CO2 Compensation Point Research Articles

Tribulus (Zygophyllaceae) as a case study for the evolution of C2 and C4 photosynthesis.

C2 photosynthesis is a photosynthetic pathway in which photorespiratory CO2 release and refixation are enhanced in leaf bundle sheath (BS) tissues. The evolution of C2 photosynthesis has been hypothesized to be a major step in the origin of C4 photosynthesis, highlighting the importance of studying C2 evolution. In this study, physiological, anatomical, ultrastructural, and immunohistochemical properties of leaf photosynthetic tissues were investigated in six non-C4 Tribulus species and four C4 Tribulus species. At 42°C, T. cristatus exhibited a photosynthetic CO2 compensation point in the absence of respiration (C*) of 21 µmol mol-1, below the C3 mean C* of 73 µmol mol-1. Tribulus astrocarpus had a C* value at 42°C of 55 µmol mol-1, intermediate between the C3 species and the C2 T. cristatus. Glycine decarboxylase (GDC) allocation to BS tissues was associated with lower C*. Tribulus cristatus and T. astrocarpus allocated 86% and 30% of their GDC to the BS tissues, respectively, well above the C3 mean of 11%. Tribulus astrocarpus thus exhibits a weaker C2 (termed sub-C2) phenotype. Increased allocation of mitochondria to the BS and decreased length-to-width ratios of BS cells, were present in non-C4 species, indicating a potential role in C2 and C4 evolution.

Photosynthetic responses of switchgrass to light and CO2 under different precipitation treatments

AbstractSwitchgrass (Panicum virgatum L.) is a prominent bioenergy crop with robust resilience to environmental stresses. However, our knowledge regarding how precipitation changes affect switchgrass photosynthesis and its responses to light and CO2 remains limited. To address this knowledge gap, we conducted a field precipitation experiment with five different treatments, including −50%, −33%, 0%, +33%, and +50% of ambient precipitation. To determine the responses of leaf photosynthesis to CO2 concentration and light, we measured leaf net photosynthesis of switchgrass under different CO2 concentrations and light levels in 2020 and 2021 for each of the five precipitation treatments. We first evaluated four light and CO2 response models (i.e., rectangular hyperbola model, nonrectangular hyperbola model, exponential model, and the modified rectangular hyperbola model) using the measurements in the ambient precipitation treatment. Based on the fitting criteria, we selected the nonrectangular hyperbola model as the optimal model and applied it to all precipitation treatments, and estimated model parameters. Overall, the model fit field measurements well for the light and CO2 response curves. Precipitation change did not influence the maximum net photosynthetic rate (Pmax) but influenced other model parameters including quantum yield (α), convexity (θ), dark respiration (Rd), light compensation point (LCP), and saturated light point (LSP). Specifically, the mean Pmax of five precipitation treatments was 17.6 μmol CO2 m−2 s−1, and the ambient treatment tended to have a higher Pmax. The +33% treatment had the highest α, and the ambient treatment had lower θ and LCP, higher Rd, and relatively lower LSP. Furthermore, precipitation significantly influenced all model parameters of CO2 response. The ambient treatment had the highest Pmax, largest α, and lowest θ, Rd, and CO2 compensation point LCP. Overall, this study improved our understanding of how switchgrass leaf photosynthesis responds to diverse environmental factors, providing valuable insights for accurately modeling switchgrass ecophysiology and productivity.

Engineering the cyanobacterial ATP-driven BCT1 bicarbonate transporter for functional targeting to C3 plant chloroplasts.

The ATP-driven bicarbonate transporter 1 (BCT1) from Synechococcus is a four-component complex in the cyanobacterial CO2-concentrating mechanism. BCT1 could enhance photosynthetic CO2 assimilation in plant chloroplasts. However, directing its subunits (CmpA, CmpB, CmpC, and CmpD) to three chloroplast sub-compartments is highly complex. Investigating BCT1 integration into Nicotiana benthamiana chloroplasts revealed promising targeting strategies using transit peptides from the intermembrane space protein Tic22 for correct CmpA targeting, while the transit peptide of the chloroplastic ABCD2 transporter effectively targeted CmpB to the inner envelope membrane. CmpC and CmpD were targeted to the stroma by RecA and recruited to the inner envelope membrane by CmpB. Despite successful targeting, expression of this complex in CO2-dependent Escherichia coli failed to demonstrate bicarbonate uptake. We then used rational design and directed evolution to generate new BCT1 forms that were constitutively active. Several mutants were recovered, including a CmpCD fusion. Selected mutants were further characterized and stably expressed in Arabidopsis thaliana, but the transformed plants did not have higher carbon assimilation rates or decreased CO2 compensation points in mature leaves. While further analysis is required, this directed evolution and heterologous testing approach presents potential for iterative modification and assessment of CO2-concentrating mechanism components to improve plant photosynthesis.

