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Sugar beet leaves can be a viable and economically interesting source of high-quality protein for the food industry. We investigated how storage conditions and leaf wounding at harvest affect the content and quality of the soluble protein. After collection, leaves were either stored intact or shredded to mimic wounding induced by commercial leaf harvesters. Leaf material was stored in small volumes at different temperatures to assess leaf physiology or in larger volumes to assess temperature development at different locations in the bins. Protein degradation was more pronounced at higher storage temperatures. Wounding accelerated the degradation of soluble protein at all temperatures. Both wounding and storage at higher temperatures greatly stimulated respiration activity and heat production. At temperatures below 5°C, ribulose-1,5-biphosphate carboxylase oxygenase (RuBisCO) in intact leaves was preserved for up to 3 weeks. At temperatures of 30–40°C, RuBisCO degradation occurred within 48 h. Degradation was more pronounced in shredded leaves. In 0.8-m3 storage bins at ambient temperature, core temperatures rapidly increased, up to 25°C in intact leaves and up to 45°C in shredded leaves within 2–3 days. Immediate storage at 5°C greatly suppressed the temperature increase in intact but not in shredded leaves. The indirect effect of excessive wounding, that is, heat production, is discussed as the pivotal factor responsible for increased degradation of protein. For optimal retention of soluble protein levels and quality in harvested sugar beet leaves, it is advised to minimize wounding and to store the material at temperatures around −5°C. Practical Application: To preserve the soluble protein content and quality for at least 3 weeks, sugar beet leaves should be harvested with minimal wounding and stored at temperatures between 1 and 5°C. When aiming to store minimally wounded leaves in larger volumes, it must be ensured that the product temperature in the core of the biomass meets the temperature criterium or the cooling strategy must be adjusted. The principles of minimal wounding and low temperature storage are transferable to other leafy crops that are harvested for food protein.

The use of alfalfa as a dietary protein source is of great interest. Membrane filtration shows potential for removal of unwanted compounds in the purification of ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCO), but fouling remains a challenge. It was hypothesized that treatments, namely centrifugation, ultracentrifugation, and heat clearing, prior to membrane processing modify the supramolecular assembly of the proteins, facilitating its filtration. The pre-treatments led to different aggregate structures. In particular, heating at 50 °C decreased the aggregate sizes by ∼500 folds relative to the raw juice, as measured by light scattering. The molar mass of RuBisCO decreased by ∼180 kDa after treatment with heat. Similarly, supernatants after ultracentrifugation also showed smaller structures. Unlike the other treatments, heating led to an efficient transmission of RuBisCO during microfiltration. These results demonstrate for the first time, the importance of controlling the supramolecular structure of RuBisCO when processing proteins from alfalfa juice. Industrial relevance: The demand for more sustainable protein sources is continuously growing. RuBisCO is of particular interest because of its favorable amino acid composition. However, improved purification strategies are needed to extract it from green biomass. The results of this study serve as a proof-of-concept for the application of an industrially relevant process, mild heat treatment in combination with microfiltration, for the purification of RuBisCO from alfalfa green biomass.

The aim of the study was to investigate the effect of co-precipitation of red clover protein and casein in combination with sulphite. Previous studies have shown antioxidation effects of introducing sulphite in the extraction of red clover protein for potential food applications. In the present study, it was hypothesized that casein co-precipitation in combination with sulphite would improve red clover's protein recovery, quality, and protein digestibility. The amount of soluble protein in the red clover juice was increased by the presence of casein, determined by both an increase in the total protein and in the amount of the photosynthetic protein RuBisCO. Casein treatment did not affect the in vitro protein digestibility, whereas the digestibility was increased with the addition of sulphite as an effect of phenolic oxidation inhibition.

The history of Earth’s carbon cycle reflects trends in atmospheric composition convolved with the evolution of photosynthesis. Fortunately, key parts of the carbon cycle have been recorded in the carbon isotope ratios of sedimentary rocks. The dominant model used to interpret this record as a proxy for ancient atmospheric CO 2 is based on carbon isotope fractionations of modern photoautotrophs, and longstanding questions remain about how their evolution might have impacted the record. Therefore, we measured both biomass (ε p ) and enzymatic (ε Rubisco ) carbon isotope fractionations of a cyanobacterial strain ( Synechococcus elongatus PCC 7942) solely expressing a putative ancestral Form 1B rubisco dating to ≫1 Ga. This strain, nicknamed ANC, grows in ambient pCO 2 and displays larger ε p values than WT, despite having a much smaller ε Rubisco (17.23 ± 0.61‰ vs. 25.18 ± 0.31‰, respectively). Surprisingly, ANC ε p exceeded ANC ε Rubisco in all conditions tested, contradicting prevailing models of cyanobacterial carbon isotope fractionation. Such models can be rectified by introducing additional isotopic fractionation associated with powered inorganic carbon uptake mechanisms present in Cyanobacteria, but this amendment hinders the ability to accurately estimate historical pCO 2 from geological data. Understanding the evolution of rubisco and the CO 2 concentrating mechanism is therefore critical for interpreting the carbon isotope record, and fluctuations in the record may reflect the evolving efficiency of carbon fixing metabolisms in addition to changes in atmospheric CO 2 .

Carboxysomes are a family of bacterial microcompartments (BMCs), present in all cyanobacteria and some proteobacteria, which encapsulate the primary CO2-fixing enzyme, Rubisco, within a virus-like polyhedral protein shell. Carboxysomes provide significantly elevated levels of CO2 around Rubisco to maximize carboxylation and reduce wasteful photorespiration, thus functioning as the central CO2-fixation organelles of bacterial CO2-concentration mechanisms. Their intriguing architectural features allow carboxysomes to make a vast contribution to carbon assimilation on a global scale. In this review, we discuss recent research progress that provides new insights into the mechanisms of how carboxysomes are assembled and functionally maintained in bacteria and recent advances in synthetic biology to repurpose the metabolic module in diverse applications.

Carbon fixation is the process by which CO2 is converted from a gas into biomass. The Calvin–Benson–Bassham cycle (CBB) is the dominant carbon-consuming pathway on Earth, driving >99.5% of the ∼120 billion tons of carbon that are converted to sugar by plants, algae, and cyanobacteria. The carboxylase enzyme in the CBB, ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco), fixes one CO2 molecule per turn of the cycle into bioavailable sugars. Despite being critical to the assimilation of carbon, rubisco's kinetic rate is not very fast, limiting flux through the pathway. This bottleneck presents a paradox: Why has rubisco not evolved to be a better catalyst? Many hypothesize that the catalytic mechanism of rubisco is subject to one or more trade-offs and that rubisco variants have been optimized for their native physiological environment. Here, we review the evolution and biochemistry of rubisco through the lens of structure and mechanism in order to understand what trade-offs limit its improvement. We also review the many attempts to improve rubisco itself and thereby promote plant growth. Expected final online publication date for the Annual Review of Biochemistry, Volume 92 is June 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Form I rubiscos evolved in Cyanobacteria ≥ 2.5 billion years ago and are enzymatically unique due to the presence of small subunits (RbcS) capping both ends of an octameric large subunit (RbcL) rubisco assembly to form a hexadecameric (L8S8) holoenzyme. Although RbcS was previously thought to be integral to Form I rubisco stability, the recent discovery of a closely related sister clade of octameric rubiscos (Form I’; L8) demonstrates that the L8 complex can assemble without small subunits (Banda et al. 2020). Rubisco also displays a kinetic isotope effect (KIE) where the 3PG product is depleted in 13C relative to 12C. In Cyanobacteria, only two Form I KIE measurements exist, making interpretation of bacterial carbon isotope data difficult. To aid comparison, we measured in vitro the KIEs of Form I’ (Candidatus Promineofilum breve) and Form I (Synechococcus elongatus PCC 6301) rubiscos and found the KIE to be smaller in the L8 rubisco (16.25 ± 1.36‰ vs. 22.42 ± 2.37‰, respectively). Therefore, while small subunits may not be necessary for protein stability, they may affect the KIE. Our findings may provide insight into the function of RbcS and allow more refined interpretation of environmental carbon isotope data.

