Air Levels Of CO2 Research Articles

Rational H2 Partial Pressure over Nickel/Ceria Crystal Enables Efficient and Durable Wide-Temperature-Zone Air-Level CO2 Methanation.

On the way to carbon neutrality, directly catalyzing atmospheric CO2 into high-value chemicals might be an effective approach to mitigate the negative impacts of rising airborne CO2 concentrations. Here, we pioneer the investigation of the influence of the H2/CO2 partial pressure ratio (PPR) on air-level CO2 methanation. Using Ni/CeO2 as a case catalyst, increasing H2/CO2 PPR significantly improves low-temperature CO2 conversion and high-temperature CH4 selectivity, i.e., from 10 of H2/CO2 PPR on, CO2 is completely methanized at 250 °C, and nearly 100% CH4 selectivity is achieved at 400 °C. 100-hour stability tests demonstrate the practical application potential of Ni/CeO2 at 250 °C and 400 °C. In-situ DRIFTS reveal that reinforced formate pathway by increasing H2/CO2 PPR is responsible for the high CH4 yield. In contrast, even though the CO pathway dominated CO2 conversion on Ni is enhanced by rising H2/CO2 PPR, but at a high reaction temperature, the promoted CO desorption still leads to lower CH4 selectivity. This work offers deep insights into the direct air-level CO2 resourceization, contributing to the achievement of airborne CO2 reductions.

Electroreduction of air-level CO2 with high conversion efficiency

The electrochemical conversion of carbon dioxide (CO 2 ) has been attracting increasingly research interest in the past decade, with the ultimate goal of utilizing electricity from renewable energy to realize carbon neutrality, as well as economic and energy benefits. Nonetheless, the capture and concentrating of CO 2 cost a substantial portion of energy, while almost all the reported researches showed CO 2 electroreduction under high concentrations of (typically pure) CO 2 reactants, and only very few recent studies have investigated the capability of applying low CO 2 concentrations (such as ~10% in flue gases). In this work, we first demonstrated the electroreduction of 0.03% CO 2 (in helium) in a homemade gas-phase electrochemical electrolyzer, using a low-cost copper (Cu) or nanoscale copper (nano-Cu) catalyst. Mixed with steam, the gas-phase CO 2 was directly delivered onto the gas-solid interface with the Cu catalyst and reduced to CO, without the need/constraint of being adsorbed by aqueous solution or alkaline electrolytes. By tuning the catalyst and experimental parameters, the conversion efficiency of CO 2 reached as high as ~95%. Furthermore, we demonstrated the direct electroreduction of 0.04% CO 2 from real air sample with an optimized conversion efficiency of ~79%, suggesting a promising perspective of the electroreduction approach toward direct CO 2 conversion. This paper realizes the direct electroreduction of 0.03% or 0.04% CO 2 in a homemade gas-phase electrochemical reactor using a low-cost copper (Cu) or nanoscale copper (nano-Cu) catalyst with high CO 2 conversion efficiency.

Modelling the pyrenoid-based CO2-concentrating mechanism provides insights into its operating principles and a roadmap for its engineering into crops

Many eukaryotic photosynthetic organisms enhance their carbon uptake by supplying concentrated CO2 to the CO2-fixing enzyme Rubisco in an organelle called the pyrenoid. Ongoing efforts seek to engineer this pyrenoid-based CO2-concentrating mechanism (PCCM) into crops to increase yields. Here we develop a computational model for a PCCM on the basis of the postulated mechanism in the green alga Chlamydomonas reinhardtii. Our model recapitulates all Chlamydomonas PCCM-deficient mutant phenotypes and yields general biophysical principles underlying the PCCM. We show that an effective and energetically efficient PCCM requires a physical barrier to reduce pyrenoid CO2 leakage, as well as proper enzyme localization to reduce futile cycling between CO2 and HCO3−. Importantly, our model demonstrates the feasibility of a purely passive CO2 uptake strategy at air-level CO2, while active HCO3− uptake proves advantageous at lower CO2 levels. We propose a four-step engineering path to increase the rate of CO2 fixation in the plant chloroplast up to threefold at a theoretical cost of only 1.3 ATP per CO2 fixed, thereby offering a framework to guide the engineering of a PCCM into land plants.

