Oxygen inhibition of dissolved inorganic carbon uptake in unicellular green algae

Unicellular green algae have a dissolved inorganic carbon (DIC) concentrating mechanism, commonly known as the DIC pump, to concentrate inorganic carbon into cells and chloroplasts. The DIC pump activity is normally measured as the K_0.5(DIC) that equals the CO₂ plus HCO₃‐ concentration at a cited pH at which the rate of DIC‐dependent photosynthetic O₂ evolution is half‐maximal, or by the amount of intra‐cellular DIC accumulation in 15–60 s, using a limited amount of NaH¹⁴CO₃, measured by the silicone oil cen‐trifugation technique. The dissolved oxygen in the assay inhibits or reduces the DIC uptake by the cells of unicellular green algae Chlamydomonas reinhardtii Dangeard, strain 137 and in a cell wall‐less marine algae Dunaliella tertiolecta Butcher. The algal cells concentrated the highest amount of DIC when little or no oxygen was present in the assay medium. The results suggest that the amount of O₂ and DIC must be carefully monitored before DIC‐pump assay.     

Responses of Two CAM Species to Different Irradiances during Growth and Susceptibility to Photoinhibition by High Light

Two CAM species, Kalanchoë daigremontiana Hamet et Perrier and Hoya carnosa (L.) R. Br., were grown under a range of five photon flux area densitites (PFD) and then characterized. Significant acclimation to shade was indicated by progressive decreases in leaf thickness, rates of respiratory O₂ uptake, light compensation point, maximum rates of photosynthetic O₂ evolution, nocturnal acid accumulation, and δ¹³C values, and increases in chlorophyll concentration and absolute levels of room temperature (25°C) and 77K fluorescence. Quantum yields (as measured by O₂ exchange) and the ratio of variable 77K fluorescence over the maximum yield (F_v/F_m) were relatively constant across the treatments. The only significant deviation from the above characteristics was in H. carnosa grown under full glasshouse PFD, where it apparently experienced photoinhibition. Following a photoinhibitory treatment, K. daigremontiana exhibited increases in the light compensation point and progressively greater reductions in the quantum yield, maximum photosynthetic rate, F_v/F_m, and the variable component of room temperature fluorescence with increasing shade during growth. Thus although Crassulacean acid metabolism plants can adjust to shaded conditions, they are susceptible to photoinhibition when exposed to higher PFD than that experienced during growth.     

Similar Photosynthetic Performance in Low Light-Grown Isonuclear Triazine-Resistant and -Susceptible Brassica napus L

Triazine-resistant plants grown under moderate to high photon flux density (PFD) conditions exhibit decreased photon yield, decreased light-saturated O₂ evolution and slower growth than triazine-susceptible plants. In this study we tested the hypothesis that the comparable growth previously observed in resistant and susceptible Brassica napus L. lines grown under low PFD was accompanied by comparable photon yield and light-saturated O₂ evolution. We measured photon yield, O₂ flash yield, fluorescence decay kinetics, fluorescence transient kinetics, and quenching components, F_v/F_m and light saturated O₂ evolution in leaf disks of low PFD-grown triazine-resistant and susceptible B. napus isogenic lines. Results indicated that slow electron transfer from the primary to secondary quinone electron acceptors of photosystem II was still present in the resistant line but photon yield and light-saturated O₂ evolution were similar in the two B. napus lines. We conclude that the alteration in the D1 protein that confers resistance does not necessarily cause decreased photosynthetic performance. Decreased photon yield in resistant plants grown at high PFD is not a direct consequence of the alteration in D1, but represents secondary damage.     

