Physiologic and metabolic responses of wheat seedlings to elevated and super-elevated carbon dioxide

The metabolic consequence of suboptimal (400 μmol mol⁻¹ or ppm), near-optimal (1500 ppm) and supra-optimal (10,000 ppm) atmospheric carbon dioxide concentrations [CO₂] was investigated in an attempt to reveal plausible underlying mechanisms for the differential physiological and developmental responses to increasing [CO₂]. Both non-targeted and targeted metabolite profiling by GC–MS and LC–MS were employed to examine primary and secondary metabolites in wheat (Triticum aestivum, cv Yocoro rojo) continuously exposed to these [CO₂] levels for 14, 21 and 28 days. Metabolite profile was altered by both [CO₂] and physiological age. In general, plants grown under high [CO₂] exhibited a metabolite profile characteristic of older plants under ambient CO₂. Elevated [CO₂] resulted in higher levels of phosphorylated sugar intermediates, though no clear trend in the content of reducing sugars was observed. Transient starch content was enhanced by increasing [CO₂] to a much greater extent at 10,000 ppm CO₂ than at 1500 ppm CO₂. The percentage increase of starch content resulting from CO₂ enrichment declined as plants develope. In contrast, elevated [CO₂] promoted the accumulation of secondary metabolites (flavonoids) progressively to a greater extent as plants became mature. Elevated [CO₂] to 1500 ppm induced a higher initial growth rate, while super-elevated [CO₂] appeared to negate such initial growth promotion. However, after 4 weeks, there was no difference in vegetative growth between 1500 and 10,000 ppm CO₂-grown plants, both elevated CO₂ levels resulted in an overall 25% increase in biomass over the control plants. More interestingly, elevated atmospheric [CO₂] reduced evapotranspiration rate (ET), but further increase to the supra-optimal level resulted in increased ET (a reversed trend), i.e. ET at 1500 ppm < ET at 10,000 ppm < ET at 400 ppm. The differential effect of elevated and super-elevated CO₂ on plants was further reflected in the nitrogen dynamics. These results provide the potential metabolic basis for the differential productivity and stomatal function of plants grown under elevated and super-elevated CO₂ levels.     

Plant responses to short- and long-term exposures to high carbon dioxide levels in closed environments.

When higher plants are exposed to elevated levels of CO2 for both short- and long-term periods photosynthetic C-gain and photoassimilate export from leaves are generally increased. Water use efficiency is increased on a leaf area basis. During long-term exposures, photosynthesis rates on leaf and whole plants bases are altered in a species specific manner. The most common pattern in C3 plants is an enhanced rate of whole plant photosynthesis in a well irradiated canopy. Nevertheless, in some herbaceous species prolonged exposure to high CO2 results in remobilization of nitrogenous reserves (i.e., leaf protein degradation) and reduced rates of mature leaf photosynthesis when assayed at ambient CO2 and O2 levels. Both short- and long-term exposures to those CO2 levels (i.e., 100 to 2,000 microliter l-1) which modify photosynthesis and export, also modify both endogenous ethylene gas (C2H4) release, and substrate, 1-aminocyclopropane-1-carboxylic acid (ACC), saturated C2H4 release rates from irradiated leaves. Photosynthetically active canopy leaves contribute most of the C2H4 released from the canopy. Prolonged growth at high CO2 results in a persistent increase in the rate of endogenous C2H4 release from leaves which can, only in part, be attributed to the increase of the endogenous pools of C2H4 pathway intermediates (e.g., methionine, M-ACC, and ACC). The capacity for increasing the rate of C2H4 release in response to short-term exposures to varying CO2 levels does not decline after prolonged growth at high CO2. When leaves, whole plants, and model canopies of tomato plants are exposed to exogenous C2H4 a reduction in the rate of photosynthesis can, in each case, be attributed to the classical effects of C2H4 on plant development and morphology. The effect of C2H4 on CO2 gas exchange of plant canopies is shown to be dependent on the canopy leaf area index.     

Effect of lamp type and temperature on development, carbon partitioning and yield of soybean.