A CRISPR/dCas9-based transcription activated system developed in marine microalga Nannochloropsis oceanica

Nannochloropsis is one of the most important microalgae utilized as the feedstuff in aquaculture and is emerging model oleaginous microalgae. Recently, different genetic tools such as RNA interference, gene overexpression and gene editing have been developed in Nannochloropsis. However, the available genetic modification methods based on CRISPR technology are rather limited for modulating gene expression. Here, we firstly explored a transcriptional activation system based on CRISPR/dCas9 in N. oceanica IMET1. The result demonstrated that this construct of CRISPRa system (dCas9 protein fused with transcriptional activator VP64) could alter efficiently gene expression in N. ocanica. The expression of targeted gene (an enzyme responsible for DNA or RNA methylation as a methyltransferase) was increased by ~2–6 fold at the transcriptional level. Moreover, the growth and photosynthetic parameter (Fv/Fm) of mutants was increased by 23% and 12% compared to wild type under air-level CO2 respectively. Thus, this system will open new avenue for genome engineering of Nannochloropsis.

Identification of the AG afterglow thermoluminescence band in the cyanobacterium Synechocystis PCC 6803.

The so-called afterglow, AG, thermoluminescence (TL) band is a useful indicator of the presence of cyclic electron flow (CEF), which is mediated by the NADH dehydrogenase-like (NDH) complex in higher plants. Although NDH-dependent CEF occurs also in cyanobacteria, the AG band has previously not been found in these organisms. In the present study, we tested various experimental conditions and could identify a TL component with ca. +40°C peak temperature in Synechocystis PCC 6803 cells, which were illuminated by far-red (FR) light at around -10°C. The +40°C band could be observed when WT cells were grown under ambient air level CO2 , but was absent in the M55 mutant, which is deficient in the NDH-1 complex. These experimental observations match the characteristics of the AG band of higher plants. Therefore, we conclude that the newly identified +40°C TL component in Synechocystis PCC 6803 is the cyanobacterial counterpart of the plant AG band and originates from NDH-1-mediated CEF. The cyanobacterial AG band was most efficiently induced when FR illumination was applied at -10°C and its contribution to the total TL intensity declined when cells were illuminated above and below this temperature. Based on this phenomenon we also conclude that CEF is blocked by low temperatures at two different sites in Synechocystis PCC 6803: (1) Below -10°C at the level of NDH-1 and (2) below -30°C at the donor or acceptor side of Photosystem I.

LCI1, a Chlamydomonas reinhardtii plasma membrane protein, functions in active CO2 uptake under low CO2.

In response to high CO2 environmental variability, green algae, such as Chlamydomonas reinhardtii, have evolved multiple physiological states dictated by external CO2 concentration. Genetic and physiological studies demonstrated that at least three CO2 physiological states, a high CO2 (0.5-5% CO2 ), a low CO2 (0.03-0.4% CO2 ) and a very low CO2 (< 0.02% CO2 ) state, exist in Chlamydomonas. To acclimate in the low and very low CO2 states, Chlamydomonas induces a sophisticated strategy known as a CO2 -concentrating mechanism (CCM) that enables proliferation and survival in these unfavorable CO2 environments. Active uptake of Ci from the environment is a fundamental aspect in the Chlamydomonas CCM, and consists of CO2 and HCO3- uptake systems that play distinct roles in low and very low CO2 acclimation states. LCI1, a putative plasma membrane Ci transporter, has been linked through conditional overexpression to active Ci uptake. However, both the role of LCI1 in various CO2 acclimation states and the species of Ci , HCO3- or CO2 , that LCI1 transports remain obscure. Here we report the impact of an LCI1 loss-of-function mutant on growth and photosynthesis in different genetic backgrounds at multiple pH values. These studies show that LCI1 appears to be associated with active CO2 uptake in low CO2 , especially above air-level CO2 , and that any LCI1 role in very low CO2 is minimal.

