Photosynthetic Carbon Fixation Research Articles

Physiological control on carbon isotope fractionation in marine phytoplankton

Abstract. One of the great challenges in biogeochemical research over the past half a century has been to quantify and understand the mechanisms underlying stable carbon isotope fractionation (εp) in phytoplankton in response to changing CO2 concentrations. This interest is partly grounded in the use of fossil photosynthetic organism remains as a proxy for past atmospheric CO2 levels. Phytoplankton organic carbon is depleted in 13C compared to its source because of kinetic fractionation by the enzyme RubisCO during photosynthetic carbon fixation, as well as through physiological pathways upstream of RubisCO. Moreover, other factors such as nutrient limitation, variations in light regime as well as phytoplankton culturing systems and inorganic carbon manipulation approaches may confound the influence of aquatic CO2 concentrations [CO2] on εp. Here, based on experimental data compiled from the literature, we assess which underlying physiological processes cause the observed differences in εp for various phytoplankton groups in response to C-demand/C-supply, i.e., particulate organic carbon (POC) production / [CO2]) and test potential confounding factors. Culturing approaches and methods of carbonate chemistry manipulation were found to best explain the differences in εp between studies, although day length was an important predictor for εp in haptophytes. Extrapolating results from culturing experiments to natural environments and for proxy applications therefore require caution, and it should be carefully considered whether culture methods and experimental conditions are representative of natural environments.

Physiological and proteomic responses of Chlamydomonas reinhardtii to arsenate and lead mixtures

Arsenic (As) and lead (Pb) are frequently emitted from various sources into environment, but microbial responses to their combined toxicity have not been systematically investigated. In this study, Chlamydomonas reinhardtii was exposed to two levels of arsenate (As (V), 50, 500 μg/L), Pb (II) (500, 5000 μg/L) and their mixture (50 μg/L As (V) + 500 μg/L Pb (II); 500 μg/L As (V) + 5000 μg/L Pb (II)). The growth of C. reinhardtii was inhibited more remarkably by As (V) than by Pb (II). The As stress was alleviated by Pb in the 50 μg/L As (V) + 500 μg/L Pb (II) treatment, but was enhanced upon the 500 μg/L As (V) + 5000 μg/L Pb (II) exposure, with more pronounced changes in a number of physiological parameters of the algal cells. Proteomic results showed that 71 differently expressed proteins (DEPs) in the treatment of 50 μg/L As (V) + 500 μg/L Pb (II), and 167 DEPs were identified in that of 500 μg/L As (V) + 5000 μg/L Pb (II). These proteins were involved in energy metabolism, photosynthetic carbon fixation, reactive oxygen scavenging and defense, and amino acid synthesis. Taken together, these physiological and proteomic data demonstrated that C. reinhardtii could resist the As (V) and Pb (II) combined treatments through extracellular complexation and intracellular pathways.

Genetic and Biochemical Investigation of Seed Fatty Acid Accumulation in Arabidopsis.

As a vegetable oil, consisting principally of triacylglycerols, is the major storage form of photosynthetically-fixed carbon in oilseeds which are of significant agricultural and industrial value. Photosynthesis in chlorophyll-containing green seeds, along with photosynthesis in leaves and other green organs, generates ATP and reductant (NADPH and NADH) needed for seed fatty acid production. However, contribution of seed photosynthesis to fatty acid accumulation in seeds have not been well-defined. Here, we report the contribution of seed-photosynthesis to fatty acid production by probing segregating green (photosynthetically-competent) and non-green or yellow (photosynthetically-non-competent) seeds in siliques of an Arabidopsis chlorophyll synthase mutant. Using this mutant, we found that yellow seeds lacking photosynthetic capacity reached 80% of amounts of oil in green seeds at maturity. Combining this with studies using shaded siliques, we determined that seed-photosynthesis accounts for 20% and silique and leaf/stem photosynthesis each account for ~40% of the ATP and reductant for seed oil production. Transmission electron microscopy (TEM) and pyridine nucleotides and ATP analyses revealed that seed photosynthesis provides ATP and reductant for oil production mostly during early development, as evidenced by delayed oil accumulation in non-green seeds. Transcriptomic analyses suggests that the oxidative pentose phosphate pathway could be the source of carbon, energy and reductants required for fatty acid synthesis beyond the early stages of seed development.

