CO2-limited Photosynthesis Research Articles

Light and carbon limited photosynthesis of Chlorella sorokiniana

Carbon dioxide (CO2) and light are essential for high photosynthetic rates of microalgal cultures. Microalgal photosynthetic behavior at low CO2 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 CO2 concentration. We used a novel CO2-based photosynthesis monitor originating from leaf research but equipped with an aquatic chamber sparged with CO2 enriched air. Photosynthesis was measured by employing a steady-state CO2 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 CO2 partial pressure into aqueous CO2 concentration to evaluate the CO2 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 CO2 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. CO2 response measurements demonstrated severe CO2 limitation at dissolved CO2 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 % O2 (air) or 2 % O2 indicated the existence of photorespiration in Chlorella. Moreover, the magnitude of photorespiration first increased and then decreased again while reduced dissolved CO2 levels, indicating the activation of a carbon concentrating mechanism at low CO2. Simultaneous measurement of the quantum yield of PSII linear electron transport (ϕPSII) and CO2 uptake revealed the transition of CO2 limited photosynthesis towards light-limited photosynthesis.

Diversity in photosynthetic electron transport under [CO2]-limitation: the cyanobacterium Synechococcus sp. PCC 7002 and green alga Chlamydomonas reinhardtii drive an O2-dependent alternative electron flow and non-photochemical quenching of chlorophyll fluorescence during CO2-limited photosynthesis.

Some cyanobacteria, but not all, experience an induction of alternative electron flow (AEF) during CO2-limited photosynthesis. For example, Synechocystis sp. PCC 6803 (S. 6803) exhibits AEF, but Synechococcus elongatus sp. PCC 7942 does not. This difference is due to the presence of flavodiiron 2 and 4 proteins (FLV2/4) in S. 6803, which catalyze electron donation to O2. In this study, we observed a low-[CO2] induced AEF in the marine cyanobacterium Synechococcus sp. PCC 7002 that lacks FLV2/4. The AEF shows high affinity for O2, compared with AEF mediated by FLV2/4 in S. 6803, and can proceed under extreme low [O2] (about a few µM O2). Further, the transition from CO2-saturated to CO2-limited photosynthesis leads a preferential excitation of PSI to PSII and increased non-photochemical quenching of chlorophyll fluorescence. We found that the model green alga Chlamydomonas reinhardtii also has an O2-dependent AEF showing the same affinity for O2 as that in S. 7002. These data represent the diverse molecular mechanisms to drive AEF in cyanobacteria and green algae. In this paper, we further discuss the diversity, the evolution, and the physiological function of strategy to CO2-limitation in cyanobacterial and green algal photosynthesis.

Evidence for a Role for NAD(P)H Dehydrogenase in Concentration of CO2 in the Bundle Sheath Cell of Zea mays.

Prior studies with Nicotiana and Arabidopsis described failed assembly of the chloroplastic NDH [NAD(P)H dehydrogenase] supercomplex by serial mutation of several subunit genes. We examined the properties of Zea mays leaves containing Mu and Ds insertions into nuclear gene exons encoding the critical o- and n-subunits of NDH, respectively. In vivo reduction of plastoquinone in the dark was sharply diminished in maize homozygous mutant compared to normal leaves but not to the extreme degree observed for the corresponding lesions in Arabidopsis. The net carbon assimilation rate (A) at high irradiance and saturating CO2 levels was reduced by one-half due to NDH mutation in maize although no genotypic effect was evident at very low CO2 levels. Simultaneous assessment of chlorophyll fluorescence and A in maize at low (2% by volume) and high (21%) O2 levels indicated the presence of a small, yet detectable, O2-dependent component of total linear photosynthetic electron transport in 21% O2 This O2-dependent component decreased with increasing CO2 level indicative of photorespiration. Photorespiration was generally elevated in maize mutant compared to normal leaves. Quantification of the proportion of total electron transport supporting photorespiration enabled estimation of the bundle sheath cell CO2 concentration (Cb) using a simple kinetic model of ribulose bisphosphate carboxylase/oxygenase function. The A versus Cb relationships overlapped for normal and mutant lines consistent with occurrence of strictly CO2-limited photosynthesis in the mutant bundle sheath cell. The results are discussed in terms of a previously reported CO2 concentration model [Laisk A, Edwards GE (2000) Photosynth Res 66: 199-224].

