Accumulation Of Carbon Reserves Research Articles

Azospirillum baldaniorum improves acclimation, lipid productivity and oxidative response of a microalga under salt stress

There is a growing interest in using microalgae for a variety of purposes, including production of pharmaceuticals, wastewater treatment, CO2 capture from flue gases, and biomass as a feedstock for biofuels and feed/food supplements. In addition to environmental conditions, the local availability of suitable water for massive production microalgal biomass often compromises biomass productivity, and thus, techno-economic feasibility towards implementation. Although salinity tends to promote accumulation of carbon reserves in some freshwater microalgae, which could be advantageous for producing feedstocks for biofuels, the negative effect on growth usually off-sets this benefit, and results in a lower productivity of the target. The aim of this study was to analyse whether inoculation with a plant growth-promoting bacterium could ameliorate the detrimental effect of NaCl on growth of a microalga. We showed that inoculation with Azospirillum baldaniorum Sp245 improved the early response of Scenedesmus obliquus CS1 to NaCl addition, and its overall growth performance. As a result, co-cultures attained higher biomass and lipids productivities than microalgal axenic cultures. Lipid productivity raised up due to both: an increased biomass production and a higher lipids enrichment in the biomass. Similar results were obtained under constant light and temperature or under climate-simulated conditions in environmental photobioreactors (Phenometrics™). Inoculation with the bacterium lowered the levels of reactive oxygen species, SH groups, chlorophyll and carotenoids in the algal cells, as well as cell damage, induced by salt stress. Inoculation with Azospirillum strains genetically modified to produce contrasting levels of IAA, suggested that although under non-stress conditions most of the algal growth promotion is IAA-dependent, under salt stress other still-unidentified factor(s) might play a more prominent role in algal stimulation.This study increased the current understanding of the mechanisms underpinning bacterial-microalgal associations, and helps to design alternative strategies to increase algal productivity under suboptimal growth conditions, as most applications demand.

Prediction of the radiation characteristics and the light absorption rate of Chlamydomonas reinhardtii cultivated under a progressive nitrogen starvation and accumulating carbon reserves

This paper aims to develop a new method to predict the radiation characteristics of the green microalgae C. reinhardtii during progressive nitrogen starvation that accounts for the physiological changes induced by nutritional stress. First, the radiation characteristics of C. reinhardtii cultivated under progressive nitrogen starvation were predicted in the photosynthetically active radiation (PAR) region based on Lorenz–Mie theory using effective spectral optical properties of cells accounting for pigment contents, cessation of cell division, cell volume and size, and carbon reserve storage in the form of starch. The predictions of the spectral mass absorption and scattering cross-sections agreed very well with experimental measurements for a wide range of starch mass fractions. Second, the predicted and measured radiation characteristics were used to determine the local rate of photon absorption (LRPA) in the PAR region using the two-flux approximation and simplified two-flux approximation. The LRPA PAR-averaged relative difference between experimental calculation and numerical predictions was less than 9% for all starch mass content considered. In addition, the influence of the mass scattering cross-section on the PAR-averaged LRPA was found to be negligible despite the starch accumulation in microalgae cells. Finally, the numerically-validated tools developed in this study could be used for generating a database of radiation characteristics helpful in order to model growth and carbon reserve accumulation kinetics in microalgae.

TOR signaling in the green picoalga Ostreococcus tauri

Target of rapamycin (TOR) is a master regulator that controls growth and metabolism by integrating external and internal signals. Although there was a great progress in the study of TOR in plants and in the model alga Chlamydomonas, scarce data are available in other green algae. Thus, in this work we studied TOR signaling in Ostreococcus tauri, the smallest free-living eukaryote described to date. This picoalga is particularly important because it has a key site at the base of the green lineage and is part of the marine phytoplankton, contributing to global photosynthesis. We investigated OtTOR complex in silico and experimentally, by using first- and second-generation TOR inhibitors, such as rapamycin and PP242. We analyzed the effect of TOR down-regulation on cell growth and on the accumulation of carbon reserves. The results showed that O. tauri responds to TOR inhibitors more similarly to plants than to Chlamydomonas, being PP242 a valuable tool to study this pathway. Besides, Ottor expression analysis revealed that the kinase is dynamically regulated under nutritional stress. Our data indicate that TOR signaling is conserved in O. tauri and we propose this alga as a good and simple model for studying TOR kinase and its regulation.

