Effect of Sulfide on Nitrogen Fixation in a Stream Sediment-Water System

Nitrogen fixation (C₂H₂ reduction) in a sediment-water system was studied under anaerobic incubation conditions. Sodium sulfide at low concentrations stimulated activity, with a twofold increase in C₂H₄ production occurring in the presence of 8 μmol of S²⁻ per ml of stream water. Sodium sulfide at concentrations of 16 μmol of S²⁻ per ml or greater inhibited nitrogen fixation, with 64 μmol of S²⁻ per ml being completely inhibitory. Sulfide at levels of 16 μmol/ml or above inhibited CO₂ production, and the degree of inhibition increased with increasing concentration of sulfide. Titanium (III) citrate (used to modify Eh levels) stimulated both nitrogen fixation and CO₂ production, but could not duplicate, at any concentration tested, the twofold increase in nitrogen fixation caused by 8 μmol of S²⁻ per ml. Sulfide additions caused pH changes in the sediment, and when the sediment was adjusted and maintained at pH 7.0 all concentrations of sulfide inhibited nitrogen fixation activity. From considerations of the redox equilibria of H₂, H₂S, and other sulfur species at various pH values, it appeared that H₂S was the toxic entity and that HS⁻ was less toxic. The observed stimulation of activity was apparently due to a pH change coupled with the concurrent production of HS⁻ from H₂S.     

Role of Sulfate Reduction Versus Methanogenesis in Terminal Carbon Flow in Polluted Intertidal Sediment of Waimea Inlet, Nelson, New Zealand

An investigation of the terminal anaerobic processes occurring in polluted intertidal sediments indicated that terminal carbon flow was mainly mediated by sulfate-reducing organisms in sediments with high sulfate concentrations (>10 mM in the interstitial water) exposed to low loadings of nutrient (equivalent to <10² kg of N · day⁻¹) and biochemical oxygen demand (<0.7 × 10³ kg · day⁻¹) in effluents from different pollution sources. However, in sediments exposed to high loadings of nutrient (>10² kg of N · day⁻¹) and biochemical oxygen demand (>0.7 × 10³ kg · day⁻¹), methanogenesis was the major process in the mediation of terminal carbon flow, and sulfate concentrations were low (≤2 mM). The respiratory index [¹⁴CO₂/(¹⁴CO₂ + ¹⁴CH₄)] for [2-¹⁴C]acetate catabolism, a measure of terminal carbon flow, was ≥0.96 for sediment with high sulfate, but in sediments with sulfate as little as 10 μM in the interstitial water, respiratory index values of ≤0.22 were obtained. In the latter sediment, methane production rates as high as 3 μmol · g⁻¹ (dry weight) · h⁻¹ were obtained, and there was a potential for active sulfate reduction.     

Inorganic Carbon Source for Photosynthesis in the Seagrass Thalassia hemprichii (Ehrenb.) Aschers

Photosynthetic carbon uptake of the tropical seagrass Thalassia hemprichii (Ehrenb.) Aschers was studied by several methods. Photosynthesis in buffered seawater in media in the range of pH 6 to pH 9 showed an exponentially increasing rate with decreasing pH, thus indicating that free CO₂ was a photosynthetic substrate. However, these experiments were unable to determine whether photosynthesis at alkaline pH also contained some component due to HCO₃⁻ uptake. This aspect was further investigated by studying photosynthetic rates in a number of media of varying pH (7.8-8.61) and total inorganic carbon (0.75-13.17 millimolar). In these media, photosynthetic rate was correlated with free CO₂ concentration and was independent of the HCO₃⁻ concentration in the medium. Short time-course experiments were conducted during equilibration of free CO₂ and HCO₃⁻ after injection of ¹⁴C labeled solution at acid or alkaline pH. High initial photosynthetic rates were observed when acidic solutions (largely free CO₂) were used but not with alkaline solutions. The concentration of free CO₂ was found to be a limiting factor for photosynthesis in this plant.     