On the role of orientation in reducing photoinhibitory damage in photosynthetic-twig desert shrubs

Quantum yields for O₂ evolution were measured in photosynthetic twigs and leaves of Hymenoclea salsola and Senecio douglasii, two common shrubs of the deserts of western North America. When exposed to long-term drought, quantum yields of leaves and twigs remained constant up to the point of leaf abscision in both species. As water stress developed further, quantum yields began to decline in twigs; the extent of this quantum yield reduction was dependent on incident photon flux density. The reduction in quantum yield in twigs, which have a near-vertical orientation, was greatly accelerated when twigs were reoriented to the near-horizontal inclination typical of leaves. The reductions in quantum yield were not rapidly reversible and are interpreted as indicating photoinhibitory damage. The results are discussed in terms of the role that a near-vertical orientation might serve in maintaining photosynthetic structures through a drought period.     

Effect of dehydration and high light on photosynthesis of two C3 plants (Phaseolus vulgaris L. and Elatostema repens (Lour.) Hall f.)

The effect of drought on the photosynthetic functioning of two C₃ plants, Phaseolus vulgaris and Elatostema repens, has been examined. Leaf net CO₂ uptake measured in normal air was negligible at a leaf water deficit of about 30% while the calculated leaf intercellular CO₂ concentration (Ci) was unchanged. However, both the maximal photosynthetic capacity (CO₂-dependent O₂ evolution) and apparent quantum yield, measured in the presence of saturating CO₂ levels (5 to 14%), only started to decrease within the range of 25 to 30% leaf water deficit. This shows that the drought-induced inhibition seen in normal air is not caused by an inhibition of the photosynthetic mechanism, and that in this case Ci values can be misleading. Both 77 K and room-temperature fluorescence measurements indicate that the functioning of the photosystem-II reaction centre is hardly modified by water shortage. Furthermore, an analysis of photochemical chlorophyll fluorescence quenching shows, in the absence of CO₂, that O₂ can be an efficient acceptor of photosynthetic energy, even in severly dehydrated plants which do not show net CO₂ uptake in normal air. In these plants, O₂ is probably reduced mainly via Mehler-type reactions. High-light treatment given at low O₂ increases photoinhibition as measured by the decrease of apparent quantum yield in dehydrated P. vulgaris, whereas, interestingly, 1% O₂ protects dehydrated E. repens against high-light damage. The two plants could have different protective mechanisms depending upon the O₂ level or different photoinhibitory sites or mechanisms.Abbreviations and symbols Ca, Ci ambient and calculated intercellular CO₂ concentration - Fm, Fo, Fv maximum, initial and variable fluorescence emission - LWD leaf water deficit - PPFD photosynthetic photon flux density - PSII photosystem II - qQ photochemical quenching of chlorophyll fluorescence     

Kok effect and the quantum yield of photosynthesis : light partially inhibits dark respiration

The linear response of photosynthesis to light at low photon flux densities is known to change abruptly in the vicinity of the light compensation point so that the quantum yield seems to decrease as radiation increases. We studied this `Kok effect' in attached sunflower (Helianthus annuus L. cv IS894) leaves using gas exchange techniques. The effect was present even though respiration was constant in the dark. It was observed at a similar photon flux density (7 to 11 micromole photons per square meter per second absorbed photosynthetically active radiation) despite a wide range of light compensation points as well as rates of photosynthesis. The effect was not apparent when photorespiration was inhibited at low pO₂ (1 kilopascal), but this result was complicated because dark respiration was quite O₂-sensitive and was partially suppressed under these conditions. The Kok effect was observed at saturating pCO₂ and, therefore, could not be explained by a change in photorespiration. Instead, the magnitude of the effect varied as dark respiration varied in a single leaf, and was minimized when dark respiration was minimized, indicating that a partial suppression of dark respiration by light is responsible. Quantum yields measured at photon flux densities between 0 and 7 to 11 micromole photons per square meter per second, therefore, represent the combined yields of photosynthesis and of the suppression of a component of dark respiration by light. This leads to an overestimate of the quantum yield of photosynthesis. In view of these results, quantum yields of photosynthesis must be measured (a) when respiration is constant in the dark, and (b) when dark respiration has been inhibited either at low pO₂ to eliminate most of the light-induced suppression of dark respiration or at photon flux densities above that required to saturate the light-induced suppression of dark respiration. Significant errors in quantum yields of photosynthesis can result in leaves exhibiting this respiratory behavior if these principles are not followed.     