Effects of biological crusts on soil organic carbon in sandlands under a precipitation gradient

Biological crusts (Biocrusts) are important surface active coverings in arid and semi-arid regions, which affect the content of soil organic carbon (SOC), SOC labile fractions and stability of SOC through photosynthetic carbon fixation. At present, studies on the variation characteristics of SOC, SOC labile fractions and the stability of SOC in biocrusts are rather limited. In this study, two types of typical biocrusts (moss crusts and algae crusts) were selected along a precipitation gradient from northwest to southeast in the Mu Us Sandland (straight line distance 188 km) by measuring soil organic carbon (SOC), soil microbial biomass carbon (MBC), water soluble carbon (DOC), particulate carbon (POC), easily oxidizable carbon (ROC). We aimed to explore the effects of biocrusts on the stability of SOC and carbon decomposition across the precipitation gradient. Results showed that:1) Two types of biocrusts significantly increased the contents of SOC, MBC, DOC, POC, ROC and stability of SOC. Moss crusts increased SOC contents by 1.6 to 2.6 times as that of algae crusts. 2) The lowest SOC contents of the two types of biocrusts were 6.43 g·kg-1 and 14.50 g·kg-1 respectively, which showed an increasing trend with increasing precipitation along the gradient. 3) With the increases of precipitation, the decomposition time of moss litters gradually decreased. The decomposition coefficient of moss litters during the study period (From July to Feb-ruary of the next year) ranged from 0.010 to 0.014, which was significantly lower than that of vascular plants. The carbon release of moss litters from northwest to southeast was 8.09, 10.89, 12.88 g·kg-1, respectively. 4) Results of canonical correspondence analysis showed that water vapor partial pressure, actual evapotranspiration, annual average temperature, subsurface short-wave radiation, potential evapotranspiration and vapor pressure difference were the key climate factors affecting the content of SOC and its active components. Silt content was the main soil factor affecting the content of SOC and its active components.

Mitochondrial phosphoenolpyruvate carboxylase contributes to carbon fixation in the diatom Phaeodactylum tricornutum at low inorganic carbon concentrations.

Photosynthetic carbon fixation is often limited by CO2 availability, which led to the evolution of CO2 concentrating mechanisms (CCMs). Some diatoms possess CCMs that employ biochemical fixation of bicarbonate, similar to C4 plants, but whether biochemical CCMs are commonly found in diatoms is a subject of debate. In the diatom Phaeodactylum tricornutum, phosphoenolpyruvate carboxylase (PEPC) is present in two isoforms, PEPC1 in the plastids and PEPC2 in the mitochondria. We used real-time quantitative polymerase chain reaction, Western blots, and enzymatic assays to examine PEPC expression and PEPC activity, under low and high concentrations of dissolved inorganic carbon (DIC). We generated and analyzed individual knockout cell lines of PEPC1 and PEPC2, as well as a PEPC1/2 double-knockout strain. While we could not detect an altered phenotype in the PEPC1 knockout strains at ambient, low or high DIC concentrations, PEPC2 and the double-knockout strains grown under ambient air or lower DIC availability conditions showed reduced growth and photosynthetic affinity for DIC while behaving similarly to wild-type (WT) cells at high DIC concentrations. These mutants furthermore exhibited significantly lower 13 C/12 C ratios compared to the WT. Our data imply that in P. tricornutum at least parts of the CCM rely on biochemical bicarbonate fixation catalyzed by the mitochondrial PEPC2.

Comparative analyses of nutritional strategies among the species within the genus Paragymnodinium (Gymnodiniales, Dinophyceae).