Comparing statistical and mechanistic models to identify the drivers of mortality within a rear-edge beech population

Since several studies have been reporting an increase in the decline of forests, a major issue in ecology is to better understand and predict tree mortality. The interactions between the different factors and the physiological processes giving rise tree mortality, as well as the inter-individual variability in mortality risk, still need to be better assessed. This study investigates mortality in a rear-edge population of European beech (Fagus sylvatica L.) using a combination of statistical and process-based modelling approaches. Based on a survey of 4323 adult beeches since 2002 within a natural reserve, we first used statistical models to quantify the effects of competition, tree growth, size, defoliation and fungi presence on mortality. Secondly, we used an ecophysiological process-based model (PBM) to separate out the different mechanisms giving rise to temporal and inter-individual variations in mortality by simulating depletion of carbon stocks, loss of hydraulic conductance and damage due to late frosts in response to climate. The combination of all these simulated processes was associated with the temporal variations in the population mortality rate. The individual probability of mortality decreased with increasing mean growth, and increased with increasing crown defoliation, earliness of budburst, fungi presence and increasing competition, in the statistical model. Moreover, the interaction between tree size and defoliation was significant, indicating a stronger increase in mortality associated to defoliation in smaller than larger trees. Finally, the PBM predicted a higher conductance loss together with a higher level of carbon reserves for trees with earlier budburst, while the ability to defoliate the crown was found to limit the impact of hydraulic stress at the expense of the accumulation of carbon reserves. We discuss the convergences and divergences obtained between statistical and process-based approaches and we highlight the importance of combining them to characterize the different processes underlying mortality, and the factors modulating individual vulnerability to mortality.

Physiological Profiling of Embryos and Dormant Seeds in Two Arabidopsis Accessions Reveals a Metabolic Switch in Carbon Reserve Accumulation.

In flowering plants, sugars act as carbon sources providing energy for developing embryos and seeds. Although most studies focus on carbon metabolism in whole seeds, knowledge about how particular sugars contribute to the developmental transitions during embryogenesis is scarce. To develop a quantitative understanding of how carbon composition changes during embryo development, and to determine how sugar status contributes to final seed or embryo size, we performed metabolic profiling of hand-dissected embryos at late torpedo and mature stages, and dormant seeds, in two Arabidopsis thaliana accessions with medium [Columbia-0 (Col-0)] and large [Burren-0 (Bur-0)] seed sizes, respectively. Our results show that, in both accessions, metabolite profiles of embryos largely differ from those of dormant seeds. We found that developmental transitions from torpedo to mature embryos, and further to dormant seeds, are associated with major metabolic switches in carbon reserve accumulation. While glucose, sucrose, and starch predominantly accumulated during seed dormancy, fructose levels were strongly elevated in mature embryos. Interestingly, Bur-0 seeds contain larger mature embryos than Col-0 seeds. Fructose and starch were accumulated to significantly higher levels in mature Bur-0 than Col-0 embryos, suggesting that they contribute to the enlarged mature Bur-0 embryos. Furthermore, we found that Bur-0 embryos accumulated a higher level of sucrose compared to hexose sugars and that changes in sucrose metabolism are mediated by sucrose synthase (SUS), with SUS genes acting non-redundantly, and in a tissue-specific manner to utilize sucrose during late embryogenesis.

Taxonomy, together with ontogeny and growing conditions, drives needleleaf species' sensitivity to climate in boreal North America.

Currently, there is no consensus regarding the way that changes in climate will affect boreal forest growth, where warming is occurring faster than in other biomes. Some studies suggest negative effects due to drought-induced stresses, while others provide evidence of increased growth rates due to a longer growing season. Studies focusing on the effects of environmental conditions on growth-climate relationships are usually limited to small sampling areas that do not encompass the full range of environmental conditions; therefore, they only provide a limited understanding of the processes at play. Here, we studied how environmental conditions and ontogeny modulated growth trends and growth-climate relationships of black spruce (Picea mariana) and jack pine (Pinus banksiana) using an extensive dataset from a forest inventory network. We quantified the long-term growth trends at the stand scale, based on analysis of the absolutely dated ring-width measurements of 2,266 trees. We assessed the relationship between annual growth rates and seasonal climate variables and evaluated the effects of various explanatory variables on long-term growth trends and growth-climate relationships. Both growth trends and growth-climate relationships were species-specific and spatially heterogeneous. While the growth of jack pine barely increased during the study period, we observed a growth decline for black spruce which was more pronounced for older stands. This decline was likely due to a negative balance between direct growth gains induced by improved photosynthesis during hotter-than-average growing conditions in early summers and the loss of growth occurring the following year due to the indirect effects of late-summer heatwaves on accumulation of carbon reserves. For stands at the high end of our elevational gradient, frost damage during milder-than-average springs could act as an additional growth stressor. Competition and soil conditions also modified climate sensitivity, which suggests that effects of climate change will be highly heterogeneous across the boreal biome.

