At the beginning of the experiments, rubisco increased followed by intense degradation. Lhcb proteins exhibited monomeric isoforms during the first 24 to 48 h, and D1 displayed sensitivity under S-depletion. Rubisco mutants exhibited a significant decrease in O2 evolution compared with the control.
Although the S-depleted medium was much more suitable than its complete counterpart for H2 production, hydrogen release was observed also in sealed S-repleted cultures of rubisco mutated cells under low-moderate light conditions. In particular, the rubisco mutant Y67A accounted for fold higher hydrogen production than the wild type under the same conditions and also displayed divergent metabolic parameters. These results indicate that rubisco is a promising target for improving hydrogen production rates in engineered microalgae.
Surveying the expanding prokaryotic Rubisco multiverse. The universal, but catalytically modest, CO2-fixing enzyme Rubisco is currently experiencing intense interest by researchers aiming to enhance crop photosynthesis. These efforts are mostly focused on the highly conserved hexadecameric enzyme found in land plants. In comparison, prokaryotic organisms harbor a far greater diversity in Rubisco forms.
Recent work towards improving our appreciation of microbial Rubisco properties and harnessing their potential is surveyed. New structural models are providing informative glimpses into catalytic subtleties and diverse oligomeric states.
Ongoing characterization is informing us about the conservation of constraints, such as sugar phosphate inhibition and the associated dependence on Rubisco activase helper proteins. Prokaryotic Rubiscos operate under a far wider range of metabolic contexts than the photosynthetic function of higher plant enzymes.
Relaxed selection pressures may have resulted in the exploration of a larger volume of sequence space than permitted in organisms performing oxygenic photosynthesis. To tap into the potential of microbial Rubiscos , in vivo selection systems are being used to discover functional metagenomic Rubiscos.
Various directed evolution systems to optimize their function have been developed. It is anticipated that this approach will provide access to biotechnologically valuable enzymes that cannot be encountered in the higher plant Rubisco space.
For permissions, please e-mail: journals. A function-based screen for seeking RubisCO active clones from metagenomes: novel enzymes influencing RubisCO activity. Although research on RubisCO genes and enzymes in plants, cyanobacteria and bacteria has been ongoing for years, still little is understood about its regulation and activation in bacteria.
Even more so, hardly any information exists about the function of metagenomic RubisCOs and the role of the enzymes encoded on the flanking DNA owing to the lack of available function-based screens for seeking active RubisCOs from the environment.
Here we present the first solely activity-based approach for identifying RubisCO active fosmid clones from a metagenomic library.
We constructed a metagenomic library from hydrothermal vent fluids and screened fosmid clones. Twelve clones exhibited RubisCO activity and the metagenomic fragments resembled genes from Thiomicrospira crunogena.
One of these clones was further analyzed. It contained a Knockouts of twelve genes and two intergenic regions on this metagenomic fragment demonstrated that the RubisCO activity was significantly impaired and was attributed to deletions in genes encoding putative transcriptional regulators and those believed to be vital for RubisCO activation.
Our new technique revealed a novel link between a poorly characterized gene and RubisCO activity. This screen opens the door to directly investigating RubisCO genes and respective enzymes from environmental samples. Rubisco catalytic properties of wild and domesticated relatives provide scope for improving wheat photosynthesis. Rubisco is a major target for improving crop photosynthesis and yield, yet natural diversity in catalytic properties of this enzyme is poorly understood.
Rubisco from 25 genotypes of the Triticeae tribe, including wild relatives of bread wheat Triticum aestivum , were surveyed to identify superior enzymes for improving photosynthesis in this crop. Rubisco large subunit genes rbcL were sequenced, and predicted corresponding amino acid differences analysed in relation to the corresponding catalytic properties. The effect of replacing native wheat Rubisco with counterparts from closely related species was analysed by modelling the response of photosynthesis to varying CO2 concentrations.
Thus, under otherwise identical conditions, catalytic variation in the Rubiscos analysed is predicted to improve photosynthetic rates at physiological CO2 concentrations. Naturally occurring Rubiscos with superior properties amongst the Triticeae tribe can be exploited to improve wheat photosynthesis and crop productivity.
