Ultrastructure of Dunaliella tertiolecta Cells Grown under Low and High CO2 Concentrations1

The cells of Dunaliella tertiolecta grown under ordinary air(low-CO₂ cells) had a well developed pyrenoid with many morestarch granules than those grown under air enriched with CO₂(high-CO₂ cells). The chloroplast was located close to the plasmamembranein low-CO₂ cells, while that in high-CO₂ cells was located inthe inner area of the cells. Chloroplast envelope was electronicallydenser in low-CO₂ cells than in high-CO₂ cells, while the oppositeeffect of CO₂ was observed for the plasmamembrane. ²On leave from Institute of Biology, University of Novi Sad,Novi Sad, Yugoslavia. (Received November 7, 1985; Accepted March 5, 1986)     

The Effect of High CO2 and Low O2 Concentrations in Simulated Landfill Gas on the Growth and Nodule Activity of Leucaena leucocephala

When roots of Leucaena leucocephala seedlings (White Popinac,a tropical legume tree belongs to the Family Mimosaceae) werefumigated with simulated landfill gas (CO₂ above 10% and O₂from 10% to atmospheric level), the stem elongation rate andstomatal conductance were inhibited at the absence of any apparentleaf water deficit. When compared with a treatment where rootsystem was flooded, the effect of gas fumigation on the shootphysiology was relatively mild and appeared later. On the otherhand, nodule activity (measured as rate of acetylene reductionactivity, ARA) was much more severely inhibited by gas fumigation.Although nodule dry weight and carbohydrate storage in noduleswere reduced, the inhibition was not likely a result of theshortage of carbohydrate reserve in the nodules. This was becausethe ARA of untreated fresh nodules was also inhibited immediatelyfollowing exposure to the simulated landfill gas. In furtherexperiments where CO₂ and O₂ were manipulated separately, althougha reduction of O₂ concentration to half of the atmospheric levelmight account for up to 30% loss of ARA with considerable variation,the high CO₂ alone showed a much more severe inhibition. ThisCO₂-induced inhibition was not reversible one hour after thehigh CO₂ gas was removed. There was some recovery of activity5 day after plants were fumigated, suggesting that the legumeplant can maintain some nitrogen-fixation activity under theinfluence of landfill gas. (Received April 10, 1995; Accepted August 22, 1995)     

The Photosynthetic Response to Elevated CO2 in High Altitude Potato Species (Solanum curtilobum)

Plants of Solanum curtilobum (from high altitude) and Solanum tuberosum (from low altitude) were grown in open-top chambers in a greenhouse at either ambient (AC, 360 µmol mol^–1) or ca. twice ambient (EC, 720 µmol mol^–1) CO₂ concentrations for 30 d. CO₂ treatments started at the reproductive stage of the plants. There were similar patterns in the physiological response to CO₂ enrichment in the two species. Stomatal conductance was reduced by 59 % in S. tuberosum and by 55 % in S. curtilobum, but such a reduction did not limit the net photosynthetic rate (P _N), which was increased by approximately 56 % in S. curtilobum and 53 % in S. tuberosum. The transpiration rate was reduced by 16 % in both potato species while instantaneous transpiration efficiency increased by 80 % in S. tuberosum and 90 % in S. curtilobum. Plants grown under EC showed 36 and 66 % increment in total dry biomass, whereas yields (dry mass of tubers) were increased by 40 and 85 % in S. tuberosum and S. curtilobum, respectively. EC promoted productivity by increasing P _N. Thus S. tuberosum, cultivated around the world at low altitudes, and S. curtilobum, endemic of the highland Andes, respond positively to EC during the tuberisation stage.     

Acclimation of Two Tomato Species to High Atmospheric CO(2): I. Sugar and Starch Concentrations

Lycopersicon esculentum Mill. cv Vedettos and Lycopersicon chmielewskii Rick, LA 1028, were exposed to two CO₂ concentrations (330 or 900 microliters per liter) for 10 weeks. Tomato plants grown at 900 microliters per liter contained more starch and more sugars than the control. However, we found no significant accumulation of starch and sugars in the young leaves of L. esculentum exposed to high CO₂. Carbon exchange rates were significantly higher in CO₂-enriched plants for the first few weeks of treatment but thereafter decreased as tomato plants acclimated to high atmospheric CO₂. This indicates that the long-term decline of photosynthetic efficiency of leaf 5 cannot be attributed to an accumulation of sugar and/or starch. The average concentration of starch in leaves 5 and 9 was always higher in L. esculentum than in L. chmielewskii (151.7% higher). A higher proportion of photosynthates was directed into starch for L. esculentum than for L. chmielewskii. However, these characteristics did not improve the long-term photosynthetic efficiency of L. chmielewskii grown at high CO₂ when compared with L. esculentum. The chloroplasts of tomato plants exposed to the higher CO₂ concentration exhibited a marked accumulation of starch. The results reported here suggest that starch and/or sugar accumulation under high CO₂ cannot entirely explain the loss of photosynthetic efficiency of high CO₂-grown plants.     

