FROM AUTHOR]

Red coralline algae are projected to be sensitive to ocean acidification, particularly in polar oceans. As important ecosystem engineers, their potential sensitivity has broad implications, and understanding their carbon acquisition mechanisms is necessary for making reliable predictions. Therefore, we investigated the localized carbonate chemistry at the surface of Arctic coralline algae using microsensors. We report for the first time carbonate ion concentration and pH measurements ([CO32-]) at and above the algal surface in the microenvironment. We show that surface pH and [CO32-] are higher than the bulk seawater in the light, and even after hours of darkness. We further show that three species of Arctic coralline algae have efficient carbon concentrating mechanisms including direct bicarbonate uptake and indirect bicarbonate use via a carbonic anhydrase enzyme. Our results suggest that Arctic corallines have strong biological control over their surface chemistry, where active calcification occurs, and that net dissolution in the dark does not occur. We suggest that the elevated pH and [CO32-] in the dark could be explained by a high rate of light independent carbon fixation that reduces respiratory CO2 release. This mechanism could provide a potential adaptation to ocean acidification in Arctic coralline algae, which has important implications for future Arctic marine ecosystems.

Allometric relationships among the dimensions of leaves and their cells hold across diverse eudicotyledons, but have remained untested in the leaves of grasses. We hypothesised that geometric (proportional) allometries of cell sizes across tissues and of leaf dimensions would arise due to the coordination of cell development and that of cell functions such as water, nutrient and energy transport, and that cell sizes across tissues would be associated with light-saturated photosynthetic rate. We tested predictions across 27 globally distributed C 3 and C 4 grass species grown in a common garden. We found positive relationships among average cell sizes within and across tissues, and of cell sizes with leaf dimensions. Grass leaf anatomical allometries were similar to those of eudicots, with exceptions consistent with the fewer cell layers and narrower form of grass leaves, and the specialised roles of epidermis and bundle sheath in storage and leaf movement. Across species, mean cell sizes in each tissue were associated with light-saturated photosynthetic rate per leaf mass, supporting the functional coordination of cell sizes. These findings highlight the generality of evolutionary allometries within the grass lineage and their interlinkage with coordinated development and function., (© 2023 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Carbon concentrating mechanism (CCM) and photorespiration (PR) are interlinked and co-regulated in Chlamydomonas reinhardtii, but conditions where co-regulation alters are not sufficiently explored. Here, we uncover that PR gene transcripts, like CCM transcripts, are induced even in the dark when both processes are not active. Such diurnal cycles show that transcript levels peak in the middle of 12 h day, decline by early part of 12-h dark followed by their onset again at mid-dark. Interestingly, the onset in the mid-dark phase is sensitive to high CO2, implying that the active carbon sensing mechanism operates even in the dark. The rhythmic alterations of both CCM and PR transcript levels are unlinked to circadian clock: the "free-running state" reveals no discernible rhythmicity in transcript changes. Only continuous light leads to high transcript levels but no detectable transcripts were observed in continuous dark. Asynchronous continuous light cultures, upon shifting to low from high CO2 exhibit only transient induction of PR transcripts/proteins while CCM transcript induction is stable, indicating the loss of co-regulation between PR and CCM gene transcription. Lastly, we also describe that both CCM and PR transcripts/proteins are induced in low CO2 even in mixotrophic cultures, but only in high light, the same being attenuated in high CO2, implying that high light is a mandatory "trigger" for CCM and PR induction in low CO2 mixotrophy. Our study provides comprehensive analyses of conditions where CCM and PR were differently regulated, setting a paradigm for a detailed mechanistic probing of these responses. [ABSTRACT FROM AUTHOR]