Comparative research on inorganic carbon acquisition by the macroalgae Ulva prolifera (Chlorophyta) and Pyropia yezoensis (Rhodophyta)

The red macroalga Pyropia/Porphyra is one of the most important marine crops in the world; Pyropia cultivation is severely affected by the green macroalga Ulva spp., a fouling organism. In this study, growth competition and acquisition of dissolved inorganic carbon (Ci) by Ulva prolifera and Pyropia yezoensis were investigated to understand the physiological characteristics of the two species. Competition experimental results showed that the relative growth rate of U. prolifera was not affected, whereas the growth rate of P. yezoensis was significantly inhibited. U. prolifera exhibited a competitive advantage when these two species were cultured together. The two algal species displayed high pH compensation points, suggesting that U. prolifera and P. yezoensis can photosynthesize by using HCO3 −; the utilization ability of U. prolifera is stronger than that of P. yezoensis. The net photosynthetic rates of P. yezoensis and U. prolifera were significantly inhibited by the carbonic anhydrase (CA) inhibitors acetazolamide and 6-ethoxyzolamide indicating that CA is implicated in carbon-concentrating mechanisms (CCM). 4,4′-diisothiocyano-stilbene-2,2′-disulfonate (DIDS), another inhibitor that prevents direct HCO3 − uptake, did not significantly affect U. prolifera. Conversely, DIDS can sharply decrease the photosynthetic rate of P. yezoensis, particularly at high pH. U. prolifera and P. yezoensis also showed CO2-limited photosynthesis by which the half-saturating concentration of Ci exceeds that of seawater. U. prolifera did not show active HCO3 − uptake; instead, U. prolifera used HCO3 − via extracellular CA for photosynthetic carbon fixation. U. prolifera also utilizes extracellular CA-mediated HCO3 − absorption to a higher extent than P. yezoensis. By contrast, P. yezoensis shows active HCO3 − uptake and extracellular CCMs for photosynthetic carbon fixation. Therefore, these two algal species can survive in changing environments with high pH and low Ci concentration because of the action of CCMs.

FLAVODIIRON2 and FLAVODIIRON4 proteins mediate an oxygen-dependent alternative electron flow in Synechocystis sp. PCC 6803 under CO2-limited conditions.

This study aims to elucidate the molecular mechanism of an alternative electron flow (AEF) functioning under suppressed (CO2-limited) photosynthesis in the cyanobacterium Synechocystis sp. PCC 6803. Photosynthetic linear electron flow, evaluated as the quantum yield of photosystem II [Y(II)], reaches a maximum shortly after the onset of actinic illumination. Thereafter, Y(II) transiently decreases concomitantly with a decrease in the photosynthetic oxygen evolution rate and then recovers to a rate that is close to the initial maximum. These results show that CO2 limitation suppresses photosynthesis and induces AEF. In contrast to the wild type, Synechocystis sp. PCC 6803 mutants deficient in the genes encoding FLAVODIIRON2 (FLV2) and FLV4 proteins show no recovery of Y(II) after prolonged illumination. However, Synechocystis sp. PCC 6803 mutants deficient in genes encoding proteins functioning in photorespiration show AEF activity similar to the wild type. In contrast to Synechocystis sp. PCC 6803, the cyanobacterium Synechococcus elongatus PCC 7942 has no FLV proteins with high homology to FLV2 and FLV4 in Synechocystis sp. PCC 6803. This lack of FLV2/4 may explain why AEF is not induced under CO2-limited photosynthesis in S. elongatus PCC 7942. As the glutathione S-transferase fusion protein overexpressed in Escherichia coli exhibits NADH-dependent oxygen reduction to water, we suggest that FLV2 and FLV4 mediate oxygen-dependent AEF in Synechocystis sp. PCC 6803 when electron acceptors such as CO2 are not available.

