Photoinhibition of respiratory CO2 release in the green alga Scenedesmus

¹⁴CO₂ evolution of prelabeled Scenedesmus obliquus Kütz, has been followed in the dark and in the light. In the light, no carbon dioxide is evolved. Addition of unlabeled NaHCO, leads to ¹⁴CO₂ release attaining 20 to 30% of the dark rate. Double-reciprocal plots of NaHCO₃ concentrations vs ¹⁴CO₂ release results in a straight line, indicative of competition between exogenously supplied bicarbonate and endogenously evolved carbon dioxide. With this method, it is possible to measure CO₂ evolved by respiration in the light and to show that true photoinhibition of respiration occurs in Scenedesmus . In the light. DCMU substantially increases ¹⁴CO₂ evolution; in the presence of the uncoupler carbonyl cyanide- m -chlorophenylhydrazone. ¹⁴CO₂ evolution is comparable to that in the dark. ¹⁴CO₂ release and oxygen uptake in the dark are only slightly affected by cyanide, indicative of a cyanide-resistant respiration and/or fermentation as the essential CO₂-yielding processes in the presence of cyanide. These results, compared with concurrent ATP levels, lead us to assume that energy charge is not the only factor responsible for photoinhibition of respiration.     

CO2 and intracellular pH

Abstract The experimental determination of cytoplasmic and vacuolar pH values is discussed. Despite variation in these values evidence indicates that intracellular pH values are normally regulated within narrow limits. The regulatory mechanisms proposed involve the metabolic consumption of OH^& and the active efflux of H ⁺. The evidence for intracellular pH modification in response to CO₂ hydration and the production of HCO^?₃ and H⁺ is examined. Theoretical calculations and experimental data indicate that CO₂ concentrations as high as 5% will lower intracellular pH. Conversely, variation in CO₂ levels around atmospheric concentrations is unlikely to perturb intracellular pH. High CO₂ levels are found in bulky tissues, and flooded root systems. Evidence is presented that the slow diffusion of dissolved CO₂ compared to gaseous CO₂ results in its accumulation. It is proposed that the accumulation of respiratory CO₂ may reduce intracellular pH values when plant tissues, cells or protoplasts are maintained in a liquid culture medium. Finally, the possible role of dark CO₂ fixation and organic acid synthesis in the regulation of intracellular pH is examined.     

Respiratory responses of higher plants to atmospheric CO2 enrichment

Although the respiratory response of native and agricultural plants to atmospheric CO₂ enrichment has been reported over the past 75 years, only recently have these effects emerged as prominent measures of plant and ecosystem response to the earth's changing climate. In this review we discuss this rapidly expanding field of study and propose that both increasing and decreasing rates of leaf and whole-plant respiration are likely to occur in response to rising CO₂ concentrations. While the stimulatory effects of CO₂ on respiration are consistent with our knowledge of leaf carbohydrate status and plant metabolism, we wish to emphasize the rather surprising short-term inhibition of leaf respiration by elevated CO₂ and the reported effects of long-term CO₂ exposure on growth and maintenance respiration. As is being found in many studies, it is easier to document the respiratory response of higher plants to elevated CO₂ than it is to assign a mechanistic basis for the observed effects. Despite this gap in our understanding of how respiration is affected by CO₂ enrichment, data are sufficient to suggest that changes in leaf and whole-plant respiration may be important considerations in the carbon dynamics of terrestrial ecosystems as global CO₂ continues to rise. Suggestions for future research that would enable these and other effects of CO₂ on respiration to be unravelled are presented.     