Red coralline algae are projected to be sensitive to ocean acidification, particularly in polar oceans. As important ecosystem engineers, their potential sensitivity has broad implications, and understanding their carbon acquisition mechanisms is necessary for making reliable predictions. Therefore, we investigated the localized carbonate chemistry at the surface of Arctic coralline algae using microsensors. We report for the first time carbonate ion concentration and pH measurements ([CO32-]) at and above the algal surface in the microenvironment. We show that surface pH and [CO32-] are higher than the bulk seawater in the light, and even after hours of darkness. We further show that three species of Arctic coralline algae have efficient carbon concentrating mechanisms including direct bicarbonate uptake and indirect bicarbonate use via a carbonic anhydrase enzyme. Our results suggest that Arctic corallines have strong biological control over their surface chemistry, where active calcification occurs, and that net dissolution in the dark does not occur. We suggest that the elevated pH and [CO32-] in the dark could be explained by a high rate of light independent carbon fixation that reduces respiratory CO2 release. This mechanism could provide a potential adaptation to ocean acidification in Arctic coralline algae, which has important implications for future Arctic marine ecosystems. [ABSTRACT FROM AUTHOR]

The response of marine phytoplankton to the ongoing increase in atmospheric pCO reflects the consequences of both increased CO concentration and decreased pH in surface seawater. In the model diatom Thalassiosira weissflogii, we explored the effects of varying pCO and pH, independently and in concert, on photosynthesis and respiration by incubating samples in water enriched in H O. In long-term experiments (~6-h) at saturating light intensity, we observed no effects of pH or pCO 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 pCO 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 CO-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 pCO predicted for the twenty-first century. [ABSTRACT FROM AUTHOR]

Aquatic photosynthetic microorganisms, cyanobacteria and microalgae, account for almost half of the world's photosynthesis. They absorb carbon di oxide (CO2) as the major substrate to support photosynthesis, the beginning of energy flow into living organisms and one of the primary processes comprising the global carbon cycle. Among all photosynthetic mechanisms, inorganic carbon transport into ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is one of the major limiting steps in photosynthetic carbon fixation which involves active transport of HCO3−, CO2and/or H+, or an energized biochemical mechanism. In fact, a unique system “carbon concentrating mechanism (CCM)” manages the inorganic carbon assimilation, accumulation of CO2around RuBisCO, and utilization in algal cells. However, the information on mechanism of CO2uptake and fixation inside the algal cells is limited. In order to make strategies for enhancement of CO2fixation, understanding of CCM is crucial. Thus, this review provides an overview of advances in CCM research, the comparative state of the art and reports on the CO2uptake model in cyanobacteria and microalgae. The review also discusses the challenges and future perspectives associated with algal CCM research. [ABSTRACT FROM AUTHOR]

Transgenic potato ( Solanum tuberosum L. cv. Desiree) plants expressing components of a novel cyanobacterial photorespiratory glycolate catabolism pathway were developed. Transgenic plant expressing glcD1 (glycolate dehydrogenase I) gene was referred to as synGDH and transgenic plants expressing gcl (glyoxylate carboligase) and tsr (tartronic semialdehyde reductase) genes simultaneously were designated as synGT. Both synGDH and synGT plants showed stable gene transformation, integration and expression. Enhanced glyoxylate contents in synGDH plants were detected as compared to synGT and non-transgenic (NT) plants. Phenotypic evaluation revealed that synGDH plants accumulated 11 % higher dry weight, while, tuber weight was 38 and 16 % higher than NT and synGT, respectively. Upon challenging the plants in high temperature and high light conditions synGDH plants maintained higher Fv/ Fm and showed less bleaching of chlorophyll as compared to synGT and NT plants. These results indicate that genetic transformation of complete pathway in one plant holds promising outcomes in terms of biomass accumulation to meet future needs for food and energy. [ABSTRACT FROM AUTHOR]