Carbon flow from volcanic CO2 into soil microbial communities of a wetland mofette

Effects of extremely high carbon dioxide (CO₂) concentrations on soil microbial communities and associated processes are largely unknown. We studied a wetland area affected by spots of subcrustal CO₂ degassing (mofettes) with focus on anaerobic autotrophic methanogenesis and acetogenesis because the pore gas phase was largely hypoxic. Compared with a reference soil, the mofette was more acidic (ΔpH ∼0.8), strongly enriched in organic carbon (up to 10 times), and exhibited lower prokaryotic diversity. It was dominated by methanogens and subdivision 1 Acidobacteria, which likely thrived under stable hypoxia and acidic pH. Anoxic incubations revealed enhanced formation of acetate and methane (CH₄) from hydrogen (H₂) and CO₂ consistent with elevated CH₄ and acetate levels in the mofette soil. ¹³CO₂ mofette soil incubations showed high label incorporations with ∼512 ng ¹³C g (dry weight (dw)) soil⁻¹ d⁻¹ into the bulk soil and up to 10.7 ng ¹³C g (dw) soil⁻¹ d⁻¹ into almost all analyzed bacterial lipids. Incorporation of CO₂-derived carbon into archaeal lipids was much lower and restricted to the first 10 cm of the soil. DNA-SIP analysis revealed that acidophilic methanogens affiliated with Methanoregulaceae and hitherto unknown acetogens appeared to be involved in the chemolithoautotrophic utilization of ¹³CO₂. Subdivision 1 Acidobacteriaceae assimilated ¹³CO₂ likely via anaplerotic reactions because Acidobacteriaceae are not known to harbor enzymatic pathways for autotrophic CO₂ assimilation. We conclude that CO₂-induced geochemical changes promoted anaerobic and acidophilic organisms and altered carbon turnover in affected soils.     

The Stoichiometry between CO(2) and H Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

The pH of the medium during CO₂ uptake into the intracellular inorganic carbon (C_i) pool of a high CO₂-requiring mutant (E₁) and wild type of Anacystis nidulans R2 was measured. Experiments were performed under conditions where photosynthetic CO₂ fixation is inhibited. There was an acidification of the medium during CO₂ uptake in the light and an alkalization during CO₂ efflux after darkening. A one to one stoichiometry existed between the amounts of H⁺ appearing in the medium and CO₂ taken up into the intracellular C_i pool, regardless of the carbon species transported. The results indicate that (a) CO₂ is taken up simultaneously with an efflux of equimolar H⁺, probably produced as a result of CO₂ hydration during transport and (b) HCO₃⁻ produced by hydration of CO₂ in the medium was transported into the cells without accompanying net flux of H⁺ or OH⁻. The influx and efflux of C_i during C_i transport produced nonequilibrium between CO₂ and HCO₃⁻ in the medium, with the concentration of HCO₃⁻ being higher than that expected under equilibrium conditions. The nonequilibrium was present even under the conditions where the influx of C_i is compensated by its efflux. The direction of this nonequilibrium suggested that efflux of HCO₃⁻ occurs during uptake of C_i.     

Forms of Dissolved Carbon Dioxide Required for Photosystem II Activity in Chloroplast Membranes

High concentrations of both bicarbonate and formate inhibit photosynthetic O₂ evolution at pH 8.0. At this pH, only 2.4% of the total dissolved carbon dioxide exists as CO₂. At pH 7.3, where 11% of the total dissolved carbon dioxide exists as CO₂, HCO₃⁻ no longer inhibits. While formate still inhibits O₂ evolution at pH 7.3, its effect can be partially overcome if CO₂ is also present. The rate of binding of added ¹⁴C-labeled inorganic carbon is nearly 10-fold more rapid when the internal pH of thylakoid membranes is at 6.0 than when it is at 7.8. These observations suggest that CO₂, not HCO₃⁻, is initially bound to the photosystem II reaction center and that the location of the binding site is on the inside thylakoid surface. However, additional data presented here suggest that, after binding, CO₂ is hydrated to HCO₃⁻ + H⁺ in a pH-dependent reaction. Two possible explanations of the “bicarbonate effect” are presented.     