C4 monocots and C4 dicots exhibit rapid photosynthetic induction response in contrast to C3 plants.

Considering the prevalence of ever-changing conditions in the natural world, investigation of photosynthetic responses in C4 plants under fluctuating light is needed. Here, we studied the effect of dynamic illumination on photosynthesis in totally 10 C3, C3-C4 intermediate, C4-like and C4 dicots and monocots at CO2 concentrations of 400 and 800 μmol mol-1. C4 and C4-like plants had faster photosynthetic induction and light-induced stomatal dynamics than C3 plants at 400 μmol mol-1, but not at 800 μmol mol-1 CO2, at which the CO2 supply rarely limits photosynthesis. C4 and C4-like plants had a higher water use efficiency than C3 plants at both CO2 concentrations. There were positive correlations between photosynthetic induction and light-induced stomatal response, together with CO2 compensation point, which was a parameter of the CO2-concentrating mechanism of C4 photosynthesis. These results clearly show that C4 photosynthesis in both monocots and dicots adapts to fluctuating light conditions more efficiently than C3 photosynthesis. The rapid photosynthetic induction response in C4 plants can be attributed to the rapid stomatal dynamics, the CO2-concentrating mechanism or both.

Integrative Physiological, Transcriptome, and Proteome Analyses Provide Insights into the Photosynthetic Changes in Maize in a Maize-Peanut Intercropping System.

Intercropping is a traditional and sustainable planting method that can make rational use of natural resources such as light, temperature, fertilizer, water, and CO2. Due to its efficient resource utilization, intercropping, in particular, maize and legume intercropping, is widespread around the world. However, the molecular details of these pathways remain largely unknown. In this study, physiological, transcriptome, and proteome analyses were compared between maize monocropping and maize-peanut intercropping. The results show that an intercropping system enhanced the ability of carbon fixation and carboxylation of maize leaves. Apparent quantum yield (AQY), the light-saturated net photosynthetic rate (LSPn), the light saturation point (LSP), and the light compensation point (LCP) were increased by 11.6%, 9.4%, 8.9%, and 32.1% in the intercropping system, respectively; carboxylation efficiency (CE), the CO2 saturation point (Cisat), the Rubisco maximum carboxylation rate (Vcmax), the maximum electron transfer rate (Jmax), and the triose phosphate utilization rate (TPU) were increased by 28.5%, 7.3%, 18.7%, 29.2%, and 17.0%, respectively; meanwhile, the CO2 compensation point (Γ) decreased by 22.6%. Moreover, the transcriptome analysis confirmed the presence of 588 differentially expressed genes (DEGs), and the numbers of up-regulated and down-regulated genes were 383 and 205, respectively. The DEGs were primarily concerned with ribosomes, plant hormone signal transduction, and photosynthesis. Furthermore, 549 differentially expressed proteins (DEPs) were identified in the maize leaves in both the maize monocropping and maize-peanut intercropping systems. Bioinformatics analysis revealed that 186 DEPs were related to 37 specific KEGG pathways in each of the two treatment groups. Based on the physiological, transcriptome, and proteome analyses, it was demonstrated that the photosynthetic characteristics in maize leaves can be improved by maize-peanut intercropping. This may be related to PS I, PS II, cytochrome b6f complex, ATP synthase, and photosynthetic CO2 fixation, which is caused by the improved CO2 carboxylation efficiency. Our results provide a more in-depth understanding of the high yield and high-efficiency mechanism in maize and peanut intercropping.