Microalgae can tolerate different CO 2 concentrations and its mechanism of rapid CO 2 fixation is key to understanding the relationship between intracellular substance and energy conversion, as well as to optimize the production of microalgae bioproducts. The present study conducted a transcriptome-based analysis of gene expression of an industrial Chlorella pyrenoidosa strain, FACHB-9, with high oil production ability under different CO 2 concentrations. Transcriptome-based gene expression analysis indicated the redirection of central metabolism under CO 2 deprivation. The C3 pathway served as the main metabolic pathway for C. pyrenoidosa , which was subjected to a high CO 2 environment in the present study. Similar to C4 plants wherein limited CO 2 activates CO 2 -concentrating mechanism to compensate for the low activity of RuBisCO in the Calvin cycle, C. pyrenoidosa undergoes CO 2 compensation by active transport to ensure sufficient amounts of Ci for its growth and metabolism. Comparative analysis has allowed the identification of several candidate genes for further strain improvement. These genes encode proteins that might be CAs, as indicated by its localization to the cell membrane or chloroplast membrane, or act as Ci transporters that assist Ci transmembrane transportation. Certain types of ABC transport proteins and CCCH-type zinc finger proteins also showed significant changes in gene expression. These findings indicate that these may be promising targets for functional and genetic modification studies. The results not only reveal the significance of the mechanism of carbon sequestration to eukaryotic green algae, but also provide a basis for further construction of the energy microalgae cell factory with high efficiency carbon biofixation capacity. [ABSTRACT FROM AUTHOR]

Photorespiration and oxygenic photosynthesis are intimately linked processes. It has been shown that under the present day atmospheric conditions cyanobacteria and all eukaryotic phototrophs need functional photorespiration to grow autotrophically. The question arises as to when this essential partnership evolved, i.e. can we assume a coevolution of both processes from the beginning or did photorespiration evolve later to compensate for the generation of 2-phosphoglycolate (2PG) due to Rubisco's oxygenase reaction? This question is mainly discussed here using phylogenetic analysis of proteins involved in the 2PG metabolism and the acquisition of different carbon concentrating mechanisms (CCMs). The phylogenies revealed that the enzymes involved in the photorespiration of vascular plants have diverse origins, with some proteins acquired from cyanobacteria as ancestors of the chloroplasts and others from heterotrophic bacteria as ancestors of mitochondria in the plant cell. Only phosphoglycolate phosphatase was found to originate from Archaea. Notably glaucophyte algae, the earliest branching lineage of Archaeplastida, contain more photorespiratory enzymes of cyanobacterial origin than other algal lineages or land plants indicating a larger initial contribution of cyanobacterial-derived proteins to eukaryotic photorespiration. The acquisition of CCMs is discussed as a proxy for assessing the timing of periods when photorespiratory activity may have been enhanced. The existence of CCMs also had marked influence on the structure and function of photorespiration. Here, we discuss evidence for an early and continuous coevolution of photorespiration, CCMs and photosynthesis starting from cyanobacteria via algae, to land plants. [ABSTRACT FROM AUTHOR]

The gammaproteobacterium Thiomicrospira crunogena XCL-2 is an aerobic sulfur-oxidizing hydrothermal vent chemolithoautotroph that has a CO concentrating mechanism (CCM), which generates intracellular dissolved inorganic carbon (DIC) concentrations much higher than extracellular, thereby providing substrate for carbon fixation at sufficient rate. This CCM presumably requires at least one active DIC transporter to generate the elevated intracellular concentrations of DIC measured in this organism. In this study, the half-saturation constant ( K) for purified carboxysomal RubisCO was measured (276 ± 18 µM) which was much greater than the K of whole cells (1.03 µM), highlighting the degree to which the CCM facilitates CO fixation under low CO conditions. To clarify the bioenergetics powering active DIC uptake, cells were incubated in the presence of inhibitors targeting ATP synthesis (DCCD) or proton potential (CCCP). Incubations with each of these inhibitors resulted in diminished intracellular ATP, DIC, and fixed carbon, despite an absence of an inhibitory effect on proton potential in the DCCD-incubated cells. Electron transport complexes NADH dehydrogenase and the bc complex were found to be insensitive to DCCD, suggesting that ATP synthase was the primary target of DCCD. Given the correlation of DIC uptake to the intracellular ATP concentration, the ABC transporter genes were targeted by qRT-PCR, but were not upregulated under low-DIC conditions. As the T. crunogena genome does not include orthologs of any genes encoding known DIC uptake systems, these data suggest that a novel, yet to be identified, ATP- and proton potential-dependent DIC transporter is active in this bacterium. This transporter serves to facilitate growth by T. crunogena and other Thiomicrospiras in the many habitats where they are found. [ABSTRACT FROM AUTHOR]

