Ribulose Bisphosphate Research Articles

Achieving High Expression of Cry in Green Tissues and Negligible Expression in Endosperm Simultaneously via rbcS Gene Fusion Strategy in Rice.

To allay excessive public concern about the safety of transgenic foods, and to optimize insect-resistant genes expression to delay the evolution of resistance in pests, we developed a promising strategy to fuse the GOI (gene of interest) with OsrbcS (rice small subunit of ribulose bisphosphate carboxylase/oxygenase) in transgenic rice, which acted as a carrier, driven by the OsrbcS native promoter to sequester its expression in green tissues. Using eYFP as a trial, we reported a high-level accumulation of eYFP in green tissue and almost none in the seed and root of the fused construct compared to the non-fused construct. After applying this fusion strategy in insect-resistant rice breeding, recombinant OsrbcS-Cry1Ab/Cry1Ac expressed rice plants conferred high resistance to leaffolders and striped stem borers, among which two single-copy lines possessed normal agronomic performance in the field. Specifically, Cry1Ab/Cry1Ac protein levels in single-copy construct transgenic lines ranged from 1.8 to 11.5 µg g-1 in the leaf, higher than the Actin I promoter-driven control, T51-1, about 1.78 µg g-1 in the leaf, but negligible (only 0.00012-0.00117 µg g-1) in endosperm by ELISA analysis. Our study provided a novel approach to creating Cry1Ab/Cry1Ac-free endosperm rice with a high level of insect-resistant protein in green tissues through the simultaneous usage of the OsrbcS promoter and OsrbcS as a fusion partner.

CpDNA Sekansları Temelinde Trifolium L. Türlerinin Filogenetik İlişkileri

DNA barcoding is an important molecular approach in the determination of species diversity, evaluation of phylogenetic relationships and identification of taxonomically problematic species because of insufficient diagnostic characters. It has great importance to determine the barcoding regions that will give the best result in the evaluation of phylogenetic relationships in species like Trifolium that spread over wide geographical regions in the world and show high variation accordingly. For this aim, matK (maturase K) and rbcL (ribulose bisphosphate carboxylase gene) regions belonging to cpDNA(chloroplast DNA) was used to determine the barcoding ability and evaluate the taxonomy of the genus Trifolium. 63 taxa from matK region and 47 taxa from rbcL region were determined and used in this study. It was observed that transitional substitutions for matK and rbcL regions are higher than transversional substitutions with the rate of 51.52 % and 70.69 %, respectively. It can be stated that both of barcoding regions are valuable to reveal the phylogenetic relationships, in addition to their grouping ability the species as taxonomically. However, especially the using of matK sequence informations that have high variable sites (158) and grouping ability clearly for all taxa is strongly recommended.

Characterization of a novel thermophilic cyanobacterium within Trichocoleusaceae, Trichothermofontia sichuanensis gen. et sp. nov., and its CO2-concentrating mechanism.

Thermophiles from extreme thermal environments have shown tremendous potential regarding ecological and biotechnological applications. Nevertheless, thermophilic cyanobacteria remain largely untapped and are rarely characterized. Herein, a polyphasic approach was used to characterize a thermophilic strain, PKUAC-SCTB231 (hereafter B231), isolated from a hot spring (pH 6.62, 55.5°C) in Zhonggu village, China. The analyses of 16S rRNA phylogeny, secondary structures of 16S-23S ITS and morphology strongly supported strain B231 as a novel genus within Trichocoleusaceae. Phylogenomic inference and three genome-based indices further verified the genus delineation. Based on the botanical code, the isolate is herein delineated as Trichothermofontia sichuanensis gen. et sp. nov., a genus closely related to a validly described genus Trichocoleus. In addition, our results suggest that Pinocchia currently classified to belong to the family Leptolyngbyaceae may require revision and assignment to the family Trichocoleusaceae. Furthermore, the complete genome of Trichothermofontia B231 facilitated the elucidation of the genetic basis regarding genes related to its carbon-concentrating mechanism (CCM). The strain belongs to β-cyanobacteria according to its β-carboxysome shell protein and 1B form of Ribulose bisphosphate Carboxylase-Oxygenase (RubisCO). Compared to other thermophilic strains, strain B231contains a relatively low diversity of bicarbonate transporters (only BicA for HCO3- transport) but a higher abundance of different types of carbonic anhydrase (CA), β-CA (ccaA) and γ-CA (ccmM). The BCT1 transporter consistently possessed by freshwater cyanobacteria was absent in strain B231. Similar situation was occasionally observed in freshwater thermal Thermoleptolyngbya and Thermosynechococcus strains. Moreover, strain B231 shows a similar composition of carboxysome shell proteins (ccmK1-4, ccmL, -M, -N, -O, and -P) to mesophilic cyanobacteria, the diversity of which was higher than many thermophilic strains lacking at least one of the four ccmK genes. The genomic distribution of CCM-related genes suggests that the expression of some components is regulated as an operon and others in an independently controlled satellite locus. The current study also offers fundamental information for future taxogenomics, ecogenomics and geogenomic studies on distribution and significance of thermophilic cyanobacteria in the global ecosystem.