Nonuniform salinity regulate leaf characteristics and improve photosynthesis of cherry tomatoes under high salinity

Cherry tomato (Lycopersicon esculentum var. cerasiforme) is a relatively salt-tolerant fruit and vegetable widely planted worldwide. In this study, the growth and physiological responses and the different regulatory of cherry tomatoes under high, low salinity, and partial root zone salinity conditions were investigated. Cherry tomatoes grown under nonuniform salinity (0/20, 0/100 mM NaCl) conditions in a split-root pot were compared with seedlings grown under uniform salinity (20, 100 mM NaCl) conditions, and their growth and physiological parameters were determined. Our results showed that the growth was not hindered under a low salinity (20 mM NaCl), and the fruit quality was improved without a yield reduction; this was caused by promoted root growth and better leaf function maintenance. The relief effects under the higher salt stress (100 mM NaCl) conditions were more significant than those under a lower salt stress condition (20 mM NaCl). Compared with uniform high salinity conditions, nonuniform salinity conditions could improve photosynthetic characteristics and maintain better leaf function. The apparent quantum efficiency (AQE), carboxylation efficiency (CE) and light saturation irradiance (LSP) increased, while the light compensation point (LCP) and CO2 compensation point (CCP) decreased; this enhanced the light energy and CO2 utilization. Chlorophyll content (Chl), including Chl a and b and the carotenoids (Car), and the chlorophyll fluorescence parameters, including the maximum quantum efficiency of photosystem II (PSII) reaction centers (Fv/Fm), the quantum efficiency of PSII (Y(II)) and electron transport rate through PSII (ETR), were increased; this significantly promoted the net photosynthetic rate (Pn) and water use efficiency (WUE). Compared with a uniform high salt stress treatment, the leaf relative water content (RWC) and water potential (Ψ) increased by 22.57% and 13.71%, respectively, the electrolyte leakage rate decreased by 36.13%, and the final yield increased by 2.89 times under a nonuniform high salinity treatment. These results indicated that nonuniform salinity conditions regulated the leaf water relations and the antioxidant system and maintained better anatomical structure to improve photosynthesis in cherry tomato under a high salinity environment. Finally, our results can aid in the improvement of tomato productivity from salt-affected lands.

Photosynthesis governed by nanoparticulate titanium dioxide. The Pisum sativum L. case study

Wide availability of anthropogenic TiO2 nanoparticles facilitates their penetration into environment and prompts interactions with plants. They alter plants growth and change their nutritional status. In particular, metabolic processes are affected. In this work the effect of nanometric TiO2 on photosynthesis efficiency in green pea (Pisum sativum L.) was studied. Hydroponic cultivations with three Ti levels (10; 50 and 100 mg L−1) were applied. At all concentrations nanoparticles penetrated into plant tissues and were detected by the single particle ICP-MS/MS method. Nanoparticles altered the CO2 assimilation rate and gas exchange parameters (i.e. transpiration, stomatal conductance, sub-stomatal CO2 concentration). The most pronounced effects were observed for Ti 50 mg L−1 cultivation where photosynthesis efficiency, transpiration and stomatal conductance were increased by 14.69%, 4.58% and 8.92%, respectively. They were further confirmed by high maximum ribulose 1,5-bisphosphate carboxylation rate (27.40% increase), maximum electron transport rate (21.51% increase) and the lowest CO2 compensation point (45.19% decrease). Furthermore, concentrations of Cu, Mn, Zn, Fe, Mg, Ca, K and P were examined with the most pronounced changes observed for elements directly involved in photosynthesis (Cu, Zn, Mn, and Fe). The Cu concentrations in roots, stems and leaves for Ti 50 mg L−1 cultivation were below the control by 33.15%, 38.28% and 10.76%, respectively. The Zn content in analogous treatment and organs decreased by 30.24%, 26.69% and 13.35%. The Mn and Fe levels in leaves were increased by 72.22% and 50.32%, respectively. Our results indicated that plant defence mechanisms which restrain the water uptake have been overcome in pea by photocatalytic activity of nanoparticulate TiO2 which stimulated photosynthesis. On the contrary to the substantial stomatal conductance, the transpiration has been reduced because exceptional part of water flow was already consumed in chloroplasts and could not have been freed to the atmosphere.