Bicarbonate species in the aqueous phase is the primary source for CO for the growth of microalgae. The potential of carbon dioxide (CO) fixation by Chlorella pyrenoidosa in enriched bicarbonate medium was evaluated. In the present study, effects of parameters such as pH, sodium bicarbonate concentration and inoculum size were assessed for the removal of CO by C. pyrenoidosa under mixotrophic condition. Central composite design tool from response surface methodology was used to validate statistical methods in order to study the influence of these parameters. The obtained results reveal that the maximum removal of CO was attained at pH 8 with sodium bicarbonate concentration of 3.33 g/l, and inoculum size of 30 %. The experimental results were statistically significant with R value of 0.9527 and 0.960 for CO removal and accumulation of chlorophyll content, respectively. Among the various interactions, interactive effects between the parameters pH and inoculum size was statistically significant ( P < 0.05) for CO removal and chlorophyll accumulation. Based on the studies, the application of C. pyrenoidosa as a potential source for carbon dioxide removal at alkaline pH from bicarbonate source is highlighted. [ABSTRACT FROM AUTHOR]

Carbon dioxide (CO 2) and light are essential for high photosynthetic rates of microalgal cultures. Microalgal photosynthetic behavior at low CO 2 concentrations has not been revealed yet at the same level of detail as leaf photosynthesis. In the present study, we investigated the short-term photosynthetic response of suspended Chlorella sorokiniana to limiting light intensity and CO 2 concentration. We used a novel CO 2 -based photosynthesis monitor originating from leaf research but equipped with an aquatic chamber sparged with CO 2 enriched air. Photosynthesis was measured by employing a steady-state CO 2 mass balance over the chamber. Light and carbon response curves were determined under constant pH, temperature, dissolved oxygen, light intensity or dissolved carbon dioxide. We determined the volumetric mass transfer coefficient of the aquatic chamber to accurately convert gaseous CO 2 partial pressure into aqueous CO 2 concentration to evaluate the CO 2 response measurements. Light response measurements on dilute algal cultures revealed a high photosynthetic capacity on a time scale of minutes which by far exceeded the average CO 2 uptake on a time scale of hours (cell growth). Light response measurements with dense and fully absorbing cultures provided accurate insight into the response of microalgal mass culture to changing incident irradiance, including the efficiency of photosynthesis of the algal culture as a whole. CO 2 response measurements demonstrated severe CO 2 limitation at dissolved CO 2 levels of <20 μM giving a concrete target for optimization of CO2 supply in large-scale cultivation systems. Measurements at a background of either 21 % O 2 (air) or 2 % O 2 indicated the existence of photorespiration in Chlorella. Moreover, the magnitude of photorespiration first increased and then decreased again while reduced dissolved CO 2 levels, indicating the activation of a carbon concentrating mechanism at low CO 2. Simultaneous measurement of the quantum yield of PSII linear electron transport (ϕ PSII) and CO 2 uptake revealed the transition of CO 2 limited photosynthesis towards light-limited photosynthesis. • Algal photosynthetic response to limiting light intensity and CO 2 concentration (81 characters) • Photosynthetic response measurement requires apt choice of biomass concentration (82 characters). • CO 2 response at different O 2 concentrations suggested photorespiration in Chlorella (85 characters). • Activation of carbon concentrating mechanism amid measurements was indicated (78 characters). • Monod equation is too simple to describe algal photosynthetic responses precisely (83 characters). [ABSTRACT FROM AUTHOR]

