Crassulacean acid metabolism in the context of other carbon-concentrating mechanisms in freshwater plants: a review

Inorganic carbon can be in short supply in freshwater relative to that needed by freshwater plants for photosynthesis because of a large external transport limitation coupled with frequent depleted concentrations of CO(2) and elevated concentrations of O(2). Freshwater plants have evolved a host of avoidance, exploitation and amelioration strategies to cope with the low and variable supply of inorganic carbon in water. Avoidance strategies rely on the spatial variation in CO(2) concentrations within and among lakes. Exploitation strategies involve anatomical and morphological features that take advantage of sources of CO(2) outside of the water column such as the atmosphere or sediment. Amelioration strategies involve carbon-concentrating mechanisms based on uptake of bicarbonate, which is widespread, C(4)-fixation, which is infrequent, and crassulacean acid metabolism (CAM), which is of intermediate frequency. CAM enables aquatic plants to take up inorganic carbon in the night. Furthermore, daytime inorganic carbon uptake is generally not inhibited and therefore CAM is considered to be a carbon-conserving mechanism. CAM in aquatic plants is a plastic mechanism regulated by environmental variables and is generally downregulated when inorganic carbon does not limit photosynthesis. CAM is regulated in the long term (acclimation during growth), but is also affected by environmental conditions in the short term (response on a daily basis). In aquatic plants, CAM appears to be an ecologically important mechanism for increasing inorganic carbon uptake, because the in situ contribution from CAM to the C-budget generally is high (18-55%).     

The stoichiometry of photorespiration during C3-photosynthesis is not fixed: evidence from combined physical and stereochemical methods

The stoichiometry of photorespiration, S, is defined as the fraction of glycolate carbon photorespired. It is postulated that under steady-state conditions there are two determinants of the ratio of photorespiration to net photosynthesis: the partitioning of ribulose bisphosphate between oxidation and carboxylation, and the partitioning of glycolate between reactions leading to complete oxidation to CO2 (S = 100%) and those yielding CO2 plus serine (S = 25%). S may be calculated using two independent probes of the system. The physical probe, using an infrared gas analyzer, measured photorespiration and net photosynthesis, and hence their ratio PR/NPS = pn(phys). The metabolic probe employed tracer (3R)-D-[3-3H1,3-14C]glyceric acid to determine r, the fraction of 3H retained in the triose phosphates leaving the chloroplasts. It is deduced from the postulated model that S = pn(phys) . r/(1 - r). Experiments have been performed with illuminated tobacco leaf discs (inverted) under varying concentrations of O2 and CO2. Increasing O2 at constant CO2 increased pn(phys) and decreased r, whereas increasing CO2 at constant O2 had the opposite effect. S more than doubled at 32 degrees C on going from 16 to 40% O2 (340 microliters CO2/liter) and decreased 40% on going from 200 to 700 microliters CO2/liter (21% O2). For discs in normal air S was somewhat greater than 27%. It is suggested that net photosynthesis, and therefore crop yields, could be increased by selecting for crop plants with reduced photorespiration stoichiometry.     

pH Dependence of Photosynthesis and Photorespiration in Soybean Leaf Cells

The effect of pH on the kinetics of photosynthesis, O(2) inhibition of photosynthesis, and photorespiration was examined with mesophyll cells isolated from soybean (Glycine max [L.] Merr.) leaves. At constant, subsaturating bicarbonate concentration (0.5 mm), O(2) inhibition of photosynthesis increased with increasing pH because high pH shifts the CO(2)-bicarbonate equilibrium toward bicarbonate, thereby reducing the CO(2) concentration. At constant, substrating CO(2) concentrations, cell photorespiration decreased with increasing pH. This was indicated by decreases in the CO(2) compensation concentration, O(2) inhibition of photosynthesis, and glycine synthesis. Km(CO(2)) values for isolated cell photosynthesis and in vitro ribulose-1, 5-diphosphate carboxylase activity decreased with increasing pH, while the Ki(O(2)) for both systems was similar at all pH values. The responses to pH of the corresponding kinetic constants of cell photosynthesis and in vitro RuDP carboxylase with respect to CO(2) and O(2) were identical. This provides additional evidence that the relative rates of photosynthesis and photorespiration in C(3) plants are determined by the kinetic properties of RuDP carboxylase.     