Integrating Transient Heterogeneity of Non-Photochemical Quenching in Shade-Grown Heterobaric Leaves of Avocado (Persea americana L.): Responses to CO2 Concentration, Stomatal Occlusion, Dehydration and Relative Humidity

Long-lived shade leaves of avocado had extremely low rates of photosynthesis. Gas exchange measurements of photosynthesis were of limited use, so we resorted to Chl fluorescence imaging (CFI) and spot measurements to evaluate photosynthetic electron transport rates (ETRs) and non-photochemical quenching (NPQ). Imaging revealed a remarkable transient heterogeneity of NPQ during photosynthetic induction in these hypostomatous, heterobaric leaves, but was adequately integrated by spot measurements, despite long-lasting artifacts from repeated saturating flashes during assays. Major veins (mid-vein, first- and second-order veins) defined areas of more static large-scale heterogeneous NPQ, with more dynamic small-scale heterogeneity most strongly expressed in mesophyll cells between third- and fourth-order veins. Both responded to external CO2 concentration ([CO2]), occlusion of stomata with Vaseline™, leaf dehydration and relative humidity (RH). We interpreted these responses in terms of independent behavior of stomata in adjacent areoles that was largely expressed through CO2-limited photosynthesis. Heterogeneity was most pronounced and prolonged in the absence of net CO2 fixation in 100 p.p.m. [CO2] when respiratory and photorespiratory CO2 cycling constrained the inferred ETR to ~75% of values in 400 or 700 p.p.m. [CO2]. Likewise, sustained higher NPQ under Vaseline™, after dehydration or at low RH, also restricted ETR to ~75% of control values. Low NPQ in chloroplast-containing cells adjacent to major veins but remote from stomata suggested internal sources of high [CO2] in these tissues.

Responses of leaf photosynthesis and plant growth to altered source–sink balance in a Japanese and a Dutch tomato cultivar

We investigated the effects of altered source/sink ratio by leaf or fruit pruning on leaf photosynthetic characteristics and whole-plant growth of ‘Momotaro York’, a Japanese cultivar, and ‘Dundee’, a Dutch cultivar and verified a hypothesis for sink-limitation of plant growth proposed by Tanaka and Fujita (1974). Plants were grown hydroponically with a high-wire system in a greenhouse for 11 weeks. Light-saturated photosynthesis of young, fully expanded leaves measured at atmospheric CO 2 partial pressure of 37 Pa and at an intercellular CO 2 partial pressure of 20 Pa was not influenced by alteration of source/sink ratio for either cultivar. Although soluble sugars were accumulated in leaves under high source/sink conditions, the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase, a rate-limiting factor for CO 2-limited photosynthesis, was not affected by the altered source–sink balance. Net assimilation rate of a whole plant increased with decreasing source/sink ratio, but this can be accounted for by the changes in light interception per unit leaf area, without taking the source–sink relationship into consideration. It was concluded that the altered source/sink ratio did not change leaf photosynthetic capacity and the sink-limitation hypothesis cannot be applied to either cultivar under the conditions of the present study.

Kranz anatomy is not essential for terrestrial C4 plant photosynthesis.

An important adaptation to CO2-limited photosynthesis in cyanobacteria, algae and some plants was development of CO2-concentrating mechanisms (CCM). Evolution of a CCM occurred many times in flowering plants, beginning at least 15-20 million years ago, in response to atmospheric CO2 reduction, climate change, geological trends, and evolutionary diversification of species. In plants, this is achieved through a biochemical inorganic carbon pump called C4 photosynthesis, discovered 35 years ago. C4 photosynthesis is advantageous when limitations on carbon acquisition are imposed by high temperature, drought and saline conditions. It has been thought that a specialized leaf anatomy, composed of two, distinctive photosynthetic cell types (Kranz anatomy), is required for C4 photosynthesis. We provide evidence that C4 photosynthesis can function within a single photosynthetic cell in terrestrial plants. Borszczowia aralocaspica (Chenopodiaceae) has the photosynthetic features of C4 plants, yet lacks Kranz anatomy. This species accomplishes C4 photosynthesis through spatial compartmentation of photosynthetic enzymes, and by separation of two types of chloroplasts and other organelles in distinct positions within the chlorenchyma cell cytoplasm.

Relationship between the suppression of photosynthesis and starch accumulation in the pod-removed bean

Effects of pod removal on the photosynthetic rates and their under lying biochemical properties were examined in leaves of bean (Phaseolus vulgaris L.). Pod removal led to decreases in the photosynthetic gas- exchange rate, stomatal conductance and intercellular CO2 pressure. H owever, these decreases in photosynthesis were not asso-ciated with changes in the amounts of ribulose-1,5-bisphosphate carb oxylase (Rubisco) and other several photo-synthetic components, but were associated with an increase in starch content. In addition, such a decrease in the photosynthetic rate was not observed when measured with O2 evolution under conditions of saturating CO2. Similarly, although a decrease in CO2-limited photosynthesis for a given Rubisco content was found in depodded plants, neither the speci fic activity nor activation state of Rubisco decreased . The decrease in the photosynthetic gas-exchange rate by pod removal was rec overed by shading the plants. CO2-limited photosynthesis per unit of Rubisco protein was also completely recovered at the same l evel in the plants with pods. At the same time, starch accumu-lation was also decreased by shading. From these results, we concluded that the suppression of photosynthesis observed in depodded beans was caused by the hindrance of CO2 diffusion due to starch accumulated in the chloroplast.