Ethylene: Response of Fruit Dehiscence to CO(2) and Reduced Pressure

These studies were conducted to determine whether ethylene serves as a natural regulator of fruit wall dehiscence, a major visible feature of ripening in some fruits. We employed treatments to inhibit ethylene action or remove ethylene and observed their effect on fruit dehiscence. CO₂ (13%), a competitive inhibitor of ethylene action in many systems, readily delayed dehiscence of detached fruits of cotton (Gossypium hirsutum L.), pecan (Carya illinoensis [Wang.] K. Koch), and okra (Hibiscus esculentus L.). The CO₂ effect was duplicated by placing fruits under reduced pressure (200 millimeters mercury), to promote the escape of ethylene from the tissue. Dehiscence of detached fruits of these species as well as attached cotton fruits was delayed. The delay of dehiscence of cotton and okra by both treatments was achieved with fruit harvested at intervals from shortly after anthesis until shortly before natural dehiscence. Pecan fruits would not dehisce until approximately 1 month before natural dehiscence, and during that time, CO₂ and reduced pressure delayed dehiscence. CO₂ and ethylene were competitive in their effects on cotton fruit dehiscence. All of the results are compatible with a hypothetical role of ethylene as a natural regulator of dehiscence, a dominant aspect of ripening of cotton, pecan, and some other fruits.     

The Occurrence of High CO2-Compensation Points in Amaranthus Species

When the CO₂-compensation points of detached leaves of Amaranthusedulis plants grown undera range of controlled environmentswere determined, they were all found to be near zero. However,similar plants grown in a greenhouse occasionally yielded high-compensation-pointleaves. The occurrence of these could not be correlated withleaf age, moisture stress, or the prevailing environmental conditions.These high-compensation-point leaves still showed a typicalKranz anatomy. Starch was found external to the bundle sheathin both high- and low-compensation-point leaves, but the distributionof the starch appeared to vary with light intensity.     

Soil microorganisms at different CO2 and O2 tensions

Microbial biomass and activity were determined in cambisol incubated under ambient and increased (up to 2.23 mmol/L) CO₂ concentrations. An immediate negative response of the soil microbial community to [CO]₂ increase was observed during the first day with respect to microbial biomass, soil respiration and specific respiration activity (both expressed as CO₂ evolution). In contrast, O₂ consumption was not affected but anabolic utilization of available substrate increased. These phenomena were observed under conditions of increased CO₂ tension but without any change in O₂ concentration.     

Photosynthetic Carbon Metabolism in Photoautotrophic Cell Suspension Cultures Grown at Low and High CO(2)

Photosynthetic carbon metabolism was characterized in four photoautotrophic cell suspension cultures. There was no apparent difference between two soybean (Glycine max) and one cotton (Gossypium hirsutum) cell line which required 5% CO₂ for growth, and a unique cotton cell line that grows at ambient CO₂ (660 microliters per liter). Photosynthetic characteristics in all four lines were more like C₃ mesophyll leaf cells than the cell suspension cultures previously studied. The pattern of ¹⁴C-labeling reflected the high ratio of ribulosebisphosphate carboxylase to phosphoenolpyruvate carboxylase activity and showed that CO₂ fixation occurred primarily by the C₃ pathway. Photorespiration occurred at 330 microliters per liter CO₂, 21% O₂ as indicated by the synthesis of high levels of ¹⁴C-labeled glycine and serine in a pulse-chase experiment and by oxygen inhibition of CO₂ fixation. Short-term CO₂ fixation in the presence and absence of carbonic anhydrase showed CO₂, not HCO₃⁻, to be the main source of inorganic carbon taken up by the low CO₂-requiring cotton cells. The cells did not have a CO₂-concentrating mechanism as indicated by silicone oil centrifugation experiments. Carbonic anhydrase was absent in the low CO₂-requiring cotton cells, present in the high CO₂-requiring soybean cell lines, and absent in other high CO₂ cell lines examined. Thus, the presence of carbonic anhydrase is not an essential requirement for photoautotrophy in cell suspension cultures which grow at either high or low CO₂ concentrations.     