Differential expression of rubisco in sporophytes and gametophytes of some marine macroalgae. In this study, the differential expression of Rubisco in sporophytes and gametophytes of four seaweed species--Porphyra yezoensis, P. Results indicated that both the Rubisco content and the initial carboxylase activity were notably higher in algal gametophytes than in the sporophytes, which suggested that the Rubisco content and the initial carboxylase activity were related to the ploidy of the generations of the four algal species.
Transgenic tobacco plants with improved cyanobacterial Rubisco expression but no extra assembly factors grow at near wild-type rates if provided with elevated CO2. Introducing a carbon-concentrating mechanism and a faster Rubisco enzyme from cyanobacteria into higher plant chloroplasts may improve photosynthetic performance by increasing the rate of CO2 fixation while decreasing losses caused by photorespiration.
We previously demonstrated that tobacco plants grow photoautotrophically using Rubisco from Synechococcus elongatus, although the plants exhibited considerably slower growth than wild-type and required supplementary CO2. Because of concerns that vascular plant assembly factors may not be adequate for assembly of a cyanobacterial Rubisco , prior transgenic plants included the cyanobacterial chaperone RbcX or the carboxysomal protein CcmM Furthermore, by altering the gene regulatory sequences on the Rubisco transgenes, cyanobacterial Rubisco expression was enhanced and the transgenic plants grew at near wild-type growth rates, although still requiring elevated CO2.
We performed detailed kinetic characterization of the enzymes produced with and without the RbcX and CcmM35 cyanobacterial proteins. These transgenic plants exhibit photosynthetic characteristics that confirm the predicted benefits of introduction of non-native forms of Rubisco with higher carboxylation rate constants in vascular plants and the potential nitrogen-use efficiency that may be achieved provided that adequate CO2 is available near the enzyme. Photosynthetic fuel for heterologous enzymes.
Recent work on the metabolic engineering of photosynthetic organisms has shown that the electron carriers such as ferredoxin and flavodoxin can be used to couple heterologous enzymes to photosynthetic reducing power. Because these proteins have a plethora of interaction partners and rely A handful of examples demonstrate channeling of photosynthetic electrons to drive the activity of heterologous enzymes , and these focus mainly on hydrogenases and cytochrome Ps.
We discuss specific approaches to address these bottlenecks and ensure high productivity of such enzymes in a photosynthetic A short history of RubisCO : the rise and fall? Although RubisCO has been intensively studied, its evolutionary origins and rise as Nature's most dominant carbon dioxide CO 2 -fixing enzyme still remain in the dark.
In this review we will bring together biochemical, structural, physiological, microbiological, as well as phylogenetic data to speculate on the evolutionary roots of the CO 2 -fixation reaction of RubisCO , the emergence of RubisCO-based autotrophic CO 2 -fixation in the context of the Calvin-Benson-Bassham cycle, and the further evolution of RubisCO into the 'RubisCOsome', a complex of various proteins assembling and interacting with the enzyme to improve its operational capacity functionality under different biological and environmental conditions.
Published by Elsevier Ltd.. Photosynthetic limitations in two Antarctic vascular plants: importance of leaf anatomical traits and Rubisco kinetic parameters.
Particular physiological traits allow the vascular plants Deschampsia antarctica Desv. The photosynthetic performance of these species was evaluated in situ, focusing on diffusive and biochemical constraints to CO2 assimilation. Leaf gas exchange, Chl a fluorescence, leaf ultrastructure, and Rubisco catalytic properties were examined in plants growing on King George and Lagotellerie islands. In spite of the species- and population-specific effects of the measurement temperature on the main photosynthetic parameters, CO2 assimilation was highly limited by CO2 diffusion.
In particular, the mesophyll conductance gm -estimated from both gas exchange and leaf chlorophyll fluorescence and modeled from leaf anatomy-was remarkably low, restricting CO2 diffusion and imposing the strongest constraint to CO2 acquisition.
Rubisco presented a high specificity for CO2 as determined in vitro, suggesting a tight co-ordination between CO2 diffusion and leaf biochemistry that may be critical ultimately to optimize carbon balance in these species. Interestingly, both anatomical and biochemical traits resembled those described in plants from arid environments, providing a new insight into plant functional acclimation to extreme conditions.
Understanding what actually limits photosynthesis in these species is important to anticipate their responses to the ongoing and predicted rapid warming in the Antarctic Peninsula. Exploiting transplastomically modified Rubisco to rapidly measure natural diversity in its carbon isotope discrimination using tuneable diode laser spectroscopy.