Soybeans grown in controlled environments are commonly taller than field-grown plants. In controlled environments, including liquid hydroponics, height of the dwarf cultivar "Hoyt" was reduced from 46 to 33 cm when plants were grown under metal halide lamps compared to high pressure sodium lamps at the same photosynthetic photon flux. Metal halide lamps reduced total biomass 14% but did not significantly reduce seed yield. Neither increasing temperature nor altering the difference between day/night temperature affected plant height. Increasing temperature from 21 to 27 degrees C increased yield 32%. High temperature significantly increased carbon partitioning to stems and increased harvest index.     

Effects of different carbon dioxide and LED lighting levels on the anti-oxidative capabilities of Gynura bicolor DC

Gynura bicolor DC is not only an edible plant but also a kind of traditional Chinese herbal medicine. G. bicolor DC grown in controlled environmental chambers under 3 CO₂ concentrations [450 (ambient), 1500 (elevated), 8000 (super-elevated) μmol mol⁻¹] and 3 LED lighting conditions [white (WL), 85% red + 15% blue (RB15), 70% red + 30% blue (RB30) ] were investigated to reveal plausible antioxidant anabolic responses to CO₂ enrichment and LED light quality. Under ambient and elevated CO₂ levels, blue light increasing from 15% to 30% was conducive to the accumulation of anthocyanins and total flavonoids, and the antioxidant activity of extract was also increased, but plant biomass was decreased. These results demonstrated that the reinforcement of blue light could induce more antioxidant of secondary metabolites, but depress the effective growth of G. bicolor DC under ambient and elevated CO₂ levels. In addition, compared with the ambient and elevated CO₂ levels, the increased anthocyanins, total flavonoids contents and antioxidant enzyme activities of G. bicolor DC under super-elevated CO₂ level could serve as important components of antioxidative defense mechanism against CO₂ stress. Hence, G. bicolor DC might have higher tolerance to CO₂ stress.     

Carbon dioxide exchange of lettuce plants under hypobaric conditions.

Growth of plants in a Controlled Ecological Life Support System (CELSS) may involve the use of hypobaric pressures enabling lower mass requirements for atmospheres and possible enhancement of crop productivity. A controlled environment plant growth chamber with hypobaric capability designed and built at Ames Research Center was used to determine if reduced pressures influence the rates of photosynthesis (Ps) and dark respiration (DR) of hydroponically grown lettuce plants. The chamber, referred to as a plant volatiles chamber (PVC), has a growing area of about 0.2 m2, a total gas volume of about 0.7 m3, and a leak rate at 50 kPa of <0.1%/day. When the pressure in the chamber was reduced from ambient to 51 kPa, the rate of net Ps increased by 25% and the rate of DR decreased by 40%. The rate of Ps increased linearly with decreasing pressure. There was a greater effect of reduced pressure at 41 Pa CO2 than at 81 Pa CO2. This is consistent with reports showing greater inhibition of photorespiration (Pr) in reduced O2 at low CO2 concentrations. When the partial pressure of O2 was held constant but the total pressure was varied between 51 and 101 kPa, the rate of CO2 uptake was nearly constant, suggesting that low pressure enhancement of Ps may be mainly attributable to lowered partial pressure of O2 and the accompanying reduction in Pr. The effects of lowered partial pressure of O2 on Ps and DR could result in substantial increases in the rates of biomass production, enabling rapid throughput of crops or allowing flexibility in the use of mass and energy resources for a CELSS.     

Carbon dioxide exchange of lettuce plants under hypobaric conditions.

