Photoreduction of O(2) Primes and Replaces CO(2) Assimilation

A mass spectrometer with a membrane inlet system was used to monitor directly gaseous components in a suspension of algae. Using labeled oxygen, we observed that during the first 20 seconds of illumination after a dark period, when no net O₂ evolution or CO₂ uptake was observed, O₂ evolution was normal but completely compensated by O₂ uptake. Similarly, when CO₂ uptake was totally or partially inhibited, O₂ evolution proceeded at a high (near maximal) rate. Under all conditions, O₂ uptake balanced that fraction of the O₂ evolution which could not be accounted for by CO₂ uptake.     

Influence of Leaf Starch Concentration on CO(2) Assimilation in Soybean

Net photosynthetic rate, CO₂ compensation concentration, and starch and soluble sugar concentrations were measured in soybean (Glycine max [L.] Merrill) leaves in an attempt to evaluate the effect of carbohydrate concentration on rate of CO₂ assimilation.     

CO(2) Assimilation by Etiolated Hordeum vulgare Seedlings during the Onset of Photosynthesis

The assimilation of CO₂ by etiolated Hordeum vulgare seedlings during an illumination period indicates a conversion of the organisms to autotrophy.

After 1 hour illumination, increases in the photo-assimilation of CO₂ are observed and the distribution of C¹⁴ in the soluble fraction of the plants is predominantly in intermediates of the Calvin cycle.

     

Photosynthetic Assimilation of NO(3) by Intact Cells of the Cyanobacterium Anacystis nidulans: Influence of NO(3) and NH(4) Assimilation on CO(2) Fixation

Illuminated suspensions of Anacystis nidulans, supplied with saturating concentrations of CO₂ evolved O₂ at a greater rate when nitrate was simultaneously present. The extent of the stimulation of noncyclic electron flow induced by nitrate was dependent on light intensity, being maximal under light saturating conditions. Accordingly, nitrate depressed the rate of CO₂ fixation at limiting but not at saturating light, this depression reflecting the competition between both processes for assimilatory power. In contrast, ammonium stimulated CO₂ fixation at any light intensity assayed, the stimulation being dependent on the incorporation of ammonium to carbon skeletons. The positive effect of ammonium on CO₂ fixation also appeared to occur when nitrate was the nitrogen source, since with either nitrogen source an increase in the incorporation of newly fixed carbon into acid-soluble metabolites took place. From these results, the in vivo partitioning of assimilatory power between photosynthetic nitrogen and carbon assimilation and the quantitative and qualitative effects of inorganic nitrogen assimilation on CO₂ fixation are discussed.     

Acclimation of CO(2) Assimilation in Cotton Leaves to Water Stress and Salinity

Cotton (Gossypium hirsutum L. cv Acala SJ2) plants were exposed to three levels of osmotic or matric potentials. The first was obtained by salt and the latter by withholding irrigation water. Plants were acclimated to the two stress types by reducing the rate of stress development by a factor of 4 to 7. CO₂ assimilation was then determined on acclimated and nonacclimated plants. The decrease of CO₂ assimilation in salinity-exposed plants was significantly less in acclimated as compared with nonacclimated plants. Such a difference was not found under water stress at ambient CO₂ partial pressure. The slopes of net CO₂ assimilation versus intercellular CO₂ partial pressure, for the initial linear portion of this relationship, were increased in plants acclimated to salinity of −0.3 and −0.6 megapascal but not in nonacclimated plants. In plants acclimated to water stress, this change in slopes was not significant. Leaf osmotic potential was reduced much more in acclimated than in nonacclimated plants, resulting in turgor maintenance even at −0.9 megapascal. In nonacclimated plants, turgor pressure reached zero at approximately −0.5 megapascal. The accumulation of Cl⁻ and Na⁺ in the salinity-acclimated plants fully accounted for the decrease in leaf osmotic potential. The rise in concentration of organic solutes comprised only 5% of the total increase in solutes in salinity-acclimated and 10 to 20% in water-stress-acclimated plants. This acclimation was interpreted in light of the higher protein content per unit leaf area and the enhanced ribulose bisphosphate carboxylase activity. At saturating CO₂ partial pressure, the declined inhibition in CO₂ assimilation of stress-acclimated plants was found for both salinity and water stress.     