Brassicaceae display variation in efficiency of photorespiratory carbon-recapturing mechanisms.

Carbon-concentrating mechanisms enhance the carboxylase efficiency of Rubisco by providing supra-atmospheric concentrations of CO2 in its surroundings. Beside the C4 photosynthesis pathway, carbon concentration can also be achieved by the photorespiratory glycine shuttle which requires fewer and less complex modifications. Plants displaying CO2 compensation points between 10 ppm and 40 ppm are often considered to utilize such a photorespiratory shuttle and are termed 'C3-C4 intermediates'. In the present study, we perform a physiological, biochemical, and anatomical survey of a large number of Brassicaceae species to better understand the C3-C4 intermediate phenotype, including its basic components and its plasticity. Our phylogenetic analysis suggested that C3-C4 metabolism evolved up to five times independently in the Brassicaceae. The efficiency of the pathway showed considerable variation. Centripetal accumulation of organelles in the bundle sheath was consistently observed in all C3-C4-classified taxa, indicating a crucial role for anatomical features in CO2-concentrating pathways. Leaf metabolite patterns were strongly influenced by the individual species, but accumulation of photorespiratory shuttle metabolites glycine and serine was generally observed. Analysis of phosphoenolpyruvate carboxylase activities suggested that C4-like shuttles have not evolved in the investigated Brassicaceae. Convergent evolution of the photorespiratory shuttle indicates that it represents a distinct photosynthesis type that is beneficial in some environments.

Mass screening of rice mutant populations at low CO2 for identification of lowered photorespiration and respiration rates.

Identifying rice (Oryza sativa) germplasm with improved efficiency of primary metabolism is of utmost importance in order to increase yields. One such approach can be attained through screening genetically diverse populations under altered environmental conditions. Growth or treatment under low carbon dioxide (CO2) concentrations can be used as a means of revealing altered leaf photorespiration, respiration and other metabolic variants. We developed a pipeline for very high throughput treatment of gamma- and ethyl methanesulfonate- (EMS) induced mutant populations of IR64 rice seedlings at very low CO2 for 7 days. 1050 seedlings per batch at 5th leaf stage were exposed to 60 ppm CO2 for the first day and 30 ppm for the remaining three days. Following this, putative candidates were identified by measuring chlorophyll depletion using SPAD. Screening results showed a distinct difference between the mutants and the WTs. The mean chlorophyll loss in WTs ranged from 65% to 11% respectively, whereas in the mutant lines chlorophyll loss ranged from 0 to 100%, suggesting considerable phenotypic variation. Rice mutants with a reduced chlorophyll reduction (<10%) were identified as 'Chlorophyll retention mutants' (CRMs) under low CO2 stress. In total, 1909 mutant lines (14,000 seedlings) were screened for chlorophyll content under 30 ppm CO2, with 26 lines selected for detailed screening. These 26 putative candidates were self-seeded to produce an M5 generation, used to determine the genetic control of the altered response to low CO2. Gas exchange of light and CO2 response revealed that there were significant variations among photosynthetic properties in two selected rice mutants. The CO2 compensation points in the absence of photorespiration and leaf respiration rates were lower than the WTs and anatomical analyses showed that CRM 29 had improved mesophyll cell area. We propose that this approach is useful for generating new material for breeding rice with improved primary metabolism.

Loss of peroxisomal NAD kinase 3 (NADK3) affects photorespiration metabolism in Arabidopsis

Nicotinamide adenine dinucleotides (NAD+ and NADP+) are electron mediators involved in various metabolic pathways. NADP(H) are produced by NAD kinase (NADK) through the phosphorylation of NAD(H). The Arabidopsis NADK3 (AtNADK3) is reported to preferentially phosphorylate NADH to NADPH and is localized in the peroxisome. To elucidate the biological function of AtNADK3 in Arabidopsis, we compared metabolites of nadk1, nadk2 and nadk3 Arabidopsis T-DNA inserted mutants. Metabolome analysis revealed that glycine and serine, which are intermediate metabolites of photorespiration, both increased in the nadk3 mutants. Plants grown for 6 weeks under short-day conditions showed increased NAD(H), indicating a decrease in the phosphorylation ratio in the NAD(P)(H) equilibrium. Furthermore, high CO2 (0.15%) treatment induced a decrease in glycine and serine in nadk3 mutants. The nadk3 showed a significant decrease in post-illumination CO2 burst, suggesting that the photorespiratory flux was disrupted in the nadk3 mutant. In addition, an increase in CO2 compensation points and a decrease in CO2 assimilation rate were observed in the nadk3 mutants. These results indicate that the lack of AtNADK3 causes a disruption in the intracellular metabolism, such as in amino acid synthesis and photorespiration.