Growth of plants in a Controlled Ecological Life Support System (CELSS) may involve the use of hypobaric pressures enabling lower mass requirements for atmospheres and possible enhancement of crop productivity. A controlled environment plant growth chamber with hypobaric capability designed and built at Ames Research Center was used to determine if reduced pressures influence the rates of photosynthesis (Ps) and dark respiration (DR) of hydroponically grown lettuce plants. The chamber, referred to as a plant volatiles chamber (PVC), has a growing area of about 0.2 m2, a total gas volume of about 0.7 m3, and a leak rate at 50 kPa of <0.1%/day. When the pressure in the chamber was reduced from ambient to 51 kPa, the rate of net Ps increased by 25% and the rate of DR decreased by 40%. The rate of Ps increased linearly with decreasing pressure. There was a greater effect of reduced pressure at 41 Pa CO2 than at 81 Pa CO2. This is consistent with reports showing greater inhibition of photorespiration (Pr) in reduced O2 at low CO2 concentrations. When the partial pressure of O2 was held constant but the total pressure was varied between 51 and 101 kPa, the rate of CO2 uptake was nearly constant, suggesting that low pressure enhancement of Ps may be mainly attributable to lowered partial pressure of O2 and the accompanying reduction in Pr. The effects of lowered partial pressure of O2 on Ps and DR could result in substantial increases in the rates of biomass production, enabling rapid throughput of crops or allowing flexibility in the use of mass and energy resources for a CELSS.     

Cometary origin of carbon and water on the terrestrial planets.

An early high-temperature phase of the protosolar accretion disk is implied by at least three different telltales in chondrites and confirmed by peculiarities in the dust grains of comet Halley. The existence this high-temperature phase implies a large accretion rate hence a massive early disk. This clarifies the origin of the Kuiper Belt and of the Oort cloud, those two cometary populations of different symmetry that subsist today. Later, when the dust sedimented and was removed from the thermal equilibrium with the gas phase, a somewhat lower temperature of the disk explains the future planets' densities as well as the location beyond 2.6 AU of the carbonaceous chondrite chemistry. This lower temperature remains however large enough to require an exogenous origin for all carbon and all water now present in the Earth. The later orbital diffusion of planetesimals, which is required by protoplanelary growth, is needed to explain the origin of the terrestrial biosphere (atmosphere, oceans, carbonates and organic compounds) by a veneer mostly made of comets.     

Effects of carbon ions on primary cultures of mouse brain cells.

Primary mixed cultures of astrocytes and microglia were obtained from neonatal mice, and were irradiated with high-LET carbon ions. Immunohistochemical staining showed astrocytes survived more prominently than microglia. Tagged with specific antibodies, astrocytes and microglia surviving after irradiation were counted by flow cytometry. Decreases in the number of microglia and astrocytes were detected at a dose as small as 2 Gy when Day 5 cultures were irradiated with 13 keV/micrometer carbon ions. When the cultures were irradiated on Day 10, the dose-dependent decrease of microglia was more prominent for 13 keV/micrometer carbon ions than 70 keV/micrometer carbon ions. Astrocytes showed a marginal decrease at Day 10 and Day 14. We concluded that microglia are more sensitive than astrocytes to carbon ions and X-rays, and that the radiosensitivity of microglia depends on both differentiation/proliferation status and radiation quality.     

The influence of radiation quality on the formation of DNA breaks.

We have aimed to present a comprehensive review of our understanding to date of the formation of DNA strand breaks induced by high LET radiation. We have discussed data obtained from DNA in solution as well as from the formation and "repair" of strand breaks in cell DNA. There is good agreement, qualitatively, between these two systems. Results were evaluated for two parameters: (1) effectivity per particle, the cross section (sigma) in micrometers 2/particle; and (2) the strand break induction frequency as number of breaks per Gy per unit DNA (bp or dalton). A series of biological effects curves (one for each Z-number) is obtained in effectivity versus LET plots. The relationships between induction frequencies of single-strand breaks, or double-strand breaks, or the residual "irrepairable" breaks and LET-values have been evaluated and discussed for a wide spectrum of heavy ions, both for DNA in solution and for DNA in the cell. For radiation induced total breaks in cell DNA, the RBE is less than one, while the RBE for the induction of DSBs can be greater than one in the 100-200 keV/micrometers range. The level of irrepairable strand breaks is highest in this same LET range and may reach 25 percent of the initial break yield. The data presented cover results obtained for helium to uranium particles, covering a particle incident energy range of about 2 to 900 MeV/u with a corresponding LET range of near 16 to 16000 keV/micrometers.     