The genus Paragymnodinium exhibits various nutritional strategies despite the fact that each species has pigmented plastids. This provides an excellent opportunity to investigate the evolution of the mode of nutrition and the function of the plastids through comparative studies. In this study, we analyzed the growth, pigment composition, absorption spectra, variable chlorophyll a fluorescence, and photosynthetic carbon fixation capabilities of P. stigmaticum, P. asymmetricum, and P. inerme. The autotrophic species P. asymmetricum and P. inerme without resorting to any nutrition from prey organisms displayed high PSII activity and carbon fixation rates. The pigment compositions of these two species were identical to those of other typical peridinin-containing type dinoflagellates. On the contrary, the phagotrophic species P. stigmaticum showed heterotrophic growth, and the variable chlorophyll a fluorescence properties and carbon fixation rates indicated significantly lower photosynthetic competence relative to those of the above two species. Paragymnodinium stigmaticum also contained peridinin, but pigment content ratios of peridinin, chlorophyll c2, and β-carotene were significantly different from those of other two species. The absorption spectrum analysis revealed a redshift in the peak of the Qy band of chlorophyll a in P. stigmaticum, presumably due to a change in chlorophyll-protein complexes. We conclude that such distinct differences in nutritional strategies between members of the genus Paragymnodinium would provide a platform for the hypothetical loss of photosynthetic function leading to colorless dinoflagellates.

Point mutations that boost aromatic amino acid production and CO2 assimilation in plants.

Aromatic compounds having unusual stability provide high-value chemicals and considerable promise for carbon storage. Terrestrial plants can convert atmospheric CO2 into diverse and abundant aromatic compounds. However, it is unclear how plants control the shikimate pathway that connects the photosynthetic carbon fixation with the biosynthesis of aromatic amino acids, the major precursors of plant aromatic natural products. This study identified suppressor of tyra2 (sota) mutations that deregulate the first step in the plant shikimate pathway by alleviating multiple effector-mediated feedback regulation in Arabidopsis thaliana. The sota mutant plants showed hyperaccumulation of aromatic amino acids accompanied by up to a 30% increase in net CO2 assimilation. The identified mutations can be used to enhance plant-based, sustainable conversion of atmospheric CO2 to high-energy and high-value aromatic compounds.

Effect of Culture pH on Properties of Exopolymeric Substances from Synechococcus PCC7942: Implications for Carbonate Precipitation

The role of culture conditions on the production of exopolymeric substances (EPS) by Synechococcus strain PCC7942 was investigated. Carbonate mineral precipitation in these EPS was assessed in forced precipitation experiments. Cultures were grown in HEPES-buffered medium and non-buffered medium. The pH of buffered medium remained constant at 7.5, but in non-buffered medium it increased to 9.5 within a day and leveled off at 10.5. The cell yield at harvest was twice as high in non-buffered medium than in buffered medium. High molecular weight (>10 kDa) and low molecular weight (3–10 kDa) fractions of EPS were obtained from both cultures. The cell-specific EPS production in buffered medium was twice as high as in non-buffered medium. EPS from non-buffered cultures contained more negatively charged macromolecules and more proteins than EPS from buffered cultures. The higher protein content at elevated pH may be due to the induction of carbon-concentrating mechanisms, necessary to perform photosynthetic carbon fixation in these conditions. Forced precipitation showed smaller calcite carbonate crystals in EPS from non-buffered medium and larger minerals in polymers from buffered medium. Vaterite formed only at low EPS concentrations. Experimental results are used to conceptually model the impact of pH on the potential of cyanobacterial blooms to produce minerals. We hypothesize that in freshwater systems, small crystal production may benefit the picoplankton by minimizing the mineral ballast, and thus prolonging the residence time in the photic zone, which might result in slow sinking rates.

Phytoplankton community composition, carbon sequestration, and associated regulatory mechanisms in a floodplain lake system