C4 Photosynthesis (The CO2-Concentrating Mechanism and Photorespiration)

Despite previous reports of no apparent photorespiration in C4 plants based on measurements of gas exchange under 2 versus 21% O2 at varying [CO2], photosynthesis in maize (Zea mays) shows a dual response to varying [O2]. The maximum rate of photosynthesis in maize is dependent on O2 (approximately 10%). This O2 dependence is not related to stomatal conductance, because measurements were made at constant intercellular CO2 concentration (Ci); it may be linked to respiration or pseudocyclic electron flow. At a given Ci, increasing [O2] above 10% inhibits both the rate of photosynthesis, measured under high light, and the maximum quantum yield, measured under limiting light ([phi]CO2). The dual effect of O2 is masked if measurements are made under only 2 versus 21% O2. The inhibition of both photosynthesis and [phi]CO2 by O2 (measured above 10% O2) with decreasing Ci increases in a very similar manner, characteristically of O2 inhibition due to photorespiration. There is a sharp increase in O2 inhibition when the Ci decreases below 50 [mu]bar of CO2. Also, increasing temperature, which favors photorespiration, causes a decrease in [phi]CO2 under limiting CO2 and 40% O2. By comparing the degree of inhibition of photosynthesis in maize with that in the C3 species wheat (Triticum aestivum) at varying Ci, the effectiveness of C4 photosynthesis in concentrating CO2 in the leaf was evaluated. Under high light, 30[deg]C, and atmospheric levels of CO2 (340 [mu]bar), where there is little inhibition of photosynthesis in maize by O2, the estimated level of CO2 around ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the bundle sheath compartment was 900 [mu]bar, which is about 3 times higher than the value around Rubisco in mesophyll cells of wheat. A high [CO2] is maintained in the bundle sheath compartment in maize until Ci decreases below approximately 100 [mu]bar. The results from these gas exchange measurements indicate that photorespiration occurs in maize but that the rate is low unless the intercellular [CO2] is severely limited by stress.     

Quantum yield variation across the three pathways of photosynthesis: not yet out of the dark

The convergent quantum yield hypothesis (CQY) assumes that thermodynamics and natural selection jointly limit variation in the maximum energetic efficiency of photosynthesis in low light under otherwise specified conditions (e.g. temperature and CO(2) concentration). A literature survey of photosynthetic quantum yield (phi) studies in terrestrial plants from C(3), C(4), and CAM photosynthetic types was conducted to test the CQY hypothesis. Broad variation in phi values from C(3) plants could partially be explained by accounting for whether the measuring conditions were permissive or restrictive for photorespiration. Assimilatory quotients (AQ), calculated from the CO(2) phi:O(2) phi ratios, indicated that 49% and 29% of absorbed light energy was allocated to carbon fixation and photorespiration in C(3) plants, respectively. The unexplained remainder (22%) may represent diversion to various other energy-demanding processes (e.g. starch synthesis, nitrogen assimilation). Individual and cumulative effects of these other processes on photosynthetic efficiency are poorly quantified. In C(4) plants, little variation in phi values was observed, consistent with the fact that C(4) plants exhibit little photorespiration. As before, AQ values indicate that 22% of absorbed light energy cannot be accounted for by carbon fixation in C(4) plants. Among all three photosynthetic types, the phi of photosynthesis in CAM plants is the least studied, appears to be highly variable, and may present the greatest challenge to the CQY hypothesis. The high amount of energy diverted to processes other than carbon fixation in C(3) and C(4) plants and the poor characterization of photosynthetic efficiency in CAM plants are significant deficiencies in our otherwise robust understanding of the energetics of terrestrial photoautotrophy.     