Stable carbon isotopes: possible clues to early life on Mars.

Organic and inorganic carbon in terrestrial near-surface environments are characterized by a marked difference in their 13C/12C ratios which can be traced back in the Earth's sedimentary record over almost 4 billion years. There is no doubt that the bias in favour of 12C displayed by biogenic matter derives, for the most part, from the isotope-selecting properties of the carbon-fixing enzyme (ribulose-1,5-bisphosphate carboxylase) that is operative in the principal photosynthetic pathway and promotes most of the carbon transfer from the non-living to the living realm. Postulating a universality of biological principles in analogy to the proven universality of the laws of physics and chemistry, we may expect enzymatic reactions in exobiological systems to be beset with B similar kinetic fractionation effects. Hence, the retrieval from the oldest Martian sediments of isotopic fractionations between reduced and oxidized (carbonate) carbon may substantially constrain current conjectures on the possible existence of former life on Mars.     

Proximate nutritional composition of CELSS crops grown at different CO2 partial pressures.

Two CELSS candidate crops, soybean (Glycine max) and potato (Solanum tuberosum), were grown hydroponically in controlled environments maintained at carbon dioxide (CO2) partial pressures ranging from 0.05 to 1.00 kPa (500 to 10,000 ppm at 101 kPa atmospheric pressure). Plants were harvested at maturity (90 days for soybean and 105 days for potato) and all tissues analyzed for proximate nutritional composition (i.e. protein, fat, carbohydrate, crude fiber, and ash content). Soybean seed ash and crude fiber were higher and carbohydrate was lower than values reported for field-grown seed. Potato tubers showed little difference from field-grown tubers. With the exception of increased crude fiber of soybean seed with increased CO2, no trends were apparent with regard to CO2 effects on proximate composition of soybean seed and potato tubers. Crude fiber of soybean stems and leaves increased with increased CO2, as did soybean leaf protein (total nitrogen). Potato leaf and stem (combined) protein levels also increased with increased CO2, while leaf and stem carbohydrates decreased. Values for leaf and stem protein and ash were higher than values generally reported for field-grown plants for both species. Results suggest that CO2 partial pressure should have little influence on proximate composition of potato tubers or soybean seed, but that high ash and protein levels might be expected from leaves and stems of crops grown in controlled environments of a CELSS.     

Effects of air velocity on photosynthesis of plant canopies under elevated CO2 levels in a plant culture system.

To obtain basic data for adequate air circulation for promoting plant growth in closed plant production modules in bioregenerative life support systems in space, effects of air velocities ranging from 0.1 to 0.8 m s-1 on photosynthesis in tomato seedlings canopies were investigated under atmospheric CO2 concentrations of 0.4 and 0.8 mmol mol-1. The canopy of tomato seedlings on a plug tray (0.4 x 0.4 m2) was set in a wind-tunnel-type chamber (0.6 x 0.4 x 0.3 m3) installed in a semi-closed-type assimilation chamber (0.9 x 0.5 x 0.4 m3). The net photosynthetic rate in the plant canopy was determined with the differences in CO2 concentrations between the inlet and outlet of the assimilation chamber multiplied by the volumetric air exchange rate of the chamber. Photosynthetic photon flux (PPF) on the plant canopy was kept at 0.25 mmol m-2 s-1, air temperature at 23 degrees C and relative humidity at 55%. The leaf area indices (LAIs) of the plant canopies were 0.6-2.5 and plant heights were 0.05-0.2 m. The net photosynthetic rate of the plant canopy increased with increasing air velocities inside plant canopies and saturated at 0.2 m s-1. The net photosynthetic rate at the air velocity of 0.4 m s-1 was 1.3 times that at 0.1 m s-1 under CO2 concentrations of 0.4 and 0.8 mmol mol-1. The net photosynthetic rate under CO2 concentrations of 0.8 mmol mol-1 was 1.2 times that under 0.4 mmol mol-1 at the air velocity ranging from 0.1 to 0.8 m s-1. The results confirmed the importance of controlling air movement for enhancing the canopy photosynthesis under an elevated CO2 level as well as under a normal CO2 level in the closed plant production modules.     