The PII superfamily consists of widespread signal transduction proteins found in all domains of life. In addition to canonical PII proteins involved in C/N sensing, structurally similar PII-like proteins evolved to fulfill diverse, yet poorly understood cellular functions. In cyanobacteria, the bicarbonate transporter SbtA is co-transcribed with the conserved PII-like protein, SbtB, to augment intracellular inorganic carbon levels for efficient CO 2 fixation. We identified SbtB as a sensor of various adenine nucleotides including the second messenger nucleotides cyclic AMP (cAMP) and c-di-AMP. Moreover, many SbtB proteins possess a C-terminal extension with a disulfide bridge of potential redox-regulatory function, which we call R-loop. Here, we reveal an unusual ATP/ADP apyrase (diphosphohydrolase) activity of SbtB that is controlled by the R-loop. We followed the sequence of hydrolysis reactions from ATP over ADP to AMP in crystallographic snapshots and unravel the structural mechanism by which changes of the R-loop redox state modulate apyrase activity. We further gathered evidence that this redox state is controlled by thioredoxin, suggesting that it is generally linked to cellular metabolism, which is supported by physiological alterations in site-specific mutants of the SbtB protein. Finally, we present a refined model of how SbtB regulates SbtA activity, in which both the apyrase activity and its redox regulation play a central role. This highlights SbtB as a central switch point in cyanobacterial cell physiology, integrating not only signals from the energy state (adenyl-nucleotide binding) and the carbon supply via cAMP binding but also from the day/night status reported by the C-terminal redox switch.

The transition of C3, via C2 towards C4 photosynthesis is an important example of stepwise evolution of a complex genetic trait. A common feature that was gradually emphasized during this trajectory is the evolution of a CO2 concentration mechanism around Rubisco. In C2 plants, this mechanism is based on tissue- specific accumulation of glycine decarboxylase (GDC) in bundle sheath (BS) cells, relative to global expression in the cells of C3 leaves. This limits photorespiratory CO2 release to BS cells. Because BS cells are surrounded by photosynthetically active mesophyll cells, this arrangement enhances the probability of re-fixation of CO2. The restriction of GDC to BS cells was mainly achieved by confinement of its P-subunit (GLDP). Here, we provide a mechanism for the establishment of C2-type gene expression by studying the upstream sequences of C3 Gldp genes in Arabidopsis thaliana. Deletion of 59 bp in the upstream region of AtGldp1 restricted expression of a reporter gene to BS cells and the vasculature without affecting diurnal variation. This region was named the 'M box'. Similar results were obtained for the AtGldp2 gene. Fusion of the M box to endogenous or exogenous promoters supported mesophyll expression. Nucleosome densities at the M box were low, suggesting an open chromatin structure facilitating transcription factor binding. In silico analysis defined a possible consensus for the element that was conserved across the Brassicaceae, but not in Moricandia nitens, a C2 plant. Collective results provide evidence that a simple mutation is sufficient for establishment of C2-specific gene expression in a C3 plant. [ABSTRACT FROM AUTHOR]

Inorganic carbon (C i ) is the major sink for photosynthetic reductant in organisms capable of oxygenic photosynthesis. In the absence of abundant C i , the cyanobacterium Synechocystis sp. strain PCC6803 expresses a high affinity C i acquisition system, the CO 2 -concentrating mechanisms (CCM), controlled by the transcriptional regulator CcmR and the metabolites NADP + and α-ketoglutarate, which act as co-repressors of CcmR by modulating its DNA binding. The CCM thus responds to internal cellular redox changes during the transition from C i -replete to C i -limited conditions. However, the actual changes in the metabolic state of the NADPH/NADP + system that occur during the transition to C i -limited conditions remain ill-defined. Analysis of changes in the redox state of cells experiencing C i limitation reveals systematic changes associated with physiological adjustments and a trend towards the quinone and NADP pools becoming highly reduced. A rapid and persistent increase in F 0 was observed in cells reaching the C i -limited state, as was the induction of photoprotective fluorescence quenching. Systematic changes in the fluorescence induction transients were also observed. As with Chl fluorescence, a transient reduction of the NADPH pool (‘M’ peak), is assigned to State 2→ State 1 transition associated with increased electron flow to NADP + . This was followed by a characteristic decline, which was abolished by C i limitation or inhibition of the Calvin –Benson–Bassham (CBB) cycle and is thus assigned to the activation of the CBB cycle. The results are consistent with the proposed regulation of the CCM and provide new information on the nature of the Chl and NADPH fluorescence induction curves. [ABSTRACT FROM AUTHOR]