Analysis of inhibition of photosynthesis due to water stress in the C3 species Hordeum vulgare and Vicia faba: Electron transport, CO2 fixation and carboxylation capacity

A C3 monocot, Hordeum vulgare and C3 dicot, Vicia faba, were studied to evaluate the mechanism of inhibition of photosynthesis due to water stress. The net rate of CO2 fixation (A) and transpiration (E) were measured by gas exchange, while the true rate of O2 evolution (J O2) was calculated from chlorophyll fluorescence analysis through the stress cycle (10 to 11 days). With the development of water stress, the decrease in A was more pronounced than the decrease in J O2 resulting in an increased ratio of Photosystem II activity per CO2 fixed which is indicative of an increase in photorespiration due to a decrease in supply of CO2 to Rubisco. Analyses of changes in the J O2 A ratios versus that of CO2 limited photosynthesis in well watered plants, and RuBP pool/RuBP binding sites on Rubisco and RuBP activity, indicate a decreased supply of CO2 to Rubisco under both mild and severe stress is primarily responsible for the decrease in CO2 fixation. In the early stages of stress, the decrease in C i (intercellular CO2) due to stomatal closure can account for the decrease in photosynthesis. Under more severe stress, CO2 supply to Rubisco, calculated from analysis of electron flow and CO2 exchange, continued to decrease. However, C i, calculated from analysis of transpiration and CO2 exchange, either remained constant or increased which may be due to either a decrease in mesophyll conductance or an overestimation of C i by this method due to patchiness in conductance of CO2 to the intercellular space. When plants were rewatered after photosynthesis had dropped to 10-30% of the original rate, both species showed near full recovery within two to four days.

Perspectives on photoinhibition and photorespiration in the field: quintessential inefficiencies of the light and dark reactions of photosynthesis?

Abstract Taking the long-held view that photoinhibition embraces several processes leading to a reduction in the efficiency of light utilization in photosynthesis, and that photorespiration embraces several processes associated with O2 uptake in the light, photoinhibition and photorespiration now can be considered as inevitable, but essential inefficiencies of photosynthesis which help preserve photosynthetic competence in bright light. Photorespiratory O2 uptake via Rubisco, and O2 uptake via the Mehler reaction, both promote non-assimilatory electron transport, and stimulate photon utilization during CO2-limited photosynthesis in bright light. Although fluorescence studies show that the proportion of total photon use via oxygenase photorespiration in air may decline to only about 10% in full sunlight, mass spectrometer studies show that O2 uptake in Mehler reaction photorespiration in C3 and CAM plants can still account for up to 50% of electron flow in saturating CO2 and light. The Mehler-ascorbate peroxidase reaction has an additional role in sustaining membrane energization which promotes dynamic photoinhibition and photon protection (rapidly reversible decrease in PSII efficiency involving dissipation of the energy of excess photons in the antennae). Net CO2 and O2 exchange studies evidently underestimate the extent of total electron transport, and hence overestimate the extent of photon excess in bright light, leading to overestimates of the role of energy-dependent photon dissipation through dynamic photo-inhibition. Nevertheless, in C3 plants in air all of these processes help to mitigate chronic photoinhibition and photon damage (slowly reversible decrease in PSII efficiency involving loss of reaction centre function). The possibility remains that residual electron transport to O2 from intermediates in the vicinity of PSII may also lead to reactive O2 species that potentiate this photon damage.

Effects of Growth Temperature on the Responses of Ribulose-1,5-Biphosphate Carboxylase, Electron Transport Components, and Sucrose Synthesis Enzymes to Leaf Nitrogen in Rice, and Their Relationships to Photosynthesis.