While knowing the fractionation by enzymes is pivotal to fully understanding plant carbon metabolism, little is known about variation in the discrimination factor of Rubisco b as it is difficult to measure using existing in vitro methodologies. This study used this technique to estimate b in vivo in five tobacco Nicotiana tabacum L. For transplastomic tobacco producing Rhodospirillum rubrum Rubisco b was Transplastomic tobacco producing chimeric Rubisco comprising tobacco Rubisco small subunits and the catalytic large subunits from either the C4 species Flaveria bidentis or the C3-C4 species Flaveria floridana had b-values of These values were not significantly different from tobacco Rubisco.
Over-expressing the C3 photosynthesis cycle enzyme Sedoheptulose Bisphosphatase improves photosynthetic carbon gain and yield under fully open air CO2 fumigation FACE. Background Biochemical models predict that photosynthesis in C3 plants is most frequently limited by the slower of two processes, the maximum capacity of the enzyme Rubisco to carboxylate RuBP Vc,max , or the regeneration of RuBP via electron transport J.
At current atmospheric [CO2] levels Rubisco is not saturated; consequently, elevating [CO2] increases the velocity of carboxylation and inhibits the competing oxygenation reaction which is also catalyzed by Rubisco. In the future, leaf photosynthesis A should be increasingly limited by RuBP regeneration, as [CO2] is predicted to exceed ppm by Results We tested the hypothesis that tobacco transformed to overexpressing SBPase will exhibit greater stimulation of A than wild type WT tobacco when grown under field conditions at elevated [CO2] ppm under fully open air fumigation.
Growth under elevated [CO2] stimulated instantaneous A and the diurnal photosynthetic integral A' more in transformants than WT. There was evidence of photosynthetic acclimation to elevated [CO2] via downregulation of Vc,max in both WT and transformants. Nevertheless, greater carbon assimilation and electron transport rates J and Jmax for transformants led to greater yield increases than WT at elevated [CO2] compared to ambient grown plants.
Conclusion These results provide proof of concept that increasing content and activity of a single photosynthesis enzyme can enhance carbon assimilation and yield of C3 crops grown at [CO2] expected by the middle of the 21st century.
Highly conserved small subunit residues influence rubisco large subunit catalysis. With a deeper understanding of its structure-function relationships and competitive inhibition by O 2 , it may be possible to engineer an increase in agricultural productivity and renewable energy.
To further define the role of the small subunit in Rubisco function, the 10 most conserved residues in all small subunits were substituted with alanine by transformation of a Chlamydomonas reinhardtii mutant that lacks the small subunit gene family. All the mutant strains were able to grow photosynthetically , indicating that none of the residues is essential for function.
Role of the Rubisco Small Subunit. However, it is a slow enzyme , and O 2 competes with CO 2 at the active site. Oxygenation initiates the photorespiratory pathway, which also results in the loss of CO 2. If carboxylation could be increased or oxygenation decreased, an increase in net CO 2 fixation would be realized.
Because Rubisco provides the primary means by which carbon enters all life on earth, there is much interest in engineering Rubisco to increase the production of food and renewable energy.
Rubisco is located in the chloroplasts of plants, and it is comprised of two subunits. Much is known about the chloroplast-gene-encoded large subunit rbcL gene , which contains the active site, but much less is known about the role of the nuclear-gene-encoded small subunit in Rubisco function rbcS gene. Both subunits are coded by multiple genes in plants, which makes genetic engineering difficult. In the eukaryotic, green alga Chlamydomonas reinhardtii, it has been possible to eliminate all the Rubisco genes.
These Rubisco -less mutants can be maintained by providing acetate as an alternative carbon source. In this project, focus has been placed on determining whether the small subunit might be a better genetic-engineering target for improving Rubisco. X-ray crystal structures of engineered chimeric-loop enzymes have indicated that additional residues and regions of the small subunit may also contribute to Rubisco function. Structural dynamics of the small-subunit carboxyl terminus was also investigated.