Microalgae are a diverse group of photosynthetic organisms that can be exploited for the production of different compounds, ranging from crude biomass and biofuels to high value-added biochemicals and synthetic proteins. Traditionally, algal biotechnology relies on bioprospecting to identify new highly productive strains and more recently, on forward genetics to further enhance productivity. However, it has become clear that further improvements in algal productivity for biotechnology is impossible without combining traditional tools with the arising molecular genetics toolkit. We review recent advantages in developing high throughput screening methods, preparing genome-wide mutant libraries, and establishing genome editing techniques. We discuss how algae can be improved in terms of photosynthetic efficiency, biofuel and high value-added compound production. Finally, we critically evaluate developments over recent years and explore future potential in the field. [ABSTRACT FROM AUTHOR]

As all plastids that have been investigated so far can be traced back to endosymbiotic uptake of cyanobacteria by heterotrophic host cells, they accordingly show a high similarity regarding photosynthesis, which includes both the photosystems and the biochemical reactions around the CO 2 fixation via the Calvin–Bassham cycle. Major differences between the different algal and plant groups may include the presence or absence of carbon concentrating mechanisms, pyrenoids, Rubisco activases, carbonic anhydrases as well as differences in the regulation of the Calvin–Bassham cycle. This review describes the diversity of primary carbon fixation steps in algae and plants and the respective regulatory mechanisms. [ABSTRACT FROM AUTHOR]

Agricultural yields have kept pace with the rising demands in the recent past as a result of the breeding and improved farming practices, but these practices alone will not be able to meet the demands of the future. The focus is now on the enhancement of the photosynthetic machinery. In photosynthesis, the rate limiting step is the one catalyzed by RuBisCO- Ribulose-1,5-bisphosphate carboxylase/oxygenase (4.1.1.39), which, because of its loose specificity and low turnover rate, is the primary target of most research programs directed towards improved photosynthesis. The other avenues of photosynthetic machinery that are under investigation to enhance it include-improved stomatal regulation and membrane permeability, RuBisCO with high specificity for CO and higher catalytic turnover; bypass of photorespiration and introduction of carbon concentrating mechanism (CCM) into the C plants. Carbon concentrating mechanisms cause accumulation of carbon dioxide in vicinity of RuBisCO producing a high CO/O ratio and hence an environment more suitable for carboxylation reactions than oxygenation reactions. This article includes the basic details of the major naturally occurring CCMs in various photosynthetic organisms to identify the knowledge gaps in each which could help study the prospects of its possible introduction into a non-native system as C plants which are devoid of any CCM. [ABSTRACT FROM AUTHOR]

The photosynthetic, unicellular green alga, Chlamydomonas reinhardtii, lives in environments that often contain low concentrations of CO and HCO, the utilizable forms of inorganic carbon (C). C. reinhardtii possesses a carbon concentrating mechanism (CCM) which can provide suitable amounts of C for growth and development. This CCM is induced when the CO concentration is at air levels or lower and is comprised of a set of proteins that allow the efficient uptake of C into the cell as well as its directed transport to the site where Rubisco fixes CO into biomolecules. While several components of the CCM have been identified in recent years, the picture is still far from complete. To further improve our knowledge of the CCM, we undertook a mutagenesis project where an antibiotic resistance cassette was randomly inserted into the C. reinhardtii genome resulting in the generation of 22,000 mutants. The mutant collection was screened using both a published PCR-based approach (Gonzalez-Ballester et al. ) and a phenotypic growth screen. The PCR-based screen did not rely on a colony having an altered growth phenotype and was used to identify colonies with disruptions in genes previously identified as being associated with the CCM-related gene. Eleven independent insertional mutations were identified in eight different genes showing the usefulness of this approach in generating mutations in CCM-related genes of interest as well as identifying new CCM components. Further improvements of this method are also discussed. [ABSTRACT FROM AUTHOR]