The overexpression of the RuBisCO (rbc) gene has recently become an achievable strategy for increasing cyanobacterial biomass and overcoming the biocompound production restriction. We successfully constructed two rbc-overexpressing Synechocystis sp. PCC 6803 strains (OX), including a strain overexpressing a large subunit of RuBisCO (OXrbcL) and another strain overexpressing all large, chaperone, and small subunits of RuBisCO (OXrbcLXS), resulting in higher and faster growth than wild type under sodium bicarbonate supplementation. This increased biomass of OX strains significantly contributed to the higher polyhydroxybutyrate (PHB) production induced by nutrient-deprived conditions, in particular nitrogen (N) and phosphorus (P). As a result of higher PHB contents in OX strains occurring at days 7 and 9 of nutrient deprivation, this enhancement was apparently made possible by cells preferentially maintaining their internal lipids while accumulating less glycogen. The OXrbcLXS strain, with the highest level of PHB at about 39 %w/dry cell weight (DCW) during 7 days of BG11-NP treatment, contained a lower glycogen level (31.9 %w/DCW) than wild type control (40 %w/DCW). In contrast, the wild type control strain exposed to N- and NP-stresses tended to retain lipid levels and store more glycogen than PHB. In this model, we, for the first time, implemented a RuBisCO-overexpressing cyanobacterial factory for overproducing PHB, destined for biofuel and biomaterial biotechnology.

Duckweeds (Lemnaceae) are representative producers in fresh aquatic ecosystems and also yield sustainable biomass for animal feeds, human foods, and biofuels, and contribute toward effective wastewater treatment; thus, enhancing duckweed productivity is a critical challenge. Plant-growth-promoting bacteria (PGPB) can improve the productivity of terrestrial plants; however, duckweed–PGPB interactions remain unclear and no previous study has investigated the molecular mechanisms underlying duckweed–PGPB interaction. Herein, a PGPB, Ensifer sp. strain SP4, was newly isolated from giant duckweed (Spirodela polyrhiza), and the interactions between S. polyrhiza and SP4 were investigated through physiological, biochemical, and metabolomic analyses. In S. polyrhiza and SP4 coculture, SP4 increased the nitrogen (N), chlorophyll, and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) contents and the photosynthesis rate of S. polyrhiza by 2.5-, 2.5-, 2.7-, and 2.4-fold, respectively. Elevated photosynthesis increased the relative growth rate and biomass productivity of S. polyrhiza by 1.5- and 2.7-fold, respectively. Strain SP4 significantly altered the metabolomic profile of S. polyrhiza, especially its amino acid profile. N stable isotope analysis revealed that organic N compounds were transferred from SP4 to S. polyrhiza. These N compounds, particularly glutamic acid, possibly triggered the increase in photosynthetic and growth activities. Accordingly, we propose a new model for the molecular mechanism underlying S. polyrhiza growth promotion by its associated bacteria Ensifer sp. SP4, which occurs through enhanced N compound metabolism and photosynthesis. Our findings show that Ensifer sp. SP4 is a promising PGPB for increasing biomass yield, wastewater purification activity, and CO2 capture of S. polyrhiza. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Two-Spotted Spider Mites (TSSMs, Tetranychus urticae Koch 1836 (Acari: Tetranychidae)) is one of the most important pests in many crop plants, and their feeding activity is based on sucking leaf cell contents. The purpose of this study was to evaluate the interaction between TSSMs and their host Lima bean (Phaseolus lunatus) by analyzing the metabolomics of leaf pigments and the transcriptomics of TSSM guanine production. We also used epifluorescence, confocal laser scanning, and transmission electron microscopies to study the morphology and structure of TSSMs and their excreta. Finally, we evaluated the potential photosynthetic ability of TSSMs and the activity and content of Ribulose-1,5-bisphosphate Carboxylase/Oxigenase (RubisCO). We found that TSSMs express several genes involved in guanine production, including Guanosine Monophosphate Synthetase (GMPS) and decoyinine (DCY), a potential inhibitor of GMPS, was found to reduce TSSMs proliferation in infested Lima bean leaves. Despite the presence of intact chloroplasts and chlorophyll in TSSMs, we demonstrate that TSSMs do not retain any photosynthetic activity. Our results show for the first time the transcriptomics of guanine production in TSSMs and provide new insight into the catabolic activity of TSSMs on leaf chlorophyll and carotenoids. Finally, we preliminary demonstrate that DCY has an acaricidal potential against TSSMs.

Background Anaerobic Saccharomyces cerevisiae cultures require glycerol formation to re-oxidize NADH formed in biosynthetic processes. Introduction of the Calvin-cycle enzymes phosphoribulokinase (PRK) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) has been shown to couple re-oxidation of biosynthetic NADH to ethanol production and improve ethanol yield on sugar in fast-growing batch cultures. Since growth rates in industrial ethanol production processes are not constant, performance of engineered strains was studied in slow-growing cultures. Results In slow-growing anaerobic chemostat cultures (D = 0.05 h−1), an engineered PRK/RuBisCO strain produced 80-fold more acetaldehyde and 30-fold more acetate than a reference strain. This observation suggested an imbalance between in vivo activities of PRK/RuBisCO and formation of NADH in biosynthesis. Lowering the copy number of the RuBisCO-encoding cbbm expression cassette from 15 to 2 reduced acetaldehyde and acetate production by 67% and 29%, respectively. Additional C-terminal fusion of a 19-amino-acid tag to PRK reduced its protein level by 13-fold while acetaldehyde and acetate production decreased by 94% and 61%, respectively, relative to the 15 × cbbm strain. These modifications did not affect glycerol production at 0.05 h−1 but caused a 4.6 fold higher glycerol production per amount of biomass in fast-growing (0.29 h−1) anaerobic batch cultures than observed for the 15 × cbbm strain. In another strategy, the promoter of ANB1, whose transcript level positively correlated with growth rate, was used to control PRK synthesis in a 2 × cbbm strain. At 0.05 h−1, this strategy reduced acetaldehyde and acetate production by 79% and 40%, respectively, relative to the 15 × cbbm strain, without affecting glycerol production. The maximum growth rate of the resulting strain equalled that of the reference strain, while its glycerol production was 72% lower. Conclusions Acetaldehyde and acetate formation by slow-growing cultures of engineered S. cerevisiae strains carrying a PRK/RuBisCO bypass of yeast glycolysis was attributed to an in vivo overcapacity of PRK and RuBisCO. Reducing the capacity of PRK and/or RuBisCO was shown to mitigate this undesirable byproduct formation. Use of a growth rate-dependent promoter for PRK expression highlighted the potential of modulating gene expression in engineered strains to respond to growth-rate dynamics in industrial batch processes.

Fusarium crown rot (FCR), typically caused by Fusarium pseudograminearum, is a severe soil-borne disease that, in recent years, has become an emerging threat to Chinese wheat crops. For the first time in this study, we investigated and compared the proteomic characteristics of two Chinese wheat varieties (04 Zhong 36 and Xinmai 26) at 24, 48, and 72 h post-inoculation using label-free quantitative proteomic analysis. A total of 9 234 proteins were successfully quantified, of which 783 were differentially expressed after inoculation. These proteins were mainly involved in metabolic, single-organism, and cellular processes. Thirty-three proteins associated with defense, cell wall formation, photosynthesis, etc., showed consistently different expression between the two genotypes at multiple time points. In particular, chitinase, which degrades chitin in the fungal cell wall and limits fungal growth, was exclusively and consistently upregulated in 04 Zhong 36 across the three time points. Other proteins such as flavonoid O-methyltransferase, glycosyltransferase, and peroxidase were only upregulated in 04 Zhong 36, and proteins, including the berberine bridge enzyme and rubisco large subunit-binding protein, were specifically downregulated in Xinmai 26. The expression of transcripts encoding eight selected proteins through qRT-PCR analysis supported the proteomic profiles. Overall, the results of this study allow us to understand FCR resistance in wheat at the protein level. Some proteins and their corresponding genes may be useful resources for the genetic improvement of FCR resistance in wheat.