Photosynthesis and Inorganic Carbon Accumulation in the Acidophilic Alga Cyanidioschyzon merolae

The intracellular pH and membrane potential were determined in the acidophilic algae Cyanidoschyzon merolae as a function of extracellular pH. The alga appear to be capable of maintaining the intracellular pH at the range of 6.35 to 7.1 over the extracellular pH range of 1.5 to 7.5. The membrane potential increase from −12 millivolts (negative inside) to −71 millivolts and thus ΔH⁺ decreased from −300 to −47 millivolts over the same range of extracellular pH. It is suggested that the ΔH⁺ may set the upper and lower limits of pH for growth. Photosynthetic performance was also determined as a function of pH. The cells appeared to utilize CO₂ from the medium as the apparent K_m(co2) was 2 to 3 micromolar CO₂ over the pH range of 1.5 to 7.5 C. merolae appear to possess a `CO₂ concentrating' mechanism.     

Carbon gain and photosynthetic response of chrysanthemum to photosynthetic photon flux density cycles

Most models of carbon gain as a function of photosynthetic irradiance assume an instantaneous response to increases and decreases in irradiance. High- and low-light-grown plants differ, however, in the time required to adjust to increases and decreases in irradiance. In this study the response to a series of increases and decreases in irradiance was observed in Chrysanthemum × morifolium Ramat. “Fiesta” and compared with calculated values assuming an instantaneous response. There were significant differences between high- and low-light-grown plants in their photosynthetic response to four sequential photosynthetic photon flux density (PPFD) cycles consisting of 5-minute exposures to 200 and 400 micromoles per square meter per second (μmol m⁻²s⁻¹). The CO₂ assimilation rate of high-light-grown plants at the cycle peak increased throughout the PPFD sequence, but the rate of increase was similar to the increase in CO₂ assimilation rate observed under continuous high-light conditions. Low-light leaves showed more variability in their response to light cycles with no significant increase in CO₂ assimilation rate at the cycle peak during sequential cycles. Carbon gain and deviations from actual values (percentage carbon gain over- or underestimation) based on assumptions of instantaneous response were compared under continuous and cyclic light conditions. The percentage carbon gain overestimation depended on the PPFD step size and growth light level of the leaf. When leaves were exposed to a large PPFD increase, the carbon gain was overestimated by 16 to 26%. The photosynthetic response to 100 μmol m⁻² s⁻¹ PPFD increases and decreases was rapid, and the small overestimation of the predicted carbon gain, observed during photosynthetic induction, was almost entirely negated by the carbon gain underestimation observed after a decrease. If the PPFD cycle was 200 or 400 μmol m⁻² s⁻¹, high- and low-light leaves showed a carbon gain overestimation of 25% that was not negated by the underestimation observed after a light decrease. When leaves were exposed to sequential PPFD cycles (200-400 μmol m⁻² s⁻¹), carbon gain did not differ from leaves exposed to a single PPFD cycle of identical irradiance integral that had the same step size (200-400-200 μmol m⁻² s⁻¹) or mean irradiance (200-300-200 μmol m⁻² s⁻¹).     

Inorganic Carbon Uptake during Photosynthesis : II. Uptake by Isolated Asparagus Mesophyll Cells during Isotopic Disequilibrium

The species of inorganic carbon (CO₂ or HCO₃⁻) taken up a source of substrate for photosynthetic fixation by isolated Asparagus sprengeri mesophyll cells is investigated. Discrimination between CO₂ or HCO₃⁻ transport, during steady state photosynthesis, is achieved by monitoring the changes (by ¹⁴C fixation) which occur in the specific activity of the intracellular pool of inorganic carbon when the inorganic carbon present in the suspending medium is in a state of isotopic disequilibrium. Quantitative comparisons between theoretical (CO₂ or HCO₃⁻ transport) and experimental time-courses of ¹⁴C incorporation, over the pH range of 5.2 to 7.5, indicate that the specific activity of extracellular CO₂, rather than HCO₃⁻, is the appropriate predictor of the intracellular specific activity. It is concluded, therefore, that CO₂ is the major source of exogenous inorganic carbon taken up by Asparagus cells. However, at high pH (8.5), a component of net DIC uptake may be attributable to HCO₃⁻ transport, as the incorporation of ¹⁴C during isotopic disequilibrium exceeds the maximum possible incorporation predicted on the basis of CO₂ uptake alone. The contribution of HCO₃⁻ to net inorganic carbon uptake (pH 8.5) is variable, ranging from 5 to 16%, but is independent of the extracellular HCO₃⁻ concentration. The evidence for direct HCO₃⁻ transport is subject to alternative explanations and must, therefore, be regarded as equivocal. Nonlinear regression analysis of the rate of ¹⁴C incorporation as a function of time indicates the presence of a small extracellular resistance to the diffusion of CO₂, which is partially alleviated by a high extracellular concentration of HCO₃⁻.     