Phytoplankton contribute approximately 50% to the global photosynthetic carbon (C) fixation. However, our understanding of the corresponding C sequestration capacity and driving mechanisms associated with each individual phytoplankton taxonomic group is limited. Particularly in the hydrologically dynamic system with highly complex surface hydrological processes (floodplain lake systems). Through investigating seasonal monitoring data in a typical floodplain lake system and estimation of primary productivity of each phytoplankton taxonomic group individually using novel equations, this study proposed a phytoplankton C fixation model. Results showed that dominant phytoplankton communities had a higher gross carbon sequestration potential (CSP) (9.50 ± 5.06 Gg C each stage) and gross primary productivity (GPP) (65.46 ± 25.32 mg C m−2 d−1), but a lower net CSP (−1.04 ± 0.79 Gg C each stage) and net primary productivity (NPP) (−5.62 ± 4.93 mg C m−3 d−1) than rare phytoplankton communities in a floodplain lake system. Phytoplanktonic GPP was high (317.94 ± 73.28 mg C m−2 d−1) during the rainy season and low (63.02 ± 9.65 mg C m−2 d−1) during the dry season. However, their NPP reached the highest during the rising-water stage and the lowest during the receding-water stage. Findings also revealed that during the rainy season, high water levels (p = 0.56**) and temperatures (p = 0.37*) as well as strong solar radiation (p = 0.36*) will increase photosynthesis and accelerate metabolism and respiration of dominant phytoplankton communities, then affect primary productivity and CSP. Additionally, water level fluctuations drive changes in nutrients (p = −0.57*) and metals (p = −0.68*) concentrations, resulting in excessive nutrients and metals slowing down phytoplankton growth and reducing GPP. Compared with the static water lake system, the floodplain lake system with a lower net CSP became a heterotrophic C source.

Characterization of the Wave Phenomenon in Flash-Induced Fluorescence Relaxation and Its Application to Study Cyclic Electron Pathways in Microalgae.

Photosynthesis is a series of redox reactions, in which several electron transport processes operate to provide the energetic balance of light harvesting. In addition to linear electron flow, which ensures the basic functions of photosynthetic productivity and carbon fixation, alternative electron transport pathways operate, such as the cyclic electron flow (CEF), which play a role in the fine tuning of photosynthesis and balancing the ATP/NADPH ratio under stress conditions. In this work, we characterized the electron transport processes in microalgae species that have high relevance in applied research and industry (e.g., Chlorella sorokiniana, Haematococcus pluvialis, Dunaliella salina, Nannochloropsis sp.) by using flash-induced fluorescence relaxation kinetics. We found that a wave phenomenon appeared in the fluorescence relaxation profiles of microalgae to different extents; it was remarkable in the red cells of H. pluvialis, D. salina and C. sorokiniana, but it was absent in green cells of H. pluvialis and N. limnetica. Furthermore, in microalgae, unlike in cyanobacteria, the appearance of the wave required the partial decrease in the activity of Photosystem II, because the relatively high Photosystem II/Photosystem I ratio in microalgae prevented the enhanced oxidation of the plastoquinone pool. The wave phenomenon was shown to be related to the antimycin A-sensitive pathway of CEF in C. sorokiniana but not in other species. Therefore, the fluorescence wave phenomenon appears to be a species-specific indicator of the redox reactions of the plastoquinone pool and certain pathways of cyclic electron flow.

Photosynthetic Carbon Fixation and Sucrose Metabolism Supplemented by Weighted Gene Co-expression Network Analysis in Response to Water Stress in Rice With Overlapping Growth Stages.

Drought stress at jointing and booting phases of plant development directly affects plant growth and productivity in rice. Limited by natural factors, the jointing and booting stages in rice varieties are known to overlap in high-latitude areas that are more sensitive to water deficit. However, the regulation of photosynthetic carbon fixation and sucrose metabolism in rice leaves under different degrees of drought stress remains unclear. In this study, rice plants were subjected to three degrees of drought stress (−10, −25, −and 40 kPa) for 15 days during the jointing-booting stage, we investigated photosynthetic carbon sequestration and sucrose metabolism pathways in rice leaves and analyzed key genes and regulatory networks using transcriptome sequencing in 2016. And we investigated the effects of drought stress on the growth periods of rice with overlapping growth periods in 2016 and 2017. The results showed that short-term drought stress promoted photosynthetic carbon fixation. However, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity significantly decreased, resulting in a significant decrease in photosynthetic rate. Drought stress increased the maximum activity of fructose-1,6-bisphosphate aldolase (FBA). FBA maintains the necessary photosynthetic rate during drought stress and provides a material base after the resumption of irrigation in the form of controlling the content of its reaction product triose phosphate. Drought stress significantly affected the activities of sucrose synthase (SuSase) and sucrose phosphate synthase (SPS). Vacuoles invertase (VIN) activity increased significantly, and the more severe the drought, the higher the VIN activity. Severe drought stress at the jointing-booting stage severely restricted the growth process of rice with overlapping growth stages and significantly delayed heading and anthesis stages. Transcriptome analysis showed that the number of differentially expressed genes was highest at 6–9 days after drought stress. Two invertase and four β-amylase genes with time-specific expression were involved in sucrose-starch metabolism in rice under drought stress. Combined with weighted gene co-expression network analysis, VIN and β-amylase genes up-regulated throughout drought stress were regulated by OsbZIP04 and OsWRKY62 transcription factors under drought stress. This study showed that any water deficit at the jointing-booting stage would have a serious effect on sucrose metabolism in leaves of rice with overlapping growth stages.