Phenotypic Evaluation of Traits and Barcode Identification using Plastid-specific Ribulose Bisphosphate Carboxylase (Rbcl) Analysis on Lablab Beans [Lablab purpureus (L.) Sweet

Background: The importance of grain legumes in world food sustainability cannot be overemphasized. Minor legumes on the other hand, contribute to agricultural strengthening, especially in the context of sustainable crop and livestock production systems. In the context of assessing different legume species for crop improvement and identification, twenty (20) accessions of Lablab purpureus (L.) Sweet collected to determine the degree of trait variability and to identify accessions with potentials, which can contribute to grain and forage production. Methods: The present study involved the evaluation of twenty (20) accessions of Lablab purpureus (L.) Sweet from IITA, Ibadan. The accessions were grown in the experimental plot and DNA from young and healthy leaves were isolated for determination of genetic diversity using ribulose bisphosphate carboxylase (rbcL). The accessions were also categorized into groups based on the performances and the highest discriminating traits that accounted for greater variability using principal component analysis (PCA) and cluster analysis (CA) respectively. Result: Cluster analysis grouped all the twenty accessions into three clusters, where cluster I, cluster II and cluster III comprised of eight, seven and five accessions respectively. The PCA revealed hundred seed-weights, number of pod per plant and seed length as the most discriminating trait that accounted for greater variability in the Lablab accessions considered and they should be considered in the hyacinth bean improvement programs. Based on diverse level of phenotypic traits, accessions from each cluster were selected for barcode identification and comparison with other cultivars from other region. Results of the molecular analysis showed two clusters and authenticated the use of universal primers, ribulose bisphosphate carboxylase (rbcL) for DNA barcoding for successful amplification, identification and discrimination of Lablab purpureus (L). This revealed diversity of the considered accessions and provided information on relatedness to other accessions from other region of the world.

Diatom DNA metabarcoding to assess the effect of natural radioactivity in mineral springs on ASV of benthic diatom communities

Little is still known about the low dose effects of radiation on the microbial communities in the environment. Mineral springs are ecosystems than can be affected by natural radioactivity. These extreme environments are, therefore, observatories for studying the influence of chronic radioactivity on the natural biota. In these ecosystems we find diatoms, unicellular microalgae, playing an essential role in the food chain. The present study aimed to investigate, using DNA metabarcoding, the effect of natural radioactivity in two environmental compartments (i.e. spring sediments and water) on the genetic richness, diversity and structure of diatom communities in 16 mineral springs in the Massif Central, France. Diatom biofilms were collected during October 2019, and a 312 bp region of the chloroplast gene rbcL (coding for the Ribulose Bisphosphate Carboxylase) used as a barcode for taxonomic assignation. A total of 565 amplicon sequence variants (ASV) were found. The dominant ASV were associated with Navicula sanctamargaritae, Gedaniella sp., Planothidium frequentissimum, Navicula veneta, Diploneis vacillans, Amphora copulata, Pinnularia brebissonii, Halamphora coffeaeformis, Gomphonema saprophilum, and Nitzschia vitrea, but some of the ASVs could not be assigned at the species level. Pearson correlation failed to show a correlation between ASV’ richness and radioactivity parameters. Non-parametric MANOVA analysis based on ASVs occurrence or abundances revealed that geographical location was the main factor influencing ASVs distribution. Interestingly, 238U was the second factor that explained diatom ASV structure. Among the ASVs in the mineral springs monitored, ASV associated with one of the genetic variants of Planothidium frequentissimum was well represented in the springs and with higher levels of 238U, suggesting its high tolerance to this particular radionuclide. This diatom species may therefore represent a bio-indicator of high natural levels of uranium.