Relationship between Net CO(2) Assimilation and Dry Weight Accumulation in Field-Grown Tobacco

To assess the variability of net photosynthetic CO₂ exchange per unit leaf area and to construct budgets for stands of field-grown tobacco (Nicotiana tabacum, Connecticut Broadleaf), a number of short-time measurements were made on all available leaf positions on two varieties using a hand-held transparent chamber for conducting gas exchange measurements on leaves. Measurements of net CO₂ exchange were carried out on 18 separate days during a 35-day period, beginning 22 days after the seedlings were transplanted to the field. Gas exchange assays on leaves were conducted under ambient conditions of temperature and light intensity at all times of day. Solar radiation was monitored throughout the period, and losses of respiratory CO₂ from stems, roots, and leaves (in the dark) were estimated. A simple model was proposed to relate daily total CO₂ input to irradiance and total leaf area. The total leaf area was assumed to be a function of day number. Dark respiratory losses accounted for 41% to 47% of total CO₂ assimilation. Analysis of variance indicated that the two varieties were not significantly different in whole plant rate of CO₂ fixation per unit of leaf area. CO₂ input was closely associated with leaf area within each variety. Throughout the experiment, the difference between the two varieties in total leaf area per plant was the largest single factor in determining net CO₂ inputs. The cumulative dry weight increase for each variety was similar to the prediction of net dry matter input obtained by gas exchange measurements, thus confirming the close relationship between total plant net CO₂ assimilation and dry weight yield.     

Contribution of Metabolites of Photosynthesis to Postillumination CO(2) Assimilation in Response to Lightflects

In the shade plant Alocasia macrorrhiza grown in low light, photosynthetic CO₂ assimilation during a 5 second lightfleck plus postillumination CO₂ assimilation can allow up to 60% more photosynthesis than that which occurs during 5 seconds of steady state light of the same intensity (RL Chazdon, RW Pearcy 1986 Oecologia. 69: 524-531). Metabolites of photosynthesis were measured to determine if the pool of ribulose 1,5-bisphosphate (RuBP) could account for all of the postillumination CO₂ assimilation following a lightfleck in Alocasia. It was found that the pool of triose-P was much larger than that of RuBP and could account for five times more postillumination CO₂ assimilation than could RuBP. The same trend was seen in the sun plant Phaseolus vulgaris when it was grown in the shade. In contrast, sun-grown Alocasia and Phasiolus did not have a large pool of triose-P relative to RuBP following a lightfleck. In sun plants, carbon may rapidly be converted to RuBP in the light whereas in shade plants there may be a restriction in the path between the triose-P and RuBP pools. It is hypothesized that in shade plants the buildup of triose-P rather than RuBP during the lightfleck prevents inhibition of electron transport which may otherwise occur because of competition for ATP between the two kinases of the photosynthetic carbon reduction cycle. Utilization of the triose-P for postillumination CO₂ fixation would require the capacity for significant postillumination ATP synthesis. The extensive grana stacking and large intrathylakoid space which accompanies the high level of chlorophyll in low-light-grown Alocasia could be an important contributing factor to postillumination ATP formation.     

Effect of Triacontanol on Chlamydomonas: I. Stimulation of Growth and Photosynthetic CO(2) Assimilation

