Effects of Nitrogen Supplementation Status on CO2 Biofixation and Biofuel Production of the Promising Microalga Chlorella sp. ABC-001.

The use of microalgal biomass as feedstock for biofuels has been discussed for decades as it provides a sustainable approach to producing fuels for the future. Nonetheless, its feasibility has not been established yet and various aspects of biomass applications such as CO2 biofixation should also be explored. Therefore, in this study, the CO2 biofixation and lipid/carbohydrate production potential of Chlorella sp. ABC-001 were examined under various nitrogen concentrations. The highest biomass productivity and CO2 biofixation rate of 0.422 g/l/d and 0.683 g/l/d, respectively, were achieved under a nitrogen-rich condition (15 mM nitrate). Carbohydrate content was generally proportional to initial nitrate concentration and showed the highest value of 41.5% with 15 mM. However, lipid content showed an inverse relationship with nitrogen supplementation and showed the highest value of 47.4% with 2.5 mM. In consideration as feedstock for biofuels (bioethanol, biodiesel, and biogas), the sum of carbohydrate and lipid contents were examined and the highest value of 79.6% was achieved under low nitrogen condition (2.5 mM). For lipid-based biofuel production, low nitrogen supplementation should be pursued. However, considering the lower feasibility of biodiesel, pursuing CO2 biofixation and the production of carbohydrate-based fuels under nitrogenrich condition might be more rational. Thus, nitrogen status as a cultivation strategy must be optimized according to the objective, and this was confirmed with the promising alga Chlorella sp. ABC-001.

Steel slag amendment impacts on soil microbial communities and activities of rice (Oryza sativa L.).

With the increase in iron/steel production, the higher volume of by-products (slag) generated necessitates its efficient recycling. Because the Linz-Donawitz (LD) slag is rich in silicon (Si) and other fertilizer components, we aim to evaluate the impact of the LD slag amendment on soil quality (by measuring soil physicochemical and biological properties), plant nutrient uptake, and strengthens correlations between nutrient uptake and soil bacterial communities. We used 16 S rRNA illumine sequencing to study soil bacterial community and APIZYM assay to study soil enzymes involved in C, N, and P cycling. The LD slag was applied at 2 Mg ha-1 to Japonica and Indica rice cultivated under flooded conditions. The LD slag amendment significantly improved soil pH, plant photosynthesis, soil nutrient availability, and the crop yield, irrespective of cultivars. It significantly increased N, P, and Si uptake of rice straw. The slag amendment enhanced soil microbial biomass, soil enzyme activities and enriched certain bacterial taxa featuring copiotrophic lifestyles and having the potential role for ecosystem services provided to the benefit of the plant. The study evidenced that the short-term LD slag amendment in rice cropping systems is useful to improve soil physicochemical and biological status, and the crop yield.

Metabolic versatility of freshwater sedimentary archaea feeding on different organic carbon sources.

Members of the phylum Bathyarchaeota and the class Thermoplasmata are widespread in marine and freshwater sediments where they have been recognized as key players in the carbon cycle. Here, we tested the responsiveness of archaeal communities on settled plant debris and sediment from a karstic lake to different organic carbon amendments (amino acids, plant-derived carbohydrates, and aromatics) using a lab-scale microcosm. Changes in the composition and abundance of sediment and biofilm archaeal communities in both DNA and RNA fractions were assessed by 16S rRNA gene amplicon sequencing and qPCR, respectively, after 7 and 30 days of incubation. Archaeal communities showed compositional changes in terms of alpha and beta diversity in relation to the type of carbon source (amino acids vs. plant-derived compounds), the nucleic acid fraction (DNA vs. RNA), and the incubation time (7 vs. 30 days). Distinct groups within the Bathyarchaeota (Bathy-15 and Bathy-6) and the Thermoplasmata (MBG-D) differently reacted to carbon supplements as deduced from the analysis of RNA libraries. Whereas Bathyarchaeota in biofilms showed a long-term positive response to humic acids, their counterparts in the sediment were mainly stimulated by the addition of tryptophan, suggesting the presence of different subpopulations in both habitats. Overall, our work presents an in vitro assessment of the versatility of archaea inhabiting freshwater sediments towards organic carbon and introduces settled leaf litter as a new habitat for the Bathyarchaeota and the Thermoplasmata.