Assessing exposure to unconventional natural gas development: using an air pollution dispersal screening model to predict new-onset respiratory symptoms

Various exposure estimates have been used to assess health impact of unconventional natural gas development (UNGD). The purpose of this study was to (1) use an air pollution dispersal screening model and wind direction to characterize the air emissions from UNGD facilities at each residence and (2) assess association of this exposure estimate with respiratory symptoms. Respiratory symptoms were abstracted from health records of a convenience sample of 104 adults from one county in southwestern PA who had completed a standard clinical interview with a nurse practitioner. Using publicly available air emission data, we applied a “box” air pollution dispersion screening model to estimate the median ambient air level of CO, NOx, PM 2.5, VOCs, and formaldehyde at the residence during the year health symptoms were reported. Sources and median emissions were categorized as north, south, east, or west of the residence to account for the effect of wind direction on dispersion. Binary logistic regression was performed for each respiratory symptom. Number of sources had varying magnitudes of association with some symptoms (i.e., cough, shortness of breath, and “any respiratory symptom”) and no association with others (i.e., sore throat, sinus problems, wheezing). Air emissions were not associated with any symptom.

Flavodiiron proteins 1-to-4 function in versatile combinations in O2 photoreduction in cyanobacteria.

Flavodiiron proteins (FDPs) constitute a group of modular enzymes widespread in Bacteria, Archaea and Eukarya. Synechocystis sp. PCC 6803 has four FDPs (Flv1-4), which are essential for the photoprotection of photosynthesis. A direct comparison of light-induced O2 reduction (Mehler-like reaction) under high (3% CO2, HC) and low (air level CO2, LC) inorganic carbon conditions demonstrated that the Flv1/Flv3 heterodimer is solely responsible for an efficient steady-state O2 photoreduction under HC, with flv2 and flv4 expression strongly down-regulated. Conversely, under LC conditions, Flv1/Flv3 acts only as a transient electron sink, due to the competing withdrawal of electrons by the highly induced NDH-1 complex. Further, in vivo evidence is provided indicating that Flv2/Flv4 contributes to the Mehler-like reaction when naturally expressed under LC conditions, or, when artificially overexpressed under HC. The O2 photoreduction driven by Flv2/Flv4 occurs down-stream of PSI in a coordinated manner with Flv1/Flv3 and supports slow and steady-state O2 photoreduction.

Enhancing photosynthetic biomass productivity of industrial oleaginous microalgae by overexpression of RuBisCO activase

Eukaryotic microalgae are promising cell factories for photosynthesis-driven CO2 sequestration and oil production, however one key challenge has been to genetically enhance their biomass productivity. Here we showed that the biomass productivity of a model industrial oleaginous microalga Nannochloropsis oceanica can be elevated by overexpression of a nuclear-encoded, cbbX-homologous, candidate RuBisCO activase (g1915 or ‘nNoRca-like’) that is transcriptionally induced under air-level CO2 (i.e., 0.04% v/v) as compared to 5% CO2 in wild-type cells. In the mutants, under air-level CO2, growth rate was enhanced by ~32%, biomass accumulation by ~46% and lipid productivity by ~41%, which were also accompanied by ~28% elevation in photosynthesis; furthermore, abundance of RuBisCO large subunit protein increased by ~45%, suggesting nNoRca-like overexpression enhanced microalgal photosynthesis by upregulating the level and activity of RuBisCO. Interestingly, under 5% CO2, no significant enhancement of RuBisCO protein expression was observed. Thus a genetic strategy that activates RuBisCO by augmenting its functional partners was proposed to increase biomass production of industrial microalgae. Our work expands the reservoir of potential photosynthetic gene targets for plant biotechnology and provides a basis for constructing advanced microalgal cell factories for carbon sequestration and biofuel production.

Cyanobacterial flv4-2 Operon-Encoded Proteins Optimize Light Harvesting and Charge Separation in Photosystem II