Treatment of Chlamydomonas reinhardtii cells, cultured at 5% CO₂, with 1 to 1000 micrograms triacontanol (TRIA) per liter resulted in 21 to 35% increases in cell density, 7 to 31% increases in total chlorophyll, and 20 to 100% increases in photosynthetic CO₂ assimilation. The increase in CO₂ fixation with TRIA treatment occurred before, and was independent of, increases in total chlorophyll or cell number. Chlamydomonas cells responded to a broad range of TRIA concentrations that were at least one order of magnitude above the optimum concentration established for higher plants. The necessity for larger concentrations of TRIA may be due to destabilizing effects of Ca²⁺ and K⁺ present in the Chlamydomonas growth medium. These ions caused flocculation of the colloidally dispersed TRIA in apparent competition with binding of [¹⁴C]TRIA to Chlamydomonas cells. Octacosanol inhibited the effect of TRIA on photosynthetic CO₂ assimilation. TRIA treatment did not alter the distribution of ¹⁴C-label among photosynthetic products. The effect of TRIA on photosynthetic CO₂ assimilation increased with time after treatment up to 3 days. Chlamydomonas cells that had been grown at low-CO₂ (air) did not respond to TRIA, and transfer of high-CO₂ (5%) grown cells that had responded to TRIA to a low-CO₂ atmosphere resulted in a loss of the effect of TRIA. The effect of pH on photosynthetic CO₂ assimilation indicated that CO₂ is probably the species of inorganic carbon utilized by control and TRIA-treated Chlamydomonas cells.     

On Autotropism in Botrytis: Measurement Technique and Control by CO(2)

The mutual orientation of the germination of nearby pairs of Botrytis spores growing in a simple, dilute medium was studied. If the medium is equilibrated with room air, they show a very strong tendency to germinate both toward each other and in a cis arrangement, i.e., toward the same side of the line joining their centers. If the medium is equilibrated with air enriched with 0.3% or 3% CO₂, (i.e., 10 or 100 times the normal CO₂ concentration) then the cells show an equally strong tendency to germinate away from each other. The interaction shows little dependence upon pH.     

Demonstration of Both a Photosynthetic and a Nonphotosynthetic CO(2) Requirement for NH(4) Assimilation in the Green Alga Selenastrum minutum

Nitrogen-limited and nitrogen-sufficient cell cultures of Selenastrum minutum (Naeg.) Collins (Chlorophyta) were used to investigate the dependence of NH₄⁺ assimilation on exogenous CO₂. N-sufficient cells were only able to assimilate NH₄⁺ maximally in the presence of CO₂ and light. Inhibition of photosynthesis with 3-(3,4-dichlorophenyl)-1,1-dimethylurea, diuron also inhibited NH₄⁺ assimilation. These results indicate that NH₄⁺ assimilation by N-sufficient cells exhibited a strict requirement for photosynthetic CO₂ fixation. N-limited cells assimilated NH₄⁺ both in the dark and in the light in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea, diuron, indicating that photosynthetic CO₂ fixation was not required for NH₄⁺ assimilation. Using CO₂ removal techniques reported previously in the literature, we were unable to demonstrate CO₂-dependent NH₄⁺ assimilation in N-limited cells. However, employing more stringent CO₂ removal techniques we were able to show a CO₂ dependence of NH₄⁺ assimilation in both the light and dark, which was independent of photosynthesis. The results indicate two independent CO₂ requirements for NH₄⁺ assimilation. The first is as a substrate for photosynthetic CO₂ fixation, whereas the second is a nonphoto-synthetic requirement, presumably as a substrate for the anaplerotic reaction catalyzed by phosphoenolpyruvate carboxylase.     

Primary Productivity and Water Balance of Grassland Vegetation on Three Soils in a Continuous CO2 Gradient: Initial Results from the Lysimeter CO2 Gradient Experiment

