June 24, 2008
Photosynthetic Utilization of Bicarbonate in Zostera Marina Is Reduced By Inhibitors of Mitochondrial ATPase and Electron Transport1
By Carr, Herman Axelsson, Lennart
When Zostera marina was irradiated after a period of darkness, initiation of photosynthetic O2 evolution occurred in two phases. During a lag phase, lasting 4 to 5 min, photosynthesis was supported by a diffusive entry of CO2. Photosynthesis then rapidly increased to its full rate. Tris buffer, at a concentration of 50 mM, completely inhibited this increase without affecting CO2-supported photosynthesis during the lag phase. These results verify that the increase in photosynthesis after the lag phase depended on an activation of bicarbonate (HCO^sub 3^^sup -^) utilization through acid zones generated by proton pumps located to the outer cell membrane. In similar experiments, 6.25 [mu]M of the mitochondrial ATPase blocker oligomycin inhibited photosynthetic HCO^sub 3^^sup - ^ utilization by more than 60%. Antimycin A, a selective blocker of mitochondrial electron transport, caused a similar inhibition of HCO^sub 3^^sup -^ utilization. Measurements at elevated CO2 concentrations verified that neither oligomycin nor antimycin interfered with linear photosynthetic electron transport or with CO2 fixation. Thus, a major part of the ATP used for the generation of acid zones involved in HCO^sub 3^^sup -^ utilization in Z. marina was derived from mitochondrial respiration. The prerequisites for photosynthetic carbon uptake differ considerably between marine and terrestrial habitats. Unlike the atmosphere, the oceans contain ionic carbon species in addition to CO2. At the normal pH of seawater (8.0-8.2), the dominant inorganic carbon species is bicarbonate (HCO^sub 3^^sup -^), while CO2 is present at very low concentrations. The availability of CO2 is therefore a potential limiting factor for marine photosynthesizers (Raven et al., 1990). However, the majority of the aquatic plants can make use of HCO^sub 3^^sup -^ as a source of CO2 (Raven, 1997; Axelsson and Beer, 2001). The utilization of HCO^sub 3^^sup -^ can occur via either nonenergy- requiring processes or a CO2-concentrating mechanism, comprising at least one energy-consuming step (Badger et al., 1994; Giordano et al., 2005). The inorganic carbon can enter both through direct uptake of HCO^sub 3^^sup -^ and after a transformation of HCO^sub 3^^sup -^ to CO2 outside the plasma membrane; CO2 then diffuses or is actively transported into the cell (Su? ltemeyer, 1997). The HCO^sub 3^^sup -^ that is transported into the cell can be transformed to CO2 in the proximity of Rubisco (Raven, 2003).
Land plants, including angiosperms, have evolved from algae. The closest living aquatic relatives to terrestrial plants are the Charophytes (Karol et al., 2001), which are assumed to have a CO2- concentrating mechanism located to their outer cell membranes. Thus, the Charophyte Chara corallina has pericellular acid zones where HCO^sub 3^^sup -^ is transformed to CO2 for use in photosynthesis (Price and Badger, 1985). These acid zones are accompanied by alkaline zones where calcification takes place, creating protective crusts (McConnaughey, 1991). Internodal cells of C. corallina seem to be dependent on plasma membrane H1-ATPase both for the dehydration of HCO^sub 3^^sup -^ to CO2 in acid zones and for direct uptake of HCO^sub 3^^sup -^, especially at high pH values (Mimura et al., 1993). Chara tomentosa is likely to have ATP-driven proton pumps, which are involved in the photosynthetic utilization of HCO^sub 3^^sup -^ (Ray et al., 2003). Although at least some species of unicellular eustigmatophyte algae appear to need mitochondrial ATP for their HCO^sub 3^^sup -^ utilization (Huertas et al., 2002a, 2002b), photosynthesis is generally assumed to be the ATP source for active HCO^sub 3^^sup -^ utilization in algae (Kaplan and Reinhold, 1999). However, the ATP source for maintenance of the acid zones in C. corallina is not known. Although land plants have evolved in a completely different environment, they have maintained the ability to create acid zones by the release of protons, which aids in their nutrient uptake (Raven, 2000; Shen et al., 2004). As nutrient uptake occurs in nonphotosynthetic parts, these acid zones are obviously supported by mitochondrial ATP.