Underwater photosynthesis and respiration in leaves of submerged wetland plants: gas films improve CO2 and O2 exchange.

Many wetland plants have gas films on submerged leaf surfaces. We tested the hypotheses that leaf gas films enhance CO(2) uptake for net photosynthesis (P(N)) during light periods, and enhance O(2) uptake for respiration during dark periods. Leaves of four wetland species that form gas films, and two species that do not, were used. Gas films were also experimentally removed by brushing with 0.05% (v/v) Triton X. Net O(2) production in light, or O(2) consumption in darkness, was measured at various CO(2) and O(2) concentrations. When gas films were removed, O(2) uptake in darkness was already diffusion-limited at 20.6 kPa (critical O(2) pressure for respiration, COP(R)>/= 284 mmol O(2) m(-3)), whereas for some leaves with gas films, O(2) uptake declined only at approx. 4 kPa (COP(R) 54 mmol O(2) m(-3)). Gas films also improved CO(2) uptake so that, during light periods, underwater P(N) was enhanced up to sixfold. Gas films on submerged leaves enable continued gas exchange via stomata and thus bypassing of cuticle resistance, enhancing exchange of O(2) and CO(2) with the surrounding water, and therefore underwater P(N) and respiration.     

Submergence-induced morphological, anatomical, and biochemical responses in a terrestrial species affect gas diffusion resistance and photosynthetic performance

Gas exchange between the plant and the environment is severely hampered when plants are submerged, leading to oxygen and energy deficits. A straightforward way to reduce these shortages of oxygen and carbohydrates would be continued photosynthesis under water, but this possibility has received only little attention. Here, we combine several techniques to investigate the consequences of anatomical and biochemical responses of the terrestrial species Rumex palustris to submergence for different aspects of photosynthesis under water. The orientation of the chloroplasts in submergence-acclimated leaves was toward the epidermis instead of the intercellular spaces, indicating that underwater CO(2) diffuses through the cuticle and epidermis. Interestingly, both the cuticle thickness and the epidermal cell wall thickness were significantly reduced upon submergence, suggesting a considerable decrease in diffusion resistance. This decrease in diffusion resistance greatly facilitated underwater photosynthesis, as indicated by higher underwater photosynthesis rates in submergence-acclimated leaves at all CO(2) concentrations investigated. The increased availability of internal CO(2) in these "aquatic" leaves reduced photorespiration, and furthermore reduced excitation pressure of the electron transport system and, thus, the risk of photodamage. Acclimation to submergence also altered photosynthesis biochemistry as reduced Rubisco contents were observed in aquatic leaves, indicating a lower carboxylation capacity. Electron transport capacity was also reduced in these leaves but not as strongly as the reduction in Rubisco, indicating a substantial increase of the ratio between electron transport and carboxylation capacity upon submergence. This novel finding suggests that this ratio may be less conservative than previously thought.     