The Calvin-Benson-Bassham cycle and Wood-Ljungdahl pathway are two well-known ones in the six carbon sequestration pathways, but the current knowledge of their occurrence in different layers of agricultural soil profiles is poor. In this study, the diversities of three genes encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), i.e., genes encoding the green-like (cbbLG) and red-like (cbbLR) forms of RubisCO I and encoding RubisCO II (cbbM), and the gene encoding carbon monoxide dehydrogenase large subunit (coxL) from five paddy soils in southern China were investigated by real-time quantitative polymerase chain reaction, restriction fragment length polymorphism (RFLP) analysis, and clone library. The abundances of the four genes rang ed from 107 to 109 copies g–1 soil, and the cbbLR gene outnumbered the other three genes in all soil samples, suggesting important roles they play in carbon dioxide (CO2) fixation. In addition, it was found that the copy numbers of cbbLR and cbbLG decreased with increasing soil depth, while the copy numbers of cbbM and coxL decreased in the shallow depths but increased with increasing soil depth. The results of RFLP showed a larger Shannon index (H) in the deeper soil layers among the four gene clone libraries, indicating that the community diversity in these soil layers was greater. The cbbLG gene had relatively low diversity (at genus level), and most of the sequences were classified as Sideroxydans and Thiobacillus. In contrast, the highly diverse groups were found in the other three gene clone libraries (cbbLR, cbbM, and coxL), most of which were distantly related to known sequences, even forming separate clusters. In summary, this study provides a new insight into CO2 fixers along agricultural soil profiles by comparing four bacterial genes.

Angiotensin I converting enzyme (ACE) inhibition is one of the key factors for cardiovascular disease, and inhibition of ACE activity is related to the prevention of high blood pressure. Until now, some ACE inhibitory peptides have been found in the protein hydrolysates of red algae, and most of them were derived from phycobiliproteins and rubisco. In this study, we modified the preparation method to evaluate the potential of ACE inhibitory activity from protein hydrolysate of red algaMazzaella japonicaexcept for phycobiliproteins and rubisco. As a result, we identified 11 peptides (YRD, VSEGLD, TIMPHPR, GGPAT, SSNDYPI, SRIYNVKSNG, VDAHY, CPYDWV, YGDPDHY, NLGN and DFGVPGHEP) in the hydrolysate by the reversed phase-HPLC and MALDI-TOF/MS/MS. Among them, YRD would be derived from phycoerythrin alpha-subunit. However, the others would be from other proteins encoded in chloroplast or nuclear genomes. This study revealed that not only phycobiliproteins and rubisco but also the other proteins in red algae have the potential for the source of ACE inhibitory peptide.

Interrogative proteome analyses are used to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in heat-stressed plants. The objectives of the study were to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in chickpea (Cicer arietinum L.) heat-tolerant (Acc#7) and sensitive genotype (Acc#8) from a field study. Proteomic and gene ontological analyses showed an upregulation in proteins related to protein synthesis, intracellular traffic, defence and transport in the heat-tolerant genotype compared to the susceptible one at the warmer site. Results from KEGG analyses indicate the involvement of probable sucrose-phosphate synthase (EC 2.4.1.14) and sucrose-phosphate phosphatase (EC 3.1.3.24) proteins, that were upregulated in the heat-tolerant genotype at the warmer site, in the starch and sucrose pathway. The presence of these differentially regulated proteins including HSP70, ribulose bisphosphate carboxylase/oxygenase activase, plastocyanin and protoporphyrinogen oxidase suggests their potential role in heat tolerance, at flowering growth stage, in field-grown chickpea. This observation supports unaltered physiological and biochemical performance of the heat-tolerant genotypes (Acc#7) relative to the susceptible genotype (Acc#8) in related studies (Makonya et al. 2019). Characterisation of the candidate proteins identified in the current study as well as their specific roles in the tolerance to heat stress in chickpea are integral to further crop improvement initiatives.

Most microalgae overcome the difficulty of acquiring inorganic carbon (Ci) in aquatic environments by inducing a CO2-concentrating mechanism (CCM). In the green alga Chlamydomonas reinhardtii, two distinct photosynthetic acclimation states have been described under CO2-limiting conditions (low-CO2 [LC] and very low-CO2 [VLC]). LC-inducible protein B (LCIB), structurally characterized as carbonic anhydrase, localizes in the chloroplast stroma under CO2-supplied and LC conditions. In VLC conditions, it migrates to aggregate around the pyrenoid, where the CO2-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase is enriched. Although the physiological importance of LCIB localization changes in the chloroplast has been shown, factors necessary for the localization changes remain uncertain. Here, we examined the effect of pH, light availability, photosynthetic electron flow, and protein synthesis on the localization changes, along with measuring Ci concentrations. LCIB dispersed or localized in the basal region of the chloroplast stroma at 8.3–15 µM CO2, whereas LCIB migrated toward the pyrenoid at 6.5 µM CO2. Furthermore, LCIB relocated toward the pyrenoid at 2.6–3.4 µM CO2, even in cells in the dark or treated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea and cycloheximide in light. In contrast, in the mutant lacking CCM1, a master regulator of CCM, LCIB remained dispersed even at 4.3 µM CO2. Meanwhile, a simultaneous expression of LCIC, an interacting protein of LCIB, induced the localization of several speckled structures at the pyrenoid periphery. These results suggest that the localization changes of LCIB require LCIC and are controlled by CO2 concentration with ∼7 µM as the boundary.

Chloroplasts are the sites for photosynthesis, and two Golden2-like factors act as transcriptional activators of chloroplast development in rice (Oryza sativa L.) and maize (Zea mays L.). Rice OsGLK1 and OsGLK2 are orthologous to maize ZmGLK1 (ZmG1) and ZmGLK2 (ZmG2), respectively. However, while rice OsGLK1 and OsGLK2 act redundantly to regulate chloroplast development in mesophyll cells, maize ZmG1 and ZmG2 are functionally specialized and expressed in different cell-specific manners. To boost rice chloroplast development and photosynthesis, we generated transgenic rice plants overexpressing ZmG1 and ZmG2, individually or simultaneously, with constitutive promoters (pZmUbi::ZmG1 and p35S::ZmG2) or maize promoters (pZmG1::ZmG1, pZmG2::ZmG2, and pZmG1::ZmG1/pZmG2::ZmG2). Both ZmG1 and ZmG2 genes were highly expressed in transgenic rice leaves. Moreover, ZmG1 and ZmG2 showed coordinated expression in pZmG1::ZmG1/pZmG2::ZmG2 plants. All Golden2-like (GLK) transgenic plants had higher chlorophyll and protein contents, Rubisco activities and photosynthetic rates per unit leaf area in flag leaves. However, the highest grain yields occurred when maize promoters were used; pZmG1::ZmG1, pZmG2::ZmG2, and pZmG1::ZmG1/pZmG2::ZmG2 transgenic plants showed increases in grain yield by 51%, 47%, and 70%, respectively. In contrast, the pZmUbi::ZmG1 plant produced smaller seeds without yield increases. Transcriptome analysis indicated that maize GLKs act as master regulators promoting the expression of both photosynthesis-related and stress-responsive regulatory genes in both rice shoot and root. Thus, by promoting these important functions under the control of their own promoters, maize GLK1 and GLK2 genes together dramatically improved rice photosynthetic performance and productivity. A similar approach can potentially improve the productivity of many other crops.

The effect of an elevated concentration of СО2 (800 ppm) on the growth rate, efficiency of photosystem I (PS I) and photosystem II (PS II), content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPc), as well as proline and sodium and potassium ions, was investigated in the shoots of С4-halophyte Kochia prostrata (L.) Schrad. An elevated concentration of СО2 caused a decrease in accumulation of dry biomass (1.2 times), a rise in the content of proline (1.2 times) and potassium ions (1.3 times), and a shortening of time required to reach the maximum P700 oxidation level (PS I) (1.7 times). The content of sodium ions and water in the shoots and efficiency of PS II (Fv/Fm) did not change. Immunoblotting showed that, at the elevated concentration of СО2, the ratio between Rubisco and PEPc proteins rose from 1.3 to 3.4. The shortening of time required to reach the maximum oxidation level of PS I indirectly points to a reduction in the activity of cyclic transport of electrons responsible for efficient operation of the С4-carbon-concentrating mechanism. The high concentration of СО2 is suggested to activate the С3-pathway of photosynthesis in С4-halophyte K. prostrata.