Seagrasses, like Zostera marina, descend from flowering land plants that have adapted to submerged conditions. These plants have maintained many characteristics of land plants, such as roots and rhizosomes, and a gas phase in the lacunae (Hemminga and Duarte, 2000). Z. marina and Zostera noltii are two marine angiosperms that depend on active HCO^sub 3^^sup -^ utilization for expressing full photosynthetic rates. This utilization is assumed to occur via an excretion of protons through the outer cell membrane, thus creating acid regions, or compartments, where the conversion of HCO^sub 3^^sup -^ to CO2 is favored (Hellblom et al., 2001; Mercado et al., 2003). The CO2 formed there may then be taken up actively or, more likely, diffuse through the plasma membrane. A similar mechanism occurs in many aquatic angiosperms with leaves featuring pH polarity, for example, Potamogeton lucens (Prins et al., 1982; van Ginkel and Prins, 1997). The brackish angiosperm Ruppia cirrhosa has been reported to have substantial photosynthetic rates at pH values as high as 9.5, where almost all carbon is present as HCO^sub 3^^sup -^ and carbonate. Also in this plant, the mechanism for HCO^sub 3^^sup -^ utilization depends on acid compartments, although external carbonic anhydrase activity is involved in the HCO^sub 3^^sup -^ dehydration (Hellblom and Axelsson, 2003). The same mechanism has been suggested for the brown macroalga Laminaria saccharina (Axelsson et al., 2000; Mercado et al., 2006).
Z. marina has characteristics of both terrestrial plants (from which they evolved) and marine algae (with which they share the marine environment). We therefore thought it relevant to examine the role of mitochondrial activity for maintaining photosynthetic rates in Z. marina and in particular in relation to their utilization of HCO^sub 3^^sup -^.
Leaves of Z. marina, dark acclimatized for more than 1 h, were irradiated in natural seawater (NSW; dissolved inorganic carbon, approximately 2.0 mM; pH, approximately 8.4). After an initial period of 4 to 5 min of constant low rate photosynthesis (lag phase), there was a rapid increase to full photosynthetic rate (Fig. 1A, NSW). The duration of this lag phase differed slightly between plants collected at different occasions. In one experiment, the lag phase lasted 4.4 +- 0.23 min, while the half-time of the increase to full photosynthesis was only 1.62 +- 0.48 min (mean 6 SD, n 5 6; compare Fig. 1A, NSW). When the leaves were irradiated in CO2- sufficient medium (NSW buffered to pH 6.15), there was no lag at the onset of photosynthesis but only an induction period with a half- time of the increase to full photosynthetic rates of 2.63 +- 0.15 min (mean +- SD, n 5 6; compare Fig. 1A, MES).
In NSW adjusted to different pH values, the photosynthetic rate during the lag phase was highest at the lowest pH (Fig. 1B). This rate was proportional to the calculated CO2 partial pressure of the medium, suggesting that photosynthesis during the lag phase depended on a diffusive uptake of CO2 (Fig. 2). The maximal rates of photosynthesis varied little with pH and were more or less proportional to the HCO^sub 3^^sup -^ concentration of the medium, except for a slightly higher rate at the lowest pH (6.9). These results indicate that photosynthesis at higher pH was limited by the CO2 supply from the HCO^sub 3^^sup -^ utilization mechanism and that the higher CO2 concentration at pH 6.9 was sufficient to abolish this limitation. Despite large differences in photosynthetic rates, the duration of the lag phase was almost the same (no significant differences, compare Fig. 1B). In this experiment, the lag phase lasted between 6.4 6 0.7 min (pH 7.5) and 5.6 6 0.4 min (pH 8.9; mean 6SD, n5 5). The presence of Tris buffer reduced photosynthesis to the same level as during the lag phase in nonbuffered seawater of the same pH (Figs. 1A and 3); thus, the difference in rates between lag phase and fully activated photosynthesis disappeared. These results strongly support the earlier conclusion that Tris buffer really acts as an efficient inhibitor of HCO^sub 3^^sup -^ utilization in Z. marina (Hellblom et al., 2001).