Effect of heavy ions on neuro-endocrine regulations.

During American and Russian short and long-term space flights neuroimmune dysregulations have been observed in man and rats for up to three months after the return. During Extra-Vehicular Activity, radiation exposure risk is greater to elicit short and/or long-term deleterious effects on the functional capacity of the neuroimmune system. In order to assess the effects of high LET events on neuroimmune networks, our preliminary ground-based study was to investigate brain inflammatory responses in mouse after low dose radiation exposure with high LET particles (12C, 95MeV/u, 42 mGy). Plasma corticosterone levels were rapidly (6 hours) increased by two-fold, then decreased 24 hours post-irradiation. At 3 days plasma corticosterone and ACTH concentrations were also two- to three-fold increased. Plasma ACTH levels were still elevated up to seven days to two months. Furthermore immune functions are under current assessment. The results of this study should allow a greater understanding of the effects of high LET particles on neuroimmune system.     

Consequences of cometary aurora on the carbon chemistry at comet P/Halley.

Various experimental data acquired during the visit of Halley's comet in 1986 have shown that the amount of carbon produced due to photodissociation of parent carbon bearing species is not ample enough to explain the observations. This requires the presence of an additional source of atomic carbon. One of the possible source could be auroral-type activities resulting from the precipitation of high-energy "auroral electrons" of solar wind origin, the evidence of which have been inferred from many observations at comet Halley. We have developed a coupled chemistry-transport model to study the role of auroral and photoelectron impact as well as of chemistry on the modelling of carbon in the inner coma (< or = 10(4) km) of comet Halley. Our study suggest that electron impact dissociation of CO is the major source of carbon production in the inner coma, not the recombination of CO+ as suggested by earlier workers, while transport is the main loss process.     

Stable carbon isotope fractionation in the search for life on early Mars.

Isotopic measurements and, more specifically, ratios of 13C to 12C in organic relative to inorganic deposits, are useful in reconstructing past biological activity on Earth. Organic matter has a lower ratio of 13C to 12C due largely to the preferential fixation of 12C over the heavier isotope by the major carbon-fixation enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase, although other factors (e.g., availability of source carbon, fixation by other carboxylating enzymes and diagenesis of organic material) also contribute to fractionation. Would carbon isotope discrepancies between inorganic and organic carbon indicate past biological activity on Mars? In order to answer this question, we analyse what is known about terrestrial biologic and abiologic carbon fixation and its preservation in the fossil record, and suggest what the isotope discrimination during possible biologic and abiologic carbon fixation on Mars might have been like. Primarily because isotopic signatures of abiotically fixed carbon overlap with those of biotic fixation, but also because heterotrophy does not significantly alter the isotopic signature of ingested carbon, fractionation alone would not be definitive evidence for life. However, a narrow range of fractionation, including no fractionation, would suggest biotic processes. Never-the-less, isotopic ratios in organic deposits on Mars would be extremely useful in analysing prebiotic, if not biotic, carbon transformations on Mars.     

The nutritional adequacy of a limited vegan diet for a Controlled Ecological Life-Support System.

Purdue University, as well as the Johnson and Kennedy Space Centers and NASA Ames Research Center, are investigating approximately 5-10 plants that will be grown hydroponically to provide not only the energy and nutrients, but also the oxygen for humans habitating in Mars and lunar bases. The growth and nutritional status of rats fed either a control diet (adequate in all macro- and micronutrients) or a strict vegetarian diet consisting of 5 (vegan-5) or 10 (vegan-10) candidate crop species were investigated. In addition, vegan-10 diets were supplemented with mineral and/or vitamin mix at a level similar to the control diets to assess the effect of supplementation on nutrient status. The assessment of inedible plant material as an alternative food source was also investigated. Results of this study demonstrated that consumption of the vegan-10 diet significantly improved weight gain of rats compared to that for rats fed the vegan-5 diet. Mineral supplementation, at a level present in the control diet, to the vegan-10 diet improved growth and nutrient status, but growth was significantly lower compared to the control-fed rats. Inclusion of inedible plant material, high in ash content, improved some indices of nutrient status, without improving growth.     