CO2 Uptake and Fixation by Endosymbiotic Chemoautotrophs from the Bivalve Solemya velum

Chemoautotrophic symbioses, in which endosymbiotic bacteria are the major source of organic carbon for the host, are found in marine habitats where sulfide and oxygen coexist. The purpose of this study was to determine the influence of pH, alternate sulfur sources, and electron acceptors on carbon fixation and to investigate which form(s) of inorganic carbon is taken up and fixed by the gamma-proteobacterial endosymbionts of the protobranch bivalve Solemya velum. Symbiont-enriched suspensions were generated by homogenization of S. velum gills, followed by velocity centrifugation to pellet the symbiont cells. Carbon fixation was measured by incubating the cells with ¹⁴C-labeled dissolved inorganic carbon. When oxygen was present, both sulfide and thiosulfate stimulated carbon fixation; however, elevated levels of either sulfide (>0.5 mM) or oxygen (1 mM) were inhibitory. In the absence of oxygen, nitrate did not enhance carbon fixation rates when sulfide was present. Symbionts fixed carbon most rapidly between pH 7.5 and 8.5. Under optimal pH, sulfide, and oxygen conditions, symbiont carbon fixation rates correlated with the concentrations of extracellular CO₂ and not with HCO₃⁻ concentrations. The half-saturation constant for carbon fixation with respect to extracellular dissolved CO₂ was 28 ± 3 μM, and the average maximal velocity was 50.8 ± 7.1 μmol min⁻¹ g of protein⁻¹. The reliance of S. velum symbionts on extracellular CO₂ is consistent with their intracellular lifestyle, since HCO₃⁻ utilization would require protein-mediated transport across the bacteriocyte membrane, perisymbiont vacuole membrane, and symbiont outer and inner membranes. The use of CO₂ may be a general trait shared with many symbioses with an intracellular chemoautotrophic partner.     

Selective and Reversible Inhibition of Active CO(2) Transport by Hydrogen Sulfide in a Cyanobacterium

The active transport of CO₂ in the cyanobacterium Synechococcus UTEX 625 was inhibited by H₂S. Treatment of the cells with up to 150 micromolar H₂S + HS⁻ at pH 8.0 had little effect on Na⁺-dependent HCO₃⁻ transport or photosynthetic O₂ evolution, but CO₂ transport was inhibited by more than 90%. CO₂ transport was restored when H₂S was removed by flushing with N₂. At constant total H₂S + HS⁻ concentrations, inhibition of CO₂ transport increased as the ratio of H₂S to HS⁻ increased, suggesting a direct role for H₂S in the inhibitory process. Hydrogen sulfide does not appear to serve as a substrate for transport. In the presence of H₂S and Na⁺ -dependent HCO₃⁻ transport, the extracellular CO₂ concentration rose considerably above its equilibrium level, but was maintained far below its equilibrium level in the absence of H₂S. The inhibition of CO₂ transport, therefore, revealed an ongoing leakage from the cells of CO₂ which was derived from the intracellular dehydration of HCO₃⁻ which itself had been recently transported into the cells. Normally, leaked CO₂ is efficiently transported back into the cell by the CO₂ transport system, thus maintaining the extracellular CO₂ concentration near zero. It is suggested that CO₂ transport not only serves as a primary means of inorganic carbon acquisition for photosynthesis but also serves as a means of recovering CO₂ lost from the cell. A schematic model describing the relationship between the CO₂ and HCO₃⁻ transport systems is presented.     