Additions of oligomycin (an inhibitor of mitochondrial ATP formation) during the dark acclimation period caused a more than 60% (61.3% +- 10.5%, mean +- SD, n 5 6) inhibition of the maximal photosynthetic rate during the following irradiation phase, with no significant effect on the lag phase (Figs. 1Aand 4A). Similar results were obtained with antimycin (an inhibitor ofmitochondrial electron transport; Vianello et al., 1997; Table II; Fig. 4B). The small decrease in the lagphasephotosynthetic rate after the additionof the inhibitors appears to be related to a stimulation of the respiration caused by the solvent (ethanol). Such an increase in respiration was 2.32 +- 0.31 [mu]mol O2 g^sup -1^ h^sup -1^ (per fresh weight average +- SD, n 5 3, compare Fig. 1A). Both inhibitors are lipophilic and comparatively large molecules (Mr 548-775); thus, the inhibition required a long period of time to be established. At a concentration of 6.25 [mu]M, no effect by oligomycin could be seen 20 min after the addition, although full effect was obtained 4 h later. Neither prolonged treatment nor increased oligomycin concentrations caused any further inhibition (Table I), suggesting that complete inhibition of ATP generation via oxidative phosphorylation caused only a partial inhibition of the HCO^sub 3^^sup -^ utilization. For oligomycin, at least 1 h of treatment was required, while the full effect of antimycin was obtained after about 45 min. Buffering the seawater to pH 6.15 (MES buffer, 25 mM), which causes a considerable increase in the CO2 concentration (from 1,000 [mu]M), restored full photosynthesis both in the oligomycin (Fig. 4A; Table I) and the antimycin-treated (Fig. 4B) leaves. These results verify that both inhibitors inhibited a process necessary for the utilization of the HCO^sub 3^^sup -^ pool from the seawater and that both CO2 fixation and photosynthetic electron transport could operate in the presence of these inhibitors. The effect of the inhibitors (obviously still present in the plant tissue) was again visible if the buffered medium was replaced by NSW (which included repeated rinsing of the electrode chamber and sample with NSW; exemplified for oligomycin in Table I). Neither antimycin nor oligomycin caused any significant inhibition of respiratory rates in darkness (measured as oxygen consumption). To investigate if a block in the mitochondrial electron transport would cause a switch from normal to alternative respiration, the combined effect of antimycin and salicylhydroxamic acid (SHAM; an inhibitor of alternative respiration; Mikulska et al., 1998; Serikawa et al., 2000) was tested. While SHAM alone caused only a small inhibition (11% +- 9%; Table II), the presence of antimycin resulted in a pronounced increase in the inhibition (from 11% to more than 45%; Table II). The inhibition by SHAM was similar at the different concentrations of antimycin. The strong synergetic effect of SHAM and antimycin verifies that antimycin actually inhibits respiration and that the capacity for alternative respiration is substantial in Z. marina.
Upon irradiation of Z. marina acclimatized to darkness, initiation of photosynthetic O2 evolution occurred as two phases in our experiments. During a lag phase, there was an almost constant rate of photosynthesis, and this rate was proportional to the actual CO2 concentration of the medium. Photosynthesis during the lag phase must consequently be assumed to depend on CO2 uptake. The lag phase was followed by a rapid increase in the photosynthetic rate until steady state was reached. This increase was completely inhibited by Tris buffer. Further, Tris buffer did not lower the rate of photosynthesis of the plants during the lag phase, and full photosynthesis rate in the presence of Tris buffer was similar to the photosynthesis rate of the control during the lag phase. These results support the assumption that Tris buffer is an efficient inhibitor of HCO^sub 3^^sup -^ utilization in Z. marina (Hellblom et al., 2001) and also that the increase to full photosynthesis after the lag phase reflects the onset of HCO^sub 3^^sup -^ utilization. A similar situation, with an initial phase of CO2 uptake followed by an onset of HCO^sub 3^^sup -^ utilization, occurs in many aquatic angiosperms featuring different pH values on the upper and lower sides of their leaves (Prins et al., 1980, 1982). The described properties of the photosynthetic induction by Z. marina made it possible to check if an inhibitor of photosynthesis acted specifically on the mechanism for utilization of HCO^sub 3^^sup -^. The general mechanism behind the inhibition of HCO^sub 3^^sup -^ utilization by buffers like Tris has been described before (Price and Badger, 1985; Hellblom et al., 2001; Mercado et al., 2006). The buffer anion competes with HCO^sub 3^^sup -^ for the protons excreted through the cell membrane, thereby preventing the formation of acid zones with high local concentrations of CO2. The fast inhibition obtained by Tris addition and the fact that inhibition disappears immediately when changing to Tris-free medium suggest that the inhibition occurs extracellularly.
Mitochondrial ATP Supports HCO^sub 3^^sup -^ Utilization
At low concentrations, antimycin and oligomycin can be used to selectively inhibit mitochondrial ATP production, targeting different processes; the electron transport chain and the ATPase, respectively. These inhibitors caused a pronounced and similar inhibition of the HCO^sub 3^^sup -^ utilization without any effect on the CO2-supported photosynthesis exhibited in the first phase following irradiation. Because a blockage in ATP synthesis of the chloroplast would also stop fixation of CO2 in the Calvin cycle, the high photosynthetic rates observed at high CO2 concentrations (NSW buffered to pH 6.16) in the presence of oligomycin and antimycin verifies that these inhibitors did not interfere with ATP synthesis in the chloroplast (with a possible exception for antimycin at the highest concentration). As the inhibition of HCO^sub 3^^sup -^ utilization was identical over a range of concentrations (0.1- 1.0 [mu]M for antimycin and 6-19 [mu]M for oligomycin), maximal inhibition of the mitochondrial ATP production must have been obtained. Thus, our results show that ATP supply from mitochondria is important for the photosynthetic utilization of HCO^sub 3^^sup - ^ in Z. marina.