Ribulose-1,5-bisphosphate regeneration in the Calvin-Benson-Bassham cycle: Focus on the last three enzymatic steps that allow the formation of Rubisco substrate.

The Calvin-Benson-Bassham (CBB) cycle comprises the metabolic phase of photosynthesis and is responsible for carbon fixation and the production of sugar phosphates. The first step of the cycle involves the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) which catalyzes the incorporation of inorganic carbon into 3-phosphoglyceric acid (3PGA). The following steps include ten enzymes that catalyze the regeneration of ribulose-1,5-bisphosphate (RuBP), the substrate of Rubisco. While it is well established that Rubisco activity acts as a limiting step of the cycle, recent modeling studies and experimental evidence have shown that the efficiency of the pathway is also impacted by the regeneration of the Rubisco substrate itself. In this work, we review the current understanding of the structural and catalytic features of the photosynthetic enzymes that catalyze the last three steps of the regeneration phase, namely ribose-5-phosphate isomerase (RPI), ribulose-5-phosphate epimerase (RPE), and phosphoribulokinase (PRK). In addition, the redox- and metabolic-based regulatory mechanisms targeting the three enzymes are also discussed. Overall, this review highlights the importance of understudied steps in the CBB cycle and provides direction for future research aimed at improving plant productivity.

Molecular mechanism of Rubisco activase: Dynamic assembly and Rubisco remodeling.

Ribulose-1,5-bisphosphate (RuBP) carboxylase-oxygenase (Rubisco) enzyme is the limiting step of photosynthetic carbon fixation, and its activation is regulated by its co-evolved chaperone, Rubisco activase (Rca). Rca removes the intrinsic sugar phosphate inhibitors occupying the Rubisco active site, allowing RuBP to split into two 3-phosphoglycerate (3PGA) molecules. This review summarizes the evolution, structure, and function of Rca and describes the recent findings regarding the mechanistic model of Rubisco activation by Rca. New knowledge in these areas can significantly enhance crop engineering techniques used to improve crop productivity.

Quantifying photosynthetic metabolites permeability across bacterial microcompartments using computational microscopy

Carboxysomes are Bacterial microcompartments, found in cyanobacteria that locally concentrate carbon dioxide (CO2) to improve the efficiency of the enzyme RuBisCO, a key step towards photosynthetic carbon fixation by the Calvin-Benson-Bassham (CBB) cycle. The carboxysome shell is made up of proteins that encapsulate RuBisCO and carbonic anhydrase. Carbonic anhydrase converts soluble bicarbonate to CO2, increasing the local concentration of CO2 for RuBisCO. In addition to CO2, other metabolites such as oxygen (O2), bicarbonate (HCO3−), 3-phosphoglyceric acid (3-PGA) and ribulose-1,5-bisphosphate (RuBP) also need to permeate through the carboxysome shell to efficiently perform CO2 fixation. Quantifying the permeability of the shell for these metabolites currently remains a challenge and one of the critical design features related to the design of synthetic bacterial microcompartments for metabolic and sustainablebioengineering applications. Leveraging a high resolution cryo-electron microscopy structure of a synthetic beta-carboxysome shell and new graphics processing unit (GPU)-resident molecular simulation engines, we determine the permeability of photosynthetic metabolites across synthetic carboxysome shell through unbiased molecular simulation at all-atom resolution. We find that the carboxysome itself is not selectively permeable to bicarbonate over carbon dioxide, as originally hypothesized. Instead, the carboxysome shell proteins form a general barrier to maintain the carbon dioxide gradient generated by carbonic anhydrase activity within the carboxysome, compensating for inefficiencies of endogenous RuBisCO, specifically slow turnover rate, and photorespiration. The results from all-atom molecular simulations presented here provide a detailed mechanistic picture, using the shell permeabilities to engineer smart synthetic bacterial microcompartments and to improve photosynthetic efficiency with a wide range of applications in sustainable, metabolic, agricultural, and biomedical engineering.