Respiration of crop species under CO2 enrichment

Respiratory characteristics of wheat (Triticum aestivum L. cvs Gabo and WW15), mung bean (Vigna radiata L. Wilczek cv. Celera) and sunflower (Helianthus annuus L. cv. Sunfola) were studied in plants grown under a normal CO₂ concentration and in air containing an additional 340 (or 250) μl l^?1 CO₂. Such an increase in global atmospheric CO₂ concentration has been forecast for about the middle of the next century. The aim was to measure the effect of high CO₂ on respiration and its components. Polarographic and, with wheat, CO₂ exchange techniques were used. The capacity of the alternative pathway of respiration in roots was determined polarographically in the presence of 0.1 mM KCN. The actual rate of alternative pathway respiration was assessed by reduction in oxygen consumption caused by 10 mM salicylhydroxamic acid. Each species responded differently. In wheat, growth in high atmospheric CO₂ was associated with up to 45% reduction in respiration by both roots and whole plants. Use of respiratory inhibitors in polarographic measurements on wheat roots implicated reduction in the degree of engagement of the alternative pathway as a major contributor to this reduced respiratory activity of high-CO₂ plants. No change was found in the total sugar content per unit wheat root dry weight as a result of high CO₂. In none of the species was there an increase in the absolute, or relative, contribution by the alternative pathway to total respiration of the root systems. Thus the improved photosynthetic assimilate supply of plants grown in high CO₂ did not lead to increased diversion of carbon through the non-phosphorylating alternative pathway of respiration in the root. On the contrary, in wheat grown in high CO₂ the reduced loss of carbon through that route must have contributed to their larger dry weight.     

Refixation of xylem sap CO2 in Populus deltoides

Vascular plants have respiring tissues which are perfused by the transpiration stream, allowing solubilization of respiratory CO₂ in the xylem sap. The transpiration stream could provide a conduit for the internal delivery of respiratory CO₂ to leaves. Trees have large amounts of respiring tissues in the root systems and stems, and may have elevated levels of CO₂ in the xylem sap which could be delivered to and refixed by the leaves. Xylem sap from the shoots of three Populus deltoides trees had mean dissolved inorganic carbon concentrations (CO₂+H₂CO₃+HCO^?₃) ranging from 0. 5 to 0. 9 mM. When excised leaves were allowed to transpire 1 mM[¹⁴C]NaHCO₃, 99. 6% of the label was fixed in the light. Seventy-seven percent of the label was fixed in major veins and the remainder was fixed in the minor veins. Autoradiography confirmed that label was confined to the vasculature. In the dark, approximately 80% of the transpired label escaped the leaf, the remainder was fixed in the major veins, slightly elevating dark respiration measurements. This indicates that the vascular tissue in P. deltoides leaves is supplied with a carbon source distinct from the atmospheric source fixed by interveinal lamina. However, the contribution of CO₂ delivered to the leaves in the transpiration stream and fixed in the veins was only 0. 5% of atmospheric CO₂ uptake. In the light 90% of the label was found in sugar, starch and protein, a pattern similar to that found for atmospheric uptake of[¹⁴C]CO₂. Compared with leaves labelled in the light, leaves labelled in the dark had more label in organic acid, amino acid and protein and less label in sugar and starch. After a 5-s pulse the majority of the label fed to petioles in both the light and the dark was found in malate. The majority of the label was found in malate at 120 s in the dark; only 2% of the label was found in phosphorylated compounds at 120 s. The proportion of label found in phosphorylated compounds increased from 17% at 5 s to 80% at 120 s in the light. This suggests that CO₂ delivered to leaves in the light via the transpiration stream is fixed in the veins, a small portion through dark fixation into malate, the remainder by C-3 photosynthesis.     

Respiration in a future, higher-CO2 world

Abstract. Apart from its impact on global warming, the annually increasing atmospheric [CO₂] is of interest to plant scientists primarily because of its direct influence on photosynthesis and photorespiration in C₃ species. But in addition, 'dark' respiration, another major component of the carbon budget of higher plants, may be affected by a change in [CO₂] independent of an increase in temperature. Literature pertaining to an impact of [CO₂] on respiration rate is reviewed. With an increase in [CO₂], respiration rate is increased in some cases, but decreased in others. The effects of [CO₂] on respiration rate may be direct or indirect. Mechanisms responsible for various observations are proposed. These proposed mechanisms relate to changes in: (1) levels of nonstructural carbohydrates, (2) growth rate and structural phytomass accumulation, (3) composition of phytomass, (4) direct chemical interactions between CO₂ and respiratory enzymes, (5) direct chemical interactions between CO₂ and other cellular components, (6) dark CO₂ fixation rate, and (7) ethylene biosynthesis rate. Because a range-of (possibly interactive) effects exist, and present knowledge is limited, the impact of future [CO₂] on respiration rate cannot be predicted. Theoretical considerations and types of experiments that can lead to an increase in the understanding of this issue are outlined.     