The selective action of the inhibitors suggested by the above results is in line with the literature. Although there have been reports on effects of oligomycin on ATPase in, for example, isolated chloroplast envelopes from spinach (Spinacia oleracea; Wu and Berkowitz, 1992) and in pea (Pisum sativum) protoplasts at high concentrations (Romanowska et al., 2005), oligomycin at sufficiently low concentrations is a well-known selective inhibitor of mitochondrial ATPase with no effect on chloroplast ATP levels (Kromer and Heldt, 1991; Igamberdiev et al., 1998). While oligomycin binds strongly to mitochondria, ATPases in chloroplasts are quite insensitive to this compound (Kro?mer and Heldt, 1991). Thus, oligomycin, at the concentration used on whole leaves in our experiments, should be regarded as a selective inhibitor of oxidative phosphorylation. For antimycin, there was a slightly higher inhibition of the HCO^sub 3^^sup -^ utilization at 10 [mu]M (as compared to the lower concentrations). This high concentration of antimycin probably also caused a limited inhibition of cyclic electron transport around PSI (Joet et al., 2001; Li et al., 2006). The inhibition of respiration by SHAM in the presence of antimycin was similar for all concentrations of this substance. Higher concentrations of antimycin thus did not have any further effect on respiration. Such findings are additional support for the suggestions that the higher inhibition of the HCO^sub 3^^sup -^ utilization at the highest antimycin concentration was partly caused by inhibition of cyclic electron transport.
ATP produced in the mitochondria is likely to play a major role in providing energy for active utilization of HCO^sub 3^^sup -^ by driving proton pumps at the plasma membrane. Although NADPH is produced in the chloroplast, chloroplasts have been reported to communicate with, and transport energy to, mitochondria in the light. Malate has, for example, been suggested as an energy carrier between chloroplasts and mitochondria in terrestrial angiosperms (Raghavendra et al., 1994). This would be in agreement with results on leaves of the freshwater angiosperm Elodea, where hyperpolarization decreased at high inorganic carbon concentrations and at a lowered NADPH to NADP1 ratio (Elzenga et al., 1989). Inorganic carbon uptake appears to also depend on respiration for some species of eustigmatophyte microalgae (Huertas et al., 2002a). This dependency occurs regardless of the inorganic carbon species utilized, which is an obvious difference from the situation for Z. marina, in which only HCO^sub 3^^sup -^ utilization depends on mitochondrial respiration.
Studies on pea protoplasts and leaves show that mitochondria are involved both in the activation of the Calvin cycle (Padmasree and Raghavendra, 1999a, 2001a, 2001b) and the activation of key chloroplast enzymes (Padmasree and Raghavendra, 2001a). There is, thus, a clear difference between terrestrial and aquatic plants regarding the relation between photosynthesis and mitochondrial ATP production. This is well illustrated by the effect of CO2 concentration on the inhibition of photosynthesis by oligomycin. In terrestrial angiosperms, the inhibition of photosynthetic rates was maximal when there was abundant CO2 (Padmasree and Raghavendra, 1999b), while in Z. marina the degree of inhibition instead decreased with increasing CO2 concentrations. The inhibition was also much more pronounced in Z. marina. The supply of ATP for HCO^sub 3^^sup -^ utilization is thus an additional function of mitochondria in angiosperms that is exclusive for aquatic plants and differs from some of the mitochondrial functions reported in terrestrial angiosperms.