SlMYC2 mediates jasmonate-induced tomato leaf senescence by promoting chlorophyll degradation and repressing carbon fixation

Leaf senescence occurs as the last developmental phase of leaf. The initiation and progression of leaf senescence is highly regulated by a plethora of internal developmental signals and environmental stimuli. Being an important class of phytohormones, jasmonates (JAs) are shown to induce premature leaf senescence in tomato (Solanum lycopersicum), nevertheless, the underlying mechanisms remain enigmatic. Here, we report that tomato MYC2, a key factor in the JA signal transduction, functions in JA-induced tomato leaf senescence by promoting chlorophyll degradation and inhibiting photosynthetic carbon fixation. We found that exogenous application of MeJA reduced chlorophyll content, decreased carbon assimilation rates and disrupted membrane integrity. We further demonstrated using SlMYC2-RNAi tomato plants that SlMYC2 enhanced the expression of SlPAO, which encodes a chlorophyll degradation enzyme, but suppressed the expression of SlRCA and SlSBPASE, both of which are required for photosynthesis and growth in plants. Dual-luciferase assay confirmed that SlMYC2 activated the transcription of SlPAO, but inhibited the transcription of SlRCA and SlSBPASE. Furthermore, repression of SlRCA led to typical features associated with leaf senescence in tomato. Taken together, these results favor that tomato MYC2 acts positively in the regulation of JA-dependent tomato leaf senescence. The results extend our mechanistic understanding of JA-induced senescence in an important horticultural crop.

Molecular Mechanism of Organic Pollutant-Induced Reduction of Carbon Fixation and Biomass Yield in Oryza sativa L.

Photosynthetic carbon fixation is fundamental for plant growth and is a key process driving the global carbon cycle. This study explored the mechanism of disturbed carbon fixation in Oryza sativa L. by organic pollutants 2,3,4,5-tetrachlorobiphenyl (CB 61), 4'-hydroxy-2,3,4,5-tetrachlorobiphenyl (4'-OH-CB 61), 2,2',4,4'-tetrabromo diphenyl ether (BDE 47), tricyclazole (TRI), and pyrene. The biomass of rice exposed to 4'-OH-CB 61, TRI, and BDE 47 was on average 80.63% of that of the control (p < 0.05), and the inhibition of net photosynthetic rate was 59.15% by 4'-OH-CB 61. Proteomics confirmed that 4'-OH-CB 61 significantly downregulated the enzymes in the photosynthetic carbon fixation pathway, which was attributed to the decrease in ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the rate-limiting enzyme in the Calvin cycle. In detail, decreased Rubisco activity (6.96-33.44%) and downregulated OsRBCS2-5 encoding small Rubisco subunits (-6.80 < log2FC < -2.13) by 4'-OH-CB 61, TRI, and BDE 47 were in line with biomass yield reduction. Molecular docking and dynamic simulation suggested that the three pollutants potentially competed with CO2 for binding to the active sites in Rubisco, leading to reduced CO2 capture efficiency. These results revealed the molecular mechanism of organic pollution-induced rice yield reduction, contributing to improving the understanding of crop growth and carbon sequestration capacity of organics-contaminated soils globally.