Cytosolic Glyceraldehyde-3-Phosphate Dehydrogenase Is Phosphorylated during Seed Development.

Cytosolic glyceraldehyde-3-phosphate dehydrogenase (NAD-GAPDH) is involved in a critical energetic step of glycolysis and also has many important functions besides its enzymatic activity. The recombinant wheat NAD-GAPDH was phosphorylated in vitro at Ser205 by a SNF1-Related protein kinase 1 (SnRK1) from wheat heterotrophic (but not from photosynthetic) tissues. The S205D mutant enzyme (mimicking the phosphorylated form) exhibited a significant decrease in activity but similar affinity toward substrates. Immunodetection and activity assays showed that NAD-GAPDH is phosphorylated in vivo, the enzyme depicting different activity, abundance and phosphorylation profiles during development of seeds that mainly accumulate starch (wheat) or lipids (castor oil seed). NAD-GAPDH activity gradually increases along wheat seed development, but protein levels and phosphorylation status exhibited slight changes. Conversely, in castor oil seed, the activity slightly increased and total protein levels do not significantly change in the first half of seed development but both abruptly decreased in the second part of development, when triacylglycerol synthesis and storage begin. Interestingly, phospho-NAD-GAPDH levels reached a maximum when the seed switch their metabolism to mainly support synthesis and accumulation of carbon reserves. After this point the castor oil seed NAD-GAPDH protein levels and activity highly decreased, and the protein stability assays showed that the protein would be degraded by the proteasome. The results presented herein suggest that phosphorylation of NAD-GAPDH during seed development would have impact on the partitioning of triose-phosphate between different metabolic pathways and cell compartments to support the specific carbon, energy and reducing equivalent demands during synthesis of storage products.

Whole Genome Re-Sequencing Identifies a Quantitative Trait Locus Repressing Carbon Reserve Accumulation during Optimal Growth in Chlamydomonas reinhardtii.

Microalgae have emerged as a promising source for biofuel production. Massive oil and starch accumulation in microalgae is possible, but occurs mostly when biomass growth is impaired. The molecular networks underlying the negative correlation between growth and reserve formation are not known. Thus isolation of strains capable of accumulating carbon reserves during optimal growth would be highly desirable. To this end, we screened an insertional mutant library of Chlamydomonas reinhardtii for alterations in oil content. A mutant accumulating five times more oil and twice more starch than wild-type during optimal growth was isolated and named constitutive oil accumulator 1 (coa1). Growth in photobioreactors under highly controlled conditions revealed that the increase in oil and starch content in coa1 was dependent on light intensity. Genetic analysis and DNA hybridization pointed to a single insertional event responsible for the phenotype. Whole genome re-sequencing identified in coa1 a >200 kb deletion on chromosome 14 containing 41 genes. This study demonstrates that, 1), the generation of algal strains accumulating higher reserve amount without compromising biomass accumulation is feasible; 2), light is an important parameter in phenotypic analysis; and 3), a chromosomal region (Quantitative Trait Locus) acts as suppressor of carbon reserve accumulation during optimal growth.

Ecophysiological responses of two dominant subalpine tree species Betula albo-sinensis and Abies faxoniana to intra- and interspecific competition under elevated temperature

Seedlings of tree species Abies faxoniana and Betula albo-sinensis were grown under different competition regimes (single plant, monoculture or mixture) at ambient temperature (AT) and elevated temperature (ET) to evaluate the impact of inter- and intraspecific competition on photosynthesis capacity, N resorption efficiency, N uptake patterns, water use efficiency (foliar carbon isotope composition, δ13C) and root carbon reserve accumulation. Our results showed that ET did not have significant effects on relative height growth rate (RGRH), relative diameter growth rate (RGRD), chlorophyll fluorescence parameters (Fv/Fm, ΦPSII), light-saturated photosynthetic rate (A1400) and nitrogen uptake in A. faxoniana, but it significantly reduced the values of these parameters in B. albo-sinensis. On the other hand, RGRH of A. faxoniana was not influenced by intraspecific competition irrespective of temperature regimes, while in B. albo-sinensis intraspecific competition was severer under AT than under ET. Decreased photosynthetic capacity and reduced plant size detected in B. albo-sinensis under ET likely contribute to its low intraspecific competitive ability. However, we found positive effects of interspecific competition on A. faxoniana but negative effects on B. albo-sinensis under ET, as A. faxoniana exposed to interspecific competition showed higher RGRH, RGRD, A1400, chlorophyll fluorescence parameters, foliar δ13C, foliar N, and percentages of 15N-ammonium and 15N-nitrate recovery than did A. faxoniana exposed to intraspecific competition or no competition. The results indicate that A. faxoniana is a better competitor than B. albo-sinensis under ET. Further, A. faxoniana altered resource use patterns and had a high plasticity in its ammonium uptake in mixtures compared with monocultures under ET, while B. albo-sinensis did not change its N use pattern. In all, our results showed that high temperature stress tolerance (e.g., higher root carbon accumulation), high ability to keep carbon balance (e.g., higher water use efficiency, lower leaf respiration rate, higher Fv/Fm), as well as a better N acquisition ability make A. faxoniana a better competitor when compared to B. albo-sinensisg under ET.