Photosystem II (PSII) complexes drive the water-splitting reaction necessary to transform sunlight into chemical energy. However, too much light can damage and disrupt PSII. In cyanobacteria, the flv4-2 operon encodes three proteins (Flv2, Flv4, and Sll0218), which safeguard PSII activity under air-level CO2 and in high light conditions. However, the exact mechanism of action of these proteins has not been clarified yet. We demonstrate that the PSII electron transfer properties are influenced by the flv4-2 operon-encoded proteins. Accelerated secondary charge separation kinetics was observed upon expression/overexpression of the flv4-2 operon. This is likely induced by docking of the Flv2/Flv4 heterodimer in the vicinity of the QB pocket of PSII, which, in turn, increases the QB redox potential and consequently stabilizes forward electron transfer. The alternative electron transfer route constituted by Flv2/Flv4 sequesters electrons from QB− guaranteeing the dissipation of excess excitation energy in PSII under stressful conditions. In addition, we demonstrate that in the absence of the flv4-2 operon-encoded proteins, about 20% of the phycobilisome antenna becomes detached from the reaction centers, thus decreasing light harvesting. Phycobilisome detachment is a consequence of a decreased relative content of PSII dimers, a feature observed in the absence of the Sll0218 protein.

Assessment of air quality and erythrocyte oxidative stress indicators of storekeepers of laboratory/industrial chemicals in owerri, Imo state, Nigeria

The present study ascertained air quality (AQ) of chemical stores and level of alterations in erythrocyte oxidative stress biomarkers of storekeepers exposed to laboratory/industrial chemicals in Owerri, Imo State, Nigeria. Erythrocyte percentage methaemoglobin (%MetHb), reduced glutathione (GSH) concentration, thiobarbituric acid reactive substances (TBARS) index and H2O2 induced erythrocyte haemolysis of participants were measured by spectrophotometric methods. The air levels of CO, NH3 and NO2 in the chemical shops were below Environmental Protection Agency (EPA) minimum standards. Particulate matters (PM) of both control and test air samples were within EPA standards. Erythrocyte %MetHb of test participants represented an increase of 61.81% compared to the control participants ( p &gt; 0.05), whereas erythrocyte GSH concentration of test participants showed significant difference ( p &lt; 0.05). Levels of H2O2 induced erythrocyte haemolysis was significantly (p &lt; 0.05) higher than those of the corresponding control participants. The ratio of erythrocyte TBARS index of the test to the control participants was 1.4:1 approx.; p &lt; 0.05. The present study showed that the gas substances in chemical stores in Owerri compromised AQ with attendant potentials to cause oxidative stress and hazardous consequence to the storekeepers.

Operon flv4-flv2 Provides Cyanobacterial Photosystem II with Flexibility of Electron Transfer

Synechocystis sp PCC 6803 has four genes encoding flavodiiron proteins (FDPs; Flv1 to Flv4). Here, we investigated the flv4-flv2 operon encoding the Flv4, Sll0218, and Flv2 proteins, which are strongly expressed under low inorganic carbon conditions (i.e., air level of CO(2)) but become repressed at elevated CO(2) conditions. Different from FDP homodimers in anaerobic microbes, Synechocystis Flv2 and Flv4 form a heterodimer. It is located in cytoplasm but also has a high affinity to membrane in the presence of cations. Sll0218, on the contrary, resides in the thylakoid membrane in association with a high molecular mass protein complex. Sll0218 operates partially independently of Flv2/Flv4. It stabilizes the photosystem II (PSII) dimers, and according to biophysical measurements opens up a novel electron transfer pathway to the Flv2/Flv4 heterodimer from PSII. Constructed homology models suggest efficient electron transfer in heterodimeric Flv2/Flv4. It is suggested that Flv2/Flv4 binds to thylakoids in light, mediates electron transfer from PSII, and concomitantly regulates the association of phycobilisomes with PSII. The function of the flv4-flv2 operon provides many β-cyanobacteria with a so far unknown photoprotection mechanism that evolved in parallel with oxygen-evolving PSII.

Interplay between Flavodiiron Proteins and Photorespiration in Synechocystis sp. PCC 6803