Carboxysomes in cyanobacteria enclose the enzymes Rubisco and carbonic anhydrase to optimize photosynthetic carbon fixation. Understanding carboxysome assembly has implications in agricultural biotechnology. Here we analyzed the role of the scaffolding protein CcmM of the β-cyanobacterium Synechococcus elongatus PCC 7942 in sequestrating the hexadecameric Rubisco and the tetrameric carbonic anhydrase, CcaA. We find that the trimeric CcmM, consisting of γCAL oligomerization domains and linked small subunit-like (SSUL) modules, plays a central role in mediation of pre-carboxysome condensate formation through multivalent, cooperative interactions. The γCAL domains interact with the C-terminal tails of the CcaA subunits and additionally mediate a head-to-head association of CcmM trimers. Interestingly, SSUL modules, besides their known function in recruiting Rubisco, also participate in intermolecular interactions with the γCAL domains, providing further valency for network formation. Our findings reveal the mechanism by which CcmM functions as a central organizer of the pre-carboxysome multiprotein matrix, concentrating the core components Rubisco and CcaA before β-carboxysome shell formation., Biochemical, biophysical and structural analysis reveals how the scaffolding protein CcmM recruits the enzymes Rubisco and carbonic anhydrase into a condensate for encapsulation into carboxysomes—microcompartments in cyanobacteria that serve to optimize CO2 assimilation.

Here, we describe a model of C3, C3–C4 intermediate, and C4 photosynthesis that is designed to facilitate quantitative analysis of physiological measurements. The model relates the factors limiting electron transport and carbon metabolism, the regulatory processes that coordinate these metabolic domains, and the responses to light, carbon dioxide, and temperature. It has three unique features. First, mechanistic expressions describe how the cytochrome b6f complex controls electron transport in mesophyll and bundle sheath chloroplasts. Second, the coupling between the mesophyll and bundle sheath expressions represents how feedback regulation of Cyt b6f coordinates electron transport and carbon metabolism. Third, the temperature sensitivity of Cyt b6f is differentiated from that of the coupling between NADPH, Fd, and ATP production. Using this model, we present simulations demonstrating that the light dependence of the carbon dioxide compensation point in C3–C4 leaves can be explained by co-occurrence of light saturation in the mesophyll and light limitation in the bundle sheath. We also present inversions demonstrating that population-level variation in the carbon dioxide compensation point in a Type I C3–C4 plant, Flaveriachloraefolia, can be explained by variable allocation of photosynthetic capacity to the bundle sheath. These results suggest that Type I C3–C4 intermediate plants adjust pigment and protein distributions to optimize the glycine shuttle under different light and temperature regimes, and that the malate and aspartate shuttles may have originally functioned to smooth out the energy supply and demand associated with the glycine shuttle. This model has a wide range of potential applications to physiological, ecological, and evolutionary questions.

Aim. To isolate and purify protein complexes – ATP synthase and RuBisCO – from pea leaf chloroplasts and study the effect of a microbiological fertilizer “Extracon” and sulfonamide inhibitors acetazolamide and ethoxyzolamide on the enzymatic activity of these proteins.Materials and methods. Chloroplasts were isolated from the leaves of two-week-old pea sprouts, protein complexes of purified thylakoid membranes were solubilized with digitonin (10 mg of digitonin per 1 mg of protein), the protein concentration was determined according to Lowry. Native electrophoresis with displacement of the charge of the soluble protein fraction from the chloroplast stroma, as well as membrane proteins, was carried out in the modified system of Anderson et al., Kolisnichenko et al. A modified Lemmley system was applied to the protein electrophoresis in the polyacrylamide gel in the presence of sodium dodecyl sulfate. The methods of Alain and Hintsik, as well as Gomorrah were used to determine the ATPase activity in the polyacrylamide gel. Visualization of the carbonic anhydrase activity in the polyacrylamide gel was performed by the method of Edwards and Petton. Results and discussion. Using physicochemical methods of potentiometry, spectrophotometry the ATPase, carbonic anhydrase and esterase activities of the enzymes were studied. The results obtained indicate that specific carbonic anhydrase inhibitors (acetazolamide and ethoxyzolamide) also block the esterase and ATPase activity of the enzyme complexes. “Extracon” (a multifunctional microbiological preparation) almost 1.5 times increases the activity of the enzymes, showing a complex activating effect of the fertilizer on both light and dark reactions of photosynthesis.Conclusions. The method of identification and isolation of RuBisCO and ATP synthase on the basis of two-dimensional electrophoresis and electrophoretic elution has been proposed. It allows determining the presence of certain enzyme activity of complexes at first in SDS plates (express analysis) and further to study the effect of various factors of endogenous and exogenous origin on the enzymatic properties of electrophoretically pure enzymes. The use of two-dimensional electrophoresis as a tool for assessing the impact of various factors of endogenous and exogenous origin on the plant cell and the plant as a whole through constant monitoring of the work and activity of enzyme systems of the plant cell is promising.

Domestication involves dramatic phenotypic and physiological diversifications due to successive selection by breeders toward high yield and quality. Although photosynthetic nitrogen use efficiency (PNUE) is a major trait for understanding leaf nitrogen economy, it is unclear whether PNUE of cotton has been improved under domestication. Here, we investigated the effect of domestication on nitrogen allocation to photosynthetic machinery and PNUE in 25 wild and 37 domesticated cotton genotypes. The results showed that domesticated genotypes had higher nitrogen content per mass (Nm), net photosynthesis under saturated light (Asat), and PNUE but similar nitrogen content per area (Na) compared with wild genotypes. As expected, in both genotypes, PNUE was positively related to Asat but negatively correlated with Na. However, the relative contribution of Asat to PNUE was greater than the contribution from Na. Domesticated genotypes had higher nitrogen allocation to light-harvesting (NL, nitrogen in light-harvesting chlorophyll-protein complex), to bioenergetics (Nb, total nitrogen of cytochrome f, ferredoxin NADP reductase, and the coupling factor), and to Rubisco (Nr) than wild genotypes; however, the two genotype groups did not differ in PNUEp, the ratio of Asat to Np (itself the sum of NL, Nb, and Nr). Our results suggest that more nitrogen allocation to photosynthetic machinery has boosted Asat under cotton domestication. Improving the efficiency of nitrogen use in photosynthetic machinery might be future aim to enhance Asat of cotton.

The complete genome sequence of the thermophilic purple sulfur bacterium Thermochromatium tepidum strain MCT (DSM 3771T) is described and contrasted with that of its mesophilic relative Allochromatium vinosum strain D (DSM 180T) and other Chromatiaceae. The Tch. tepidum genome is a single circular chromosome of 2,958,290 base pairs with no plasmids and is substantially smaller than the genome of Alc. vinosum. The Tch. tepidum genome encodes two forms of RuBisCO and contains nifHDK and several other genes encoding a molybdenum nitrogenase but lacks a gene encoding a protein that assembles the Fe–S cluster required to form a functional nitrogenase molybdenum-iron cofactor, leaving the phototroph phenotypically Nif–. Tch. tepidum contains genes necessary for oxidizing sulfide to sulfate as photosynthetic electron donor but is genetically unequipped to either oxidize thiosulfate as an electron donor or carry out assimilative sulfate reduction, both of which are physiological hallmarks of Alc. vinosum. Also unlike Alc. vinosum, Tch. tepidum is obligately phototrophic and unable to grow chemotrophically in darkness by respiration. Several genes present in the Alc. vinosum genome that are absent from the genome of Tch. tepidum likely contribute to the major physiological differences observed between these related purple sulfur bacteria that inhabit distinct ecological niches.