Soil CO2 efflux in a tropical forest in the central Amazon

This study investigated the spatial and temporal variation in soil carbon dioxide (CO₂) efflux and its relationship with soil temperature, soil moisture and rainfall in a forest near Manaus, Amazonas, Brazil. The mean rate of efflux was 6.45±0.25 SE μmol CO₂ m^?2s^?1 at 25.6±0.22 SE°C (5 cm depth) ranging from 4.35 to 9.76 μmol CO₂ m^?2s^?1; diel changes in efflux were correlated with soil temperature (r²=0.60). However, the efflux response to the diel cycle in temperature was not always a clear exponential function. During period of low soil water content, temperature in deeper layers had a better relationship with CO₂ efflux than with the temperature nearer the soil surface. Soil water content may limit CO₂ production during the drying‐down period that appeared to be an important factor controlling the efflux rate (r²=0.39). On the other hand, during the rewetting period microbial activity may be the main controlling factor, which may quickly induce very high rates of efflux. The CO₂ flux chamber was adapted to mimic the effects of rainfall on soil CO₂ efflux and the results showed that efflux rates reduced 30% immediately after a rainfall event. Measurements of the CO₂ concentration gradient in the soil profile showed a buildup in the concentration of CO₂ after rain on the top soil. This higher CO₂ concentration developed shortly after rainfall when the soil pores in the upper layers were filled with water, which created a barrier for gas exchange between the soil and the atmosphere.     

The response of stomata to CO2 relates to its effect on respiration and ATP level

External ATP enhanced stomatal opening of Commelina communis L. differently from EDTA. ATP was more effective in opening stomata than EDTA, when both were applied in amounts yielding equivalent free Ca²⁺ concentration. The stimulation by ATP depended upon its de-phosphorylation and was not due to the P₁ released. Hence an energetical contribution of external ATP appears possible. Increase in CO₂ concentration increased the stimulation of stomatal opening by ATP and diminished the internal ATP level, ATP/(ADP+AMP) ratio and respiration rate.     

Interactions of tropospheric CO2 and O3 enrichments and moisture variations on microbial biomass and respiration in soil

Soil microbial biomass C (C_mic) is a sensitive indicator of trends in organic matter dynamics in terrestrial ecosystems. This study was conducted to determine the effects of tropospheric CO₂ or O₃ enrichments and moisture variations on total soil organic C (C_org), mineralizable C fraction (C_Min), C_mic, maintenance respiratory (qCO₂) or C_mic death (qD) quotients, and their relationship with basal respiration (BR) rates and field respiration (FR) fluxes in wheat‐soybean agroecosystems. Wheat (Triticum aestivum L.) and soybean (Glycine max. L. Merr) plants were grown to maturity in 3‐m dia open‐top field chambers and exposed to charcoal‐filtered (CF) air at 350 μL CO₂ L^?1; CF air + 150 μL CO₂ L^?1; nonfiltered (NF) air + 35 nL O₃ L^?1; and NF air + 35 nL O₃ L^?1 + 150 μL CO₂ L^?1 at optimum (? 0.05 MPa) and restricted soil moisture (? 1.0 ± 0.05 MPa) regimes. The + 150 μL CO₂ L^?1 additions were 18 h d^?1 and the + 35 nL O₃ L^?1 treatments were 7 h d^?1 from April until late October. While C_org did not vary consistently, C_Min, C_mic and C_mic fractions increased in soils under tropospheric CO₂ enrichment (500 μL CO₂ L^?1) and decreased under high O₃ exposures (55 ± 6 nL O₃ L^?1 for wheat; 60 ± 5 nL O₃ L^?1 for soybean) compared to the CF treatments (25 ± 5 nL O₃ L^?1). The qCO₂ or qD quotients of C_mic were also significantly decreased in soils under high CO₂ but increased under high O₃ exposures compared to the CF control. The BR rates did not vary consistently but they were higher in well‐watered soils. The FR fluxes were lower under high O₃ exposures compared to soils under the CF control. An increase in C_mic or C_mic fractions and decrease in qCO₂ or qD observed under high CO₂ treatment suggest that these soils were acting as C sinks whereas, reductions in C_mic or C_mic fractions and increase in qCO₂ or qD in soils under elevated tropospheric O₃ exposures suggest the soils were serving as a source of CO₂.     