Flavodiiron (Flv) proteins are involved in detoxification of O(2) and NO in anaerobic bacteria and archaea. Cyanobacterial Flv proteins, on the contrary, function in oxygenic environment and possess an extra NAD(P)H:flavin oxidoreductase module. Synechocystis sp. PCC 6803 has four genes (sll1521, sll0219, sll0550, and sll0217) encoding Flv proteins (Flv1, Flv2, Flv3, and Flv4). Previous in vitro studies with recombinant Flv3 protein from Synechocystis provided evidence that it functions as a NAD(P)H:oxygen oxidoreductase, and subsequent in vivo studies with Synechocystis confirmed the role of Flv1 and Flv3 proteins in the Mehler reaction (photoreduction of O(2) to H(2)O). Interestingly, homologous proteins to Flv1 and Flv3 can be found also in green algae, mosses, and Selaginella. Here, we addressed the function of Flv1 and Flv3 in Synechocystis using the Δflv1, Δflv3, and Δflv1/Δflv3 mutants and applying inorganic carbon (C(i))-deprivation conditions. We propose that only the Flv1/Flv3 heterodimer form is functional in the Mehler reaction in vivo. (18)O(2) labeling was used to discriminate between O(2) evolution in photosynthetic water splitting and O(2) consumption. In wild type, ∼20% of electrons originated from water was targeted to O(2) under air level CO(2) conditions but increased up to 60% in severe limitation of C(i). Gas exchange experiments with Δflv1, Δflv3, and Δflv1/Δflv3 mutants demonstrated that a considerable amount of electrons in these mutants is directed to photorespiration under C(i) deprivation. This assumption is in line with increased transcript abundance of photorespiratory genes and accumulation of photorespiratory intermediates in the WT and to a higher extent in mutant cells under C(i) deprivation.

Cross‐talk between photomixotrophic growth and CO2‐concentrating mechanism inSynechocystissp. strain PCC 6803

Simultaneous catabolic and anabolic glucose metabolism occurs in the same compartment during photomixotrophic growth of the model cyanobacterium Synechocystis sp. PCC 6803. The presence of glucose is stressful to the cells; it is reflected in the high frequency of suppression mutations in glucose-sensitive mutants. We show that glucose affects many cellular processes. It stimulates respiration and the rate of photosynthesis and quantum yield in low- but not high-CO(2) -grown cells. Fluorescence and thermoluminescence parameters of photosystem II are also affected but the results did not lend support to sustained glucose driven over reduction in the light. Glucose-sensitive mutants such as ΔpmgA (impaired in photomixotrophic growth) and Δhik31 (lacking histidine kinase 31) are far more susceptible under high than low air level of CO(2) . A glycine to tryptophan mutation in position 354 in NdhF3, involved in the high-affinity CO(2) uptake, rescued ΔpmgA. A rise in the apparent photosynthetic affinity to external inorganic carbon is observed in high-CO(2) -grown wild-type cells after the addition of glucose, but not in mutant ΔpmgA. This is attributed to upregulation of certain low-CO(2) -induced genes, involved in inorganic carbon uptake, in the wild type but not in ΔpmgA. These data uncovered a new level of interaction between CO(2) fixation (and the CO(2) -concentrating mechanism) and photomixotrophic growth in cyanobacteria.

Flavodiiron Proteins in Oxygenic Photosynthetic Organisms: Photoprotection of Photosystem II by Flv2 and Flv4 in Synechocystis sp. PCC 6803

BackgroundFlavodiiron proteins (FDPs) comprise a group of modular enzymes that function in oxygen and nitric oxide detoxification in Bacteria and Archaea. The FDPs in cyanobacteria have an extra domain as compared to major prokaryotic enzymes. The physiological role of cyanobacteria FDPs is mostly unknown. Of the four putative flavodiiron proteins (Flv1–4) in the cyanobacterium Synechocystis sp. PCC 6803, a physiological function in Mehler reaction has been suggested for Flv1 and Flv3.Principal FindingsWe demonstrate a novel and crucial function for Flv2 and Flv4 in photoprotection of photosystem II (PSII) in Synechocystis. It is shown that the expression of Flv2 and Flv4 is high under air level of CO2 and negligible at elevated CO2. Moreover, the rate of accumulation of flv2 and flv4 transcripts upon shift of cells from high to low CO2 is strongly dependent on light intensity. Characterization of FDP inactivation mutants of Synechocystis revealed a specific decline in PSII centers and impaired translation of the D1 protein in Δflv2 and Δflv4 when grown at air level CO2 whereas at high CO2 the Flvs were dispensable. Δflv2 and Δflv4 were also more susceptible to high light induced inhibition of PSII than WT or Δflv1 and Δflv3.SignificanceAnalysis of published sequences revealed the presence of cyanobacteria-like FDPs also in some oxygenic photosynthetic eukaryotes like green algae, mosses and lycophytes. Our data provide evidence that Flv2 and Flv4 have an important role in photoprotection of water-splitting PSII against oxidative stress when the cells are acclimated to air level CO2. It is conceivable that the function of FDPs has changed during evolution from protection against oxygen in anaerobic microbes to protection against reactive oxygen species thus making the sustainable function of oxygen evolving PSII possible. Higher plants lack FDPs and distinctly different mechanisms have evolved for photoprotection of PSII.