On the relationship between the quantum yield of Photosystem II electron transport,as determined by chlorophyll fluorescence and the quantum yield of CO2-dependent O2 evolution

We tested the two empirical models of the relationship between chlorophyll fluorescence and photosynthesis, previously published by Weis E and Berry JA 1987 (Biochim Biophys Acta 894: 198–208) and Genty B et al. 1989 (Biochim Biophys Acta 990: 87–92). These were applied to data from different species representing different states of light acclimation, to species with C₃ or C₄ photosynthesis, and to wild-type and a chlorophyll b-less chlorina mutant of barley. Photosynthesis measured as CO₂-saturated O₂ evolution and modulated fluorescence were simultaneously monitored over a range of photon flux densities. The quantum yields of O₂ evolution (ØO₂) were based on absorbed photons, and the fluorescence parameters for photochemical (q_p) and non-photochemical (q_N) quenching, as well as the ratio of variable fluorescence to maximum fluorescence during steady-state illumination (F'_v/F'_m), were determined. In accordance with the Weis and Berry model, most plants studied exhibited an approximately linear relationship between ØO₂/q_p (i.e., the yield of O₂ evolution by open Photosystem II reaction centres) and q_N, except for wild-type barley that showed a non-linear relationship. In contrast to the linear relationship reported by Genty et al. for q_p×F'_v/F'_m (i.e., the quantum yield of Photosystem II electron transport) and ØCO₂, we found a non-linear relationship between q_p×F'_v/F'_m and ØO₂ for all plants, except for the chlorina mutant of barley, which showed a largely linear relationship. The curvilinearity of wild-type barley deviated somewhat from that of other species tested. The non-linear part of the relationship was confined to low, limiting photon flux densities, whereas at higher light levels the relationship was linear. Photoinhibition did not change the overall shape of the relationship between q_p×F'_v/F'_m and ØO₂ except that the maximum values of the quantum yields of Photosystem II electron transport and photosynthetic O₂ evolution decreased in proportion to the degree of photoinhibition. This implies that the quantum yield of Photosystem II electron transport under high light conditions may be similar for photoinhibited and non-inhibited plants. Based on our experimental results and theoretical analyses of photochemical and non-photochemical fluoresce quenching processes, we conclude that both models, although not universal for all plants, provide useful means for the prediction of photosynthesis from fluorescence parameters. However, we also discuss that conditions which alter one or more of the rate constants that determine the various fluorescence parameters, as well as differential light penetration in assays for oxygen evolution and fluorescence emission, may have direct effect on the relationships of the two models.Abbreviations F₀ and F'₀ fluorescence when all Photosystem II reaction centres are open in dark- and light-acclimated leaves, respectively - F_m and F'_m fluorescence when all Photosystem II reaction centres are closed in dark and light, respectively - F_v variable fluorescence equal to F_m-F₀ - F_s steady state level of fluorescence in light - F'_v and F'_m variable (F'_m-F'₀) and maximum fluorescence under steady state light conditions - HEPES N-2-hydroxyethylpiperazine-N-2-ethane-sulphonic acid - Q_A the primary, stabile quinone acceptor of Photosystem II - q_N non-photochemical quenching of fluorescence - q_p photochemical quenching of fluorescence - ØO₂ quantum yield of CO₂-saturated O₂ evolution based on absorbed photons     

Investigation of the CO(2) Dependence of Quantum Yield and Respiration in Eucalyptus pauciflora