CAM photosynthesis in submerged aquatic plants

Crassulacean acid metabolism (CAM) is a CO₂-concentrating mechanism selected in response to aridity in terrestrial habitats, and, in aquatic environments, to ambient limitations of carbon. Evidence is reviewed for its presence in five genera of aquatic vascular plants, includingIsoëtes, Sagittaria, Vallisneria, Crassula, andLittorella. Initially, aquatic CAM was considered by some to be an oxymoron, but some aquatic species have been studied in sufficient detail to say definitively that they possess CAM photosynthesis. CO₂-concentrating mechanisms in photosynthetic organs require a barrier to leakage; e.g., terrestrial C₄ plants have suberized bundle sheath cells and terrestrial CAM plants high stomatal resistance. In aquatic CAM plants the primary barrier to CO₂ leakage is the extremely high difrusional resistance of water. This, coupled with the sink provided by extensive intercellular gas space, generates daytime CO₂(pi) comparable to terrestrial CAM plants. CAM contributes to the carbon budget by both net carbon gain and carbon recycling, and the magnitude of each is environmentally influenced. Aquatic CAM plants inhabit sites where photosynthesis is potentially limited by carbon. Many occupy moderately fertile shallow temporary pools that experience extreme diel fluctuations in carbon availability. CAM plants are able to take advantage of elevated nighttime CO2 levels in these habitats. This gives them a competitive advantage over non-CAM species that are carbon starved during the day and an advantage over species that expend energy in membrane transport of bicarbonate. Some aquatic CAM plants are distributed in highly infertile lakes, where extreme carbon limitation and light are important selective factors. Compilation of reports on diel changes in titratable acidity and malate show 69 out of 180 species have significant overnight accumulation, although evidence is presented discounting CAM in some. It is concluded that similar proportions of the aquatic and terrestrial floras have evolved CAM photosynthesis. AquaticIsoëtes (Lycophyta) represent the oldest lineage of CAM plants and cladistic analysis supports an origin for CAM in seasonal wetlands, from which it has radiated into oligotrophic lakes and into terrestrial habitats. Temperate Zone terrestrial species share many characteristics with amphibious ancestors, which in their temporary terrestrial stage, produce functional stomata and switch from CAM to C₃. Many lacustrineIsoëtes have retained the phenotypic plasticity of amphibious species and can adapt to an aerial environment by development of stomata and switching to C₃. However, in some neotropical alpine species, adaptations to the lacustrine environment are genetically fixed and these constitutive species fail to produce stomata or loose CAM when artificially maintained in an aerial environment. It is hypothesized that neotropical lacustrine species may be more ancient in origin and have given rise to terrestrial species, which have retained most of the characteristics of their aquatic ancestry, including astomatous leaves, CAM and sediment-based carbon nutrition.     

CO(2)-concentrating: consequences in crassulacean acid metabolism

The consequences of CO(2)-concentrating in leaf air-spaces of CAM plants during daytime organic acid decarboxylation in Phase III of CAM (crassulacean acid metabolism) are explored. There are mechanistic consequences of internal CO(2) partial pressures, p(i)(CO(2)). These are (i) effects on stomata, i.e. high p(i)(CO(2)) eliciting stomatal closure in Phase III, (ii) regulation of malic acid remobilization from the vacuole, malate decarboxylation and refixation of CO(2) via Rubisco (ribulose bisphosphate carboxylase/oxygenase), and (iii) internal signalling functions during the transitions between Phases II and III and III and IV, respectively, in the natural day/night cycle and in synchronizing the circadian clocks of individual leaf cells or leaf patches in the free-running endogenous rhythmicity of CAM. There are ecophysiological consequences. Obvious beneficial ecophysiological consequences are (i) CO(2)-acquisition, (ii) increased water-use- efficiency, (iii) suppressed photorespiration, and (iv) reduced oxidative stress by over-energization of the photosynthetic apparatus. However, the general potency of these beneficial effects may be questioned. There are also adverse ecophysiological consequences. These are (i) energetics, (ii) pH effects and (iii) Phase III oxidative stress. A major consequence of CO(2)-concentrating in Phase III is O(2)-concentrating, increased p(i)(CO(2)) is accompanied by increased p(i)(O(2)). Do reversible shifts of C(3)/CAM-intermediate plants between the C(3)-CAM-C(3) modes of photosynthesis indicate that C(3)-photosynthesis provides better protection from irradiance stress? There are many open questions and CAM remains a curiosity.     