The Effect of pH, O(2), and Temperature on the CO(2) Compensation Point of Isolated Asparagus Mesophyll Cells

The effect of pH, O₂ concentration, and temperature on the CO₂ compensation point (Г[CO₂]) of isolated Asparagus sprengeri Regel mesophyll cells has been determined in a closed, aqueous environment by a sensitive gas-chromatographic technique. Measured values range between 10 and 100 microliters per liter CO₂ depending upon experimental conditions. The Г(CO₂) increases with increasing temperature. The rate of increase is dependent upon the O₂ concentration and is more rapid at high (250-300 micromolar), than at low (30-60 micromolar), O₂ concentrations. The differential effect of temperature on Г(CO₂) is more pronounced at pH 6.2 than at pH 8.0, but this pH-dependence is not attributable to a direct, differential effect of pH on the relative rates of photosynthesis and photorespiration, as the O₂-sensitive component of Г(CO₂) remains constant over this range. The Г(CO₂) of Asparagus cells at 25°C decreases by 50 microliters per liter when the pH is raised from 6.2 to 8.0, regardless of the prevailing O₂ concentration. It is suggested that the pH-dependence of Г(CO₂) is related to the ability of the cell to take up CO₂ from the aqueous environment. The correlation between high HCO₃⁻ concentrations and low Г(CO₂) at alkaline pH indicates that extracellular HCO₃⁻ facilitates the uptake of CO₂, possibly by increasing the flux of inorganic carbon from the bulk medium to the cell surface. The strong O₂− and temperature-dependence of Г(CO₂) indicates that isolated Asparagus mesophyll cells lack an efficient means for concentrating intracellular CO₂ to a level sufficient to reduce or suppress photorespiration.     

Coral reef calcification: carbonate,bicarbonate and proton flux under conditions of increasing ocean acidification

Data on calcification rate of coral and crustose coralline algae were used to test the proton flux model of calcification. There was a significant correlation between calcification (G) and the ratio of dissolved inorganic carbon (DIC) to proton concentration ([DIC] : [H⁺] ratio). The ratio is tightly correlated with [CO₃²⁻] and with aragonite saturation state (Ω_a). An argument is presented that correlation does not prove cause and effect, and that Ω_a and [CO₃²⁻] have no basic physiological meaning on coral reefs other than a correlation with [DIC] : [H⁺] ratio, which is the driver of G.     

Interactive Effects of Ocean Acidification and Nitrogen-Limitation on the Diatom Phaeodactylum tricornutum

Climate change is expected to bring about alterations in the marine physical and chemical environment that will induce changes in the concentration of dissolved CO₂ and in nutrient availability. These in turn are expected to affect the physiological performance of phytoplankton. In order to learn how phytoplankton respond to the predicted scenario of increased CO₂ and decreased nitrogen in the surface mixed layer, we investigated the diatom Phaeodactylum tricornutum as a model organism. The cells were cultured in both low CO₂ (390 μatm) and high CO₂ (1000 μatm) conditions at limiting (10 μmol L⁻¹) or enriched (110 μmol L⁻¹) nitrate concentrations. Our study shows that nitrogen limitation resulted in significant decreases in cell size, pigmentation, growth rate and effective quantum yield of Phaeodactylum tricornutum, but these parameters were not affected by enhanced dissolved CO₂ and lowered pH. However, increased CO₂ concentration induced higher rETR_max and higher dark respiration rates and decreased the CO₂ or dissolved inorganic carbon (DIC) affinity for electron transfer (shown by higher values for K_{1/2 DIC} or K_{1/2 CO2}). Furthermore, the elemental stoichiometry (carbon to nitrogen ratio) was raised under high CO₂ conditions in both nitrogen limited and nitrogen replete conditions, with the ratio in the high CO₂ and low nitrate grown cells being higher by 45% compared to that in the low CO₂ and nitrate replete grown ones. Our results suggest that while nitrogen limitation had a greater effect than ocean acidification, the combined effects of both factors could act synergistically to affect marine diatoms and related biogeochemical cycles in future oceans.     