Importance of Respiration
The strong synergetic effect between SHAM and antimycin shows that alternative respiration becomes highly active upon antimycin addition. The alternative respiratory pathway bypasses energy- conserving sites, resulting in very low ATP production (Siedow and Umbach, 1995),which explains the inhibition of HCO^sub 3^^sup -^ utilization with only antimycin present. Inhibition of respiratory ATP production by oligomycin is less likely to result in any decrease in the respiratory oxygen consumption. Mitochondria in both plants and animals can excrete protons through proton gates instead of passing them through ATPase (Ricquier and Bouillaud, 2000), including in the monocotyledon maize (Zea mays; Faavaro et al., 2006). Such bypasses by uncoupling proteins is believed to be important for the regulation of mitochondrial redox status and energy level (Ricquier and Bouillaud, 2000; Brandalise et al., 2003). Unfortunately, it is not possible to use SHAM to study the importance of alternative respiration for photosynthetic HCO^sub 3^^sup -^ utilization. As SHAM has a pKa close to 8 and because it is applied at comparatively high concentrations, this buffer interferes strongly with the acid zones involved in the HCO^sub 3^^sup -^ utilizationmechanism of buffersensitive plants like Z. marina, resulting in a severe inhibition of photosynthesis (compare Price and Badger, 1985; Mercado et al., 2006). The Lag Phase Depends on a Lag in HCO^sub 3^^sup -^ Utilization
In many terrestrial plants, a lag phase in the onset of photosynthesis is known to occur when darkacclimatized samples are exposed to light. Such lag phases have been ascribed to, for example, stomatal opening and, thus, the availability of CO2 (Sacher and Burian, 1994). The lag phase in some aquatic angiosperms has been related to a lag in the onset of HCO^sub 3^^sup -^ utilization (Prins et al., 1980, 1982). A coincidence between the onset of HCO^sub 3^^sup -^ utilization and the increase to full photosynthesis does not necessarily imply that the lag phase is a result of a lag in the HCO^sub 3^^sup -^ utilization. An alternative explanation could be that the lag depends on a shortage of CO2 that is needed for the activation of Rubisco in the Calvin cycle (Kursar and Coley, 1993; Jensen, 2004) or for light activation of enzymes required for restoration of photosynthetic intermediates after a dark period (Kirschbaum et al., 2005). If so, one should expect a shorter lag phase at higher CO2 concentrations as well as photosynthetic rates during the lag phase that did not depend on the CO2 concentration. However, the photosynthetic rates during the lag phase varied considerably as a function of the actual CO2 concentrations, while the length of the lag phase remained constant (compare Fig. 1B). Thus, our data suggest that the lag in the onset of photosynthesis in NSWby Z. marina depends on a lag before the onset of the activation of HCO^sub 3^^sup -^ utilization, and there could only be a minor contribution, if any at all, from a lag in the activation of enzymes or in the formation of intermediates in the Calvin cycle. At high pH values and, consequently, low CO2 concentrations in the medium, the lag phase was slightly S-shaped (Fig. 1B), implying that the Calvin cycle actually operates immediately, relying on CO2 from the respiration accumulated within the plant tissue. The CO2 fixation (and the O2 production) decrease as this internal pool of respiratory released CO2 is used up and the CO2 fixation thus becomes dependent on diffusive entry of CO2.
MATERIALS AND METHODS
Zostera marina, a marine monocotyledon, was collected at 1.0 to 2.0mdepth outside Kristineberg Marine Research Station on the Swedish west coast during May and September. The plants were allowed to acclimatize in aquaria with a flow of temperature-controlled NSW (16[degrees]C; saltwater from 35 m depth, salinity approximately 30) and at an irradiance of approximately 150 [mu]mol m^sup -2^ s^sup - 1^ (16-h photoperiod). Disregarding the youngest and oldest leaves, 20- or 25-mm-long, epiphyte-free sections were cut from the middle of leaves of similar width (5 mm) for the experiments.
Oxygen Electrode Setup
Photosynthetic O2 evolution was measured in six 3.0-mL temperaturecontrolled (15[degrees]C-17[degrees]C) O2 electrode chambers (Larsson et al., 1997). The leaf segments were inserted in a semicircle in the O2 electrode chambers and directed like a parabola facing the light source supplied through optical fibers. The approximately 300 photons [mu]mol m^sup -2^ s^sup -1^ irradiated on all samples were light saturating. Oxygen electrode tracings were recorded on a strip chart recorder (Seconic flat bed recorder).