Ethylene Biosynthesis and Accumulation under Drained and Submerged Conditions (A Comparative Study of Two Rumex Species)

A model is presented of the regulation of ethylene biosynthesis in relation to submergence and flooding resistance. It is based on time-course measurements of ethylene production, ethylene accumulation, and concentrations of free and conjugated 1-aminocyclo-propane-1-carboxylic acid (ACC) in submerged and drained flooding-resistant Rumex palustris Sm. and flooding-sensitive Rumex acetosella L. plants. From these data, in vivo reaction rates of the final steps in the ethylene biosynthetic pathway were calculated. According to our model, submergence stimulates ACC formation and inhibits conversion of ACC to ethylene in both Rumex species, and as a result, ACC accumulates. This may explain the stimulated ACC conjugation observed in submerged plants. Although submergence inhibited ethylene production, physical entrapment increased endogenous ethylene concentrations in both flooding-resistant R. palustris and flooding-sensitive R. acetosella plants. However, R. palustris plants controlled their internal ethylene levels in the long term by a negative regulation of ACC synthase induced by ethylene. In flooding-sensitive R. acetosella plants, absence of negative regulation increased internal ethylene levels to more than 20 [mu]L L-1 after 6 d of submergence. This may accelerate the process of senescence and contribute to their low level of flooding resistance.     

Response of Wheat Seedlings to Low O2 Concentrations in Nutrient Solution: I. GROWTH, O2 UPTAKE AND SYNTHESIS OF FERMENTATIVE END-PRODUCTS BY ROOT SEGMENTS

Root growth of 7-d-old wheat (Triticum aestivum cv. Gamenya)seedlings was impaired at dissolved O₂ concentrations of 0.01and 0.055 mol m^–3 O₂, while growth at 0.115 mol m^–3O₂ was the same as that in continuously aerated controls (0.26mol m^–3 O2). Oxygen uptake by apical (0–2 mm), expanding (2–4mm) and expanded (10–12 mm) tissues of the roots decreasedbelow 0.16, 0.09 and 0.05 mol m^–3 O₂, respectively. Thishierarchy is consistent with the metabolic rates of these tissues.There was a small (c. 9%) inhibition of O₂ uptake and some netsynthesis of ethanol and alanine in root apices at 0.115 molm^–3 O₂. Significant amounts of anaerobic end-productsaccumulated at 0.055 mol m^–3 O₂ and even more so at 0.01mol m^–3 O₂, indicating that oxidative phosphorylationwas strongly inhibited. Net alanine synthesis increased in fully expanded (10–16mm) tissues exposed to <0.003–0.01 mol m^–3 O₂,and this increase was accompanied either by a proportionallysmaller increase in the concentration of other free amino acidsor by a net decrease in free amino acid levels excluding alanine.This suggests that alanine was synthesized as an end-productof anaerobic catabolism and did not accumulate simply becauseof decreased net protein synthesis. Comparing the carbon flow to CO₂, ethanol, lactate and alaninein roots at 0.01 mol m^–3 O₂ with carbon loss as CO₂ inaerated roots suggests that carbon flow to products of metabolismwas not greatly enhanced due to O₂ deficiency. This infers,but does not prove that, in wheat, generation of energy duringperiods of O₂ deficiency is not enhanced due to a Pasteur effect. Key words: Anaerobic, fermentation, oxygen, wheat     

Wheat Response to CO2 Enrichment: Growth and CO2 Exchanges at Two Plant Densities

Du Cloux, H. C, André, M., Daguenet, A. and Massinuno,J. 1987. Wheat response to CO₂ enrichment: Growth and CO₂ exchangesat two plant densities.—J. exp. Bot. 38: 1421–1431. The vegetative growth of wheat (Triticum aestivum L., var. Capitole)was followed for almost 40 d after germination in controlledconditions. Four different treatments were carried out by combiningtwo air concentrations of CO₂, either normal (330 mm³ dm ³)or doubled (660 mm³ dm ³) with two plant densities, either 200plants m ² or 40 plants m ². Throughout the experiment the CO₂gas exchanges of each canopy were measured 24 h d¹. These provideda continuous growth curve for each treatment, which were comparedwith dry weights. After a small stimulation at the start (first13 d), no further effect of CO₂ enrichment was observed on relativegrowth rate (RGR). However, RGR was stimulated throughout theexperiment when plotted as a function of biomass. The finalstimulation ol dry weight at 660 mm³ dm ³ CO₂ was a factor of1·45 at high density and 1·50 at low density,contrary to other studies, no diminution of this CO₂ effecton dry weight was observed over time. Nevertheless, at low density,a transient additional enhancement of biomass (up to 1·70)was obtained at a leaf area index (LAI) below 1. This effectwas attributed to a different build up of the gain of carbonin the case of an isolated plant or a closed canopy. In theformer, the stimulation of leaf area and the net assimilationrate are both involved; in the latter the enhancement becomesindependent of the effect on leaf area because the canopy photosynthesisper unit ground area as a function of LAI reaches a plateau. Key words: Triticum aestuum, L. var. Capitole, Vegetative growth, Canopy     