Continuous Measurements of the Free Dissolved CO(2) Concentration during Photosynthesis of Marine Plants: Evidence for HCO(3) Use in Chondrus crispus

An experimental system consisting of a gas exchange column linked to an assimilation chamber has been developed to record continuously the free dissolved CO₂ concentration in seawater containing marine plants. From experiments performed on the red macroalga Chondrus crispus (Rhodophyta, Gigartinales), this measurement is in agreement with the free CO₂ concentration calculated from the resistance to CO₂ exchanges in a biphasic system (gas and liquid) as earlier reported. The response time of this apparatus is short enough to detect, in conditions of constant pH, a photosynthesis-caused gradient between free CO₂ and HCO₃⁻ pools which half-equilibrates in 25 seconds. Abolished by carbonic anhydrase, the magnitude of this gradient increases with decreasing time of seawater transit from the chamber to the column apparatus. But its maximum magnitude (0.35 micromolar CO₂) is negligible compared to the difference between air and free CO₂ (11.4 micromolar CO₂). This illustrates the extent of the physical limiting-step occurring at the air-water interface when inorganic carbon consumption in seawater is balanced by dissolving gaseous CO₂. The direction of this small free CO₂/HCO₃⁻ gradient indicates that HCO₃⁻ is consumed during photosynthesis.     

Carbon Oxysulfide Inhibition of the CO(2)-Concentrating Process of Unicellular Green Algae

Carbonyl sulfide (COS), a substrate for carbonic anhydrase, inhibited alkalization of the medium, O₂ evolution, dissolved inorganic carbon accumulation, and photosynthetic CO₂ fixation at pH 7 or higher by five species of unicellular green algae that had been air-adapted for forming a CO₂-concentrating process. This COS inhibition can be attributed to inhibition of external HCO₃⁻ conversion to CO₂ and OH⁻ by the carbonic anhydrase component of an active CO₂ pump. At a low pH of 5 to 6, COS stimulated O₂ evolution during photosynthesis by algae with low CO₂ in the media without alkalization of the media. This is attributed to some COS hydrolysis by carbonic anhydrase to CO₂. Although COS had less effect on HCO₃⁻ accumulation at pH 9 by a HCO₃⁻ pump in Scenedesmus, COS reduced O₂ evolution probably by inhibiting internal carbonic anhydrases. Because COS is hydrolyzed to CO₂ and H₂S, its inhibition of the CO₂ pump activity and photosynthesis is not accurate, when measured by O₂ evolution, by NaH¹⁴CO₃ accumulation, or by ¹⁴CO₂ fixation.     

Two Systems for Concentrating CO(2) and Bicarbonate during Photosynthesis by Scenedesmus

Scenedesmus cells grown on high CO₂, when adapted to air levels of CO₂ for 4 to 6 hours in the light, formed two concentrating processes for dissolved inorganic carbon: one for utilizing CO₂ from medium of pH 5 to 8 and one for bicarbonate accumulation from medium of pH 7 to 11. Similar results were obtained with assays by photosynthetic O₂ evolution or by accumulation of dissolved inorganic carbon inside the cells. The CO₂ pump with K_0.5 for O₂ evolution of less than 5 micromolar CO₂ was similar to that previously studied with other green algae such as Chlamydomonas and was accompanied by plasmalemma carbonic anhydrase formation. The HCO₃⁻ concentrating process between pH 8 to 10 lowered the K_0.5 (DIC) from 7300 micromolar HCO₃⁻ in high CO₂ grown Scenedesmus to 10 micromolar in air-adapted cells. The HCO₃⁻ pump was inhibited by vanadate (K_i of 150 micromolar), as if it involved an ATPase linked HCO₃⁻ transporter. The CO₂ pump was formed on low CO₂ by high-CO₂ grown cells in growth medium within 4 to 6 hours in the light. The alkaline HCO₃⁻ pump was partially activated on low CO₂ within 2 hours in the light or after 8 hours in the dark. Full activation of the HCO₃⁻ pump at pH 9 had requirements similar to the activation of the CO₂ pump. Air-grown or air-adapted cells at pH 7.2 or 9 accumulated in one minute 1 to 2 millimolar inorganic carbon in the light or 0.44 millimolar in the dark from 150 micromolar in the media, whereas CO₂-grown cells did not accumulate inorganic carbon. A general scheme for concentrating dissolved inorganic carbon by unicellular green algae utilizes a vanadate-sensitive transporter at the chloroplast envelope for the CO₂ pump and in some algae an additional vanadate-sensitive plasmalemma HCO₃⁻ transporter for a HCO₃⁻ pump.     