Effect of iodine disinfection products on higher plants.

Iodine is used to disinfect potable water on United States spacecraft. Iodinated potable water will likely be used to grow plants in space. Little is known about the effects of iodine disinfection products on plants. Seeds of select higher plants were germinated in water iodinated using the Shuttle Microbial Check Valve, and water to which measured amounts of iodide was added. Percent germination was decreased in seeds of most species germinated in iodinated water. Beans were most affected. Germination rates, determined from germination half-times, were decreased for beans germinated in iodinated water, and water to which iodide was added. Development was retarded and rootlets were conspicuously absent in bean and several other plant species germinated in iodinated water. Iodide alone did not elicit these responses. Clearly iodine disinfection products can affect higher plants. These effects must be carefully considered for plant experimentation and cultivation in space, and in design and testing of closed environmental life support systems.     

Processing of amorphous carbon grains produced in hydrogen-rich atmosphere.

Recent laboratory experiments, compared with observations, have strongly suggested that the amount of hydrogen, present in the grain formation regions, around C-rich stars, can play a major role in the definition of the spectral characteristics of the freshly formed carbon dust particles. Such spectral characteristics could also be modified, during the thermal processing, due to the shock waves and/or the strong radiation field, that the grains undergo after their formation. In this work we report about a series of laboratory tests, during which submicronic dust particles of amorphous carbon, condensed in an hydrogen-rich atmosphere, have been processed, by means of two different methods. Some samples have been annealed into an oven at different temperatures, while others have been exposed to strong ultraviolet radiation. Their spectra have been obtained before and after the processing in the full range from ultraviolet (200 nm) to far infrared (0.1 mm). The results of the experiment are presented and discussed, together with their astrophysical implications.     

Laboratory astrophysics and molecular astronomy of pure carbon molecules.

The pure carbon molecules Cn are currently of great experimental and theoretical interest. Our work in this area begins with detection of the SiC molecule, which is isovalent with C2. New infrared electronic transitions of C2 and C3 were discovered by emission spectroscopy of hydrocarbon dicharges. The C3 and C5 molecules were found by infrared vibration-rotation spectroscopy of the prototypical obscured carbon star, IRC+10216. C7 and C9 were searched for in the same source, but not found. The laboratory infrared emission spectrum of C60 was recorded to aid in a search for C60 in extraterrestrial sources.     

Effects of gamma-ray and high energy carbon ion irradiation on swimming velocity of Euglena gracilis.

The effects of gamma-ray and high energy carbon ion irradiation on the swimming velocity of the photosynthetic flagellate Euglena gracilis strain Z were studied, focusing on a dose-effect relationship. Cells were exposed to 60Co gamma-rays at 6 doses of 10, 15, 20, 40, 100 and 200 Gy for water, and also to 290 MeV/amu carbon ions from the Heavy Ion Medical Accelerator in Chiba at 7 doses (5, 10, 15, 20, 50, 100 and 200 Gy for water). The swimming velocity was measured by a biomonitoring system, called ECOTOX. The swimming velocities of Euglena gracilis cells were significantly decreased by >40 Gy gamma-rays and >5 Gy carbon ions, respectively. The 50% effective doses for inhibition, 34 +/- 4 Gy (gamma-rays) and 13 +/- 1 Gy (290 MeV/amu carbon ions), were estimated from the best fit to data of the logistic model. The relative biological effectiveness (2.6 +/- 0.4) was calculated by the ratio of 50% effective doses. The inhibition of the swimming velocity of the cells irradiated with gamma-rays was still present after 3 days, while recovery of the swimming velocity was shown in the cells exposed to 290 MeV/amu carbon ions. It is suggested that ionizing radiation inhibits ATP production and/or increases frictional drag on beating of the flagellum, thus decreasing swimming velocity.