The carbon-concentrating mechanism of the extremophilic red microalga Cyanidioschyzon merolae

Cyanidioschyzonmerolae is an extremophilic red microalga which grows in low-pH, high-temperature environments. The basis of C. merolae’s environmental resilience is not fully characterized, including whether this alga uses a carbon-concentrating mechanism (CCM). To determine if C. merolae uses a CCM, we measured CO2 uptake parameters using an open-path infra-red gas analyzer and compared them to values expected in the absence of a CCM. These measurements and analysis indicated that C. merolae had the gas-exchange characteristics of a CCM-operating organism: low CO2 compensation point, high affinity for external CO2, and minimized rubisco oxygenation. The biomass δ13C of C. merolae was also consistent with a CCM. The apparent presence of a CCM in C. merolae suggests the use of an unusual mechanism for carbon concentration, as C. merolae is thought to lack a pyrenoid and gas-exchange measurements indicated that C. merolae primarily takes up inorganic carbon as carbon dioxide, rather than bicarbonate. We use homology to known CCM components to propose a model of a pH-gradient-based CCM, and we discuss how this CCM can be further investigated.

CO&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; Demand-Supply Coordination in Photosynthesis Reflecting the Plant-Environment Interaction: Extension and Parameterization of Demand Function and Supply Function

Photosynthesis consists of a biochemical process named demand and a CO2 diffusion process named supply function. The intersection (Ci, An) at equal to the demand function and the supply function reflects a steady state of the plant subjected to the environment. The intersections of these demand-supply functions under different photosynthetically active radiation (PAR) can be fitted to a regression line (names DSF) in which slope (ΔAn/ΔCi) can be defined as dsf. We found that DSF information was embedded in both Laisk method (CO2 response curve (A/Ci) measured at three sub-saturated PARs, and their intersections were used to estimate daytime respiration (Rd), and CO2 compensation point (Ci*) and light response curve measurements, which could be used to estimate dsf values. This study investigated the relationship between dsf and the parameters related to the biochemical process and the CO2 diffusion process of photosynthesis. The results showed that dsf was negatively correlated with gs, apparent carboxylation efficiency, and apparent quantum yield. This suggests that DSF may coordinate the influence of environmental conditions (light, CO2 and water) on photosynthesis in the biochemical and CO2 diffusion process. Moreover, dsf was independent of gas exchange measurement conditions and showed species specificity. In conclusion, we speculated that dsf seems to be a comprehensive parameter that might be related to the intrinsic adaptation mechanism of plants to environmental conditions. We proposed an auxiliary line perpendicular to the DSF and used it to improve the stability of Ci* and Rd estimated from the Laisk dataset.

Overexpression of Setaria italica phosphoenolpyruvate carboxylase gene in rice positively impacts photosynthesis and agronomic traits

C4 plants have the inherent capacity to concentrate atmospheric CO2 in the vicinity of RuBisCo, thereby increasing carboxylation, and inhibiting photorespiration. Carbonic anhydrase (CA), the first enzyme of C4 photosynthesis, converts atmospheric CO2 to HCO3−, which is utilized by PEPC to produce C4 acids. Bioengineering of C4 traits into C3 crops is an attractive strategy to increase photosynthesis and water use efficiency. In the present study, we isolated the PEPC gene from the C4 plant Setaria italica and transferred it to C3 rice. Overexpression of SiPEPC resulted in a 2-6-fold increment in PEPC enzyme activity in transgenic lines with respect to non-transformed control. Photosynthetic efficiency was enhanced in transformed plants, which was associated with increased ФPSII, ETR, lower NPQ, and higher chlorophyll accumulation. Water use efficiency was increased by 16–22% in PEPC transgenic rice lines. Increased PEPC activity enhanced quantum yield and carboxylation efficiency of PEPC transgenic lines. Transgenic plants exhibited higher light saturation photosynthesis rate and lower CO2 compensation point, as compared to non-transformed control. An increase in net photosynthesis increased the yield by (23–28.9%) and biomass by (24.1–29%) in transgenic PEPC lines. Altogether, our findings indicate that overexpression of C4-specific SiPEPC enzyme is able to enhance photosynthesis and related parameters in transgenic rice.