Sensing of atmospheric CO2 by plants

Abstract. Despite recent interest in the effects of high CO₂ on plant growth and physiology, very little is known about the mechanisms by which plants sense changes in the concentration of this gas. Because atmospheric CO₂ concentration is relatively constant and because the conductance of the cuticle to CO₂ is low, sensory mechanisms are likely to exist only for intercellular CO₂ concentration. Therefore, responses of plants to changes in atmospheric CO₂ will depend on the effect of these changes on intercellular CO₂ concentration. Although a variety of plant responses to atmospheric CO₂ concentration have been reported, most of these can be attributed to the effects of intercellular CO₂ on photosynthesis or stomatal conductance. Short-term and long-term effects of CO₂ on photosynthesis and stomatal conductance are discussed as sensory mechanisms for responses of plants to atmospheric CO₂. Available data suggest that plants do not fully realize the potential increases in productivity associated with increased atmospheric CO₂. This may be because of genetic and environmental limitations to productivity or because plant responses to CO₂ have evolved to cope with variations in intercellular CO₂ caused by factors other than changes in atmospheric CO₂.     

Wood CO2 efflux in a primary tropical rain forest

The balance between photosynthesis and plant respiration in tropical forests may substantially affect the global carbon cycle. Woody tissue CO₂ efflux is a major component of total plant respiration, but estimates of ecosystem‐scale rates are uncertain because of poor sampling in the upper canopy and across landscapes. To overcome these problems, we used a portable scaffolding tower to measure woody tissue CO₂ efflux from ground level to the canopy top across a range of sites of varying slope and soil phosphorus content in a primary tropical rain forest in Costa Rica. The objectives of this study were to: (1) determine whether to use surface area, volume, or biomass for modeling and extrapolating wood CO₂ efflux, (2) determine if wood CO₂ efflux varied seasonally, (3) identify if wood CO₂ efflux varied by functional group, height in canopy, soil fertility, or slope, and (4) extrapolate wood CO₂ efflux to the forest. CO₂ efflux from small diameter woody tissue (<10 cm) was related to surface area, while CO₂ efflux from stems >10 cm was related to both surface area and volume. Wood CO₂ efflux showed no evidence of seasonality over 2 years. CO₂ efflux per unit wood surface area at 25° (F_A) was highest for the N‐fixing dominant tree species Pentaclethra macroloba, followed by other tree species, lianas, then palms. Small diameter F_A increased steeply with increasing height, and large diameter F_A increased with diameter. Soil phosphorus and slope had slight, but complex effects on F_A. Wood CO₂ efflux per unit ground area was 1.34±0.36 μmol m^?2 s^?1, or 508±135 g C m^?2 yr^?1. Small diameter wood, only 15% of total woody biomass, accounted for 70% of total woody tissue CO₂ efflux from the forest; while lianas, only 3% of total woody biomass, contributed one‐fourth of the total wood CO₂ efflux.     