In leaves of C₃ plants, the rate of nonphotorespiratory respiration appears to be higher in darkness than in the light. This change from a high to a low rate of carbon loss with increasing photon flux density leads to an increase in the apparent quantum yield of photosynthetic CO₂ assimilation at low photon flux densities (Kok effect). The mechanism of this suppression of nonphotorespiratory respiration is not understood, but biochemical evidence and the observation that a Kok effect is often not observed under low O₂, has led to the suggestion that photorespiration might be involved in some way. This hypothesis was tested with snowgum (Eucalyptus pauciflora Sieb. ex Spreng.) using gas exchange methods. The test was based on the assumption that if photorespiration were involved, then it would be expected that the intercellular partial pressure of CO₂ would also have an influence on the Kok effect. Under normal atmospheric levels of CO₂ and O₂, a Kok effect was found. Changing the intercellular partial pressure of CO₂, however, did not affect the estimate of nonphotorespiratory respiraton, and it was concluded that its decrease with increasing photon flux density did not involve photorespiration. Concurrent measurements showed that the quantum yield of net assimilation of CO₂ increased with increasing intercellular partial pressure of CO₂, and this increase agreed closely with predictions based on recent models of photosynthesis.     

Photoacoustic and fluorescence measurements of the chilling response and their relationship to carbon dioxide uptake in tomato plants

The response of tomato plants to various chilling treatments was studied using two approaches for the measurement of photosynthetic activity. One involved the use of a portable fluorometer for the measurement of in-vivo chlorophyll fluorescence, while the other employed a newly introduced photoacoustic system which allowed changes in oxygen evolution to be followed in a leaf disc. A strong correlation was found between results obtained by each system and those obtained by a conventional open gas-exchange system for the determination of CO₂ uptake. Both systems of measurements could readily distinguish between the effects of chilling in the dark (at 3° C for 18 h) and chilling at high photon flux density (2000 mol m^-2 s^-1 for 5h at 5° C). Chilling in the dark had practically no effect on the quantum yield of oxygen evolution, chlorophyll fluorescence or CO₂ uptake, while chilling at excessively high photon flux density resulted in a sharp reduction (50–70%) in the quantum yields obtained. The results support the view that photosystem II cannot be the primary site of damage by chilling in the dark, although it is significantly affected by chilling at high light intensity.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - PA photoacoustic - PFD photon flux density - PSII photosystem II     

Effects of light intensity and temperature on the photosynthetic irradiance response curves and chlorophyll fluorescence in three picocyanobacterial strains of Synechococcus

Chrococcoid cyanobacteria of the genus Synechococcus are the important component of marine and freshwater ecosystems. Picocyanobacteria comprise even 80% of total cyanobacterial biomass and contribute to 50% of total primary cyanobacterial bloom production. Chlorophyll (Chl) fluorescence and photosynthetic light response (P-I) curves are commonly used to characterize photoacclimation of Synechococcus strains. Three brackish, picocyanobacterial strains of Synechococcus (BA-132, BA-124, BA-120) were studied. They were grown under 4 irradiances [10, 55, 100, and 145 μmol(photon) m^?2 s^?1] and at 3 temperatures (15, 22.5, and 30°C). Photosynthetic rate was measured by Clark oxygen electrode, whereas the Chl fluorescence was measured using Pulse Amplitude Modulation fluorometer. Based on P-I, two mechanisms of photoacclimation were recognized in Synechococcus. The maximum value of maximum rate of photosynthesis (P _max) expressed per biomass unit at 10 μmol(photon) m^?2 s^?1 indicated a change in the number of photosynthetic units (PSU). The constant values of initial slope of photosynthetic light response curve (α) and the maximum value of P _max expressed per Chl unit at 145 μmol(photon) m^?2 s^?1 indicated another mechanism, i.e. a change in PSU size. These two mechanisms caused changes in photosynthetic rate and its parameters (compensation point, α, saturation irradiance, dark respiration, P _max) upon the influence of different irradiance and temperature. High irradiance had a negative effect on fluorescence parameters, such as the maximum quantum yield and effective quantum yield of PSII photochemistry (φ_PSII), but it was higher in case of φ_PSII.     

Oxygen dependence of photoinhibition at low temperature in intact protoplasts of Valerianella locusta L.