Molecular Genetics of Crassulacean Acid Metabolism

Most higher plants assimilate atmospheric CO2 through the C3 pathway of photosynthesis using ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). However, when CO2 availability is reduced by environmental stress conditions, the incomplete discrimination of CO2 over O2 by Rubisco leads to increased photorespiration, a process that reduces the efficiency of C3 photosynthesis. To overcome the wasteful process of photorespiration, approximately 10% of higher plant species have evolved two alternate strategies for photosynthetic CO2 assimilation, C4 photosynthesis and Crassulacean acid metabolism. Both of these biochemical pathways employ a "CO2 pump" to elevate intracellular CO2 concentrations in the vicinity of Rubisco, suppressing photorespiration and therefore improving the competitiveness of these plants under conditions of high light intensity, high temperature, or low water availability. This CO2 pump consists of a primary carboxylating enzyme, phosphoenolpyruvate carboxylase. In C4 plants, this CO2-concentrating mechanism is achieved by the coordination of two carboxylating reactions that are spatially separated into mesophyll and bundle-sheath cell types (for review, see R.T. Furbank, W.C. Taylor [1995] Plant Cell 7: 797-807;M.S.B. Ku, Y. Kano-Murakami, M. Matsuoka [1996] Plant Physiol 111: 949-957). In contrast, Crassulacean acid metabolism plants perform both carboxylation reactions within one cell type, but the two reactions are separated in time. Both pathways involve cell-specific changes in the expression of many genes that are not present in C3 plants.     

Aquatic adventitious roots of the wetland plant Meionectes brownii can photosynthesize: implications for root function during flooding

? Many wetland plants produce aquatic adventitious roots from submerged stems. Aquatic roots can form chloroplasts, potentially producing endogenous carbon and oxygen. Here, aquatic root photosynthesis was evaluated in the wetland plant Meionectes brownii, which grows extensive stem-borne aquatic roots during submergence. ? Underwater photosynthetic light and CO(2) response curves were determined for aquatic-adapted leaves, stems and aquatic roots of M. brownii. Oxygen microelectrode and (14)CO(2)-uptake experiments determined shoot inputs of O(2) and photosynthate into aquatic roots. ? Aquatic adventitious roots contain a complete photosynthetic pathway. Underwater photosynthetic rates are similar to those of stems, with a maximum net photosynthetic rate (P(max)) of 0.38 μmol O(2) m(-2) s(-1); however, this is c. 30-fold lower than that of aquatic-adapted leaves. Under saturating light with 300 mmol m(-3) dissolved CO(2), aquatic roots fix carbon at 0.016 μmol CO(2) g(-1) DM s(-1). Illuminated aquatic roots do not rely on exogenous inputs of O(2). ? The photosynthetic ability of aquatic roots presumably offers an advantage to submerged M. brownii as aquatic roots, unlike sediment roots, need little O(2) and carbohydrate inputs from the shoot when illuminated.     

Drought-stress-induced up-regulation of CAM in seedlings of a tropical cactus, Opuntia elatior, operating predominantly in the C3 mode

Immediately after unfolding, cotyledons of the tropical platyopuntoid cactus, Opuntia elatior Mill., exhibited a C(3)-type diel CO(2) exchange pattern characterized by net CO(2) uptake in the light. Significant nocturnal increases in titratable acidity typical of crassulacean acid metabolism (CAM) were not detected at this early developmental stage. As cotyledons matured and the first cladode (flattened stem) developed, features of CAM were observed and the magnitude of CAM increased. Nonetheless, in well-watered seedlings up to 10 cm tall, C(3) photosynthetic CO(2) fixation in the light remained the major pathway of carbon fixation. Reduced soil water availability led to an up-regulation of net dark CO(2) fixation and greater nocturnal increases in tissue acidity, consistent with facultative CAM. These observations demonstrate that C(3) photosynthesis, drought-stress-related facultative CAM, and developmentally controlled constitutive CAM can all contribute to the early growth of O. elatior. The strong C(3) component and facultative CAM features expressed in young O. elatior contrast with mature plants in which obligate CAM is the major pathway of carbon acquisition.     