Photosynthesis is a fundamental biosynthetic process in plants that can enhance carbon absorption and increase crop productivity. Heat stress severely inhibits photosynthetic efficiency. Melatonin is a bio-stimulator capable of regulating diverse abiotic stress tolerances. However, the underlying mechanisms of melatonin-mediated photosynthesis in plants exposed to heat stress largely remain elucidated. Our results revealed that melatonin treatment (100 μM) in tomato seedlings increased the endogenous melatonin levels and photosynthetic pigment content along with upregulated of their biosynthesis gene expression under high-temperature stress (42 °C for 24 h), whereas heat stress significantly decreased the values of gas exchange parameters. Under heat stress, melatonin boosted CO2 assimilation, i.e., Vc,max (maximum rate of ribulose-1,5-bisphosphate carboxylase, Rubisco), and Jmax (electron transport of Rubisco generation) and also enhanced the Rubisco and FBPase activities, which resulted in upregulated photosynthetic related gene expression. In addition, heat stress greatly reduced the photochemical chemistry of photosystem II (PSII) and photosystem I (PSI), particularly the maximum quantum efficiency of PSII (Fv/Fm) and PSI (Pm). Conversely, melatonin supplementation increased the chlorophyll a fluorescence parameters led to amplifying the electron transport efficiency. Moreover, heat stress decreased the actual PSII efficiency (ΦPSII), electron transport rate (ETR) and photochemical quenching coefficient (qP), while increasing nonphotochemical quenching (NPQ); however, melatonin reversed these values, which helps to fostering the dissipation of excess excitation energy. Taken together, our results provide a concrete insight into the efficacy of melatonin-mediated photosynthesis performance in a high-temperature regime.

In this study, we studied and developed the modification schemes of environmentally friendly substitutes of neonicotinoid insecticides (NNIs) along with the regulatory measures that effectively enhanced the synergistic degradation of NNIs by soil rhizobia and carbon-fixing bacteria. Firstly, the binding ability of NNIs to the two key proteins was characterized by molecular docking; secondly, the mean square deviation decision method, which is a comprehensive evaluation method, was used to investigate the binding ability of NNI molecules with the two Rubisco rate-limiting enzymes. The three-dimensional quantitative structure-activity relationship (3D-QSAR) model was established for the synergistic degradation and single effect of rhizobia and carbon-fixing bacteria. Finally, after combining the 3D-QSAR model with a contour map analysis of the synergistic degradation effect of soil rhizobia and carbon-fixing bacteria, 102 NNI derivatives were designed. Flonicamid-36 and other four NNI derivatives passed the functional and environmentally friendly evaluation. Taguchi orthogonal experiment and factorial experiment-assisted molecular dynamics method were used to simulate the effects of 32 regulation schemes on the synergistic degradation of NNIS and its derivatives by rhizobia and carbon fixing bacteria. The synergistic degradation capacity of soil rhizobia and carbon-fixing bacteria was increased to 33.32% after right nitrogen supplementation. This indicated that supplementing the correct amount of nitrogen in the soil environment was beneficial to the microbial degradation of NNIs and their derivatives.

Increasing temperature is a key component of global climate change, affecting crop growth and productivity worldwide. Wheat is a major cereal crop grown in various parts of the globe, which is affected severely by heat stress. The morphological parameters affected include germination, seedling establishment, source-sink activity, leaf area, shoot and root growth. The physiological parameters such as photosynthesis, respiration, leaf senescence, water and nutrient relation are also affected by heat. At the cellular level, heat stress leads to the generation of reactive oxygen species that disrupt the membrane system of thylakoid, chloroplast and plasma membrane. The deactivation of the photosystem, reduction in photosynthesis and inactivation of rubisco affect the production of photoassimilates and their allocation. This ultimately affects anthesis, grain filling, size, number and maturity of wheat grains, which hamper crop productivity. The interplay of various systems comprising antioxidants and hormones plays a crucial role in imparting heat stress tolerance in wheat. Thus, implementation of various omics technologies could foster in-depth insights on heat stress effects, eventually devising heat stress mitigation strategies by conventional and modern breeding to develop heat-tolerant wheat varieties. This review provides an integrative view of heat stress responses in wheat and also discusses approaches to develop heat-tolerant wheat varieties.

Jasmonate (JA)-induced plant senescence has been mainly studied with a dark/starvation-promoted system using detached leaves; yet, the induction of whole-plant senescence by JA remains largely unclear. This work reports the finding of a JA-induced whole-plant senescence of tobacco under light/non-starvation conditions and the investigation of underlying regulations. Methyl jasmonate (MeJA) treatment induces the whole-plant senescence of tobacco in a light-intensity–dependent manner, which is suppressed by silencing of NtCOI1 that encodes the receptor protein of JA-Ile (the bioactive derivative of JA). MeJA treatment could induce the senescence-specific cysteine protease gene SAG12 and another cysteine protease gene SAG-L1 to high expression levels in the detached leaf patches under dark conditions but failed to induce their expression in tobacco whole plants under light conditions. Furthermore, MeJA attenuates the RuBisCo activase (RCA) level in the detached leaves but has no effect on this protein in the whole plant under light conditions. A genome-wide transcriptional assay also supports the presence of a differential regulatory pattern of senescence-related genes during MeJA-induced whole-plant senescence under non-starvation conditions and results in the finding of a chlorophylase activity increase in this process. We also observed that the MeJA-induced senescence of tobacco whole plants is reversible, which is accompanied by a structural change of chloroplasts. This work provides novel insights into JA-induced plant senescence under non-starvation conditions and is helpful to dissect the JA-synchronized process of whole-plant senescence.

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the key CO2-fixing enzyme in photosynthesis, is notorious for its low carboxylation. We report a highly active and assembly-competent Form II Rubisco from the endosymbiont of a deep-sea tubeworm Riftia pachyptila (RPE Rubisco), which shows a 50.5% higher carboxylation efficiency than that of a high functioning Rubisco from Synechococcus sp. PCC7002 (7002 Rubisco). It is a simpler hexamer with three pairs of large subunit homodimers around a central threefold symmetry axis. Compared with 7002 Rubisco, it showed a 3.6-fold higher carbon capture efficiency in vivo using a designed CO2 capture model. The simple structure, high carboxylation efficiency, easy heterologous soluble expression/assembly make RPE Rubisco a ready-to-deploy enzyme for CO2 capture that does not require complex co-expression of chaperones. The chemosynthetic CO2 fixation machinery of chemolithoautotrophs, CO2-fixing endosymbionts, may be more efficient than previously realized with great potential for next-generation microbial CO2 sequestration platforms.

Photoprotection strategies that have evolved in plants to cope with high light (HL) stress provide plants with the ability to resist HL. However, it has not been clearly confirmed which photoprotection strategy is the major HL resistance mechanism. To reveal the major photoprotection mechanism against short-term high light (STHL), the physiological and biochemical responses of three Arabidopsis mutants (Col, chi and ans) under STHL were analyzed in this study. After STHL treatment, the most serious photosynthetic pigment damage was observed in chi plants. At the same time, the degrees of membrane and Rubisco damage in chi was the highest, followed by Col, and ans was the smallest. The results showed that ans with high antioxidant capacity showed higher resistance to STHL treatment than Col containing anthocyanins, while chi with no anthocyanin accumulation and small antioxidant capacity had the lowest resistance. In addition, the gene expression results showed that plants tend to synthesize anthocyanin precursor flavonoids with antioxidant capacity under STHL stress. To further determine the major mechanism of photoprotection under STHL, we also analyzed Arabidopsis lines (Col, CHS1, CHS2 and tt4) that had the same anthocyanin content but different antioxidant capacities. It was found that CHS2 with high antioxidant capacity had higher cell viability, smaller maximal quantum yield of PSII photochemistry (Fv/Fm) reduction and less reactive oxygen species (ROS) accumulation under HL treatment of their mesophyll protoplasts. Therefore, the antioxidant capacity provided by antioxidant substances was the major mechanism of plant photoprotection under STHL treatment.