The articles in this special issue of Photosynthesis Research arose from the presentations given at the Eighth International Symposium on Inorganic Carbon Uptake by Aquatic Photosynthetic Organisms held from May 27 to June 1, 2013 in New Orleans, Louisiana USA. The meeting covered all the aspects of CO concentrating mechanisms (CCMs) present in photosynthetic bacteria, microalgae and macrophytes, and spanned disciplines from the molecular biology of CCMs to the importance of CCMs in aquatic ecosystems. The publications in this special issue represent our current understanding of CCMs and highlight recent advances in the field. The influences of CCMs on algal biofuel production as well as recent efforts to use the CCM to improve crop plants are also explored. [ABSTRACT FROM AUTHOR]

Abstract: We investigated the catalytic activity and inhibition of the β-class carbonic anhydrase (CA, EC 4.2.1.1) CahB1, from the relict cyanobacterium Coleofasciculus chthonoplastes (previously denominated Microcoleus chthonoplastes). The enzyme showed good activity as a catalyst for the CO2 hydration, with a k cat of 2.4×105 s−1 and a k cat/K m of 6.3×107 M−1 s−1. A range of inorganic anions and small molecules were investigated as inhibitors of CahB1. Perchlorate and tetrafluoroborate did not inhibit the enzyme (K Is >200mM) whereas selenate and selenocyanide were ineffective inhibitors too, with K Is of 29.9–48.61mM. The halides, pseudohalides, carbonate, bicarbonate, trithiocarbonate and a range of heavy metal ions-containing anions were submillimolar–millimolar inhibitors (K Is in the range of 0.15–0.90mM). The best CahB1 inhibitors were N,N-diethyldithiocarbamate, sulfamate, sulfamide, phenylboronic acid and phenylarsonic acid, with K Is in the range of 8–75μM, whereas acetazolamide inhibited the enzyme with a K I of 76nM. This is the first kinetic and inhibition study of a cyanobacterial CA. As these enzymes are widespread in many cyanobacteria, being crucial for the carbon concentrating mechanism which assures substrate to RubisCO for the CO2 fixation by these organisms, a detailed kinetic/inhibition study may be essential for a better understanding of this superfamily of metalloenzymes and for potential biotechnological applications in biomimetic CO2 capture processes. [Copyright &y& Elsevier]

Threat of global warming due to carbon dioxide (CO) emissions has stimulated research into carbon sequestration and emissions reduction technologies. Alkaline scrubbing allows CO to be captured as bicarbonate, which can be photochemically fixed by microalgae. The carbon concentrating mechanism (CCM), of which external carbonic anhydrase is a key component, allows organisms to utilise this bicarbonate. In order to select a suitable strain for this application, a screening tool is required. The current method for determining carbonic anhydrase activity, the Wilbur and Anderson assay, was found to be unsuitable as a screening tool as the associated error was unacceptably large and tests on whole cells were inconclusive. This paper presents the development of a new, whole cell assay to measure inorganic carbon uptake and external carbonic anhydrase activity, based on classical pH drift experiments. Spirulina platensis was successfully used to develop a correlation between the specific carbon uptake (C) and the specific pH change (dpH). The relationship is described by the following: C[mmol C (g dry algae) h] = 0.064 × (dpH). Inhibitor and salt dissociation tests validated the activity and presence of external carbonic anhydrase and allowed correlation between the Wilbur and Anderson assay and the new whole cell assay. Screening tests were conducted on S. platensis, Scenedesmus sp., Chlorella vulgaris and Dunaliella salina that were found to have carbon uptake rates of 5.76, 5.86, 3.86 and 2.15 mmol C (g dry algae) h, respectively. These results corresponded to the species' known bicarbonate utilisation abilities and validated the use of the assay as a screening tool. [ABSTRACT FROM AUTHOR]