Tradeoff in the supply and demand for CO 2 dominates the divergence of net photosynthesis rates of functional plants in alpine ecosystems

As regional heterogeneity on the Qinghai Tibetan Plateau (QTP), the “greening rate” between alpine steppe in the west and alpine meadow ecosystems in the east is difference during the past several decades. To investigate the difference, the net photosynthetic rate (An) and the supply (mesophyll conductance ( g), stomatal conductance ( g)) and demand (the maximum rates of Rubisco carboxylase activity ( V) and photosynthetic electron transport ( J)) for CO of three plants functional types (PFTs) were measured. Other functional traits and influencing factors were compared among ecosystems along the altitudinal gradients of QTP. The An of the PFTs was simulated under potential future conditions. At high altitudes, grass was found to maintain a relatively stable An by decreasing V, J, and g, while slightly increasing g, compared with that at a low altitude. The An of sedge and shrubs increased with rising V, J and g and g values, resulting in a large increment in the An at low altitudes. Grass seemed to be less sensitive to the environment by reducing the supply of and holding onto CO , while sedge and shrub increased both. Grass and sedge should be divided into two PFTs rather than remaining as one based on their opposite physiological and morphological functions in response to climate change. The ecosystem at 3600 m was transitional. C was likely to be a more dominant factor than T in affecting the An of grass. The order of rising An in PFTs was shrub > sedge > grass and the An of alpine meadow was found to increase more under the two future climate scenarios.

EFFECT OF SALT (NaCl) ON PHOTOSYNTHESIS AND CARBOHYDRATES IN LEAVES OF TWO RAGI (ELEUSINE CORACANA (L.) GAERTN) VARIETIES

Salinity is one of the most widespread environmental threats to global crop production, especially in arid and semi-arid regions. Photosynthesis and carbohydrates were determined in two ragi (Eleusine coracana (L.) Gaertn) varieties (CO13 and PAIYUR-1), subjected to salt stress of different concentrations (0, 40, 80 and 120mM). Salinity was given as a basal dose and sampling was done in leaves on 30th Days. After Treatment (DAT). There was a marked variation in the photosynthetic rates and ribulose-1, 5-bisphosphate carboxylase activity between the two ragi varieties subjected to salt stress. Photosystem II (PSII) and sucrose phosphate synthase activities were also significantly reduced as measured by salt stressed conditions. The quantity of glucose and sucrose decreased with increasing salt stress while starch showed a reverse trend under salt-stressed conditions. The results revealed that CO-13 exhibits higher photosynthetic rates and activities of ribulose-1,5-bisphosphate carboxylase, sucrose phosphate synthase with photochemical efficiency of PSII compared to PAIYUR-1

Nitrogen Fixation in Soybean Nodules Affects Seed Protein and Oil Contents: The Suggested Mechanism from the Coordinated Changes of Seed Chemical Compositions and Phosphoenolpyruvate Carboxylase Activity Caused by Different Types of Nitrogen Fertilizer

The contents of seed storage compounds, protein and oil, determine the best use of soybean seeds, namely materials for food processing and oil production. Genetic and environmental factors could affect the chemical compositions of soybean seeds. However, the mechanisms of how the accumulation of these primary seed compounds is regulated are mostly unclear. In this chapter, we describe the different effects of nodulation on the protein and oil contents in soybean seeds and the crucial role of phosphoenolpyruvate carboxylase (PEPC) in the protein accumulation of soybean seeds. Based on our previous studies on soybean seeds, we introduce five manners deduced; (1) protein accumulation is independent of oil accumulation, (2) nitrogen fixation results in decreasing oil amount per seed and decreased seed oil content, (3) a high pseudo negative correlation between protein and oil contents in seeds is likely to be observed under less nitrogen supply from the soil, (4) nitrogen absorbed from soil during the late growth stage promote seed production, (5) plant-type PEPC, ex. Gmppc2 in soybean could play a role in amino acid biosynthesis for storage protein accumulation in seeds during the late maturation period.

Updating the steady state model of C4 photosynthesis

C4 plants play a key role in world agriculture. For example, C4 crops such as maize and sorghum are major contributors to both first and third world food production and the C4 grasses sugarcane; miscanthus and switchgrass are major plant sources of bioenergy. In the challenge to manipulate and enhance C4 photosynthesis, steady state models of leaf photosynthesis provide and important tool for gas exchange analysis and thought experiments that can explore photosynthetic pathway changes. Here the C4 photosynthetic model by von Caemmerer and Furbank (1999) has been updated with new kinetic parameterisation and temperature dependencies added. The parameterisation was derived from experiments on the C4 monocot, Setaria viridis, which for the first time provides a cohesive parametrisation. Mesophyll conductance and its temperature dependence have also been included, as this is an important step in the quantitative correlation between the initial slope of the CO2 response curve of CO2 assimilation and in vitro PEP carboxylase activity. Furthermore, the equations for chloroplast electron transport have been updated to include cyclic electron transport flow and equations have been added to calculate electron transport rate from measured CO2 assimilation rates.HighlightThe C4 photosynthesis model by von Caemmerer and Furbank (1999) has been updated. It now includes temperature dependencies and equations to calculate electron transport rate from measured CO2 assimilation rates.