Photoinhibition of photosynthesis in vivo is shown to be considerably promoted by O₂ under circumstances where energy turnover by photorespiration and photosynthetic carbon metabolism are low. Intact protoplasts of Valerianella locusta L. were photoinhibited by 30 min irradiation with 3000 mol photons · m^–2 · s^–1 at 4° C in saturating [CO₂] at different oxygen concentrations, corresponding to 2–40% O₂ in air. The photoinhibition of light-limited CO₂-dependent photosynthetic O₂ evolution increased with increasing oxygen concentration. The uncoupled photochemical activity of photosystem II, measured in the presence of the electron acceptor 1,4-benzoquinone, and maximum variable fluorescence, F_v, were strongly affected and this inhibition was closely correlated to the O₂ concentration. The effect of O₂ did not saturate at the highest concentrations applied. An increase in photoinhibitory fluorescence quenching with [O₂], although less pronounced than in protoplasts, was also observed with intact leaves irradiated at 4° C in air. Initial fluorescence, F_o, was slightly (about 10%) increased by the inhibitory treatments but not influenced by [O₂]. A long-term cold acclimation of the plants did not substantially alter the O₂-sensitivity of the protoplasts under the high-light treatment. From these experiments we conclude that oxygen is involved in the photoinactivation of photosystem II by excess light in vivo.Abbreviations and Symbols Chl chlorophyll - F_o initial fluorescence - F_M maximum fluorescence - F_v maximum variable fluorescence - PCO photorespiratory carbon oxidation - PCR photosynthetic carbon reduction - PFD photon flux density - q_N non-photochemical quenching - q_P photochemical quenching - _S quantum efficiency of electron transport of photosystem II This study was financially supported by the Deutsche Forschungs-gemeinschaft (SFB 189) and the Foundation for Fundamental Biological Research (BION), which is subsidised by the Netherlands Organization for the Advancement of Pure Research (NWO).     

Effects of Irradiance on the in Vivo CO(2):O(2) Specificity Factor in Tobacco Using Simultaneous Gas Exchange and Fluorescence Techniques

The effects of gas phase O₂ concentration (1%, 20.5%, and 42.0%, v/v) on the quantum yield of net CO₂ fixation and fluorescence yield of chlorophyll a are examined in leaf tissue from Nicotiana tabacum at normal levels of CO₂ and 25 to 30°C. Detectable decreases in nonphotochemical quenching of absorbed excitation occurred at the higher O₂ levels relative to 1% O₂ when irradiance was nearly or fully saturating for photosynthesis. Photochemical quenching was increased by high O₂ levels only at saturating irradiance. Simultaneous measurements of CO₂ and H₂O exchange and fluorescence yield permit estimation of partitioning of linear photosynthetic electron transport between net CO₂ fixation and O₂-dependent, dissipative processes such as photorespiration as a function of leaf internal CO₂ concentration. Changes in the in vivo CO₂:O₂ `specificity factor' (K_sp) with increasing irradiance are examined. The magnitude K_sp was found to decline from a value of 85 at moderate irradiance to 68 at very low light, and to 72 at saturating photon flux rates. The results are discussed in terms of the applicability of the ribulose bisphosphate carboxylase/oxygenase enzyme model to photosynthesis in vivo.     

16O2/18O2 analysis of oxygen exchange in Dunaliella tertiolecta. Evidence for the inhibition of mitochondrial respiration in the light