Effect of compost and inorganic fertilizer on organic carbon and activities of carbon cycle enzymes in aggregates of an intensively cultivated Vertisol.

BACKGROUND AND AIMS: This paper was primarily devoted to understand the interactions of soil aggregates, organic carbon (C) and carbon cycle enzymes in aggregates under different fertilization managements, aiming to identify the effects of organic and inorganic fertilizer amendments on soil organic C accumulation and the activities of carbon cycle enzymes within aggregates in Vertisol. METHODS: A Vertisol soil following 4-year compost and inorganic fertilizer amendments, i.e. no fertilizer (CK), mineral fertilizer (FR) and 60% compost N plus 40% fertilizer N (FRM), was collected to identify the dynamics of organic C, enzymes activities and their associations with macroaggregation using aggregate fractionation techniques. RESULTS: The organic C content in all FR and FRM treatments was 8.24-41.15% higher than that in CK. An increased amounts of carbon cycle enzymes in aggregates or 0-20 cm bulk soil were also observed in FRM plots. Compared to FR, FRM significantly strengthened the structural stability of macroaggregates and the intimate connection between enzyme activities and macroaggregates. CONCLUSIONS: As a recommended measure, supplementation with organic manure such as compost strengthened the process of mutual promotion between carbon cycle enzymes and macroaggregates, and the synergistic effect would be highly beneficial to soil organic C sequestration.

Pyrenoidal sequestration of cadmium impairs carbon dioxide fixation in a microalga.

Mixotrophic microorganisms are able to use organic carbon as well as inorganic carbon sources and thus, play an essential role in the biogeochemical carbon cycle. In aquatic ecosystems, the alteration of carbon dioxide (CO2 ) fixation by toxic metals such as cadmium - classified as a priority pollutant - could contribute to the unbalance of the carbon cycle. In consequence, the investigation of cadmium impact on carbon assimilation in mixotrophic microorganisms is of high interest. We exposed the mixotrophic microalga Chlamydomonas reinhardtii to cadmium in a growth medium containing both CO2 and labelled 13 C-[1,2] acetate as carbon sources. We showed that the accumulation of cadmium in the pyrenoid, where it was predominantly bound to sulphur ligands, impaired CO2 fixation to the benefit of acetate assimilation. Transmission electron microscopy (TEM)/X-ray energy dispersive spectroscopy (X-EDS) and micro X-ray fluorescence (µXRF)/micro X-ray absorption near-edge structure (µXANES) at Cd LIII- edge indicated the localization and the speciation of cadmium in the cellular structure. In addition, nanoscale secondary ion mass spectrometry (NanoSIMS) analysis of the 13 C/12 C ratio in pyrenoid and starch granules revealed the origin of carbon sources. The fraction of carbon in starch originating from CO2 decreased from 73 to 39% during cadmium stress. For the first time, the complementary use of high-resolution elemental and isotopic imaging techniques allowed relating the impact of cadmium at the subcellular level with carbon assimilation in a mixotrophic microalga.

Tracking acetate through a journey of living world: Evolution as alternative cellular fuel with potential for application in cancer therapeutics.