Storage of Shredded Cabbage under a Dynamically Controlled Atmosphere of High O2 and High CO2

Shredded cabbage (Brassica oleracea L., Capitata group) was stored under a dynamically controlled atmosphere (DCA) and modified atmosphere (MA) at 5°C. Quality factors were measured every 2 days. Browning was suppressed as the CO₂ concentration was increased (0% to 15%), with no influence from O₂ concentration (2.5% to 10%). The development of an off-odor was markedly delayed with an increase in O₂ concentration, such an odor being detected after 6 days at 2.5% O₂,8 to 10 days at 5% to 7.5% O₂, and not at all above 10 days at 10% O₂, while the off-odor development was little affected by CO₂ concentration (5% to 15%). Total sugar, polyphenolics, total ascorbate, and total microbial count were little influenced by O₂ and CO₂. These results show that shredded cabbage can be kept in good condition with a combined high O₂ and high CO₂ atmosphere. These findings are largely different from those for MA storage.     

CO(2) Inhibits Respiration in Leaves of Rumex crispus L

Curly dock (Rumex crispus L.) was grown from seed in a glasshouse at an ambient CO₂ partial pressure of about 35 pascals. Apparent respiration rate (CO₂ efflux in the dark) of expanded leaves was then measured at ambient CO₂ partial pressure of 5 to 95 pascals. Calculated intercellular CO₂ partial pressure was proportional to ambient CO₂ partial pressure in these short-term experiments. The CO₂ level strongly affected apparent respiration rate: a doubling of the partial pressure of CO₂ typically inhibited respiration by 25 to 30%, whereas a decrease in CO₂ elicited a corresponding increase in respiration. These responses were readily reversible. A flexible, sensitive regulatory interaction between CO₂ (a byproduct of respiration) and some component(s) of heterotrophic metabolism is indicated.     

High Temperature Sensitivity of Auxin-induced Ethylene Production in Mung Bean Hypocotyl Sections

High temperature sensitivities of IAA-induced and 1-aminocyclopropane-1-carboxylicacid (ACC)-dependent ethylene production in etiolated mung bean(Vigna radiata [L] Wilczek) hypocotyl sections were comparedat 30,40, 42.5°C. When ethylene production at 30°C wastaken as control, IAA-induced production at 40°C was firstenhanced and then suppressed after 3 h, whereas ACC-dependentproduction was enhanced two-fold throughout the 8 h experimentalperiod. However, when hypocotyl sections treated with 1 mM ACCat 30°C for several hours were transferred to 40°C,the ACC-dependent production rate fell below that at 30°C.An initial transient enhancement of IAA-induced ethylene productionat 40°C was supported by increased ACC synthase activityand thus by ACC content. At 42.5°C, both IAA-induced andACC-dependent production were almost completely suppressed.The results indicate that auxin-induced ethylene productionis affected by high temperatures in two different steps: a)at 40°C, the auxin action gradually deteriorates althoughconversion of ACC to ethylene is not affected at all, and at42.5°C, the conversion is nearly completely suppressed. (Received July 8, 1985; Accepted January 24, 1986)     

Two Polypeptides Inducible by Low Levels of CO2 in Soluble Protein Fractions from Chlorella regularis Grown at Low or High pH

Previous studies suggested that certain protein(s) other thancarbonic anhydrase might play an important role in the facilitatedtransport of dissolved inorganic carbon (DIC) from the mediumto the site of CO₂ fixation by ribulose-1,5-bisphosphate carboxylase/oxygenasein the unicellular green alga Chlorella regularis adapted tolow-CO₂ (ordinary air) conditions [Shiraiwa et al. (1991) Jpn.J. Phycol. 39: 355; Satoh and Shiraiwa (1992) Research in Photosynthesis,Vol. III, p. 779]. The proteins that might be involved in thisfacilitated transport of DIC were investigated by pulse-labelingof induced proteins with ³⁵S-sulfate during adaptation of cellsgrown under high-CO₂ conditions to low CO₂. Analysis by SDS-PAGErevealed that synthesis of two polypeptides, with molecularmasses of 98 and 24 kDa, respectively, was induced under low-CO₂conditions. The 24-kDa polypeptide was induced at pH 5.5 butnot at pH 8.0, whereas the 98-kDa polypeptide was induced atboth pH 5.5 and pH 8.0. The possible role of these polypeptidesin the facilitated transport of DIC in Chlorella regularis isdiscussed. (Received October 30, 1995; Accepted February 26, 1996)     