A mass spectrometric ¹⁶O₂/¹⁸O₂-isotope technique was used to analyse the rates of gross O₂ evolution, net O₂ evolution and gross O₂ uptake in relation to photon fluence rate by Dunaliella tertiolecta adapted to 0.5, 1.0, 1.5, 2.0 and 2.5 M NaCl at 25°C and pH 7.0.At concentrations of dissolved inorganic carbon saturating for photosynthesis (200 M) gross O₂ evolution and net O₂ evolution increased with increasing salinity as well as with photon fluence rate. Light compensation was also enhanced with increased salinities. Light saturation of net O₂ evolution was reached at about 1000 mol m^-2s^-1 for all salt concentrations tested. Gross O₂ uptake in the light was increased in relation to the NaCl concentration but it was decreased with increasing photon fluence rate for almost all salinities, although an enhanced flow of light generated electrons was simultaneously observed. In addition, a comparison between gross O₂ uptake at 1000 mol photons m^-2s^-1, dark respiration before illumination and immediately after darkening of each experiment showed that gross O₂ uptake in the light paralleled but was lower than mitochondrial O₂ consumption in the dark.From these results it is suggested that O₂ uptake by Dunaliella tertiolecta in the light is mainly influenced by mitochondrial O₂ uptake. Therefore, it appears that the light dependent inhibition of gross O₂ uptake is caused by a reduction in mitochondrial O₂ consumption by light.Abbreviations DCMU 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea - DHAP dihydroxy-acetonephosphate - DIC dissolved inorganic carbon - DR_a rate of dark respiration immediately after illumination - DR_b rate of dark respiration before illumination - E₀ rate of gross oxygen evolution in the light - NET rate of net oxygen evolution in the light - PFR photon fluence rate - RubP rubulose-1,5-bisphosphate - SHAM salicyl hydroxamic acid - U₀ rate of gross oxygen uptake in the light     

Photoinhibition in Vitis californica: The Role of Temperature during High-Light Treatment

Leaves of Vitis californica Benth. (California wild grape) exposed to a photon flux density (PFD) equivalent to full sun exhibited temperature-dependent reductions in the rates or efficiencies of component photosynthetic processes. During high-PFD exposure, net CO₂ uptake, photon yield of oxygen evolution, and photosystem II chlorophyll fluorescence at 77 Kelvin (F_m, F_v, and F_v/F_m) were more severely inhibited at high and low temperatures than at intermediate temperatures. Sun leaves tolerated high PFD more than growth chamber-grown leaves but exhibited qualitatively similar temperature-dependent responses to high-PFD exposures. Photosystem II fluorescence and net CO₂ uptake exhibited different sensitivities to PFD and temperature. Fluorescence and gas exchange kinetics during exposure to high PFD suggested an interaction of multiple, temperature-dependent processes, involving both regulation of energy distribution and damage to photosynthetic components. Comparison of F_v/F_m to photon yield of oxygen evolution yielded a single, curvilinear relationship, regardless of growth condition or treatment temperature, whereas the relationship between F_m (or F_v) and photon yield varied with growth conditions. This indicated that F_v/F_m was the most reliable fluorescence indicator of PSII photochemical efficiency for leaves of different growth conditions and treatments.     

Low growth temperature effects a differential inhibition of photosynthesis in spring and winter wheat

In vivo room temperature chlorophyll a fluorescence coupled with CO₂ and O₂ exchange was measured to determine photosynthetic limitation(s) for spring and winter wheat (Triticum aestivum L.) grown at cold-hardening temperatures (5°C/5°C, day/night). Plants of comparable physiological stage, but grown at nonhardening temperatures (20°C/16°C, day/night) were used in comparison. Winter wheat cultivars grown at 5°C had light-saturated rates of CO₂ exchange and apparent photon yields for CO₂ exchange and O₂ evolution that were equal to or greater than those of winter cultivars grown at 20°C. In contrast, spring wheat cultivars grown at 5°C showed 35% lower apparent photon yields for CO₂ exchange and 25% lower light-saturated rates of CO₂ exchange compared to 20°C grown controls. The lower CO₂ exchange capacity is not associated with a lower efficiency of photosystem II activity measured as either the apparent photon yield for O₂ evolution, the ratio of variable to maximal fluorescence, or the level of reduced primary quinone electron acceptor maintained at steady-state photosynthesis, and is most likely associated with carbon metabolism. The lower CO₂ exchange capacity of the spring cultivars developed following long-term exposure to low temperature and did not occur following over-night exposure of nonhardened plants to 5°C.     