Field studies of atmospheric CO₂ effects on ecosystems usually include few levels of CO₂ and a single soil type, making it difficult to ascertain the shape of responses to increasing CO₂ or to generalize across soil types. The Lysimeter CO₂ Gradient (LYCOG) chambers were constructed to maintain a linear gradient of atmospheric CO₂ (~250 to 500 μl l⁻¹) on grassland vegetation established on intact soil monoliths from three soil series. The chambers maintained a linear daytime CO₂ gradient from 263 μl l⁻¹ at the subambient end of the gradient to 502 μl l⁻¹ at the superambient end, as well as a linear nighttime CO₂ gradient. Temperature variation within the chambers affected aboveground biomass and evapotranspiration, but the effects of temperature were small compared to the expected effects of CO₂. Aboveground biomass on Austin soils was 40% less than on Bastrop and Houston soils. Biomass differences between soils resulted from variation in biomass of Sorghastrum nutans, Bouteloua curtipendula, Schizachyrium scoparium (C₄ grasses), and Solidago canadensis (C₃ forb), suggesting the CO₂ sensitivity of these species may differ among soils. Evapotranspiration did not differ among the soils, but the CO₂ sensitivity of leaf-level photosynthesis and water use efficiency in S. canadensis was greater on Houston and Bastrop than on Austin soils, whereas the CO₂ sensitivity of soil CO₂ efflux was greater on Bastrop soils than on Austin or Houston soils. The effects of soil type on CO₂ sensitivity may be smaller for some processes that are tightly coupled to microclimate. LYCOG is useful for discerning the effects of soil type on the CO₂ sensitivity of ecosystem function in grasslands. Author Contributions: PF conceived study, analyzed data, and wrote the paper. AK, AP analyzed data. DH, VJ, RJ, HJ, and WP conceived study, and conducted research.     

Estimation of Photorespiration Based on the Initial Rate of Postillumination CO(2) Release: I. A Nonsteady State Model for Measurement of CO(2) Exchange Transients

Although open systems have been used for the study of transients in leaf CO₂ exchange such as the postillumination burst, these systems frequently do not permit reliable estimates of transient rates due to their nonsteady state nature. A nonsteady state mathematical approach is described which predicts changes in CO₂ concentration in the leaf chamber and infrared gas analyzer measuring cell as a function of leaf CO₂ exchange rate in Nicotiana tabacum vars John Williams Broadleaf and Havana Seed. With the aid of a computer, a numerical formula simulates the mixing and dilution which occurs as CO₂ passes through the finite volume of the measuring cell of the analyzer. The method is presented with special relevance to photorespiration as manifested by the postillumination burst of CO₂. The latter is suggested to decline with the first order kinetics following darkening of a C₃ leaf. This approach provides a basis for reliable estimation of the initial and, hence, maximal rate of CO₂ evolution during the postillumination burst under a variety of environmental conditions.     

Measurement of CO(2) and H(2)O Vapor Exchange in Spinach Leaf Discs : Effects of Orthophosphate

A leaf chamber has been designed which allows the measurement of both CO₂ and water vapor exchange in Spinacia oleracea leaf discs. The center of the disc lies within a cylindrical gas chamber and its margins are enclosed within a cavity through which water or various metabolites can be pumped. In saturating light and normal atmospheres, the leaf discs have a relatively low resistance to H₂O vapor transfer (r_w = 1.87 seconds per centimeter) and can support high rates of photosynthesis for several hours. The abaxial surface of a disc had a higher resistance to water vapor transfer (r_w = 3.22 seconds per centimeter) than the adaxial (r_w = 2.45 seconds per centimeter) despite having a higher stomatal frequency (abaxial, 105/square millimeter; adaxial, 58/square millimeter). In 2% O₂, the discs required an internal concentration of CO₂ of 115 microliters per liter to support one-half of the maximal velocity of apparent photosynthesis (average value, 66 milligrams CO₂ per square decimeter per hour). In 20% O₂, the comparable values are 156 microliters per liter and 56 milligrams CO₂ per square decimeter per hour. In air, apparent photosynthesis saturated at intensities (750 microeinsteins per square meter per second) well below that of daylight but, when the internal CO₂ was raised to 700 to 900 microliters per liter, photosynthesis was not saturated even at daylight intensities (2025 microeinsteins per square meter per second). The distribution of Prussian blue crystals, formed after ferrocyanide feeding, showed that water entered the disc via the vasculature. When 25-minute pulses of orthophosphate were provided in the feeding solution, there were concentration-dependent increases in both r_w and r_m leading to inhibition of photosynthesis. The orthophosphate-dependent inhibitions were reversible.     