Elevated CO2 concentrations and persistent drought are predicted to inhibit the photosynthetic performance and yield of crops. Such effects vary considerably between species groups. Modern cultivated (Triticum aestivum L., AABBDD) and wild (Triticum dicoccoides Korn L., AABB and Triticum monococcum L., AA) species of wheat were subjected to elevated CO2 concentration (ambient concentration +200 μmol mol−1) and drought (well watered: 75–85% of the field water capacity; drought: 50–60% of the field water capacity) at open-top chamber experimental facilities. Elevated CO2 concentration decreased the limitation of stomatal morphology traits on stomatal conductance regulation. This could compensate the disadvantage of plants who has low stomatal density and large single stomatal area as well as low leaf water use efficiency such as modern wheat Z9023 in drought acclimation in the future CO2 rising world. Moreover, elevated CO2 concentration largely increased the dependence of light harvesting and electron transportation performance per photosynthesis system II reaction center, maximum rubisco carboxylation rate, and maximum Ribulose-1,5-bisphosphate regeneration rate on leaf nitrogen concentration across the selected wheat species and water regimes. Modern cultivated cultivars Z9023 and CH58 have higher photosynthetic performance per unit of leaf nitrogen than wild species under elevated CO2 concentrations. The increasing CO2 may present opportunities to breeders and possibly allow them to select for cultivars with better photosynthetic nitrogen use efficiency response to future CO2 rising climate.

Sugarcane (Saccharum spp.) is one of the main alternatives for the bioenergy sector due to its great potential for phytomass production, being the photosynthesis a key process related to conversion of the intercepted sunlight energy into phytomass. The aim of this study was to investigate the photosynthesis of nine sugarcane varieties originated from crosses between 1948 and 2006, as well as the species Saccharum officinarum, S. spontaneum and S. robustum. In plants grown inside a growth chamber, photosynthetic response curves to increasing air CO2 concentration were performed and key photosynthetic traits and their heritabilities were estimated. Fourteen physiological traits explained ~ 95% of the variation observed. Significant differences were found in leaf CO2 assimilation at air CO2 concentration of 1500 (A1500) and 400 (A400) μmol mol−1 among sugarcane genotypes, with S. spontaneum, IAC87-3396 and RB83-5486 presenting the highest photosynthetic rates. Sugarcane photosynthesis was strongly correlated with effective quantum yield of photosystem II (r = 0.93), non-photochemical quenching (r = − 0.74), stomatal conductance (r = 0.87), in vivo maximum carboxylation rates of PEPC (Vpmax, r = 0.83) and Rubisco (Vcmax, r = 0.91) and leaf nitrogen content (r = 0.83). The heritability varied from 0.97 (leaf CO2 assimilation) to 0.84 (leaf area). The A1500:A400 ratio higher than one for all genotypes indicates that Rubisco carboxylation sites are not saturated by CO2 and a non-efficient CO2 concentration mechanism. This study revealed a significant natural genotypic variation in sugarcane photosynthesis with high heritability of important traits, suggesting the possibility of breeding for increased photosynthetic activity.

Waterlogging caused by rainfall or improper irrigation is a serious threat that limits the growth and yield of crop plants by hypoxia stress. In the current study, the seedlings of vegetable sweet potato (Ipomoea batatas (L.) Lam.) variety ‘NC1’ (hypoxia-tolerant) and ‘C211’ (hypoxia-sensitive) were treated with oxygen content of 2 mg/L. It was found that the growth rate, net photosynthetic rate (PN), transpiration rate (E), stomatal limitation (Ls), maximum net photosynthetic rate (Amax), diurnal respiratory rate (Rd), maximum Rubisco carboxylation rate (Vcmax), maximum electron transport rate (Jmax), ABA (abscisic acid) and IAA (indole-3-acetic acid) content were significantly higher in ‘NC1’ than those of ‘C211’. It is concluded that under hypoxia stress, tolerant genotype could maintain higher photosynthetic efficiency by maintaining higher stomatal conductance and Rubisco efficiency, to maintain the growth of the root system and the canopy. This study provides the basis for further dissection of the mechanism of low-oxygen stress in sweet potato.

The success of the dwarf breeding of rice and wheat, called the Green Revolution, was due to increased source and sink capacities, which depended on large increases in N fertilizer inputs. However,...

Seaweeds are one of the ancient food supplements on Earth. Especially Asian countries use seaweeds as the fundamental ingredient in their cuisine. Seaweeds are photosynthetic organisms living in aquatic ecosystems and in the coastal territories. Seaweeds out of farm areas are frequently observed as coastal wastes. However, seaweeds are outstanding sources for bioactive substances and investigation bioactive properties of seaweed RuBisCO has never been done. RuBisCO is the most abundant protein on Earth but a vast amount of RuBisCO goes through waste. In this study, bioactive peptide prediction of frequently consumed seaweed RuBisCO proteins were analyzed in silico to identify possible bioactive peptides as substitute or support for grain, meat, and dairy based bioactive peptides. A huge portion of peptides were di-, tri- peptides with IC50 values less than 300 µM according to the comparison of BIOPEP database. Including gastric digestion, more than half of the peptides showed DDP-IV and ACE inhibitory activity followed by antioxidant properties. Also, novel antiinflammatory and anti-cancer peptides were found through in silico analysis.

Chlamydomonas reinhardtii undergoes conditional senescence when grown in batch culture due to nutrient limitation. Here, we explored plastid and photo-physiological adaptations in Chlamydomonas reinhardtii during a long-term ageing experiment by methodically sampling them over 22 weeks. Following exponential growth, Chlamydomonas entered an extended declining growth phase where cells continued to divide, although at a lower rate. Ultimately, this ongoing division was fueled by the recycling of macromolecules that was obvious in the rapidly declining protein and chlorophyll content in the cell during this phase. This process was sufficient to maintain a high level of cell viability as the culture entered stationary phase. Beyond that the cell viability starts to plummet. During the turnover of macromolecules after exponential growth that saw RuBisCO levels drop, the LHCII antenna was relatively stable. This, along with the upregulation of the light stress-related proteins (LHCSR), contributes to an efficient energy dissipation mechanism to protect the ageing cells from photooxidative stress during the senescence process. Ultimately, viability dropped to about 7% at 22 weeks in a batch culture. We anticipate that this research will help further understand the various acclimation strategies carried out by Chlamydomonas to maximize survival under conditional senescence.

Studies have reported on the regulation of clonal growth in Chinese sea buckthorn in response to environmental resource availability, but these studies have been limited to external mechanisms. In this report, we controlled irrigation to generate a soil moisture gradient in order to examine the photosynthetic physiological mechanisms regulating clonal growth in this species. The results indicated that as irrigation intensity increased, the soil water content increased vertically and tissue water content first increased and then decreased. Furthermore, Rubisco activase (RCA) and Mg-chelatase H subunit (CHLH) gene expression levels, photosynthetic capacity (net photosynthetic rate, transpiration rate, chlorophyll content, and stomatal conductance), and clonal growth (ramet growth, clonal proliferation, clonal propagation) all showed a quadratic parabolic change (i.e., first increasing and then decreasing). In addition, gene expression levels and tissue water content, photosynthetic capacity and gene expression levels, and clonal growth and photosynthetic capacity were all significantly positively correlated. When irrigation intensity (soil water content) is exceedingly low or high, the tissue water content is also low, RCA and CHLH gene expression levels are low, photosynthetic capacity is weak, clonal growth ability is inhibited, and clonal growth layout tends toward the “guerrilla type.” This type manifests as fewer and smaller clonal daughter ramets that are sparsely distributed with reduced clonal organ extension ability and branching intensity. When irrigation intensity (soil water content) is moderate, the tissue water content, gene expression levels, and photosynthetic capacity is high, clonal growth ability is completely uninhibited, and the clonal growth layout tends toward the “aggregated type.” This type is associated with numerous large clonal daughter ramets that are densely distributed with high clonal organ extension ability and branching intensity. Therefore, as irrigation intensity continuously changes from inordinately low to moderate to exceedingly high, Chinese sea buckthorn regulates clonal growth by photosynthetic capacity through photosynthetic gene expression. This results in a clonal growth layout continuum of “guerrilla-aggregated-guerrilla” that depends on irrigation intensity.