Acetate is a short chain fatty acid, comprising carbon, hydrogen and oxygen (C2H3O2-), which has emerged as a key alternative fuel for cellular metabolism. Beginning its voyage from the abiotic atmosphere, acetate has contributed to the physiology of both prokaryotes and eukaryotes. The main role of acetate includes its contribution to the global carbon cycle, bioenergetic and biosynthetic metabolic processes. Based on the ability to produce and consume acetate, organisms are categorized as acetogenic, acetate-consumers or both depending on their genetic make-up of the metabolizing enzymes' repertoire. The key molecules implicated in utilization and production of acetate include, but not limited to, monocarboxylate transporters, enzymes regulating acetate utilization like AMP-forming Acetyl CoA synthetase (ACS-AMP), Acyl-CoA short chain synthetase 1, 2 (ACSS1, 2), and production like Acetate kinase (ACK)/Phosphotransacetylase (PTA), ADP-forming acetyl CoA synthetase (ACS-ADP), Pyruvate:ferredoxin oxidoreductase, histone deacetylase and acetyl CoA hydrolase. These enzymes are utilized by the acetate homeostasis machinery in a variable manner. As malignant cells also display highly upregulated metabolic processes for rapid energy generation, they display an immense need for alternative carbon sources to fuel their metabolism. Tumor cells display over expression of transporters and enzymes implicated in their acetate utility machinery. This review also highlights mechanisms of the pro and antitumor potential of acetate depending on the genetic and metabolic makeup of neoplastic cells. The present review is a comprehensive compilation of the available literature with respect to the role of acetate in the biology of living organisms and its potential for being maneuvered in anticancer therapeutics.

One-carbon cycle support rescues sperm damage in experimentally induced varicocoele in rats.

OBJECTIVES: To investigate whether micronutrients in support of the one-carbon cycle and glutathione synthesis are effective in improving sperm damage after surgical varicocoele induction in rats and whether any effect is achieved without a rebound reductive stress as seen with oral antioxidants. MATERIALS AND METHODS: Surgical varicocoele was induced in adult male Wistar rats and resulted in significant damage to the testis and sperm cells measured at 2 and 4 months after surgery. At 2 months after surgery, rats received a 2-month oral supplementation in support of the one-carbon cycle containing B vitamins (B2, B3, B6, folic acid and B12), N-acetyl-cysteine, zinc, small amounts of vitamin E, and a natural source of betalains and quercetine (Condensyl® ; Parthenogen SAGL, Lugano, Switzerland and Nurilia SARL, Lyon, France). RESULTS: One-carbon cycle supplementation, compared to untreated controls, significantly improved the morphometric characteristics of testis (P < 0.05), sperm concentration, motility and abnormal morphology (P < 0.001), sperm chromatin condensation (aniline blue staining, P < 0.05), sperm DNA damage (acridine orange staining, P < 0.05) and sperm lipid peroxidation (BODIPY C11, P < 0.001). The improvement in both nuclear condensation and DNA damage and the lack of excessive inhibition of lipid peroxidation confirmed that no reductive stress had occurred. CONCLUSIONS: Micronutrients in support of the one-carbon cycle are effective in the treatment of surgically induced varicocoele in rats, probably by activating natural antioxidant defences and epigenetics. These results support the idea that essential micronutrients including B vitamins may also have a positive influence in clinical varicocoele, which should be tested in prospective clinical trials.

Growth on ATP Elicits a P-Stress Response in the Picoeukaryote Micromonas pusilla.

The surface waters of oligotrophic oceans have chronically low phosphate (Pi) concentrations, which renders dissolved organic phosphorus (DOP) an important nutrient source. In the subtropical North Atlantic, cyanobacteria are often numerically dominant, but picoeukaryotes can dominate autotrophic biomass and productivity making them important contributors to the ocean carbon cycle. Despite their importance, little is known regarding the metabolic response of picoeukaryotes to changes in phosphorus (P) source and availability. To understand the molecular mechanisms that regulate P utilization in oligotrophic environments, we evaluated transcriptomes of the picoeukaryote Micromonas pusilla grown under Pi-replete and -deficient conditions, with an additional investigation of growth on DOP in replete conditions. Genes that function in sulfolipid substitution and Pi uptake increased in expression with Pi-deficiency, suggesting cells were reallocating cellular P and increasing P acquisition capabilities. Pi-deficient M. pusilla cells also increased alkaline phosphatase activity and reduced their cellular P content. Cells grown with DOP were able to maintain relatively high growth rates, however the transcriptomic response was more similar to the Pi-deficient response than that seen in cells grown under Pi-replete conditions. The results demonstrate that not all P sources are the same for growth; while M. pusilla, a model picoeukaryote, may grow well on DOP, the metabolic demand is greater than growth on Pi. These findings provide insight into the cellular strategies which may be used to support growth in a stratified future ocean predicted to favor picoeukaryotes.