Medium and Inhibitors
All measurements were carried out in NSWor in artificial seawater (ASW; salinity 30; Hellblom and Axelsson, 2003) with inorganic carbon added. The NSW, with a salinity of 30 to 32, was taken from 35 m depth by means of the deep saltwater system of the Research Station. A stock solution (1 g L^sup -1^) of a mixture of oligomycin A, B, and C (Sigma chemicals), an inhibitor of mitochondrial ATPase, was prepared in 60% ethanol and was supplied to the O2 chambers at a final concentration of 5 mg L^sup -1^ (approximately 6.25 mM). Antimycin A (Sigma Chemicals) was dissolved in 96% ethanol to give three stock solutions at the concentrations of 3.3, 0.33, and 0.033 mg/mL, which were injected into the O2 chamber to generate final concentrations of approximately 1, 0.1, and 0.01 [mu]M, respectively. SHAM (Sigma Chemicals) was dissolved in 1.0 M NaOH to give a stock solution of 1.0 M and was added to a final concentration of 15 mM. As SHAM becomes oxidized in air at room temperature, the stock solutions were stored frozen at 220[degrees]C and were allowed to equilibrate at room temperature only prior to use. Tris buffer was added from a 2.0 M Tris-HCl stock solution prepared to generate the required pH when added to the medium (final Tris concentration, 50 mM). MES buffer was prepared as a stock solution of 1.0 M and supplied to give a final concentration of 25 mM and a pH of 6.15. This decrease in pH increases the available CO2 of NSW more than 100 times. Seawater media of different pH values were prepared by adding HCl, HCO^sub 3^^sup -^, and HCO^sub 3^^sup - 2^ to ASW in different proportions, as described by Hellblom and Axelsson (2003). The pH electrode (a glass electrode with Ag/AgCl reference) was calibrated on the National Bureau of Standards scale, and the concentration of different inorganic carbon species of the seawater at different pH values was calculated according to Millero (1979), using the dissociation constants from Mehrbach et al. (1973).
Measurements of the inhibitory effects on HCO^sub 3^^sup -^ utilization were carried out in the O2 electrode chambers using the desired medium as follows. The Z. marina samples were exposed to irradiation until steady-state O2 evolution was obtained. The light was then turned off, and the samples were kept in darkness until the oxygen had decreased to the same level as before irradiation (usually 2 h). The plants were then exposed to a new light/dark cycle. During the dark period of this cycle, three of the six samples were supplied with an inhibitor of mitochondrial ATP formation (oligomycin or antimycin). During the next light/dark cycle, the three remaining samples were supplied with the inhibitors. This approach made it possible to keep track of (and even correct for) any changes in the O2 traces not caused by the inhibitors. Net photosynthetic rates were calculated directly from the O2 tracings. HCO^sub 3^^sup -^ utilization was calculated by subtracting the photosynthetic rate during the lag phase from the full photosynthetic rate at saturating irradiance. SHAM is a buffer and inhibits HCO^sub 3^^sup -^ utilization in light and, consequently, affects photosynthesis in a manner similar to Tris buffer. Therefore, SHAM was added in darkness after a light period to avoid lower respiratory rates as a consequence of low photosynthetic rates. Respiration was calculated directly from the O2 tracings.
Received January 10, 2008; accepted April 15, 2008; published April 23, 2008.
1 This work was supported by the Stockholm Marine Research Centre (travel grant to H.C.).
Axelsson L, Beer S (2001) Algal adaptation to environmental stresses. In LC Rai, JP Gaur, eds, Carbon Limitations. Springer- Verlag, Berlin, pp 21-43
Axelsson L, Mercado JM, Figueroa FL (2000) Utilization of HCO^sub 3^^sup -^ at high pH by the brown macroalga Laminaria saccharina. Eur J Phycol 35: 53-59
Badger MR, Palmqvist K, Yu JW (1994) Measurement of CO2 and HCO^sub 3^^sup -^ fluxes in cyanobacteria and microalgae during steady-state photosynthesis. Physiol Plant 90: 529-536
Brandalise M, Maia ID, Borecky J, Vercesi AE, Arruda P (2003) ZmPUMP encodes a maize mitochondrial uncoupling protein that is induced by oxidative stress. Plant Sci 165: 329-335
Elzenga JTM, Staal M, Prins HBA (1989) ATPase activity of isolated plasma membrane vesicles of leaves of Elodea as affected by thiol reagents and NADH/NAD1 ratio. Physiol Plant 76: 379-385