Photosynthetic consequences of phenotypic plasticity in response to submergence: Rumex palustris as a case study

Survival and growth of terrestrial plants is negatively affected by complete submergence. This is mainly the result of hampered gas exchange between plants and their environment, since gas diffusion is severely reduced in water compared with air, resulting in O2 deficits which limit aerobic respiration. The continuation of photosynthesis could probably alleviate submergence-stress in terrestrial plants, but its potential under water will be limited as the availability of CO2 is hampered. Several submerged terrestrial plant species, however, express plastic responses of the shoot which may reduce gas diffusion resistance and enhance benefits from underwater photosynthesis. In particular, the plasticity of the flooding-tolerant terrestrial species Rumex palustris turned out to be remarkable, making it a model species suitable for the study of these responses. During submergence, the morphology and anatomy of newly developed leaves changed: 'aquatic' leaves were thinner and had thinner cuticles. As a consequence, internal O2 concentrations and underwater CO2 assimilation rates were higher at the prevailing low CO2 concentrations in water. Compared with heterophyllous amphibious plant species, underwater photosynthesis rates of terrestrial plants may be very limited, but the effects of underwater photosynthesis on underwater survival are impressive. A combination of recently published data allowed quantification of the magnitude of the acclimation response in this species. Gas diffusion resistance in terrestrial leaves underwater was about 15,000 times higher than in air. Strikingly, acclimation to submergence reduced this factor to 400, indicating that acclimated leaves of R. palustris had an approximately 40 times lower gas diffusion resistance than non-acclimated ones.     

Comparison of the photosynthetic characteristics of three submersed aquatic plants

Light- and CO(2)-saturated photosynthetic rates of the submersed aquatic plants Hydrilla verticillata, Ceratophyllum demersum, and Myriophyllum spicatum were 50 to 60 mumol O(2)/mg Chl.hr at 30 C. At air levels of CO(2), the rates were less than 5% of those achieved by terrestrial C(3) plants. The low photosynthetic rates correlated with low activities of the carboxylation enzymes. In each species, ribulose 1,5-diphosphate carboxylase was the predominant carboxylation enzyme. The apparent K(m)(CO(2)) values for photosynthesis were 150 to 170 mum at pH 4, and 75 to 95 mum at pH 8. The K(m)(CO(2)) of Hydrilla ribulose 1,5-diphosphate carboxylase was 45 mum at pH 8. Optimum temperatures for the photosynthesis of Hydrilla, Myriophyllum, and Ceratophyllum were 36.5, 35.0, and 28.5 C, respectively. The apparent ability of each species to use HCO(3) (-) ions for photosynthesis was similar, but at saturating free CO(2) levels, there was no indication of HCO(3) (-) use. Increasing the pH from 3.1 to 9.2 affected the photosynthetic rate indirectly, by decreasing the free CO(2). With saturating free CO(2) (0.5 mm), the maximum photosynthetic rates were similar at pH 4 and 8. Carbonic anhydrase activity, although much lower than in terrestrial C(3) plants, was still in excess of that required to support HCO(3) (-) utilization.Hydrilla and Ceratophyllum had CO(2) compensation points of 44 and 41 mul/l, respectively, whereas the value for Myriophyllum was 19. Relatively high CO(2) compensation points under 1% O(2) indicated that some "dark" respiration occurred in the light. The inhibition of photosynthesis by O(2) was less than with terrestrial C(3) plants. Glycolate oxidase activity was 12.3 to 27.5 mumol O(2)/mg Chl.hr, as compared to 78.4 for spinach. Light saturation of photosynthesis occurred at 600 to 700 mueinsteins/m(2).sec in each species grown under full sunlight. Hydrilla had the lowest light compensation point, and required the least irradiance to achieve the half-maximal photosynthetic rate.Field measurements in a Hydrilla mat indicated that in the afternoon, free CO(2) dropped to zero, and O(2) rose to over 200% air saturation. Most photosynthetic activity occurred in the morning when the free CO(2) was highest and O(2) and solar radiation lowest. The low light requirement of Hydrilla probably provides a competitive advantage under these field conditions.     

Stem CO2 release under illumination: corticular photosynthesis,photorespiration or inhibition of mitochondrial respiration?