The effect of growth at elevated CO2 concentrations on photosynthesis in wheat

Rising levels of atmospheric CO₂ will have profound, direct effects on plant carbon metabolism. In this study we used gas exchange measurements, models describing the instantaneous response of leaf net CO₂ assimilation rate (A) to intercellular CO₂ partial pressure (C_i), in vitro enzyme activity assay, and carbohydrate assay in order to investigate the photosynthetic responses of wheat (Triticum aestivum L., cv. Wembley) to growth under elevated partial pressures of atmospheric CO₂ (C_a). At flag leaf ligule emergence, the modelled, in vivo, maximum carboxylation velocity for RuBisCO was significantly lower in plants grown at elevated C_a than in plants grown at ambient C_a (70 Pa compared with 40 Pa). By 12 d after ligule emergence, no significant difference in this parameter was detectable. At ligule emergence, plants grown at elevated C_a exhibited reduced in vitro initial activities and activation states of RuBisCO. At their respective growth C_i values, the photosynthesis of 40-Pa-grown plants was sensitive to p(O₂) and to p(CO₂) whereas that of 70-Pa-grown plants was insensitive. Both sucrose and starch accumulated more rapidly in the leaves of plants grown at 70 Pa. At flag leaf ligule emergence, modelled non-photorespiratory respiration in the light (R_d) was significantly higher in 70-Pa-grown plants than in 40-Pa-grown plants. By 12 d after ligule emergence no significant differences in R_d were detectable.     

Ecosystem CO2 exchange and plant biomass in the littoral zone of a boreal eutrophic lake

• 1 In order to study the dynamics of primary production and decomposition in the lake littoral, an interface zone between the pelagial, the catchment and the atmosphere, we measured ecosystem/atmosphere carbon dioxide (CO₂) exchange in the littoral zone of an eutrophic boreal lake in Finland during two open water periods (1998–1999). We reconstructed the seasonal net CO₂ exchange and identified the key factors controlling CO₂ dynamics. The seasonal net ecosystem exchange (NEE) was related to the amount of carbon accumulated in plant biomass.
• 2 In the continuously inundated zones, spatial and temporal variation in the density of aerial shoots controlled CO₂ fluxes, but seasonal net exchange was in most cases close to zero. The lower flooded zone had a net CO₂ uptake of 1.8–6.2 mol m^?2 per open water period, but the upper flooded zone with the highest photosynthetic capacity and above‐ground plant biomass, had a net CO₂ loss of 1.1–7.1 mol m^?2 per open water period as a result of the high respiration rate. The excess of respiration can be explained by decomposition of organic matter produced on site in previous years or leached from the catchment.
• 3 Our results from the two study years suggest that changes in phenology and water level were the prime cause of the large interannual difference in NEE in the littoral zone. Thus, the littoral is a dynamic buffer and source for the load of allochthonous and autochthonous carbon to small lakes.

     

Effects of an induced drought on soil carbon dioxide (CO2) efflux and soil CO2 production in an Eastern Amazonian rainforest, Brazil

In the next few decades, climate of the Amazon basin is expected to change, as a result of deforestation and rising temperatures, which may lead to feedback mechanisms in carbon (C) cycling that are presently unknown. Here, we report how a throughfall exclusion (TFE) experiment affected soil carbon dioxide (CO₂) production in a deeply weathered sandy Oxisol of Caxiuanã (Eastern Amazon). Over the course of 2 years, we measured soil CO₂ efflux and soil CO₂ concentrations, soil temperature and moisture in pits down to 3 m depth. Over a period of 2 years, TFE reduced on average soil CO₂ efflux from 4.3±0.1 μmol CO₂ m⁻² s⁻¹ (control) to 3.2±0.1 μmol CO₂ m⁻² s⁻¹ (TFE). The contribution of the subsoil (below 0.5 m depth) to the total soil CO₂ production was higher in the TFE plot (28%) compared with the control plot (17%), and it did not differ between years. We distinguished three phases of drying after the TFE was started. The first phase was characterized by a translocation of water uptake (and accompanying root activity) to deeper layers and not enough water stress to affect microbial activity and/or total root respiration. During the second phase a reduction in total soil CO₂ efflux in the TFE plot was related to a reduction of soil and litter decomposers activity. The third phase of drying, characterized by a continuing decrease in soil CO₂ production was dominated by a water stress‐induced decrease in total root respiration. Our results contrast to results of a drought experiment on clay Oxisols, which may be related to differences in soil water retention characteristics and depth of rooting zone. These results show that large differences exist in drought sensitivity among Amazonian forest ecosystems, which primarily seem to be affected by the combined effects of texture (affecting water holding capacity) and depth of rooting zone.     