Differential sensitivities of photosynthetic processes and carbon loss mechanisms govern N-induced variation in net carbon assimilation rate for field-grown cotton.

Nitrogen (N) deficiency limits the net carbon assimilation rate (AN), but the relative N sensitivities of photosynthetic component processes and carbon loss mechanisms remain relatively unexplored for field-grown cotton. Therefore, the objective of the current study was to define the relative sensitivity of individual physiological processes driving N deficiency-induced declines in AN for field-grown cotton. Among the potential diffusional limitations evaluated, mesophyll conductance was the only parameter substantially reduced by N deficiency, but this did not affect CO2 availability in the chloroplast. A number of metabolic processes were negatively impacted by N deficiency, and these effects were more pronounced at lower leaf positions in the cotton canopy. Ribulose bisphosphate (RuBP) regeneration and carboxylation, AN, and gross photosynthesis were the most sensitive metabolic processes to N deficiency, whereas photosynthetic electron transport processes, electron flux to photorespiration, and dark respiration exhibited intermediate sensitivity to N deficiency. Among thylakoid-specific processes, the quantum yield of PSI end electron acceptor reduction was the most sensitive process to N deficiency. It was concluded that AN is primarily limited by Rubisco carboxylation and RuBP regeneration under N deficiency in field-grown cotton, and the differential N sensitivities of the photosynthetic process and carbon loss mechanisms contributed significantly to photosynthetic declines.

Photorespiration is the solution, not the problem

The entry of carbon dioxide from the atmosphere into the biosphere is mediated by the enzyme Rubisco, which catalyzes the carboxylation of ribulose 1,5-bisphosphate (RuBP) as the entry reaction of the Calvin Benson Bassham cycle (CBBC), leading to the formation of 2 molecules of 3-phosphoglyceric acid (3PGA) per CO2 fixed. 3PGA is reduced to triose phosphates at the expense of NADPH + H+ and ATP that are provided by the photosynthetic light reactions. Triose phosphates are the principal products of the CBBC and the precursors for almost any compound in the biosphere.

Morpho-Genetic Characterization of Abelmoschus Moench. Accessions

This study was carried out to assess the morphological and genetic variability of seven accessions of Abelmoschus L. species using morphometric analysis and ribulose bisphosphate carboxylase large chain (RBCL) molecular markers. Using a completely randomized block design, seeds of the okra accessions were planted with three replicates each. During the developmental stage, the morphological features of the accessions were observed and recorded according to the standard descriptor for the crop. Morphologically, all the accessions exhibited a degree of similarities, albeit, at maturity, the leaf, plant height, leaf colour, and leaf shape became distinct. A cluster of the phenotypic characterization was observed at a 3.74 level of coefficient of similarity with two distinct clusters, which were predicted to be A. esculentus and A. caillei. The percentage variance of the two principal components was 55.12% and 22.69% with corresponding Eigenvalues of 4.11 and 1.69, respectively. Results of the RBCL analysis revealed genetic variability at a 0.80 level of coefficient of similarity. Two distinguishable clusters were observed. Both morphometric and genotyping results suggest that variations exist among and within the seven accessions. In conclusion, there is a need to frequently evaluate plant genetic resources held in gene banks as they may not reflect the whole range of diversity inherent in the species.

β‐amino carbonyl derivatives: Synthesis, Molecular Docking, ADMET, Molecular Dynamic and Herbicidal studies.