In this study, we hypothesized that Copaifera langsdorffii Desf. seedlings do not tolerate cultivation under water deficit, but shading could minimize stressor effects of water shortage and enable fast antioxidant responses, maintaining high photosynthetic rates and favoring stressed plant recovery. Therefore, we evaluated gas exchange and antioxidant activity of C. langsdorffii seedlings submitted to intermittent water deficit and different shading and their potential for metabolic recovery after re-irrigation. Seedlings were grown under 0, 30, and 70% shading and under continuous and intermittent irrigation. The plants were evaluated at the beginning of experiment, first and second zero photosynthesis, first and second recovery, and the end of experiment. Seedlings under intermittent irrigation and 0% shading could not recover their photosynthetic metabolism at the end of experiment, showing a marked decrease in rubisco carboxylation efficiency (A/Ci) and increase in antioxidant activity. Yet, seedlings grown under shading and intermittent irrigation could recover their photosynthetic metabolism, without permanent changes and/or damages to photosystem II efficiency, proving our hypothesis.

In the current study, salicylic acid (SA) assesses the physiological and biochemical responses in overcoming the potential deleterious impacts of arsenic (As) on Brassica napus cultivar Neelam. The toxicity caused by As significantly reduced the observed growth and photosynthetic attributes and accelerated the reactive oxygen species (ROS). Plants subjected to As stress revealed a significant (p ≤ 0.05) reduction in the plant growth and photosynthetic parameters, which accounts for decreased carbon (C) and sulfur (S) assimilation. Foliar spray of SA lowered the oxidative burden in terms of hydrogen peroxide (H2O2), superoxide anion (O2•−), and lipid peroxidation in As-affected plants. Application of SA in two levels (250 and 500 mM) protected the Brassica napus cultivar from As stress by enhancing the antioxidant capacity of the plant by lowering oxidative stress. Among the two doses, 500 mM SA was most effective in mitigating the adverse effects of As on the Brassica napus cultivar. It was found that SA application to the Brassica napus cultivar alleviated the stress by lowering the accumulation of As in roots and leaves due to the participation of metal chelators like phytochelatins, enhancing the S-assimilatory pathway, carbohydrate metabolism, higher cell viability in roots, activity of ribulose 1, 5-bisphosphate carboxylase (Rubisco), and proline metabolism through the active participation of γ-glutamyl kinase (GK) and proline oxidase (PROX) enzyme. The current study shows that SA has the capability to enhance the growth and productivity of B. napus plants cultivated in agricultural soil polluted with As and perhaps other heavy metals.

Alfalfa has a high potential as a novel food protein and can contribute to meet the increasing food demand. To facilitate food grade quality, protein extraction is essential. In this study, different extractions with isoelectric focusing the protein were compared; acid precipitation (citric and lactic acids) and fermentation (endogenous and Lactobacillus salivarius). Acid precipitation resulted in the highest protein recovery (30 %) compared with L. salivarius and endogenous fermentation. The protein ribulose-1.5-bisphosphate carboxylase/oxygenase (RuBisCO) was found intact and in high concentrations in acid precipitated samples. With L. salivarius, a higher content of peptides and possible cross-linked proteins were discovered. However, for both extractions, the amino acid composition showed a promising distribution meeting the recommendation of the Food and Agriculture Organization (FAO). Overall, these results indicate that protein extracts obtained with acid precipitation might be more appropriate as a food ingredient, whereas fermented products might have potential as food supplement.

Chlamydomonas reinhardtii can grow photosynthetically using CO2 or in the dark using acetate as the carbon source. In the light in air, the CO2 concentrating mechanism (CCM) of C. reinhardtii accumulates CO2, enhancing photosynthesis. A combination of carbonic anhydrases (CAs) and bicarbonate transporters in the CCM of C. reinhardtii increases the CO2 concentration at Ribulose 1,5-bisphosphate carboxylase oxygenase (Rubisco) in the chloroplast pyrenoid. Previously, CAs important to the CCM have been found in the periplasmic space, surrounding the pyrenoid and inside the thylakoid lumen. Two almost identical mitochondrial CAs, CAH4 and CAH5, are also highly expressed when the CCM is made, but their role in the CCM is not understood. Here, we adopted an RNAi approach to reduce the expression of CAH4 and CAH5 to study their possible physiological functions. RNAi mutants with low expression of CAH4 and CAH5 had impaired rates of photosynthesis under ambient levels of CO2 (0.04% CO2 [v/v] in air). These strains were not able to grow at very low CO2 (

Photosynthesis is crucial to the reduction of carbon dioxide in the atmosphere. The key enzyme of photosynthesis, Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), has two mutably competing substrates, CO2 and O2. It has features of carboxylase and oxygenase. Rubisco performs the function of carboxylase to reduce inorganic carbon to form organic substances, which precondition is that more carbon dioxide accumulates around it. Carbon dioxide concentrating mechanisms (CCMs) are vital to cope with the limit of carbon dioxide. Various bicarbonate use pathway has a differential contribution to inorganic carbon assimilation. Bicarbonate transport, extracellular bicarbonate dehydration, or H+-ATPase-driven bicarbonate uptake, which induced CCMs, can support a considerable share of photosynthesis in photosynthetic organisms. However, CCMs in thylakoid membranes may be the most important. The CCMs occurred in the plasma membrane were secondary, evolutionary, and inducible, while CCMs coupled with photosynthetic oxygen evolution in thylakoid membranes, were primitive, major, and indispensable. A hypothetical schematic model of CCMs occurred in the plasma membrane and thylakoid membranes being proposed.

The most productive C4 food and biofuel crops, such as Saccharum officinarum (sugarcane), Sorghum bicolor (sorghum) and Zea mays (maize), all use NADP-ME-type C4 photosynthesis. Despite high productivities, these crops fall well short of the theoretical maximum solar conversion efficiency of 6%. Understanding the basis of these inefficiencies is key for bioengineering and breeding strategies to increase the sustainable productivity of these major C4 crops. Photosynthesis is studied predominantly at steady state in saturating light. In field stands of these crops light is continually changing, and often with rapid fluctuations. Although light may change in a second, the adjustment of photosynthesis may take many minutes, leading to inefficiencies. We measured the rates of CO2 uptake and stomatal conductance of maize, sorghum and sugarcane under fluctuating light regimes. The gas exchange results were combined with a new dynamic photosynthesis model to infer the limiting factors under non-steady-state conditions. The dynamic photosynthesis model was developed from an existing C4 metabolic model for maize and extended to include: (i) post-translational regulation of key photosynthetic enzymes and their temperature responses; (ii) dynamic stomatal conductance; and (iii) leaf energy balance. Testing the model outputs against measured rates of leaf CO2 uptake and stomatal conductance in the three C4 crops indicated that Rubisco activase, the pyruvate phosphate dikinase regulatory protein and stomatal conductance are the major limitations to the efficiency of NADP-ME-type C4 photosynthesis during dark-to-high light transitions. We propose that the level of influence of these limiting factors make them targets for bioengineering the improved photosynthetic efficiency of these key crops. © 2021 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

Hereditary properties of strong growth and huge accumulation of biomass in Oryza officinalis exhibit a great potential; however, the genes that code for its high photosynthesis performance are not established. This study screened eight hybrids, using biomass accumulation and photosynthesis analysis, based on the introgression lines constructed by analyzing distant hybridization patterns between Oryza officinalis and cultivars HY-8. We designed 23 primer pairs using transcriptome sequencing of Oryza officinalis and screened two types of photosynthetic enzymes: phosphoenolpyruvate carboxylase (PEPC) and pyruvate orthophosphate dikinase (PPDK), which are two related proteins of ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco), its binding protein (rubis-subs-bind), and a small subunit (rbcS). Results showed that C4 photosynthetic pathway enzymes, PEPC and PPDK, were highly expressed in hybrids and the source plant, Oryza officinalis. Homology analysis also indicated that the sequences of those two genes were different from those of the C3 and C4 plants investigated. Therefore, a better understanding of the photosynthetic characteristics of Oryza officinalis would provide clues for further isolation of valuable genes from this plant.