In illuminated stems and branches, CO2 release is often reduced. Many light-triggered processes are thought to contribute to this reduction, namely photorespiration, corticular photosynthesis or even an inhibition of mitochondrial respiration. In this study, we investigated these processes with the objective to discriminate their influence to the overall reduction of branch CO2 release in the light. CO2 gas-exchange measurements of young birch (Betula pendula Roth.) branches (< 1.5 cm) performed under photorespiratory (20% O2) and non-photorespiratory (< 2%) conditions revealed that photorespiration does not play a pre-dominant role in carbon exchange. This suppression of photorespiration was attributed to the high CO2 concentrations (C(i)) within the bark tissues (1544 +/- 227 and 618 +/- 43 micromol CO2 mol(-1) in the dark and in the light, respectively). Changes in xylem CO2 were not likely to explain the observed decrease in stem CO2 release as gas-exchange measurements before and after cutting of the branches did not effect CO2 efflux to the atmosphere. Combined fluorescence and gas-exchange measurements provided evidence that the light-dependent reduction in CO2 release can pre-dominantly be attributed to corticular refixation, whereas an inhibition of mitochondrial respiration in the light is unlikely to occur. Corticular photosynthesis was able to refix up to 97% of the CO2 produced by branch respiration, although it rarely led to a positive net photosynthetic rate.     

Underwater photosynthesis in flooded terrestrial plants: a matter of leaf plasticity

BACKGROUND: Flooding causes substantial stress for terrestrial plants, particularly if the floodwater completely submerges the shoot. The main problems during submergence are shortage of oxygen due to the slow diffusion rates of gases in water, and depletion of carbohydrates, which is the substrate for respiration. These two factors together lead to loss of biomass and eventually death of the submerged plants. Although conditions under water are unfavourable with respect to light and carbon dioxide supply, photosynthesis may provide both oxygen and carbohydrates, resulting in continuation of aerobic respiration. SCOPE: This review focuses on evidence in the literature that photosynthesis contributes to survival of terrestrial plants during complete submergence. Furthermore, we discuss relevant morphological and physiological responses of the shoot of terrestrial plant species that enable the positive effects of light on underwater plant performance. CONCLUSIONS: Light increases the survival of terrestrial plants under water, indicating that photosynthesis commonly occurs under these submerged conditions. Such underwater photosynthesis increases both internal oxygen concentrations and carbohydrate contents, compared with plants submerged in the dark, and thereby alleviates the adverse effects of flooding. Additionally, several terrestrial species show high plasticity with respect to their leaf development. In a number of species, leaf morphology changes in response to submergence, probably to facilitate underwater gas exchange. Such increased gas exchange may result in higher assimilation rates, and lower carbon dioxide compensation points under water, which is particularly important at the low carbon dioxide concentrations observed in the field. As a result of higher internal carbon dioxide concentrations in submergence-acclimated plants, underwater photorespiration rates are expected to be lower than in non-acclimated plants. Furthermore, the regulatory mechanisms that induce the switch from terrestrial to submergence-acclimated leaves may be controlled by the same pathways as described for heterophyllous aquatic plants.     

Evidence for the occurrence of photorespiration in synurophyte algae

The fluxes of CO(2) and oxygen during photosynthesis by cell suspensions of Tessellaria volvocina and Mallomonas papillosa were monitored mass spectrometrically. There was no rapid uptake of CO(2,) only a slow drawdown to compensation concentrations of 26 μM for T. volvocina and 18 μM for M. papillosa, when O(2) evolution ceased, indicating a lack of active bicarbonate uptake by the cells. Darkening of the cells after a period of photosynthesis did not cause rapid release of CO(2), indicating the absence of an intracellular inorganic carbon pool. However, upon darkening a brief burst of CO(2) was observed similar to the post-illumination burst characteristic of C(3) higher plants. Treatment of the cells of both species with the membrane-permeable carbonic anhydrase inhibitor ethoxyzolamide had no adverse effect on photosynthetic rate, but stimulated the dark CO(2) burst indicating the dark oxidation of a compound formed in the light. In the absence of any active accumulation of inorganic carbon photosynthesis in these species should be inhibited by O(2). This was investigated in four synurophyte species T. volvocina, M. papillosa, Synura petersenii, and Synura uvella: photosynthetic O(2) evolution rates in all four algae, measured by O(2) electrode, were significantly higher (40-50%) in media at low O(2) (4%) than in air-equilibrated (21% O(2)) media, indicating an O(2) inhibition of photosynthesis (Warburg effect) and thus the occurrence of photorespiration in these species.     