A search for predictive understanding of plant responses to elevated [CO2]

This paper reviews two decades of effort by the scientific community in a search for predictive understanding of plant responses to elevated [CO₂]. To evaluate the progress of research in leaf photosynthesis, plant respiration, root nutrient uptake, and carbon partitioning, we divided scientific activities into four phases: (I) initial assessments derived from our existing knowledge base to provide frameworks for experimental studies; (II) experimental tests of the initial assessments; (III) in cases where assessments were invalidated, synthesis of experimental results to stimulate alternative hypotheses and further experimentation; and (IV) formation of new knowledge. This paper suggests that photosynthetic research may have gone through all four phases, considering that (a) variable responses of photosynthesis to [CO₂] are generally explainable, (b) extrapolation of leaf-level studies to the global scale has been examined, and (c) molecular studies are under way. Investigation of plant respiratory responses to [CO₂] has reached the third phase: experimental results have been accumulated, and mechanistic approaches are being developed to examine alternative hypotheses in search for new concepts and/or new quantitative frameworks to understand respiratory responses to elevated [CO₂]. The study of nutrient uptake kinetics is still in the second phase: experimental evidence has contradicted some of the initial assessments, and more experimental studies need to be designed before generalizations can be made. It is quite unfortunate that we have not made much progress in understanding mechanisms of carbon partitioning during the past two decades. This is due in part to the fact that some of the holistic theories, such as functional balance and optimality, have not evolved into testable hypotheses to guide experimental studies. This paper urges modelers to play an increasing role in plant–CO₂ research by disassembling these existing theories into hypotheses and urges experimentalists to design experiments to examine these holistic concepts.     

Leaf gas exchange and nitrogen dynamics of N2-fixing, field-grown Alnus glutinosa under elevated atmospheric CO2

Few studies have investigated the effects of elevated CO₂ on the physiology of symbiotic N₂-fixing trees. Tree species grown in low N soils at elevated CO₂ generally show a decline in photosynthetic capacity over time relative to ambient CO₂ controls. This negative adjustment may be due to a reallocation of leaf N away from the photosynthetic apparatus, allowing for more efficient use of limiting N. We investigated the effect of twice ambient CO₂ on net CO₂ assimilation (A), photosynthetic capacity, leaf dark respiration, and leaf N content of N2-fixing Alnus glutinosa (black alder) grown in field open top chambers in a low N soil for 160 d. At growth CO₂, A was always greater in elevated compared to ambient CO₂ plants. Late season A vs. internal leaf p(CO₂) response curves indicated no negative adjustment of photosynthesis in elevated CO₂ plants. Rather, elevated CO₂ plants had 16% greater maximum rate of CO₂ fixation by Rubisco. Leaf dark respiration was greater at elevated CO₂ on an area basis, but unaffected by CO₂ on a mass or N basis. In elevated CO₂ plants, leaf N content (μg N cm^?2) increased 50% between Julian Date 208 and 264. Leaf N content showed little seasonal change in ambient CO₂ plants. A single point acetylene reduction assay of detached, nodulated root segments indicated a 46% increase in specific nitrogenase activity in elevated compared to ambient CO₂ plants. Our results suggest that N₂-fixing trees will be able to maintain high A with minimal negative adjustment of photosynthetic capacity following prolonged exposure to elevated CO₂ on N-poor soils.