The Evolution of C4 Photosynthesis in Flaveria (Asteraceae): Insights from the Flaveria linearis Complex.

Flaveria is a leading model for C4 plant evolution due to the presence of a dozen C3-C4 intermediate species, many of which are associated with a phylogenetic complex centered around Flaveria linearis. To investigate C4 evolution in Flaveria, we updated the Flaveria phylogeny and evaluated gas exchange, starch δ13C, and activity of C4 cycle enzymes in 19 Flaveria species and 28 populations within the F. linearis complex. A principal component analysis identified six functional clusters: (1) C3, (2) sub-C2, (3) full C2, (4) enriched C2, (5) sub-C4, and (6) fully C4 species. The sub-C2 species lacked a functional C4 cycle, while a gradient was present in the C2 clusters from little to modest C4 cycle activity as indicated by δ13C and enzyme activities. Three Yucatan populations of F. linearis had photosynthetic CO2 compensation points equivalent to C4 plants but showed little evidence for an enhanced C4 cycle, indicating they have an optimized C2 pathway that recaptures all photorespired CO2 in the bundle sheath (BS) tissue. All C2 species had enhanced aspartate aminotransferase activity relative to C3 species and most had enhanced alanine aminotransferase activity. These aminotransferases form aspartate and alanine from glutamate and in doing so could help return photorespiratory nitrogen (N) from BS to mesophyll cells, preventing glutamate feedback onto photorespiratory N assimilation. Their use requires upregulation of parts of the C4 metabolic cycle to generate carbon skeletons to sustain N return to the mesophyll, and thus could facilitate the evolution of the full C4 photosynthetic pathway.

Increased leaf area compensated photosynthetic downregulation in response to elevated CO2 and warming in white birch

Predicting photosynthetic acclimation to elevated CO2 and warming is difficult because they have opposite effects. We investigated physiological and morphological responses in white birch ( Betula papyrifera Marshall) to a combination of CO2 and temperature (ACT — 400 µmol·mol–1 CO2, current temperature; ECT — 750 µmol·mol–1 CO2, current + 4 °C temperature). ECT reduced photosynthesis, maximum Rubisco carboxylation ( Vcmax), maximum electron transport rate ( Jmax), photorespiration, daytime respiration, leaf N, and stomatal and mesophyll conductance, but increased biomass, height, total leaf area, electron partitioning to carboxylation and oxygenation ratio, and CO2 compensation point. The photosynthetic acclimation is consistent with the optimal carbon gain theory (carbon gain drives the coordination of carboxylation, electron transport, and respiration). While the photosynthetic acclimation was similar to acclimation to elevated CO2, ECT reduced Jmax/ Vcmax, which is consistent with the response to warming but opposite to the response to elevated CO2, suggesting that thermal acclimation may be the primary mechanism of photosynthetic acclimation to ECT and ECT probably altered N allocation between machinery for carboxylation and that for ribulose-1,5-bisphosphate regeneration. The increase in total leaf area by ECT more than offset the negative effect of photosynthetic downregulation on carbon sequestration, resulting in faster growth and greater biomass under ECT.

Evolution of an intermediate C4 photosynthesis in the non-foliar tissues of the Poaceae.