Effects of nitrogen and phosphorus additions on soil microbial biomass and community structure in two reforested tropical forests.

Elevated nitrogen (N) deposition may aggravate phosphorus (P) deficiency in forests in the warm humid regions of China. To our knowledge, the interactive effects of long-term N deposition and P availability on soil microorganisms in tropical replanted forests remain unclear. We conducted an N and P manipulation experiment with four treatments: control, N addition (15 g N m(-2)·yr(-1)), P addition (15 g P m(-2)·yr(-1)), and N and P addition (15 + 15 g N and P m(-2)·yr(-1), respectively) in disturbed (planted pine forest with recent harvests of understory vegetation and litter) and rehabilitated (planted with pine, but mixed with broadleaf returning by natural succession) forests in southern China. Nitrogen addition did not significantly affect soil microbial biomass, but significantly decreased the abundance of gram-negative bacteria PLFAs in both forest types. Microbial biomass increased significantly after P addition in the disturbed forest but not in the rehabilitated forest. No interactions between N and P additions on soil microorganisms were observed in either forest type. Our results suggest that microbial growth in replanted forests of southern China may be limited by P rather than by N, and this P limitation may be greater in disturbed forests.

Nutrient Enrichment Mediates the Relationships of Soil Microbial Respiration with Climatic Factors in an Alpine Meadow.

Quantifying the effects of nutrient additions on soil microbial respiration (R m) and its contribution to soil respiration (R s) are of great importance for accurate assessment ecosystem carbon (C) flux. Nitrogen (N) addition either alone (coded as LN and HN) or in combination with phosphorus (P) (coded as LN + P and HN + P) were manipulated in a semiarid alpine meadow on the Tibetan Plateau since 2008. Either LN or HN did not affect R m, while LN + P enhanced R m during peak growing periods, but HN + P did not affect R m. Nutrient addition also significantly affected R m /R s, and the correlations of R m /R s with climatic factors varied with years. Soil water content (Sw) was the main factor controlling the variations of R m /R s. During the years with large rainfall variations, R m /R s was negatively correlated with Sw, while, in years with even rainfall, R m/R s was positively correlated with Sw. Meanwhile, in N + P treatments the controlling effects of climatic factors on R m /R s were more significant than those in CK. Our results indicate that the sensitivity of soil microbes to climatic factors is regulated by nutrient enrichment. The divergent effects of Sw on R m /R s suggest that precipitation distribution patterns are key factors controlling soil microbial activities and ecosystem C fluxes in semiarid alpine meadow ecosystems.

Accumulation and enhanced cycling of polyphosphate by Sargasso Sea plankton in response to low phosphorus.

Phytoplankton alter their biochemical composition according to nutrient availability, such that their bulk elemental composition varies across oceanic provinces. However, the links between plankton biochemical composition and variation in biogeochemical cycling of nutrients remain largely unknown. In a survey of phytoplankton phosphorus stress in the western North Atlantic, we found that phytoplankton in the phosphorus-depleted subtropical Sargasso Sea were enriched in the biochemical polyphosphate (polyP) compared with nutrient-rich temperate waters, contradicting the canonical oceanographic view of polyP as a luxury phosphorus storage molecule. The enrichment in polyP coincided with enhanced alkaline phosphatase activity and substitution of sulfolipids for phospholipids, which are both indicators of phosphorus stress. Further, polyP appeared to be liberated preferentially over bulk phosphorus from sinking particles in the Sargasso Sea, thereby retaining phosphorus in shallow waters. Thus, polyP cycling may form a feedback loop that attenuates the export of phosphorus when it becomes scarce, contributes bioavailable P for primary production, and supports the export of carbon and nitrogen via sinking particles.

Methanogenic burst in the end-Permian carbon cycle.