Enhancement of the Stomatal Response to Blue Light by Red Light, Reduced Intercellular Concentrations of CO(2), and Low Vapor Pressure Differences

The effects of environmental parameters on the blue light response of stomata were studied by quantifying transient increases in stomatal conductance in Commelina communis following 15 seconds by 0.100 millimole per square meter per second pulses of blue light. Because conductance increases were not observed following red light pulses of the same or greater (30 seconds by 0.200 millimole per square meter per second) fluences, the responses observed could be reliably attributed to the specific blue light response of the guard cells, rather than to guard cell chlorophyll. In both Paphiopedilum harrisianum, which lacks guard cell chloroplasts, and Commelina, the blue light response was enhanced by 0.263 millimole per square meter per second continuous background red light. Thus, the blue light response and its enhancement do not require energy derived from red-light-driven photophosphorylation by the guard cell chloroplasts. In Commelina, reduction of the intercellular concentration of CO₂ by manipulation of ambient CO₂ concentrations resulted in an enhanced blue light response. In both Commelina and Paphiopedilum, the blue light response was decreased by an increased vapor pressure difference. The magnitude of blue-light-specific stomatal opening thus appears to be sensitive to environmental conditions that affect the carbon and water status of the plant.     

Photosynthesis, Ribulose-1,5-bisphosphate Carboxylase and Leaf Characteristics of Nicotiana tabacum L. Genotypes Selected by Survival at Low CO2 Concentrations

The photosynthetic characteristics (responses to CO₂ and light),ribulose-1,5-bisphosphate carboxylase (Rubisco) properties,and the size and number of cells of the mesophyll of Nicotianatabacum L. leaves of genotypes selected for survival at lowatmospheric CO₂ concentrations are described. When grown inthe greenhouse with nutrient solutions, the total dry matterproduction of the selected genotypes was 23% greater than thatof the parent genotype; this increase was related to a greaternumber of mesophyll cells of smaller size in the selected plantscompared to the parent. However, it was not related to changesin the photosynthetic characteristics nor to Rubisco properties.These results suggest that the increased dry matter accumulationof the selected genotypes is not due to a reduction in photorespirationnor an increase in the CO₂ assimilation rates. Rather, the selectionof haploid tobacco plantlets in low CO₂ has resulted in plantswith greater leaf area (shown in previous work), due to theproduction of more cells of smaller size and to lower respirationrates per unit of leaf dry mass (previous work), thus increasinglight capture, reducing the loss of assimilates and increasingtotal plant dry matter production. Key words: Photosynthesis, ribulose-1,5-bisphosphate carboxylase, leaf anatomy, tobacco, genotypes     

Maximum Quantum Yields of O2 Evolution in C4 Plants under High CO2

The quantum yields of photosynthetic O₂ evolution were measuredin 15 species of C₄ plants belonging to three different decarboxylationtypes (NADP-ME type, NAD-ME type and PEP-CK type) and 5 speciesof C₃ plants and evaluated relative to the maximum theoreticalvalue of 0.125 mol oxygen quanta^-1. At 25°C and 1% CO₂,the quantum yield in C₄ plants averaged 0.079 (differences betweensubgroups not significant) which was significantly lower thanthe quantum yield in C₃ plants (average of 0.105 for 5 species).This lower quantum yield in C₄ plants is thought to reflectthe requirement of energy in the C₄ cycle. For the C₄ NADP-MEtype plant Z. mays and NAD-ME type plant P. miliaceum, quantumyields were also measured over a range of CO₂ levels between1 and 20%. In both species maximum quantum yields were obtainedunder 10% CO₂ (0.105 O₂ quanta_-1 in Z. mays and 0.097 O₂ quanta_-1in P. miliaceum) indicating that at this CO₂ concentration thequantum yields are similar to those obtained in C₃ plants underCO₂ saturation. The high quantum yield values in C₄ plants undervery high CO₂ may be accomplished by direct diffusion of atmosphericCO₂ to bundle sheath cells, its fixation in the C₃ pathway,and feedback inhibition of the C₄ cycle by inorganic carbon. (Received June 6, 1995; Accepted August 15, 1995)