Alanine-scanning mutagenesis of the most-conserved small-subunit residues has identified a. Proceedings of the international seminar Reports on photosynthetic CO2-assimilating enzymes by the international workshops ; nendo chikyu kankyo sangyo gijutsu kaihatsu suishin jigyo kokusai seminar jigyo shiryo. Kogosei CO2 kotei koso kokusai workshop hokokusho. Described herein are the reports on photosynthetic CO2-assimilating enzymes , presented to the international symposium. These enzymes are important for assimilating CO2 in air, maintaining the environments and production of foods.
For activity regulation of PEPC, the topics include three-dimensional structures of PEPC and phosphorylation mechanisms and activity regulation therefor. For activity regulation of RuBisCO , the topics include post-translational activity of the enzymes e. For leaf photosynthesis and RuBisCO , the topics include importance of enzymes and involved in-vivo reaction steps for leaf photosynthesis CO2 assimilation reactions.
For function of PEPC, the topics include the biochemically and molecular biologically necessary and sufficient conditions for the C4 mechanism as the special photosynthesis mechanism. For transgenic approaches, the topics include procedure for allowing the RuBisCO gene of a dissimilar organism to function in the tobacco chloroplast, and introduction of enzymes involved in the C4 photosynthesis pathway in C3 plants.
Brassinosteroid-induced CO2 assimilation is associated with increased stability of redox-sensitive photosynthetic enzymes in the chloroplasts in cucumber plants. BRs upregulated the transcript levels of genes and activity of enzymes involved in the ascorbate—glutathione cycle in the chloroplasts, leading to an increased ratio of reduced GSH to oxidized GSSG glutathione in the chloroplasts. These results strongly suggest that BRs are capable of regulating the glutathione redox state in the chloroplasts through the activation of the ascorbate—glutathione cycle.
The resulting increase in the chloroplast thiol reduction state promotes CO 2 assimilation, at least in part, by enhancing the stability and activity of redox-sensitive photosynthetic enzymes through post-translational modifications.
Characterization of Rubisco activase from thermally contrasting genotypes of Acer rubrum Aceraceae. The lability of Rubisco activase function is thought to have a major role in the decline of leaf photosynthesis under moderate heat RCA1 and RCA2 proteins increased modestly in FL plants under warmer temperature, while only RCA2 protein increased in MN plants.
Rubisco large subunit RbsL protein abundance was relatively unaffected in either genotype by temperature. These results support the idea that Rubisco activase, particularly the ratio of Rubisco activase to Rubisco , may play a role in the photosynthetic heat acclimation in A.
This mechanism alone is not likely to entirely explain the thermotolerance in the FL genotype, and future research on adaptive mechanisms to high temperatures should consider activase function in a multipathway framework. Alterations in Rubisco activity and in stomatal behavior induce a daily rhythm in photosynthesis of aerial leaves in the amphibious-plant Nuphar lutea.
Nuphar lutea is an amphibious plant with submerged and aerial foliage, which raises the question how do both leaf types perform photosynthetically in two different environments. We found that the aerial leaves function like terrestrial sun-leaves in that their photosynthetic capability was high and saturated under high irradiance ca. We show that stomatal opening and Rubisco activity in these leaves co-limited photosynthesis at saturating irradiance fluctuating in a daily rhythm.
In the morning, sunlight stimulated stomatal opening, Rubisco synthesis, and the neutralization of a night-accumulated Rubisco inhibitor. Consequently, the light-saturated quantum efficiency and rate of photosynthesis increased fold by midday. During the afternoon, gradual closure of the stomata and a decrease in Rubisco content reduced the light-saturated photosynthetic rate.
However, at limited irradiance, stomatal behavior and Rubisco content had only a marginal effect on the photosynthetic rate, which did not change during the day. In contrast to the aerial leaves, the photosynthesis rate of the submerged leaves, adapted to a shaded environment, was saturated under lower irradiance. The light-saturated quantum efficiency of these leaves was much lower and did not change during the day.
Due to their low photosynthetic affinity for CO 2 35 muM and inability to utilize other inorganic carbon species, their photosynthetic rate at air-equilibrated water was CO 2 -limited. These results reveal differences in the photosynthetic performance of the two types of Nuphar leaves and unravel how photosynthetic daily rhythm in the aerial leaves is controlled. The plastid casein kinase 2 phosphorylates Rubisco activase at the Thr site but is not essential for regulation of Rubisco activation state.