Thermal responses of Symbiodinium photosynthetic carbon assimilation

The symbiosis between hermatypic corals and their dinoflagellate endosymbionts, genus Symbiodinium, is based on carbon exchange. This symbiosis is disrupted by thermally induced coral bleaching, a stress response in which the coral host expels its algal symbionts as they become physiologically impaired. The disruption of the dissolved inorganic carbon (DIC) supply or the thermal inactivation of Rubisco have been proposed as sites of initial thermal damage that leads to the bleaching response. Symbiodinium possesses a highly unusual Form II ribulose bisphosphate carboxylase/oxygenase (Rubisco), which exhibits a lower CO₂:O₂ specificity and may be more thermally unstable than the Form I Rubiscos of other algae and land plants. Components of the CO₂ concentrating mechanism (CCM), which supplies inorganic carbon for photosynthesis, may also be temperature sensitive. Here, we examine the ability of four cultured Symbiodinium strains to acquire and fix DIC across a temperature gradient. Surprisingly, the half-saturation constant of photosynthesis with respect to DIC concentration (K _P), an index of CCM function, declined with increasing temperature in three of the four strains, indicating a greater potential for photosynthetic carbon acquisition at elevated temperatures. In the fourth strain, there was no effect of temperature on K _P. Finding no evidence for thermal inhibition of the CCM, we conclude that CCM components are not likely to be the primary sites of thermal damage. Reduced photosynthetic quantum yields, a hallmark of thermal bleaching, were observed at low DIC concentrations, leaving open the possibility that reduced inorganic carbon availability is involved in bleaching.     

An inter-basin comparison of nutrient limitation and the irradiance response of pulse-amplitude modulated (PAM) fluorescence in Lake Erie phytoplankton

This research examined the application of the maximum quantum efficiency (F _v/F _m) and relative electron transport rate versus irradiance curves (rETR) as a rapid, sensitive assessment of Lake Erie phytoplankton nutrient status. I evaluated the potential benefits of the variable fluorescence parameters by comparing these parameters with chemical and physiological nutrient status assays. I tested the hypothesis that F _v/F _m and rETR curves could diagnose nutrient status in natural lake phytoplankton and be capable of discriminating which inorganic nutrient is limited temporally and spatially. F _v/F _m was on average highest in the more eutrophic west basin (WB) and lowest in the more oligotrophic central basin (CB). According to the chemical and physiological indicators, P deficiency was most severe in the CB during summer stratification and N deficiency was strongest in the WB during isothermal conditions. Like F _v/F _m, rETR at light saturation (rETR_max) and the initial slope of the rETR versus irradiance curve (α) decreased as the severity of N and P deficiency increased. Amendment with N or P stimulated increased F _v/F _m, rETR_max, and α in N- and P-limited samples, respectively, and abolished the photoinhibition apparent in rETR curves of nutrient-limited samples. These results supported the view that the N and P deficiency assays, and corresponding variations of variable fluorescence parameters, were valid indicators of widely variable N and P deficiency in the phytoplankton, and could be used to provide a promising tool in determining phytoplankton nutrient status. Contrary to my hopes, it did not appear that rETR–irradiance curves could discriminate between N and P deficiency. Identification of the most limiting nutrient still demanded additional information beyond the variable fluorescence measurements.     

Photorespiration and Carbon Limitation Determine Productivity in Temperate Seagrasses

The gross primary productivity of two seagrasses, Zostera marina and Ruppia maritima, and one green macroalga, Ulva intestinalis, was assessed in laboratory and field experiments to determine whether the photorespiratory pathway operates at a substantial level in these macrophytes and to what extent it is enhanced by naturally occurring shifts in dissolved inorganic carbon (DIC) and O₂ in dense vegetation. To achieve these conditions in laboratory experiments, seawater was incubated with U. intestinalis in light to obtain a range of higher pH and O₂ levels and lower DIC levels. Gross photosynthetic O₂ evolution was then measured in this pretreated seawater (pH, 7.8–9.8; high to low DIC:O₂ ratio) at both natural and low O₂ concentrations (adjusted by N₂ bubbling). The presence of photorespiration was indicated by a lower gross O₂ evolution rate under natural O₂ conditions than when O₂ was reduced. In all three macrophytes, gross photosynthetic rates were negatively affected by higher pH and lower DIC. However, while both seagrasses exhibited significant photorespiratory activity at increasing pH values, the macroalga U. intestinalis exhibited no such activity. Rates of seagrass photosynthesis were then assessed in seawater collected from the natural habitats (i.e., shallow bays characterized by high macrophyte cover and by low DIC and high pH during daytime) and compared with open baymouth water conditions (where seawater DIC is in equilibrium with air, normal DIC, and pH). The gross photosynthetic rates of both seagrasses were significantly higher when incubated in the baymouth water, indicating that these grasses can be significantly carbon limited in shallow bays. Photorespiration was also detected in both seagrasses under shallow bay water conditions. Our findings indicate that natural carbon limitations caused by high community photosynthesis can enhance photorespiration and cause a significant decline in seagrass primary production in shallow waters.     