High glycolate oxidase activity is required for survival of maize in normal air

A mutant in the maize (Zea mays) Glycolate Oxidase1 (GO1) gene was characterized to investigate the role of photorespiration in C4 photosynthesis. An Activator-induced allele of GO1 conditioned a seedling lethal phenotype when homozygous and had 5% to 10% of wild-type GO activity. Growth of seedlings in high CO2 (1%-5%) was sufficient to rescue the mutant phenotype. Upon transfer to normal air, the go1 mutant became necrotic within 7 d and plants died within 15 d. Providing [1-14C]glycolate to leaf tissue of go1 mutants in darkness confirmed that the substrate is inefficiently converted to 14CO2, but both wild-type and GO-deficient mutant seedlings metabolized [1-14C]glycine similarly to produce [14C]serine and 14CO2 in a 1:1 ratio, suggesting that the photorespiratory pathway is otherwise normal in the mutant. The net CO2 assimilation rate in wild-type leaves was only slightly inhibited in 50% O2 in high light but decreased rapidly and linearly with time in leaves with low GO. When go1 mutants were shifted from high CO2 to air in light, they accumulated glycolate linearly for 6 h to levels 7-fold higher than wild type and 11-fold higher after 25 h. These studies show that C4 photosynthesis in maize is dependent on photorespiration throughout seedling development and support the view that the carbon oxidation pathway evolved to prevent accumulation of toxic glycolate.     

Two photosynthetic mechanisms mediating the low photorespiratory state in submersed aquatic angiosperms

The submersed angiosperms Myriophyllum spicatum L. and Hydrilla verticillata (L.f.) Royal exhibited different photosynthetic pulse-chase labeling patterns. In Hydrilla, over 50% of the ¹⁴C was initially in malate and aspartate, but the fate of the malate depended upon the photorespiratory state of the plant. In low photorespiration Hydrilla, malate label decreased rapidly during an unlabeled chase, whereas labeling of sucrose and starch increased. In contrast, for high photorespiration Hydrilla, malate labeling continued to increase during a 2-hour chase. Thus, malate formation occurs in both photorespiratory states, but reduced photorespiration results when this malate is utilized in the light. Unlike Hydrilla, in low photorespiration Myriophyllum, ¹⁴C incorporation was via the Calvin cycle, and less than 10% was in C₄ acids.

Ethoxyzolamide, a carbonic anhydrase inhibitor and a repressor of the low photorespiratory state, increased the label in glycolate, glycine, and serine of Myriophyllum. Isonicotinic acid hydrazide increased glycine labeling of low photorespiration Myriophyllum from 14 to 25%, and from 12 to 48% with high photorespiration plants. Similar trends were observed with Hydrilla. Increasing O₂ increased the per cent [¹⁴C]glycine and the O₂ inhibition of photosynthesis in Myriophyllum. In low photorespiration Myriophyllum, glycine labeling and O₂ inhibition of photosynthesis were independent of the CO₂ level, but in high photorespiration plants the O₂ inhibition was competitively decreased by CO₂. Thus, in low but not high photorespiration plants, glycine labeling and O₂ inhibition appeared to be uncoupled from the external [O₂]/[CO₂] ratio.

These data indicate that the low photorespiratory states of Hydrilla and Myriophyllum are mediated by different mechanisms, the former being C₄-like, while the latter resembles that of low CO₂-grown algae. Both may require carbonic anhydrase to enhance the use of inorganic carbon for reducing photorespiration.