13C tracer analysis identifies extensive recycling of endogenous CO2 in vivo

Abstract13C tracing analysis is increasingly used to monitor cellular metabolism in vivo and in intact cells, but data interpretation is still the key element to unveil the complexity of metabolic activities. We have performed [U-13C]-glucose and [U-13C]-glutamine tracing in sarcoma-bearing mice (in vivo) and in cancer cell lines (in vitro). 13C enrichment of metabolites in cultured cells and tissues was determined by liquid chromatography coupled with high-resolution mass spectrometer (LC-HRMS). As expected, citrate M+2 or M+4 is the dominant mass isotopologue in vitro. However, citrate M+1 was unexpectedly the dominant isotopologue in mice receiving [U-13C]-glucose or [U-13C]-glutamine infusion. One plausible explanation is that 13CO2 produced from the oxidation of 13C tracers in vitro is negligible due to the dilution of HCO3- supplemented to cell culture when sodium bicarbonante is used and diffusible volume of CO2 in the culture incubator, while endogenous 13CO2 in vivo is substantial and is fixed into the TCA cycle, purine, and serine, resulting in M+1 isotopologues. A time course study shows the generation of high abundance citrate M+1 early in plasma, which may serve as a potent non-invasive biomarker of tissue pyruvate carboxylase activity. Altogether, our results show that recycling of endogenous CO2 is substantial in vivo and provides important insights into the experimental design and data interpretation of 13C tracing assays.

Engineering intracellular malonyl-CoA availability in microbial hosts and its impact on polyketide and fatty acid synthesis

Malonyl-CoA is an important central metabolite serving as the basic building block for the microbial synthesis of many pharmaceutically interesting polyketides, but also fatty acid–derived compounds including biofuels. Especially Saccharomyces cerevisiae, Escherichia coli, and Corynebacterium glutamicum have been engineered towards microbial synthesis of such compounds in recent years. However, developed strains and processes often suffer from insufficient productivity. Usually, tightly regulated intracellular malonyl-CoA availability is regarded as the decisive bottleneck limiting overall product formation. Therefore, metabolic engineering towards improved malonyl-CoA availability is essential to design efficient microbial cell factories for the production of polyketides and fatty acid derivatives. This review article summarizes metabolic engineering strategies to improve intracellular malonyl-CoA formation in industrially relevant microorganisms and its impact on productivity and product range, with a focus on polyketides and other malonyl-CoA-dependent products.Key Points• Malonyl-CoA is the central building block of polyketide synthesis.• Increasing acetyl-CoA supply is pivotal to improve malonyl-CoA availability.• Improved acetyl-CoA carboxylase activity increases availability of malonyl-CoA.• Fatty acid synthesis as an ambivalent target to improve malonyl-CoA supply.

Electronic Structure Benchmark Calculations of CO2 Fixing Elementary Chemical Steps in RuBisCO Using the Projector-Based Embedding Approach

<div>Ribulose 1,5-bisphosphate carboxylase-oxygenase (RuBisCO) is the main enzyme involved in atmospheric carbon dioxide (CO₂) fixation in the biosphere. This enzyme catalyses a set of five chemical steps that take place in the same active-site within magnesium (II) coordination sphere. Here, a set of electronic structure benchmark calculations have been carried out on a reaction path proposed by Gready <i>et al.</i> by means of the projector-based embedding approach. Activation and reaction energies for all main steps catalyzed by RuBisCO have been calculated at the MP2, SCS-MP2, CCSD and CCSD(T)/aug-cc-pVDZ and cc-pVDZ levels of theory. </div><div><br></div><div>The treatment of the magnesium cation with post-HF methods is explored to determine the nature of its involvement in the mechanism. With the high-level ab initio values as a reference, we tested the performance of a set of density functional theory (DFT) exchange-correlation (xc) functionals in reproducing the reaction energetics of RuBisCO carboxylase activity on a set of model fragments. Different DFT xc-functionals show large variation in activation and reaction energies. Activation and reaction energies computed at the B3LYP level are close to the reference SCS-MP2 results for carboxylation, hydration and protonation reactions.</div><div><br></div><div>However, for the carbon-carbon bond dissociation reaction, B3LYP and other functionals give results that differ significantly from the ab initio reference values. The results show the applicability of the projector-based embedding approach to metalloenzymes. This technique removes the uncertainty associated with the selection of different DFT xc-functionals and so can overcome some of inherent limitations of DFT calculations, complementing and potentially adding to modelling of enzyme reaction mechanisms with DFT methods.</div>

Electronic Structure Benchmark Calculations of CO2 Fixing Elementary Chemical Steps in RuBisCO Using the Projector-Based Embedding Approach

<div>Ribulose 1,5-bisphosphate carboxylase-oxygenase (RuBisCO) is the main enzyme involved in atmospheric carbon dioxide (CO₂) fixation in the biosphere. This enzyme catalyses a set of five chemical steps that take place in the same active-site within magnesium (II) coordination sphere. Here, a set of electronic structure benchmark calculations have been carried out on a reaction path proposed by Gready <i>et al.</i> by means of the projector-based embedding approach. Activation and reaction energies for all main steps catalyzed by RuBisCO have been calculated at the MP2, SCS-MP2, CCSD and CCSD(T)/aug-cc-pVDZ and cc-pVDZ levels of theory. </div><div><br></div><div>The treatment of the magnesium cation with post-HF methods is explored to determine the nature of its involvement in the mechanism. With the high-level ab initio values as a reference, we tested the performance of a set of density functional theory (DFT) exchange-correlation (xc) functionals in reproducing the reaction energetics of RuBisCO carboxylase activity on a set of model fragments. Different DFT xc-functionals show large variation in activation and reaction energies. Activation and reaction energies computed at the B3LYP level are close to the reference SCS-MP2 results for carboxylation, hydration and protonation reactions.</div><div><br></div><div>However, for the carbon-carbon bond dissociation reaction, B3LYP and other functionals give results that differ significantly from the ab initio reference values. The results show the applicability of the projector-based embedding approach to metalloenzymes. This technique removes the uncertainty associated with the selection of different DFT xc-functionals and so can overcome some of inherent limitations of DFT calculations, complementing and potentially adding to modelling of enzyme reaction mechanisms with DFT methods.</div>