The end-Permian extinction is associated with a mysterious disruption to Earth's carbon cycle. Here we identify causal mechanisms via three observations. First, we show that geochemical signals indicate superexponential growth of the marine inorganic carbon reservoir, coincident with the extinction and consistent with the expansion of a new microbial metabolic pathway. Second, we show that the efficient acetoclastic pathway in Methanosarcina emerged at a time statistically indistinguishable from the extinction. Finally, we show that nickel concentrations in South China sediments increased sharply at the extinction, probably as a consequence of massive Siberian volcanism, enabling a methanogenic expansion by removal of nickel limitation. Collectively, these results are consistent with the instigation of Earth's greatest mass extinction by a specific microbial innovation.

Interactive biotic and abiotic regulators of soil carbon cycling: evidence from controlled climate experiments on peatland and boreal soils.

Partially decomposed plant and animal remains have been accumulating in organic soils (i.e. >40% C content) for millennia, making them the largest terrestrial carbon store. There is growing concern that, in a warming world, soil biotic processing will accelerate and release greenhouse gases that further exacerbate climate change. However, the magnitude of this response remains uncertain as the constraints are abiotic, biotic and interactive. Here, we examined the influence of resource quality and biological activity on the temperature sensitivity of soil respiration under different soil moisture regimes. Organic soils were sampled from 13 boreal and peatland ecosystems located in the United Kingdom, Ireland, Spain, Finland and Sweden, representing a natural resource quality range of C, N and P. They were incubated at four temperatures (4, 10, 15 and 20 °C) at either 60% or 100% water holding capacity (WHC). Our results showed that chemical and biological properties play an important role in determining soil respiration responses to temperature and moisture changes. High soil C : P and C : N ratios were symptomatic of slow C turnover and long-term C accumulation. In boreal soils, low bacterial to fungal ratios were related to greater temperature sensitivity of respiration, which was amplified in drier conditions. This contrasted with peatland soils which were dominated by bacterial communities and enchytraeid grazing, resulting in a more rapid C turnover under warmer and wetter conditions. The unexpected acceleration of C mineralization under high moisture contents was possibly linked to the primarily role of fermented organic matter, instead of oxygen, in mediating microbial decomposition. We conclude that to improve C model simulations of soil respiration, a better resolution of the interactions occurring between climate, resource quality and the decomposer community will be required.

The influence of phosphorus availability and Laccaria bicolor symbiosis on phosphate acquisition, antioxidant enzyme activity, and rhizospheric carbon flux in Populus tremuloides.

Many forest tree species are dependent on their symbiotic interaction with ectomycorrhizal (ECM) fungi for phosphorus (P) uptake from forest soils where P availability is often limited. The ECM fungal association benefits the host plant under P limitation through enhanced soil exploration and increased P acquisition by mycorrhizas. To study the P starvation response (PSR) and its modification by ECM fungi in Populus tremuloides, a comparison was made between nonmycorrhizal (NM) and mycorrhizal with Laccaria bicolor (Myc) seedlings grown under different concentrations of phosphate (Pi) in sand culture. Although differences in growth between NM and Myc plants were small, Myc plants were more effective at acquiring P from low Pi treatments, with significantly lower k m values for root and leaf P accumulation. Pi limitation significantly increased the activity of catalase, ascorbate peroxidase, and guaiacol-dependent peroxidase in leaves and roots to greater extents in NM than Myc P. tremuloides. Phosphoenolpyruvate carboxylase activity also increased in NM plants under P limitation, but was unchanged in Myc plants. Formate, citrate, malonate, lactate, malate, and oxalate and total organic carbon exudation by roots was stimulated by P limitation to a greater extent in NM than Myc plants. Colonization by L. bicolor reduced the solution Pi concentration thresholds where PSR physiological changes occurred, indicating that enhanced Pi acquisition by P. tremuloides colonized by L. bicolor altered host P homeostasis and plant stress responses to P limitation. Understanding these plant-symbiont interactions facilitates the selection of more P-efficient forest trees and strategies for tree plantation production on marginal soils.