Isolation of Bundle Sheath Cell Chloroplasts from the NADP-ME Type C(4) Plant Zea mays: Capacities for CO(2) Assimilation and Malate Decarboxylation

Bundle sheath chloroplasts have been isolated from Zea mays leaves by a procedure involving enzymic digestion of mechanically prepared strands of bundle sheath cells followed by gentle breakage and filtration. The resulting crude chloroplast preparation was enriched by Percoll density layer centrifugation to yield intact chloroplasts (about 20 micrograms chlorophyll per 10-gram leaf tissue) with high metabolic activities. Based on activities of marker enzymes in the chloroplast and bundle sheath cell extracts, the chloroplasts were essentially free of contamination by other organelles and cytoplasmic material, and were generally about 70% intact. Chlorophyll a/b ratios were high (about 10). With appropriate substrates these chloroplasts displayed high rates of malate decarboxylation, measured as pyruvate formation, and CO₂ assimilation (maximum rates approximately 5 and 3 micromoles per minute per milligram chlorophyll, respectively). These activities were light dependent, linear for at least 20 minutes at 30°C, and displayed highest rates at pH 8.0. High metabolic rates were dependent on addition of an exogenous source of carbon to the photosynthetic carbon reduction cycle (3-phosphoglycerate or dihydroxyacetone phosphate) and a nucleotide (ATP, ADP, or AMP), as well as aspartate. Generally, neither malate decarboxylation nor CO₂ assimilation occurred substantially in the absence of the other activity indicating a close relationship between these processes. Presumably, NADPH required for the photosynthetic carbon reduction cycle is largely supplied during the decarboxylation of malate by NADP-malic enzyme. The results are discussed in relation to the role of bundle sheath chloroplasts in C₄ photosynthesis by species of the NADP-malic enzyme type.     

CO(2) Assimilation and Malate Decarboxylation by Isolated Bundle Sheath Chloroplasts from Zea mays

Conditions for optimal CO₂ fixation and malate decarboxylation by isolated bundle sheath chloroplasts from Zea mays were examined. The relative rates of these processes varied according to the photosynthetic carbon reduction cycle intermediate provided. Highest rates of malate decarboxylation, measured as pyruvate formation, were seen in the presence of 3-phosphoglycerate, while carbon fixation was highest in the presence of dihydroxyacetone phosphate; only low rates were measured with added ribose-5-phosphate. Chloroplasts exhibited a distinct phosphate requirement and this was optimal at a level of 2 millimolar inorganic phosphate in the presence of 2.5 millimolar 3-phosphoglycerate, dihydroxyacetone phosphate, or ribose-5-phosphate. Malate decarboxylation and CO₂ fixation were stimulated by additions of AMP, ADP, or ATP with half-maximal stimulation occurring at external adenylate concentrations of about 0.15 millimolar. High concentrations (>1 millimolar) of AMP were inhibitory. Aspartate included in the incubation medium stimulated malate decarboxylation and CO₂ assimilation. In the presence of aspartate, the apparent Michaelis constant (malate) for malate decarboxylation to pyruvate by chloroplasts decreased from 6 to 0.67 millimolar while the calculated V_max for this process increased from 1.3 to 3.3 micromoles per milligram chlorophyll. Aspartate itself was not metabolized. It was concluded that the processes mediating the transport of phosphate, 3-phosphoglycerate, and dihydroxyacetone phosphate transport on the one hand, and also of malate might differ from those previously described for chloroplasts from C₃ plants.     

Effect of High Temperature on Photosynthesis in Beans (II. CO2 Assimilation and Metabolite Contents)