Effects of growth temperature on the photosynthetic gas-exchange rates and their underlying biochemical properties were examined in young, fully expanded leaves of rice (Oryza sativa L.). The plants were grown hydroponically under day/night temperature regimes of 18/15[deg]C, 23/18[deg]C, and 30/23[deg]C and all photosynthetic measurements were made at a leaf temperature of 25[deg]C and an irradiance of 1800 [mu]mol quanta m-2 s-1. Growth temperature affected the photosynthetic CO2 response curve. The relative ratio of the initial slope to the CO2-saturated photosynthesis increased with rising growth temperature. This was caused mainly by an increase in CO2-limited photosynthesis for a given leaf nitrogen content with rising growth temperature. However, there was no difference in ribulose-1,5-bisphosphate carboxylase (Rubisco) content at any given leaf nitrogen content among temperature treatments. In addition, the activation state and catalytic turnover rate of Rubisco were not affected by growth temperature. The increase in CO2-limited photosynthesis with rising growth temperature was the result of an increase in the CO2 transfer conductance between the intercellular airspaces and the carboxylation sites. The amounts of total chlorophyll and light-harvesting chlorophyll a/b protein II increased for the same leaf nitrogen content with rising growth temperature, but the amounts of cytochrome f and coupling factor 1 and the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were the same between plants grown at 23/18[deg]C and those grown at 30/23[deg]C. Similarly, CO2-saturated photosynthesis was not different for the same leaf nitrogen content between these treatments. For the 18/15[deg]C-grown plants, a slight decrease in the amounts of cytochrome f and coupling factor 1 and an increase in the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were found, but these were not reflected in CO2-saturated photosynthesis.

Responses of Ribulose-1,5-Bisphosphate Carboxylase, Cytochrome f, and Sucrose Synthesis Enzymes in Rice Leaves to Leaf Nitrogen and Their Relationships to Photosynthesis.

The photosynthetic gas-exchange rates and various biochemical components of photosynthesis, including ribulose-1,5-bisphosphate carboxylase (Rubisco) content, cytochrome (Cyt) f content, and the activities of two sucrose synthesis enzymes, were examined in young, fully expanded leaves of rice (Oryza sativa L.) grown hydroponically in different nitrogen concentrations. The light-saturated rate of photosynthesis at an intercellular CO2 pressure of 20 Pa (CO2-limited photosynthesis) was linearly dependent on leaf nitrogen content, but curvilinearly correlated with Rubisco content. This difference was due to a greater than proportional increase in Rubisco content relative to leaf nitrogen content and the presence of a CO2 transfer resistance between the intercellular air spaces and the carboxylation sites. CO2-limited photosynthesis was proportional to Cyt f content, one of the key components of electron transport, but was not proportional to the activities of cytosolic fructose-1,6-bisphosphatase and sucrose phosphate synthase, the two regulatory enzymes of sucrose synthesis. Light-saturated photosynthesis above an intercellular CO2 pressure of 60 Pa (CO2-saturated photosynthesis) was curvilinearly dependent on leaf nitrogen content. This CO2-saturated photosynthesis was proportional to Cyt f content in the low- and normal-nitrogen leaves, and correlated better with the activities of cytosolic fructose-1,6-bisphosphatase and sucrose phosphate synthase in the high-nitrogen leaves. The increase in the activities of these two enzymes with increasing leaf nitrogen was not as great as the increase in Cyt f content. Thus, as leaf nitrogen increased, the limitation caused by the activities of sucrose synthesis enzymes came into play, which resulted in the curvilinear relationship. However, this limitation by sucrose synthesis enzymes did not affect photosynthesis under normal ambient air.

An analysis of the temperature response curves of CO2 exchange in the leaves of two temperate and one tropical grass species

The temperature dependence of carbon dioxide exchange in two temperate grass species, Lolium perenne (S.23) and Lolium multiflorum (S.22), and one tropical grass species, Cenchrus ciliaris, have been examined. It has been shown that the decline in the net CO2 exchange rate above the optimum temperature is caused, in the case of C. ciliaris, by denaturation, and, in the case of L. perenne and L. multiforum by the greater response to temperature of respiration compared with photosynthesis.Activation energies have been obtained for both photosynthetic and respiratory systems. For both systems the values fall into discrete groups, and it is postulated that each group may be characteristic of a particular rate-limiting process. The distribution of values changes with changes in experimental conditions, there being marked differences in the distribution of values obtained for light-limited and CO2-limited photosynthesis.