Rubisco activase RCA is essential for the activation of Rubisco , the carboxylating enzyme of photosynthesis. The activity of Rubisco is controlled in res Black-Right-Pointing-Pointer BRs upregulate the activity of the ascorbate-glutathione cycle in the chloroplasts.
BRs upregulated the transcript levels of genes and activity of enzymes involved in the ascorbate-glutathione cycle in the chloroplasts, leading to an increased ratio of reduced GSH to oxidized GSSG glutathione in the chloroplasts. These results strongly suggest that BRs are capable of regulating the glutathione redox state in the chloroplasts through the activation of the ascorbate-glutathione cycle. Photosynthetic and enzymatic metabolism of Schinus terebinthifolius Raddi seedlings under water deficit.
Full Text Available ABSTRACT Schinus terebinthifolius Raddi is a tree species that can be used in the recovery of degraded areas, as it exhibits rapid growth and has a very expansive root system, facilitating water uptake from the deeper layers of the soil. The objective of this study was to evaluate photosynthesis and enzymatic activity in S. At the beginning of the experiment and during the suspension of irrigation and rehydration, plants were evaluated for gas and antioxidant enzyme exchanges.
Hydric stress significantly reduced photosynthesis, stomatal transpiration conductance, carboxylation efficiency of Rubisco , and the chlorophyll content of the S. Following rehydration, plants recovered the carboxylation efficiency of Rubisco , but not the photosynthetic rate. Antioxidant enzyme activity increased in both the aerial part and the root in response to water deficit.
The temporal and species dynamics of photosynthetic acclimation in flag leaves of rice Oryza sativa and wheat Triticum aestivum under elevated carbon dioxide. Zhu, J. Chinese Academy of Sciences. State Key Lab. National Institute for Agro-Environmental Sciences. Agro-Meteorology Div. Crop Systems and Global Change Lab. Jiangsu Institute of Botany, Nanjing China. No acclimation was observed for either crop at full flag leaf expansion. However, at the mid-anthesis stage, photosynthetic acclimation in rice was associated with RuBP carboxylation and regeneration limitations, while wheat only had the carboxylation limitation.
Although an increase in non-structural carbohydrates did occur during these later stages, it was not consistently associated with changes in SPS and SS or photosynthetic acclimation. Standard growing conditions in vitro low light and CO 2 are not conducive to autotrophy. An experiment was conducted to improve photosynthesis in vitro in the hope of increasing survival in acclimatization. A factorial experiment was elaborated where CO 2 and PPFD were supplied to in vitro cultured strawberry plants in the rooting stage.
Activities of carboxylating enzymes were determined after 4 weeks of culture. The activities of non-activated and activated rubisco and PEP-Case were measured after extraction of the enzymes and a reaction with NaH 14 CO 3 followed by scintillation counting spectroscopy. There was no difference in PEP activity at low light levels. The rubisco activity was lower at and ppm CO 2. Physiological significance of high activity of PEP-Case over rubisco will be discussed.
Enzymes involved in organellar DNA replication in photosynthetic eukaryotes. Plastids and mitochondria possess their own genomes. Although the replication mechanisms of these organellar genomes remain unclear in photosynthetic eukaryotes, several organelle-localized enzymes related to genome replication, including DNA polymerase, DNA primase, DNA helicase, DNA topoisomerase, single-stranded DNA maintenance protein, DNA ligase, primer removal enzyme , and several DNA recombination-related enzymes , have been identified.
In the reference Eudicot plant Arabidopsis thaliana, the replication-related enzymes of plastids and mitochondria are similar because many of them are dual targeted to both organelles, whereas in the red alga Cyanidioschyzon merolae, plastids and mitochondria contain different replication machinery components. The enzymes involved in organellar genome replication in green plants and red algae were derived from different origins, including proteobacterial, cyanobacterial, and eukaryotic lineages.
This increases both their nitrogen and water use efficiency compared to C 3 species. C 4 plants have greater rates of CO 2 assimilation than C 3 species for a given leaf nitrogen when both parameters are expressed either on a mass or an area basis Ghannoum et al. Although the range in leaf nitrogen content per unit areas is less in C 4 compared to C 3 plants, the range in leaf nitrogen concentration per unit dry mass is similar for both C 4 and C 3 species. Even though leaf nitrogen is invested into photosynthetic components into the same fraction in both C 3 and C 4 species, C 4 plants allocate less nitrogen to Rubisco protein and more to other soluble protein and thylakoids components.