Effects of desiccation and rehydration on carbon fixation and DOC release in Sargassum thunbergii

Although macroalgae is regarded as the emerging fourth category of “blue carbon”, few studies are available on its carbon sequestration and DOC (dissolved organic carbon) release mechanism at present. In addition, tidal action is an important environmental factor in the intertidal zone, which affects the carbon sequestration and DOC release of macroalgae. Therefore, we set 4-h desiccation and 1-h rehydration to examine the effects of drying-rewetting on the photosynthetic carbon fixation capacity and DOC release mechanism of Sargassum thunbergii. The flowing seawater can provide the nutrients for the growth of marine plants, which plays a vital role in the growth and reproduction of marine plants, so a measurement system that can adjust the flow speed was designed to study the effects of 1 h-rehydration on S. thunbergii. The results showed the desiccation decreased carbon fixation capacity, but increased the protection of PSII by down-regulating Fv/Fm and ФPSII and up-regulating NPQ, thereby promoting the recovery of carbon fixation capacity during 1-h rehydration period. It was found that the DOC release rate of S. thunbergii decreased with the carbon fixation rate increase and it was much greater than the carbon fixation rate in the 1-h rehydration period. The DOC release after rehydration was somehow a passive release, which was related to the increase of osmotic pressure, cell membrane damage, and carbon accumulation caused by desiccation. This measurement system can provide a practical reference for the study of carbon sink of macroalgae.

Potassium regulates diel leaf growth of Brassica napus by coordinating the rhythmic carbon supply and water balance.

Carbon and water are two main factors limiting leaf expansion. Restriction of leaf growth by low availability of carbon or water is among the earliest visible effects of potassium (K) deficiency. It is not known how K is involved in regulating the rhythmic supply of these two substrates, which differ remarkably across the day-night cycle, affecting leaf expansion. We investigated the effects of different K regimes on the time courses of leaf expansion, carbon assimilation, carbohydrates, and hydraulic properties of Brassica napus. Potassium supply increased leaf area, predominantly by promoting night-time leaf expansion (>60%), which was mainly associated with increased availability of carbohydrates from photosynthetic carbon fixation and import from old leaves rather than improvement of leaf hydraulics. However, sufficient K improved leaf hydraulic conductance to balance diurnal evaporative water loss and increase the osmotic contribution of water-soluble carbohydrates, thereby maintaining leaf turgor and increasing the daytime expansion rate. The results also indicated an ontogenetic role of K in modifying the amplitude of circadian expansion; almost 80% of the increase in leaf area occurred before the area reached 66.9% of the mature size. Our data provide mechanistic insight into K-mediated diel coordination of rhythmic carbon supply and water balance in leaf expansion.

Improving biomass growth of Nannochloropsis oceanica with electrical treatment

In order to improve photosynthetic carbon fixation efficiency of microalgae, electrochemical responses of Nannochloropsis oceanica were investigated in terms of cell growth and physiological metabolism. A direct current with varying voltage was applied to N. oceanica during cultivation and effects on bio-regulation and electron transfer were assessed. When applied voltage increased from 0 V to 0.005 V and then to 0.5 V, the biomass dry weight first increased due to enhanced effective quantum yield of PSII photochemical energy conversion, chlorophyll a and nitrogen/orthophosphate uptake rates of microalgal cells, and then decreased due to disruption of cell structures. The optimal photosynthetic carbon fixation with 15% CO2 occurred at 0.005 V, resulting in a 32. 09% increase in microalgal biomass compared to that without electrical treatment.

The Marine Biodiversity Observation Network Plankton Workshops: Plankton Ecosystem Function, Biodiversity, and Forecasting—Research Requirements and Applications

The Marine Biodiversity Observation Network Plankton Workshops: Plankton Ecosystem Function, Biodiversity, and Forecasting—Research Requirements and Applications

Single and combined exposure to micro(nano)plastics and azithromycin disturbing the photosynthetic carbon fixation of Synechocystis sp.

The co-exposure to MPs and AZM caused greater inhibitory effects on the photosynthetic carbon fixation in Synechocystis sp. compared to the co-exposure to NPs and AZM.

Proteomic analysis of a plastid gene encoding RPS4 mutant in Chinese cabbage (Brassica campestris L. ssp. pekinensis).