Carbon concentrating mechanisms (CCMs) in plants are abaptive features that have evolved to sustain plant growth in unfavorable environments, especially at low atmospheric carbon levels and high temperatures. Uptake of CO2 and its storage in the aerenchyma tissues of Lycopsids and diurnal acidity fluctuation in aquatic plants during the Palaeozoic era (ca. 300Ma.) would represent the earliest evolution of a CCM. The CCM parts of the dark reactions of photosynthesis have evolved many times, while the light reactions are conserved across plant lineages. A C4 type CCM, leaf C4 photosynthesis is evolved in the PACMAD clade of the Poaceae family. The evolution of C4 photosynthesis from C3 photosynthesis was an abaptation. Photosynthesis in reproductive tissues of sorghum and maize (PACMAD clade) has been shown to be of a weaker C4 type (high CO2 compensation point, low carbon isotope discrimination, and lack of Rubisco compartmentalization, when compared to the normal C4 types) than that in the leaves (normal C4 type). However, this does not fit well with the character polarity concept from an evolutionary perspective. In a recent model proposed for CCM evolution, the development of a rudimentary CCM prior to the evolution of a more efficient CCM (features contrasting to a weaker C4 type, leading to greater biomass production rate) has been suggested. An intermediate crassulacean acid metabolism (CAM) type of CCM (rudimentary) was reported in the genera, Brassia, Coryanthes, Eriopsis, Peristeria, of the orchids (well-known group of plants that display the CAM pathway). Similarly, we propose here the evolution of a rudimentary CCM (C4-like type pathway) in the non-foliar tissues of the Poaceae, prior to the evolution of the C4 pathway as identified in the leaves of the C4 species of the PACMAD clade.

Catalase protects against nonenzymatic decarboxylations during photorespiration in Arabidopsis thaliana.

Photorespiration recovers carbon that would be otherwise lost following the oxygenation reaction of rubisco and production of glycolate. Photorespiration is essential in plants and recycles glycolate into usable metabolic products through reactions spanning the chloroplast, mitochondrion, and peroxisome. Catalase in peroxisomes plays an important role in this process by disproportionating H2O2 resulting from glycolate oxidation into O2 and water. We hypothesize that catalase in the peroxisome also protects against nonenzymatic decarboxylations between hydrogen peroxide and photorespiratory intermediates (glyoxylate and/or hydroxypyruvate). We test this hypothesis by detailed gas exchange and biochemical analysis of Arabidopsis thaliana mutants lacking peroxisomal catalase. Our results strongly support this hypothesis, with catalase mutants showing gas exchange evidence for an increased stoichiometry of CO2 release from photorespiration, specifically an increase in the CO2 compensation point, a photorespiratory‐dependent decrease in the quantum efficiency of CO2 assimilation, increase in the 12CO2 released in a 13CO2 background, and an increase in the postillumination CO2 burst. Further metabolic evidence suggests this excess CO2 release occurred via the nonenzymatic decarboxylation of hydroxypyruvate. Specifically, the catalase mutant showed an accumulation of photorespiratory intermediates during a transient increase in rubisco oxygenation consistent with this hypothesis. Additionally, end products of alternative hypotheses explaining this excess release were similar between wild type and catalase mutants. Furthermore, the calculated rate of hydroxypyruvate decarboxylation in catalase mutant is much higher than that of glyoxylate decarboxylation. This work provides evidence that these nonenzymatic decarboxylation reactions, predominately hydroxypyruvate decarboxylation, can occur in vivo when photorespiratory metabolism is genetically disrupted.

Effects of growth temperature and nitrogen nutrition on expression of C3-C4 intermediate traits in Chenopodium album.

Proto-Kranz plants represent an initial phase in the evolution from C3 to C3-C4 intermediate to C4 plants. The ecological and adaptive aspects of C3-C4 plants would provide an important clue to understand the evolution of C3-C4 plants. We investigated whether growth temperature and nitrogen (N) nutrition influence the expression of C3-C4 traits in Chenopodium album (proto-Kranz) in comparison with Chenopodium quinoa (C3). Plants were grown during 5weeks at 20 or 30°C under standard or low N supply levels (referred to as 20SN, 20LN, 30SN, and 30LN). Net photosynthetic rate and leaf N content were higher in 20SN and 30SN plants than in 20LN and 30LN plants of C. album but did not differ among growth conditions in C. quinoa. The CO2 compensation point (Γ) of C. album was lowest in 30LN plants (36µmolmol-1), highest in 20SN plants (51µmolmol-1), and intermediate in 20LN and 30SN plants, whereas Γ of C. quinoa did not differ among the growth conditions (51-52µmolmol-1). The anatomical structure of leaves was not considerably affected by growth conditions in either species. However, ultrastructural observations in C. album showed that the number of mitochondria per mesophyll or bundle sheath (BS) cell was lower in 20LN and 30LN plants than in 20SN and 30SN plants. Immunohistochemical observations revealed that lower accumulation level of P-protein of glycine decarboxylase (GDC-P) in mesophyll mitochondria than in BS mitochondria is the major factor causing the decrease in Γ values in C. album plants grown under low N supply and high temperature. These results suggest that high growth temperature and low N supply lead to the expression of C3-C4 traits (the reduction of Γ) in the proto-Kranz plants of C. album through the regulation of GDC-P expression.