The cultivation of Chlorella vulgaris in sludge extract (SE) from municipal excess sludge diluted with BG11 medium was investigated to achieve sludge reduction and microalgae cultivation. A maximum biomass of 1.404 ± 0.101 g/L was achieved at 15 days in a mixture of 25% BG11 and 75% sludge extract under 10,000-lx light. In early culture periods, heterotrophy dominated the process in the group with a high proportion of sludge extract of excess sludge extract containing relatively high amounts of organic material, which led to enhanced citrate synthase enzyme activities and reduced rubisco enzyme activities. Moreover, increasing the light intensity did not significantly improve biomass production due to its inhibitory effects on respiration in the early culture stages. The response surface model showed that organic matter was the main factor influencing biomass production from 0 to 2 days. The influence of light intensity on biomass production became more noticeable only after depleting the organic matter in sludge extract.

The common carbon and nitrogen currency, 2-oxoglutarate, could become a valuable resource for nitrogen assimilation and carbon centered biochemical fates. Here in this in silico study, a myriad of factors was used, namely phylogeny, sequence comparisons, and presence and location of clustered cysteines in specific plastid transporters of 2-oxoglutarate, to examine their evolution away from more generalized transporters. This transition would be to adopt the capability of internalizing 2-oxoglutarate alone or with superior specificities at the expense of malate. In phylogeny, the specific 2-oxoglutarate transporters (Cluster 1) are clustered in a separate clade away from 2 clades of general transporters (Cluster 2 and 3). The exclusivity (Cluster 1) and promiscuity of transporters (Cluster 2 and 3) compared to Arabidopsis counterparts characterized prior to this study, were used as a benchmark for my study. Within this mother clade of exclusive transporters, C4 and C3 2-oxoglutarate transporters once again form separate clusters of monophyly. Furthermore, a pattern of Cys –X-X-Cys-X(19)-Cys is conserved within the 2-oxo-glutarate only transporters that is missing in general transporters. Cysteines which are functionally key residues are inferred to be mediating intra- or inter-reactive disulfide bond formation or using a thiol (sulfhydryl) group for transport or to be forming a metal binding site. When a disulfide bond prediction tool was employed, it showed with negligible doubt that the Cys-X-X Cys-X(19) -Cys region was a strong contender for 2 separate disulfide bonds, although the middle cysteine was predicted to be involved in both. In addition, Cluster 2 general Zea mays C4 transporters are shown to be more recalcitrant to mutations of cysteines, compared to Panicum and Oryza counterparts. The study of 2-oxoglutarate and its availability in the chloroplast could play a two-prong role in C4 plants: to be a candidate for synthesis of bundle sheath cell Rubisco enzyme, which makes up ~50% of plant proteins, via ammonia assimilation, and even playing a role in carbon-centered biochemical pathways. This study could greatly facilitate choices in the tinkering of the right transporters for a future C4 rice in a climate change impacted world.

To elucidate the mechanism of succinic acid (SA) inhibition of Microcystis aeruginosa, the chlorophyll fluorescence transients, photosynthesis, photosynthetic electron transport activity, and gene expression of M. aeruginosa were evaluated under various doses of SA. The results demonstrated that, after treatment with 60 mg L-1 SA for 1 h, the chlorophyll fluorescence transients and related parameters changed significantly, indicating that the function and structure of photosynthetic apparatuses of Microcystis were seriously damaged. The initial quantum efficiency α, maximum net photosynthetic rate Pnmax, dark respiration rate Rd, and gross photosynthetic rate decreased to 57%, 49%, 49%, and 46%, respectively, relative to the control. Furthermore, photosystem II (PSII) activity (H2O→p-BQ) and the electron transport activity of H2O→MV and DPC→MV significantly decreased. Real-time PCR analysis revealed that, following incubation with 60 mg L-1 SA for 24 h, the expression level of core protein genes (psbA, psaB, psbD, and psbO) of the photosynthesis centers photosystem I (PSI) and PSII decreased significantly. However, the transcription of gene nblA encoding phycobilisome degradation protein was elevated. The downregulation of the rbcL gene, which encodes the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), resulted in the suppression of CO2 fixation and assimilation. High concentration (60 mg L-1) of SA resulted in damage to oxygen-evolving complex (OEC) and reaction center of PSII, blocking photosynthetic electron transport, thereby lowering the rate photosynthesis and inhibiting the growth of Microcystis. We concluded that inhibition of photosynthesis is an important mechanism of SA inhibition in M. aeruginosa.

This study aims at investigating the specific ion effects of Na+ and K+ on Ulva lactuca (L.) growth. U. lactuca was grown in balanced nutrient solutions with 10, 100, 300 and 600 mM NaCl or KCl. The growth was significantly higher at 300 and 600 mM NaCl compared to KCl, with the highest growth rate at 300 mM NaCl. NaCl-treated alga showed increases in the photosynthetic pigments and Rubisco protein content. However, KCl treatments adversely affected these photosynthetic attributes. U. lactuca needs adjusted, but not high K+/Na+ ratio for a proper growth, since the high K+/Na+ ratio in KCl-treated alga was associated with growth retardation. The cell wall was more extensible at high concentrations of NaCl compared to KCl. Therefore, the deleterious effect of K+ could be mainly on the cell wall and hence inhibiting the growth and perhaps the vitality of the whole cell. The transcript of plasma membrane (PM) H+-ATPase was detected only at 300 and 600 mM NaCl, implying that this gene was specifically induced by high concentrations of Na+ but not K+. The transcript of PM-Na+/K+-ATPase-like exhibited no Na+ specificity and its induction alone could not improve the growth of KCl-treated U. lactuca. The simultaneous induction of the two PM-ATPases could positively affect the algal growth at high NaCl concentrations by maintaining the proper cellular K+/Na+ ratio. Also, both PM-ATPases might contribute to energizing the plasma membrane and thereby promoting the cellular growth of U. lactuca at high Na+, but not K+, concentrations.

A state-of-the-art canopy modelling framework was used to predict changes in crop carbon uptake after Rubisco substitutions. We provide a new perspective and improve predictions from previous Rubisco modelling studies at the leaf level., Improving the performance of the CO2-fixing enzyme Rubisco is among the targets for increasing crop yields. Here, Earth system model (ESM) representations of canopy C3 and C4 photosynthesis were combined with species-specific Rubisco parameters to quantify the consequences of bioengineering foreign Rubiscos into C3 and C4 crops under field conditions. The ‘two big leaf’ (sunlit/shaded) model for canopy photosynthesis was used together with species-specific Rubisco kinetic parameters including maximum rate (Kcat), Michaelis–Menten constant for CO2 at ambient atmospheric O2 (Kc21%O2), specificity for CO2 to O2 (Sc/o), and associated heat activation (Ha) values. Canopy-scale consequences of replacing native Rubiscos in wheat, maize, and sugar beet with foreign enzymes from 27 species were modelled using data from Ameriflux and Fluxnet databases. Variation among the included Rubisco kinetics differentially affected modelled carbon uptake rates, and Rubiscos from several species of C4 grasses showed the greatest potential of >50% carbon uptake improvement in wheat, and >25% improvement in sugar beet and maize. This study also reaffirms the need for data on fully characterized Rubiscos from more species, and for better parameterization of ‘Vcmax’ and temperature response of ‘Jmax’ in ESMs.

The photosynthetic capacity of leaves is dramatically influenced by nitrogen (N) availability in the soil, as CO2 concentration in chloroplasts and photosynthetic biochemical capacity are related to leaf N content. The relationship between mesophyll conductance (gm) and leaf N content was expected to be shaped by leaf anatomical traits. However, the increased gm in mature leaves achieved by N top-dressing is unlikely to be caused by changes in leaf anatomy. Here, we assessed the impacts of N supply on leaf anatomical, biochemical, and photosynthetic features, specifically, the dynamic responses of leaf anatomy, biochemistry, and photosynthesis to N top-dressing in tobacco. Plant performance was substantially affected by soil N status. In comparison with the leaves of plants subjected to low N treatment, leaves of plants with high N treatment photosynthesized significantly more, due to higher CO2 diffusion conductance and photosynthetic biochemical capacity. The high gm in high N-treated leaves apparently related to modifications in the leaf anatomy; however, the rapid response of gm to N top-dressing cannot be fully explained by leaf anatomical modifications.