Plastids are important plant cell organelles containing a genome and bacterial-type 70S ribosomes-primarily composed of plastid ribosomal proteins and ribosomal RNAs. In this study, a chlorophyll-deficient mutant (cdm) obtained from double-haploid Chinese cabbage 'FT' was identified as a plastome mutant with an A-to-C base substitution in the plastid gene encoding the ribosomal protein RPS4. To further elucidate the function and regulatory mechanisms of RPS4, a comparative proteomic analysis was conducted between cdm and its wild-type 'FT' plants by isobaric tags and a relative and absolute quantitation (iTRAQ)-based strategy. A total of 6,245 proteins were identified, 540 of which were differentially abundant proteins (DAPs) in the leaves of cdm as compared to those of 'FT'-including 233 upregulated and 307 downregulated proteins. Upregulated DAPs were mainly involved in translation, organonitrogen compound biosynthetic process, ribosomes, and spliceosomes. Meanwhile, downregulated DAPs were mainly involved in photosynthesis, photosynthetic reaction centres, photosynthetic light harvesting, carbon fixation, and chlorophyll binding. These results indicated an important role of RPS4 in the regulation of growth and development of Chinese cabbage, possibly by regulating plastid translation activity by affecting the expression of specific photosynthesis- and cold stress-related proteins. Moreover, a multiple reaction monitoring (MRM) test and quantitative real-time polymerase chain reaction analysis confirmed our iTRAQ results. Quantitative proteomic analysis allowed us to confirm diverse changes in the metabolic pathways between cdm and 'FT' plants. This work provides new insights into the regulation of chlorophyll biosynthesis and photosynthesis in Chinese cabbage.

Proteomic and biochemical responses to different concentrations of CO2 suggest the existence of multiple carbon metabolism strategies in Phaeodactylum tricornutum

BackgroundDiatoms are well known for high photosynthetic efficiency and rapid growth rate, which are not only important oceanic primary producer, but also ideal feedstock for microalgae industrialization. Their high success is mainly due to the rapid response of photosynthesis to inorganic carbon fluctuations. Thus, an in-depth understanding of the photosynthetic carbon fixation mechanism of diatoms will be of great help to microalgae-based applications. This work directed toward the analysis of whether C4 photosynthetic pathway functions in the model marine diatom Phaeodactylum tricornutum, which possesses biophysical CO2-concentrating mechanism (CCM) as well as metabolic enzymes potentially involved in C4 photosynthetic pathway.ResultsFor P. tricornutum, differential proteome, enzyme activities and transcript abundance of carbon metabolism-related genes especially biophysical and biochemical CCM-related genes in response to different concentrations of CO2 were tracked in this study. The upregulated protein abundance of a carbonic anhydrases and a bicarbonate transporter suggested biophysical CCM activated under low CO2 (LC). The upregulation of a number of key C4-related enzymes in enzymatic activity, transcript and protein abundance under LC indicated the induction of a mitochondria-mediated CCM in P. tricornutum. Moreover, protein abundance of a number of glycolysis, tricarboxylic acid cycle, photorespiration and ornithine–urea cycle related proteins upregulated under LC, while numbers of proteins involved in the Calvin cycle and pentose phosphate pathway were downregulated. Under high CO2 (HC), protein abundance of most central carbon metabolism and photosynthesis-related proteins were upregulated.ConclusionsThe proteomic and biochemical responses to different concentrations of CO2 suggested multiple carbon metabolism strategies exist in P. tricornutum. Namely, LC might induce a mitochondrial-mediated C4-like CCM and the improvement of glycolysis, tricarboxylic acid cycle, photorespiration and ornithine–urea cycle activity contribute to the energy supply and carbon and nitrogen recapture in P. tricornutum to cope with the CO2 limitation, while P. tricornutum responds to the HC environment by improving photosynthesis and central carbon metabolism activity. These findings can not only provide evidences for revealing the global picture of biophysical and biochemical CCM in P. tricornutum, but also provide target genes for further microalgal strain modification to improve carbon fixation and biomass yield in algal-based industry.