AbstractIn the current report, 3D structures of the enzyme Lathyrus aphaca Ribulose bisphosphate carboxylase (LArbcL) was modeled using homology modeling. The structures of the synthesized β‐amino carbonyl derivatives were made by means of “Chem Draw ultra‐12.0”, ADMET prediction were conducted to compute physicochemical properties. Pharmacokinetics studies and the molecular docking study of the synthesized compounds were performed against Lathyrus aphaca Ribulose bisphosphate carboxylase. Docking experiments verified a significant Docking Score values between 6.2 to 7.3 kcal mol−1. The highest‐ranking complexes obtained from docking results were subjected to 100 ns Molecular Dynamics simulations using Gromacs program to investigate the constancy of the docked “protein–ligand complexes” as well as the oscillation and conformational variations that occur during protein–ligand interaction. The synthesized derivatives were screened for pre‐emergence and post‐emergence herbicidal activity adjacent to weed species named Lathyrus aphaca with concentrations of “0.005 M, 0.01 M and 0.02 M”, and the activity was compared with Butachlor and penoxulum which are standard herbicide. Every single synthesized compounds show good to moderate activity.

Enhanced genome-wide association reveals the role of YABBY11-NGATHA-LIKE1 in leaf serration development of Populus.

Leaf margins are complex plant morphological features that contribute to leaf shape diversity, which affects plant structure, yield, and adaptation. Although several leaf margin regulators have been identified to date, the genetic basis of their natural variation has not been fully elucidated. In this study, we profiled two distinct leaf morphology types (serrated and smooth) using the persistent homology mathematical framework (PHMF) in two poplar species (Populus tomentosa and Populus simonii, respectively). A combined genome-wide association study (GWAS) and expression quantitative trait nucleotide (eQTN) mapping were applied to create a leaf morphology control module using data from P. tomentosa and P. simonii populations. Natural variation in leaf margins was associated with YABBY11 (YAB11) transcript abundance in poplar. In P. tomentosa, PtoYAB11 carries a premature stop codon (PtoYAB11PSC), resulting in the loss of its positive regulation of NGATHA-LIKE1 (PtoNGAL-1) and RIBULOSE BISPHOSPHATE CARBOXYLASE LARGE SUBUNIT (PtoRBCL). Overexpression of PtoYAB11PSC promoted serrated leaf margins, enlarged leaves, enhanced photosynthesis, and increased biomass. Overexpression of PsiYAB11 in P. tomentosa promoted smooth leaf margins, higher stomatal density, and greater light damage repair ability. In poplar, YAB11-NGAL1 is sensitive to environmental conditions, acts as a positive regulator of leaf margin serration, and may also link environmental signaling to leaf morphological plasticity.

The determiner of photosynthetic acclimation induced by biochemical limitation under elevated CO2 in japonica rice

Photosynthetic acclimation to prolonged elevated CO2 could be attributed to the two limited biochemical capacity, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation and ribulose-1,5-bisphosphate (RuBP) regeneration, however, which one is the primary driver is unclear. To quantify photosynthetic acclimation induced by biochemical limitation, we investigated photosynthetic characteristics and leaf nitrogen allocation to photosynthetic apparatus (Rubisco, bioenergetics, and light-harvesting complex) in a japonica rice grown in open-top chambers at ambient CO2 and ambient CO2+200 μmol mol−1 (e [CO2]). Results showed that photosynthesis was stimulated under e [CO2], but concomitantly, photosynthetic acclimation obviously occurred across the whole growth stages. The content of leaf nitrogen allocation to Rubisco and biogenetics was reduced by e [CO2], while not in light-harvesting complex. Unlike the content, there was little effects of CO2 enrichment on the percentage of nitrogen allocation to photosynthetic components. Additionally, leaf nitrogen did not reallocate within photosynthetic apparatus until the imbalance of sink-source under e [CO2]. The contribution of biochemical limitations, including Rubisco carboxylation and RuBP regeneration, to photosynthetic acclimation averaged 36.2% and 63.8% over the growing seasons, respectively. This study suggests that acclimation of photosynthesis is mainly driven by RuBP regeneration limitation and highlights the importance of RuBP regeneration relative to Rubisco carboxylation in the future CO2 enrichment.