Faavaro RD, Borecky J, Colombi D, Maia IG (2006) ZmPUMP encodes a fully functional monocot plant uncoupling mitochondrial protein whose affinity to fatty acid is increased with the introduction of a His pair at the second matrix loop. Biochem Biophys Res Commun 344: 194-199
Giordano M, Beardall J, Raven JA (2005) CO2 concentrating mechanisms in algae: mechanisms, environmental modulation, and evolution. Annu Rev Plant Biol 56: 99-131
Hellblom F, Beer S, Bjork M,Axelsson L (2001)Abuffer sensitive inorganic carbon utilisation system in Zostera marina. Aquat Bot 69: 55-62
Hellblom F, Axelsson L (2003) External HCO3 2 dehydration maintained by acid zones in the plasma membrane is an important component of the photosynthetic carbon uptake in Ruppia cirrhosa. Photosynth Res 77: 173-181
Hemminga MA, Duarte CM (2000) Seagrass Ecology. Cambridge University Press, Cambridge
Huertas E, Colman B, Espie GS (2002a) Inorganic carbon acquisition and its energization in eustigmatophyte algae. Funct Plant Biol 29: 271-277
Huertas E, Colman B, Espie GS (2002b) Mitochondrial-driven bicarbonate transport supports photosynthesis in a marine microalga. Plant Physiol 130: 284-291
Igamberdiev AU, Hurry V, Kromer S, Gardestrom P (1998) The role of mitochondrial electron transport during photosynthetic induction. A study with barley (Hordeum vulgare) protoplasts incubated with rotenone and oligomycin. Physiol Plant 104: 431-439
Jensen RG (2004) Activation of Rubisco controls CO2 assimilation in light: a perspective on its discovery. Photosynth Res 82: 187- 193
Joet T, Cournac L, Horvath EM, Medgyesy P, Peltier G (2001) Increased sensitivity of photosynthesis to antimycin A induced by inactivation of the chloroplast ndhB gene. Evidence for a participation of the NADHdehydrogenase complex to cyclic electron flow around photosystem I. Plant Physiol 125: 1919-1929 Kaplan A, Reinhold L (1999) CO2 concentrating mechanisms in photosynthetic microorganisms. Annu Rev Plant Physiol Plant Mol Biol 50: 539-570
Karol KG, McCourt RM, Cimino MT, Delwiche CF (2001) The closest living relatives of land plants. Science 294: 2351-2353
Kirschbaum MUF, Oja V, Laisk A (2005) The quantum yield of CO2 fixation is reduced for several minutes after prior exposure to darkness. Exploration of the underlying causes. Plant Biol 7: 58-66
Kromer S, Heldt HW (1991) On the role of mitochondrial oxidative phosphorylation in photosynthesis metabolism as studied by the effect of oligomycin on photosynthesis in protoplasts and leaves of barley (Hordeum vulgare). Plant Physiol 95: 1270-1276
Kursar TA, Coley PD (1993) Photosynthetic inductions times in shadetolerant species with long and short-lived leaves. Oecologia 93: 165-170
Larsson C, Axelsson L, Ryberg H, Beer S (1997) Photosynthetic carbon utilization by Enteromorpha intestinalis (Chlorophyta) from a Swedish rockpool. Eur J Phycol 32: 49-54
Li XG, Xu PL, Zhao JP, Meng JJ, He QW (2006) Ferredoxin-quinone reductase benefits cyclic electron flow around photosystem 1 in tobacco leaves upon exposure to chilling stress under low irradiance. Photosynthetica 44: 349-354
McConnaughey T (1991) Calcification in Chara corallina: CO2 hydroxylation generates protons for bicarbonate assimilation. Limnol Oceanogr 36: 619-628
Mehrbach C, Culberson CH, Hawley JE, Pytkowicz RM (1973) Measurement of the apparent dissociation constants of carbonic acid in seawater at atmospheric pressure. Limnol Oceanogr 18: 897-907
Mercado JM, Andria R, Perez-Llorens JL, Vergara JJ, Axelsson L (2006) Evidence for a plasmalemma-based CO2 concentrating mechanism in Laminaria saccharina. Photosynth Res 88: 259-268
Mercado JM, Niell FX, Silva J, Santos R (2003) Use of light and inorganic carbon acquisition by two morphotypes of Zostera noltii Hornem. J Exp Mar Biol Ecol 297: 71-81
Mikulska M, Bomsel JL, Rychter AM (1998) The influence of phosphate deficiency on photosynthesis, respiration and adenine nucleotide pool in bean leaves. Photosynthetica 35: 79-88
Millero FJ (1979) The thermodynamics of the carbonate system in seawater. Geochim Cosmochim Acta 43: 1651-1661
Mimura T, Muller R, Kaiser WM, Shimmen T, Dietz KJ (1993) ATPdependent carbon transport in perfused Chara cells. Plant Cell Environ 16: 653-661