Inorganic Carbon Uptake by Chlamydomonas reinhardtii

The rates of CO₂-dependent O₂ evolution by Chlamydomonas reinhardtii, grown with either air levels of CO₂ or air with 5% CO₂, were measured at varying external pH. Over a pH range of 4.5 to 8.5, the external concentration of CO₂ required for half-maximal rates of photosynthesis was constant, averaging 25 micromolar for cells grown with 5% CO₂. This is consistent with the hypothesis that these cells take up CO₂ but not HCO₃⁻ from the medium and that their CO₂ requirement for photosynthesis reflects the K_m(CO₂) of ribulose bisphosphate carboxylase. Over a pH range of 4.5 to 9.5, cells grown with air required an external CO₂ concentration of only 0.4 to 3 micromolar for half-maximal rates of photosynthesis, consistent with a mechanism to accumulate external inorganic carbon in these cells. Air-grown cells can utilize external inorganic carbon efficiently even at pH 4.5 where the HCO₃⁻ concentration is very low (40 nanomolar). However, at high external pH, where HCO₃⁻ predominates, these cells cannot accumulate inorganic carbon as efficiently and require higher concentrations of NaHCO₃ to maintain their photosynthetic activity. These results imply that, at the plasma membrane, CO₂ is the permeant inorganic carbon species in air-grown cells as well as in cells grown on 5% CO₂. If active HCO₃⁻ accumulation is a step in CO₂ concentration by air-grown Chlamydomonas, it probably takes place in internal compartments of the cell and not at the plasmalemma.     

Chlorophyll a Fluorescence and Photosynthetic and Growth Responses of Pinus radiata to Phosphorus Deficiency, Drought Stress, and High CO(2)

Needles from phosphorus deficient seedlings of Pinus radiata D. Don grown for 8 weeks at either 330 or 660 microliters CO₂ per liter displayed chlorophyll a fluorescence induction kinetics characteristic of structural changes within the thylakoid chloroplast membrane, i.e. constant yield fluorescence (F_O) was increased and induced fluorescence ([F_P-F_I]/F_O) was reduced. The effect was greatest in the undroughted plants grown at 660 μl CO₂ L⁻¹. By week 22 at 330 μl CO₂ L⁻¹ acclimation to P deficiency had occurred as shown by the similarity in the fluorescence characteristics and maximum rates of photosynthesis of the needles from the two P treatments. However, acclimation did not occur in the plants grown at 660 μl CO₂ L⁻¹. The light saturated rate of photosynthesis of needles with adequate P was higher at 660 μl CO₂ L⁻¹ than at 330 μl CO₂ L⁻¹, whereas photosynthesis of P deficient plants showed no increase when grown at the higher CO₂ concentration. The average growth increase due to CO₂ enrichment was 14% in P deficient plants and 32% when P was adequate. In drought stressed plants grown at 330 μl CO₂ L⁻¹, there was a reduction in the maximal rate of quenching of fluorescence (R_Q) after the major peak. Constant yield fluorescence was unaffected but induced fluorescence was lower. These results indicate that electron flow subsequent to photosystem II was affected by drought stress. At 660 μl CO₂ L⁻¹ this response was eliminated showing that CO₂ enrichment improved the ability of the seedlings to acclimate to drought stress. The average growth increase with CO₂ enrichment was 37% in drought stressed plants and 19% in unstressed plants.