Main components of free organic carbon generated by obligate chemoautotrophic bacteria that inhibit their CO2 fixation.

Chemoautotrophic bacteria play an important role in combating the rise in global CO2. However, recently it was found that extracellular free organic carbon (EFOC) generated by chemoautotrophic bacteria inhibits their CO2 fixation. Although continuous-flow membrane bioreactor can remove EFOC and enrich bacteria, it may also remove beneficial bio-factors for bacterial growth. Finding out the main inhibitory factors and inhibitory mechanisms in EFOC can provide theoretical guidance for the development of targeted inhibitory component removal technology. The results show a significant negative correlation between the increasing proportion of small-molecule EFOC and the decreasing trend of CO2 fixation efficiency, and simulation experiments confirm that the small molecule organics such as amino acids and organic acids are the main components of EFOC that inhibit CO2 fixation by inhibiting ribulose bisphosphate carboxylase/oxygenase (RuBisCO) gene (cbb) transcription efficiency. Therefore, amino acids and organic acids are suggested to be recovered to promote efficient CO2 fixation of autotrophic bacteria.

A non-carboxylating pentose bisphosphate pathway in halophilic archaea

Bacteria and Eucarya utilize the non-oxidative pentose phosphate pathway to direct the ribose moieties of nucleosides to central carbon metabolism. Many archaea do not possess this pathway, and instead, Thermococcales utilize a pentose bisphosphate pathway involving ribose-1,5-bisphosphate (R15P) isomerase and ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco). Intriguingly, multiple genomes from halophilic archaea seem only to harbor R15P isomerase, and do not harbor Rubisco. In this study, we identify a previously unrecognized nucleoside degradation pathway in halophilic archaea, composed of guanosine phosphorylase, ATP-dependent ribose-1-phosphate kinase, R15P isomerase, RuBP phosphatase, ribulose-1-phosphate aldolase, and glycolaldehyde reductase. The pathway converts the ribose moiety of guanosine to dihydroxyacetone phosphate and ethylene glycol. Although the metabolic route from guanosine to RuBP via R15P is similar to that of the pentose bisphosphate pathway in Thermococcales, the downstream route does not utilize Rubisco and is unique to halophilic archaea.

Proteomics and photosynthetic apparatus response to vermicompost attenuation of salinity stress Vicia faba leaves

Crop production and growth are severely affected by salt stress. Nevertheless, the bio-fertilizer vermicompost (VC) can be participated as a potent inhibitor of salinity on plant growth and crop production by regulating photosynthetic efficiency. We investigated the effect of VC on photosynthetic performance of salt-stressed broad bean (Vicia faba L. Aspani cultivar). Seeds were grown in soil mixture; clay and sand in ratio 1:2 by volume with five different volumetric ratios of VC; 0, 2.5, 5, 10 and 15% irrigated with either water and/or 200 mM NaCl. Leaf area, Na and K contents, chlorophylls, photosystem II efficiency, Rubisco content, soluble sugars, chloroplasts’ organization and proteomics were analyzed. The imposed stress decrease leaf area, chlorophyll contents, maximum quantum efficiency (Fv/Fm), Rubisco content, increase soluble sugars and damage chloroplasts organization. Salinity upregulated glucose-1-phosphate adenylyl transferase, ribulose bisphosphate carboxylase large subunit and chloroplastic peptidyl-prolyl cis–trans isomerase. The increased leaf area, chlorophyll a, b and carotenoids, maximum quantum efficiency of photosystem II, Rubisco content, improving the degeneration of thylakoid lamellae and lessening plastoglobuli number in thylakoid membranes are the major benefits attained with vermicompost treatments under salt stress.Analysis of proteomic revealed that VC upregulated chloroplastic ferredoxin–NADP reductase, plastocyanin, polyphenol oxidase, peptidyl-prolyl cis–trans isomerase, alpha-glucan phosphorylase H isozyme and maturase expression under salt stress. The results suggest that VC controls protein expression at the level of transcriptional and translational which may conserve photosynthetic components and prevent salt-induced harmful effects in broad bean plants.