The effect of high temperatures on CO2 assimilation, metabolite content, and capacity for reducing power production in non-photorespiratory conditions has been assessed in two different bean (Phaseolus vulgarus L.) varieties, Blue Lake (commercially available in the United Kingdom) and Barbucho (a noncommercially bred Chilean variety), which are known to differ in their resistance to extreme high temperatures. Barbucho maintains its photosynthetic functions for a longer period of time under extreme heat compared with Blue Lake. The CO2 assimilation rate was increased by increases in temperature, with a decrease in ratio of rates of temperatures differing by 10[deg]C. It is suggested that limitations to CO2 assimilation are caused by metabolic restrictions that can be differentiated between those occurring in the range of 20 to 30[deg]C and 30 to 35[deg]C. It is likely that changes in the capacity for Calvin cycle regeneration and starch synthesis affect photosynthesis in the range of 20 to 30[deg]C. But following an increase in temperature from 30 to 35[deg]C, the supply of reducing power becomes limiting. From analysis of adenylate concentration, transthylakoid energization, and, indirectly, NADPH/NADP+ ratio, it was concluded that the limitation in the assimilatory power was due to an oxidation of the NADPH/NADP+ pool. In the range of 30 to 35[deg]C, the photosystem I quantum yield increased and photosystem II maintained its value. We conclude that the reorganization of thylakoids observed at 30 to 35[deg]C increased the excitation of photosystem I, inducing an increase in cyclic electron transport and a decrease in the supply of NADPH, limiting carbon assimilation.     

Leaf Conductance in Relation to Rate of CO(2) Assimilation: III. Influences of Water Stress and Photoinhibition

Rates of CO₂ assimilation and leaf conductances to CO₂ transfer were measured in plants of Zea mays during a period of 14 days in which the plants were not rewatered, and leaf water potential decreased from −0.5 to −8.0 bar. At any given ambient partial pressure of CO₂, water stress reduced rate of assimilation and leaf conductance similarly, so that intercellular partial pressure of CO₂ remained almost constant. At normal ambient partial pressure of CO₂, the intercellular partial pressure of CO₂ was estimated to be 95 microbars. This is the same as had been estimated in plants of Zea mays grown with various levels of nitrogen supply, phosphate supply and irradiance, and in plants of Zea mays examined at different irradiances.

After leaves of Phaseolus vulgaris L. and Eucalyptus pauciflora Sieb. ex Spreng had been exposed to high irradiance in an atmosphere of CO₂-free N₂ with 10 millibars O₂, rates of assimilation and leaf conductances measured in standard conditions had decreased in similar proportions, so that intercellular partial pressure of CO₂ remained almost unchanged. As the conductance of each epidermis that had not been directly irradiated had declined as much as that in the opposite, irradiated surface it was hypothesized that conductance may have been influenced by photoinhibition within the mesophyll tissue.

     

Inorganic Carbon Diffusion between C(4) Mesophyll and Bundle Sheath Cells: Direct Bundle Sheath CO(2) Assimilation in Intact Leaves in the Presence of an Inhibitor of the C(4) Pathway

Photosynthesis rates of detached Panicum miliaceum leaves were measured, by either CO₂ assimilation or oxygen evolution, over a wide range of CO₂ concentrations before and after supplying the phosphoenolpyruvate (PEP) carboxylase inhibitor, 3,3-dichloro-2-(dihydroxyphosphinoyl-methyl)-propenoate (DCDP). At a concentration of CO₂ near ambient, net photosynthesis was completely inhibited by DCDP, but could be largely restored by elevating the CO₂ concentration to about 0.8% (v/v) and above. Inhibition of isolated PEP carboxylase by DCDP was not competitive with respect to HCO₃⁻, indicating that the recovery was not due to reversal of enzyme inhibition. The kinetics of ¹⁴C-incorporation from ¹⁴CO₂ into early labeled products indicated that photosynthesis in DCDP-treated P. miliaceum leaves at 1% (v/v) CO₂ occurs predominantly by direct CO₂ fixation by ribulose 1,5-bisphosphate carboxylase. From the photosynthesis rates of DCDP-treated leaves at elevated CO₂ concentrations, permeability coefficients for CO₂ flux into bundle sheath cells were determined for a range of C₄ species. These values (6-21 micromoles per minute per milligram chlorophyll per millimolar, or 0.0016-0.0056 centimeter per second) were found to be about 100-fold lower than published values for mesophyll cells of C₃ plants. These results support the concept that a CO₂ permeability barrier exists to allow the development of high CO₂ concentrations in bundle sheath cells during C₄ photosynthesis.