Padmasree K, Raghavendra AS (1999a) Prolongation of photosynthetic induction as a consequence with mitochondrial oxidative metabolism in mesophyll protoplasts of the pea (Pisum sativum L). Plant Sci 142: 29-36
Padmasree K, Raghavendra AS (1999b) Response of photosynthetic carbon assimilation in mesophyll protoplasts to restriction on mitochondrial oxidative metabolism: metabolites related to the redox status and sucrose biosynthesis. Photosynth Res 62: 231-239
Padmasree K, Raghavendra AS (2001a) Consequence of restricted mitochondrial oxidative metabolism on photosynthetic carbon assimilation in mesophyll protoplasts: decrease in light activation of four chloroplastic enzymes. Physiol Plant 112: 582-588
Padmasree K, Raghavendra AS (2001b) Restriction of mitochondrial oxidative metabolism leads to suppression of photosythetic carbon assimilation but not of photochemical electron transport in pea mesophyll protoplasts. Curr Sci 81: 680-684
Price GD, Badger MR (1985) Inhibition by proton buffers of photosynthetic utilization of bicarbonate in Chara corallina. Aust J Plant Physiol 12: 257-267
Prins HBA, Snel JFH, Helder RJ, Zanstra PE (1980) Photosynthetic HCO^sub 3^^sup -^ utilization and OH2 excretion in aquatic angiosperms. Light induced pH changes at the leaf surface. Plant Physiol 66: 818-822
Prins HBA, Snel JFH, Zanstra PE, Helder RJ (1982) The mechanism of bicarbonate assimilation by the polar leaves of Potamogeton and Elodea. CO2 concentrations at the leaf surface. Plant Cell Environ 5: 207-214
Raghavendra AS, Padmasree K, Saradadevi K (1994) Interdependence of photosynthesis and respiration in plant cells: interactions between chloroplasts and mitochondria. Plant Sci 97: 1-14
Raven JA (1997) Inorganic carbon acquisition by marine autotrophs. Adv Bot Res 27: 85-183
Raven JA (2000) Land plant biochemistry. Philos Trans R Soc Lond B Biol Sci 355: 833-846
Raven JA (2003) Inorganic carbon concentrating mechanisms in relation to the biology of algae. Photosynth Res 77: 155-171
Raven JA, Johnston AM, Macfarlane JJ (1990) Carbon metabolism. In KM Cole, RG Sheath, eds, Biology of the Red Algae. Cambridge University Press, Cambridge, pp 171-202
Ray S, Klenell M, Choo KS, Pedersen M, Snoeijs P (2003) Carbon acquisition mechanisms in Chara tomentosa. Aquat Bot 76: 141-154
Ricquier D, Bouillaud F (2000) The uncoupling protein homologues: UCP1, UCP2, UCP3, StUCP and AtUCP. Biochem J 345: 161-179
Romanowska E, Pokorska B, SiedleckaM (2005) The effects of oligomycin on content of adenylates in mesophyll protoplasts, chloroplasts and mitochondria from Pb^sup 2+^ treated pea and barley leaves. Acta Physiol Plant 27: 29-36
Sacher R, Burian K (1994) Effects of simulated orbital light/ dark cycles on the photosynthesis of mungbean, soybean and millet. Photosynthetica 30: 215-223
Serikawa KA, Porterfield DM, Smith PJS, Mandoli DF (2000) Calcification and measurements of net proton and oxygen flux reveal subcellular domains in Acetabularia acetabulum. Planta 211: 474-483
Shen J, Tang C, Rengel Z, Zhang F (2004) Root-induced acidification and excess cation uptake by N-2-fixing Lupinus albus grown in phosphorusdeficient soil. Plant Soil 260: 69-77
Siedow JN, Umbach AL (1995) Plant mitochondrial electron transfer and molecular biology. Plant Cell 7: 821-831
Sultemeyer D (1997) Carbonic anhydrase in eukaryotic algae: characterization, regulation, and possible function during photosynthesis. Can J Bot 76: 962-972
van Ginkel LC, Prins HBA (1997) Bicarbonate utilization and pH polarity. The response of photosynthetic electron transport to carbon limitation in Putamogeton lucens leaves. Can J Bot 76: 1018- 1024
Vianello A, Braidot E, Petrussa E, Macri F (1997) ATP synthesis driven by alpha-keto acid-stimulated alternative oxidase in pea leaf mitochondria. Plant Cell Physiol 38: 1368-1374
Wu WH, Berkowitz GA (1992) K^sup +^ stimulation of ATPase activity associated with the chloroplast inner envelope. Plant Physiol 99: 553-560
Herman Carr* and Lennart Axelsson
Department of Botany, Stockholm University, SE-106 91 Stockholm, Sweden (H.C.); and Kristineberg Marine Research Station, SE-450 34 Fiskebaeckskil, Sweden (L.A.)
* Corresponding author; e-mail [email protected]
The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Herman Carr ([email protected]).
Copyright American Society of Plant Biologists Jun 2008
(c) 2008 Plant Physiology. Provided by ProQuest Information and Learning. All rights Reserved.