Halamphora minima (Catenulaceae, Bacillariophyta), new brackish diatom species from the mudflat in Hampyeong Bay, Korea

The diversity of benthic diatoms living in the tidal flats is thought to be very high but has been somewhat marginalized from our main concern. We propose a new species, Halamphora minima sp. nov. isolated as monoclonal cultures from a tidal mudflat based on light and scanning electron microscopy and molecular analysis. This species is the smallest known species of the genus Halamphora to date. Other main characteristics that distinguish this species are the biseriate separated by strongly developed costa for the majority of the valve face, and density of dorsal striae. Phylogenetic analysis of the chloroplast ribulose bisphosphate carboxylase large chain gene showed that H. minima seems to be related to H. foramina, H. sydowii and H. tumida, but the phylogenetic location of H. minima was not clear due to the low bootstrap.

Combined effects of elevated CO2 and warmer temperature on limitations to photosynthesis and carbon sequestration in yellow birch.

Elevated CO2 and warmer temperature occur simultaneously under the current climate change. However, their combined effects on the photosynthetic traits in boreal trees are not well understood. This study investigated the morphological and photosynthetic responses of yellow birch (Betula alleghaniensis Britt.) to a combined treatment of CO2 and temperature (ambient, ACT (400μmolmol-1 CO2 and current temperature) vs elevated, ECT (750μmolmol-1 CO2 and current +4 °C temperature)). It was found that ECT significantly reduced leaf-area based photosynthetic rate (An), maximum Rubisco carboxylation rate (Vcmax), photosynthetic electron transport rate (Jmax), leaf nitrogen concentration, respiration and mesophyll conductance. There were two interesting findings: first, the primary mechanism of photosynthetic limitation shifted from Ribulose-1,5-bisphosphate (RuBP) carboxylation (related to Vcmax) to RuBP regeneration (related to Jmax) in response to ECT, leading to decreased transition point (Ci-t and An-t) from RuBP carboxylation to regeneration; second, the increase in total leaf area in response to ECT more than compensated for the downregulation of leaf-area based photosynthesis, leading to greater biomass in ECT than in ACT. We proposed a new protocol for evaluating photosynthetic limitations by comparing the relative relationship between the transition point (Ci-t and An-t) and the photosynthetic rate at growth CO2 (Ci-g and An-g). Furthermore, we found that Jmax (RuBP regeneration) was the primary limitation to An under ECT.

The trajectory in catalytic evolution of Rubisco in Posidonia seagrass species differs from terrestrial plants.

The CO2-fixing enzyme Ribulose bisphosphate carboxylase-oxygenase (Rubisco) links the inorganic and organic phases of the global carbon cycle. In aquatic systems, the catalytic adaptation of algae Rubiscos has been more expansive and followed an evolutionary pathway that appears distinct to terrestrial plant Rubisco. Here, we extend this survey to differing seagrass species of the genus Posidonia to reveal how their disjunctive geographical distribution and diverged phylogeny, along with their CO2 concentrating mechanisms (CCMs) effectiveness, have impacted their Rubisco kinetic properties. The Rubisco from Posidonia species showed lower carboxylation efficiencies and lower sensitivity to O2 inhibition than those measured for terrestrial C3 and C4-plant Rubiscos. Compared with the Australian Posidonia species, Rubisco from the Mediterranean Posidonia oceanica had 1.5-2-fold lower carboxylation and oxygenation efficiencies, coinciding with effective CCMs and five Rubisco large subunit amino acid substitutions. Among the Australian Posidonia species, CCM effectiveness was higher in Posidonia sinuosa and lower in the deep-living Posidonia angustifolia, likely related to the 20%-35% lower Rubisco carboxylation efficiency in P. sinuosa and the two-fold higher Rubisco content in P. angustifolia. Our results suggest that the catalytic evolution of Posidonia Rubisco has been impacted by the low CO2 availability and gas exchange properties of marine environments, but with contrasting Rubisco kinetics according to the time of diversification among the species. As a result, the relationships between maximum carboxylation rate and CO2- and O2-affinities of Posidonia Rubiscos follow an alternative path to that characteristic of terrestrial angiosperm Rubiscos.