Single particle analysis of thylakoid proteins from Thermosynechococcus elongatus and Synechocystis 6803: Localization of the CupA subunit of NDH-1

The larger protein complexes of the cyanobacterial photosynthetic membrane of Thermosynechoccus elongatus and Synechocystis 6803 were studied by single particle electron microscopy after detergent solubilization, without any purification steps. Besides the “standard” L-shaped NDH-1L complex, related to complex I, large numbers of a U-shaped NDH-1MS complex were found in both cyanobacteria. In membranes from Synechocystis Δ cupA and Δ cupA/cupB mutants the U-shaped complexes were absent, indicating that CupA is responsible for the U-shape by binding at the tip of the membrane-bound arm of NDH-1MS. Comparison of membranes grown under air levels of CO 2 or 3% CO 2 indicates that the number of NDH-1MS particles is 30-fold higher under low-CO 2.

CO2 Sensing at Ocean Surface Mediated by cAMP in a Marine Diatom

Marine diatoms are known to be responsible for about a quarter of global primary production and their photosynthesis is sustained by inorganic carbon-concentrating mechanisms and/or C(4) metabolism. Activities of the inorganic carbon-concentrating mechanism are attenuated under enriched [CO(2)]; however, impacts of this factor on primary productivity and the molecular mechanisms of CO(2) responses in marine diatoms are unknown. In this study, transgenic cells were generated of the marine diatom Phaeodactylum tricornutum by the introduction of a beta-glucuronidase reporter gene under the control of an intrinsic CO(2)-responsive promoter, which is the sequence between -80 to +61 relative to the transcription start site of a chloroplastic-carbonic anhydrase gene, ptca1, obtained from P. tricornutum. The activity of the ptca1 promoter was effectively repressed in air-level CO(2) by treating cells with a 1.0 mm cAMP analog, dibutyryl cAMP, or a cAMP phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine. Deletion of the intrinsic cAMP-response element from the ptca1 promoter caused a lack of repression of the reporter gene uidA, even under elevated [CO(2)] and a null phenotype to the strong repressive effects of dibutyryl cAMP and 3-isobutyl-1-methylxanthine on the ptca1 promoter. Deletion of the cAMP-response element was also shown to cause derepression of the uidA reporter gene in the dark. These results indicate that the cytosolic cAMP level increases under elevated [CO(2)] and represses the ptca1 promoter. This strongly suggests the participation of cAMP metabolism, presumably at the cytosolic level, in controlling CO(2)-acquisition systems under elevated [CO(2)] at the ocean surface in a marine diatom.

Regulation of the expression of intracellular beta-carbonic anhydrase in response to CO2 and light in the marine diatom Phaeodactylum tricornutum.

Cells of the marine diatom Phaeodactylum tricornutum Bohlin (UTEX 642) grown in 5% CO(2) were transferred to air-level CO(2) in the light or dark and allowed to acclimate to air. No accumulation of the transcript of the P. tricornutum beta-carbonic anhydrase 1 (ptca1) was detected in 5% CO(2)-grown cells, but ptca1 mRNA accumulated and reached a peak after 6 h acclimation to air but decreased over the next 18 h. A similar accumulation time course was observed in cells air-acclimated in the dark, except that levels of mRNA were <50% those in the light. These results suggest that air-level [CO(2)] is required to trigger the transcription of ptca1 and that light affects the extent of acclimation. During acclimation to air for 120 h in the light, levels of ptca1 mRNA exhibited a periodic oscillation with a cycle of about 24 h, which, however, was not reflected in protein accumulation levels. A 5'-upstream region from the transcription-start site toward -1,292 bp of ptca1 was cloned by inverse polymerase chain reaction, and 5'-truncations were carried out on this fragment. The truncated promoter regions were fused with the beta-glucuronidase gene (uidA) and introduced into P. tricornutum. The promoter fragments, truncated at positions -1,292, -824, -484, -225, and -70 bp, conferred on transformants clear CO(2)-responsive beta-glucuronidase expressions. In contrast, the CO(2)-responsive regulation was severely impaired or completely abolished by truncations, respectively, at position -50 or -30 bp. These results indicate that critical cis-elements required for CO(2)-responsive transcription of ptca1 may be located between -70 and -30 bp relative to the transcription start site.