Elevated efficiency of C3 photosynthesis in bamboo grasses: A possible consequence of enhanced refixation of photorespired CO2

AbstractBamboos are productive grasses that currently yield a high‐quality wood and potentially an abundance of lignocellulose for bioenergy. All are C3 grasses of warm habitats, where they are prone to significant photorespiratory inhibition and competitive suppression by C4 grasses. Here, we investigate whether three bamboo species from the Brazilian Cerrado (Dendrocalamus asper, Guadua angustifolia, and Guadua magna) exhibit unique adaptations that suppress photorespiratory costs and enhance photosynthetic efficiency. We evaluated photosynthetic efficiency of the bamboos and rice (Oryza sativa) by measuring C*, the CO2 compensation point in the absence of mitochondrial respiration. At 25℃, C* averaged 2.81 Pa in each of the bamboo species, which is closer to a C2 plant (2.71 Pa) than the C3 plant rice (3.31 Pa). Assuming a chloroplast CO2 concentration of 200 µmol mol−1, this represents an 18% lower cost of apparent photorespiration in bamboo than rice. Light and transmission electronic microscopy of the bamboo leaves exhibited few organelles in the bundle and mestome sheath cells, and mesophyll (M) cells are deeply lobed with 99% of the cell periphery adjacent to intercellular air space covered by chloroplast and stromules. The chloroplast layer in bamboo M cells is thick, with mitochondria adjacent to or engulfed by chloroplasts. This arrangement slows CO2 efflux and facilitates refixation of photorespired CO2, which could explain the low C* in the bamboos. The bamboos also had higher water use efficiency than rice, which may reflect efficient refixation of photorespired CO2.

A low CO2-responsive mutant of Setaria viridis reveals that reduced carbonic anhydrase limits C4 photosynthesis.

In C4 species, β-carbonic anhydrase (CA), localized to the cytosol of the mesophyll cells, accelerates the interconversion of CO2 to HCO3–, the substrate used by phosphoenolpyruvate carboxylase (PEPC) in the first step of C4 photosynthesis. Here we describe the identification and characterization of low CO2-responsive mutant 1 (lcr1) isolated from an N-nitroso-N-methylurea- (NMU) treated Setaria viridis mutant population. Forward genetic investigation revealed that the mutated gene Sevir.5G247800 of lcr1 possessed a single nucleotide transition from cytosine to thymine in a β-CA gene causing an amino acid change from leucine to phenylalanine. This resulted in severe reduction in growth and photosynthesis in the mutant. Both the CO2 compensation point and carbon isotope discrimination values of the mutant were significantly increased. Growth of the mutants was stunted when grown under ambient pCO2 but recovered at elevated pCO2. Further bioinformatics analyses revealed that the mutation has led to functional changes in one of the conserved residues of the protein, situated near the catalytic site. CA transcript accumulation in the mutant was 80% lower, CA protein accumulation 30% lower, and CA activity ~98% lower compared with the wild type. Changes in the abundance of other primary C4 pathway enzymes were observed; accumulation of PEPC protein was significantly increased and accumulation of malate dehydrogenase and malic enzyme decreased. The reduction of CA protein activity and abundance in lcr1 restricts the supply of bicarbonate to PEPC, limiting C4 photosynthesis and growth. This study establishes Sevir.5G247800 as the major CA allele in Setaria for C4 photosynthesis and provides important insights into the function of CA in C4 photosynthesis that would be required to generate a rice plant with a functional C4 biochemical pathway.