Low root reserve accumulation during drought may lead to winter mortality in poplar seedlings

Climate models suggest that more frequent drought events of greater severity and length, associated with climate change, can be expected in the coming decades. Although drought-induced tree mortality has been recognized as an important factor modulating forest demography at the global scale, the mechanisms underlying drought-induced tree mortality remain contentious. Above- and below-ground growth, gas exchange, water relations and carbon reserve accumulation dynamics at the organ and whole-plant scale were quantified in Populus tremuloides and P. balsamifera seedlings in response to severe drought. Seedlings were maintained in drought conditions over one growing and one dormant winter season. Our experiment presents a detailed description of the effect of severe drought on growth and physiological variables, leading to seedling mortality after an extended period of drought and dormancy. After re-watering following the dormant period, drought-exposed seedlings did not re-flush, showing that the root system had died off. The results of this study suggest a complex series of physiological feedbacks between the measured variables in both Populus species. Further, they reveal that reduced reserve accumulation in the root system during drought decreases the conversion of starch to soluble sugars in roots, which may contribute to the root death of drought-exposed seedlings during the dormant season by compromising the frost tolerance of the root system.

A Dual-Targeted Soybean Protein Is Involved in Bradyrhizobium japonicum Infection of Soybean Root Hair and Cortical Cells

The symbiotic interaction between legumes and soil bacteria (e.g., soybean [Glycine max L.] and Bradyrhizobium japonicum]) leads to the development of a new root organ, the nodule, where bacteria differentiate into bacteroids that fix atmospheric nitrogen for assimilation by the plant host. In exchange, the host plant provides a steady carbon supply to the bacteroids. This carbon can be stored within the bacteroids in the form of poly-3-hydroxybutyrate granules. The formation of this symbiosis requires communication between both partners to regulate the balance between nitrogen fixation and carbon utilization. In the present study, we describe the soybean gene GmNMNa that is specifically expressed during the infection of soybean cells by B. japonicum. GmNMNa encodes a protein of unknown function. The GmNMNa protein was localized to the nucleolus and also to the mitochondria. Silencing of GmNMNa expression resulted in reduced nodulation, a reduction in the number of bacteroids per infected cell in the nodule, and a clear reduction in the accumulation of poly-3-hydroxybutyrate in the bacteroids. Our results highlight the role of the soybean GmNMNa gene in regulating symbiotic bacterial infection, potentially through the regulation of the accumulation of carbon reserves.

Carbon dioxide enrichment does not reduce leaf longevity or alter accumulation of carbon reserves in the woodland spring ephemeral Erythronium americanum.

Woodland spring ephemerals exhibit a relatively short epigeous growth period prior to canopy closure. However, it has been suggested that leaf senescence is induced by a reduction in the carbohydrate sink demand, rather than by changes in light availability. To ascertain whether a potentially higher net carbon (C) assimilation rate could shorten leaf lifespan due to an accelerated rate of storage, Erythronium americanum plants were grown under ambient (400 ppm) and elevated (1100 ppm) CO2 concentrations. During this growth-chamber experiment, plant biomass, bulb starch concentration and cell size, leaf phenology, gas exchange rates and nutrient concentrations were monitored. Plants grown at 1100 ppm CO2 had greater net C assimilation rates than those grown at 400 ppm CO2. However, plant size, final bulb mass, bulb filling rate and timing of leaf senescence did not differ. Erythronium americanum fixed more C under elevated than under ambient CO2 conditions, but produced plants of similar size. The similar bulb growth rates under both CO2 concentrations suggest that the bulb filling rate is dependant on bulb cell elongation rate, rather than on C availability. Elevated CO2 stimulated leaf and bulb respiratory rates; this might reduce feed-back inhibition of photosynthesis and avoid inducing premature leaf senescence.