The lower nitrogen requirement of C 4 plants results from their CO 2 -concentrating mechanism, which raises the bundle sheath CO 2 concentration, saturating Rubisco in normal air and almost eliminating photorespiration.
To attain comparable photosynthetic rates to those in C 4 plants, C 3 leaves must therefore invest more heavily in Rubisco and have a greater nitrogen requirement. Because the Rubisco specificity for CO 2 decreases with increasing temperature Long, , this difference between the C 3 and C 4 photosynthetic nitrogen-use efficiency is greatest at high temperatures Long, The high photosynthetic nitrogen-use efficiency of C 4 plants is partially offset by the nitrogen-requirement for CO 2 -concentrating mechanism enzymes, but the high maximum catalytic rate of PEP-carboxylase means that these account for only ca.
Improved leaf and plant water use efficiency in C 4 plants is due to both higher photosynthetic rates per unit leaf area and lower stomatal conductance, with the greater CO 2 assimilation contributing to a major extent Ghannoum et al. The advantages of greater nitrogen use efficiency and water use efficiency of C 4 relative to C 3 photosynthesis are fully realized at high light and temperature, where oxygenase reaction of Rubisco is greatly increased.
It is worth noting, although in C 4 plants energy loss due to photorespiration is eliminated, and additional energy is required to operate the C 4 cycle 2 ATPs per CO 2 assimilated. In dim light, when photosynthesis is linearly dependent on the radiative flux, the rate of CO 2 assimilation depends entirely on the energy requirements of carbon assimilation Long, The additional ATP required for assimilation of one CO 2 in C 4 photosynthesis, compared with C 3 photosynthesis, increases the energy requirement in C 4 plants Hatch, However, when the temperature of a C 3 leaf exceeds ca.
This is the reason why at temperatures below ca. It is interesting to note, that while global distribution of C 4 grasses is positively correlated with growing season temperature, the geographic distribution of the different C 4 subtypes is strongly correlated with rainfall Ghannoum et al. On the contrary, C 4 plants are rare to absent in cold environments.
Although there are examples of plants with C 4 metabolisms that show cold adaptation, they still require warm periods during the day in order to exist in cold habitats Sage et al.
The mechanisms explaining the lower performance of C 4 plants under cold conditions have not been clarified Sage et al. Among early plausible explanations were the low quantum yield of the C 4 relative to the C 3 pathway Ehleringer et al. Both hypothesis are insufficient since maximum quantum yield differences do not relate to conditions under which the vast majority of daily carbon is assimilated and there cold-adapted C 4 species that have cold stabled forms of PEP-carboxylase and pyruvate orthophosphate dikinase, and synthesize sufficient quantity to overcome any short term limitation Du et al.
The current hypothesis is that C 4 photosynthesis is limited by Rubisco capacity at low temperatures. Only this time humans are the drivers of these changes and not glacial-interglacial cycles. Human-caused increases in atmospheric CO 2 concentration are thought to be largely responsible for recent increases in global mean surface temperatures and are projected to increase by 1.
Regarding plants, higher atmospheric CO 2 levels tend to reduce stomatal conductance and transpiration, thereby lowering latent heat loss and causing higher leaf temperatures Bernacchi et al. Thus, in the future, plants will likely experience increases in acute heat and drought stress, which can impact ecosystem productivity Cias et al.
The sensitivity of photosynthesis to each of the environmental variables including high temperature, low water availability, vapor pressure deficit and soil salinity, associated with the inevitable rise in atmospheric CO 2 , has not been well documented in assessing plant responses to the new changing environment Reddy et al.
How plant growth responds to the rising CO 2 concentration will not only affect ecosystem productivity in the future, but also the magnitude of C sequestration by plants and, consequently, the rate of CO 2 increase in the atmosphere. C 4 plants are directly affected by all major global change parameters, often in a manner that is distinct from that of C 3 plants.
In the present chapter, we will focus on the effect of increased CO 2 , and its relation to temperature and drought, on C 4 plants. Protein Sci. Janasch, M. Download references. You can also search for this author in PubMed Google Scholar. Correspondence to Elton P. Reprints and Permissions. CO 2 fixation gets a second chance. Nat Catal 4, 94—95 Download citation.
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