Electron transport is the functional limitation of photosynthesis in field‐grown Pima cotton plants at high temperature

ABSTRACTRestrictions to photosynthesis can limit plant growth at high temperature in a variety of ways. In addition to increasing photorespiration, moderately high temperatures (35–42 °C) can cause direct injury to the photosynthetic apparatus. Both carbon metabolism and thylakoid reactions have been suggested as the primary site of injury at these temperatures. In the present study this issue was addressed by first characterizing leaf temperature dynamics in Pima cotton (Gossypium barbadense) grown under irrigation in the US desert south‐west. It was found that cotton leaves repeatedly reached temperatures above 40 °C and could fluctuate as much as 8 or 10 °C in a matter of seconds. Laboratory studies revealed a maximum photosynthetic rate at 30–33 °C that declined by 22% at 45 °C. The majority of the inhibition persisted upon return to 30 °C. The mechanism of this limitation was assessed by measuring the response of photosynthesis to CO2 in the laboratory. The first time a cotton leaf (grown at 30 °C) was exposed to 45 °C, photosynthetic electron transport was stimulated (at high CO2) because of an increased flux through the photorespiratory pathway. However, upon cooling back to 30 °C, photosynthetic electron transport was inhibited and fell substantially below the level measured before the heat treatment. In the field, the response of assimilation (A) to various internal levels of CO2 (Ci) revealed that photosynthesis was limited by ribulose‐1,5‐bisphosphate (RuBP) regeneration at normal levels of CO2 (presumably because of limitations in thylakoid reactions needed to support RuBP regeneration). There was no evidence of a ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) limitation at air levels of CO2 and at no point on any of 30 A–Ci curves measured on leaves at temperatures from 28 to 39 °C was RuBP regeneration capacity measured to be in substantial excess of the capacity of Rubisco to use RuBP. It is therefore concluded that photosynthesis in field‐grown Pima cotton leaves is functionally limited by photosynthetic electron transport and RuBP regeneration capacity, not Rubisco activity.

Inhibition of photosynthesis by heat stress: the activation state of Rubisco as a limiting factor in photosynthesis.

Although the catalytic activity of Rubisco increases with temperature, the low affinity of the enzyme for CO(2) and its dual nature as an oxygenase limit the possible increase in net photosynthesis with temperature. For cotton, comparisons of measured rates of net photosynthesis with predicted rates that take into account limitations imposed by the kinetic properties of Rubisco indicate that direct inhibition of photosynthesis occurs at temperatures higher than about 30 degrees C. Inhibition of photosynthesis by moderate heat stress (i.e. 30-42 degrees C) is generally attributed to reduced rates of RuBP regeneration caused by disruption of electron transport activity, and specifically inactivation of the oxygen evolving enzymes of photosystem II. However, measurements of chlorophyll fluorescence and metabolite levels at air-levels of CO(2) indicate that electron transport activity is not limiting at temperatures that inhibit CO(2) fixation. Instead, recent evidence shows that inhibition of net photosynthesis correlates with a decrease in the activation state of Rubisco in both C(3) and C(4) plants and that this decrease in the amount of active Rubisco can fully account for the temperature response of net photosynthesis. Biochemically, the decrease in Rubisco activation can be attributed to: (1) more rapid de-activation of Rubisco caused by a faster rate of dead-end product formation; and (2) slower re-activation of Rubisco by activase. The net result is that as temperature increases activase becomes less effective in keeping Rubisco catalytically competent. In this opinionated review, we discuss how these processes limit photosynthetic performance under moderate heat stress.