Differential Control of Xanthophylls and Light-Induced Stress Proteins, as Opposed to Light-Harvesting Chlorophyll a/b Proteins, during Photosynthetic Acclimation of Barley Leaves to Light Irradiance

Barley (Hordeum vulgare L.) plants were grown at different photon flux densities ranging from 100 to 1800 μmol m⁻² s⁻¹ in air and/or in atmospheres with reduced levels of O₂ and CO₂. Low O₂ and CO₂ partial pressures allowed plants to grow under high photosystem II (PSII) excitation pressure, estimated in vivo by chlorophyll fluorescence measurements, at moderate photon flux densities. The xanthophyll-cycle pigments, the early light-inducible proteins, and their mRNA accumulated with increasing PSII excitation pressure irrespective of the way high excitation pressure was obtained (high-light irradiance or decreased CO₂ and O₂ availability). These findings indicate that the reduction state of electron transport chain components could be involved in light sensing for the regulation of nuclear-encoded chloroplast gene expression. In contrast, no correlation was found between the reduction state of PSII and various indicators of the PSII light-harvesting system, such as the chlorophyll a-to-b ratio, the abundance of the major pigment-protein complex of PSII (LHCII), the mRNA level of LHCII, the light-saturation curve of O₂ evolution, and the induced chlorophyll-fluorescence rise. We conclude that the chlorophyll antenna size of PSII is not governed by the redox state of PSII in higher plants and, consequently, regulation of early light-inducible protein synthesis is different from that of LHCII.     

INORGANIC CARBON LIMITATION OF PHOTOSYNTHESIS IN ULVA ROTUNDATA (CHLOROPHYTA)1

A computerized oxygen electrode Astern was used to make rapid and accurate measurements of photosynthetic light and dissolved inorganic carbon (DIC) response cures with a macroalga. Ulva rotundata Blid. was grown in an outdoor, continuous flow system in seawater under sunlight or 9% of sunlight at Beaufort, North Carolina. The light compensation points in the shade- and sun-grown plants, measured in seawater, were at photon flux densities (PFDs) of 16 and 27 μmol. Photons·m^?2·s^?1, respectively but the quantum yield of O₂ evolution was not significantly different. Rates of photosynthesis in seawater per unit area of thallus under saturating light and rates of dark respiration were about 1.5-fold higher in sun- than in shade-grown plants. The concentration of DIC in seawater (approximately 2 mM) limited photosynthesis at absorbed PFDs above 60–70 μmol photons·m^?2·s^?1 Addition of 20 mM inorganic carbon had no effect on quantum yield but caused about a 1.5-fold increase in the light-saturated photosynthetic rate in both shade- and sun-grown Ulva. The effect of DIC supplementation was greatest in plants grown in October and least in plants grown in June. The light- and DIC-saturated rate of photosynthesis in seawater was similar to the maximum rate obtained by exposing Ulva to 10% CO₂, in the gas phase. The carbon isotope values (δ¹³C, reflecting the ¹³C/¹²C ratio compared to a standard) of Ulva grown in the same seawater supply were dependent on light and agitation. Samples from Beaufort Inlet were more negative (δ¹³C value, ?20.03‰) than those grown in bright light with agitation (δ¹³C value, ?17.78‰ outdoors; ?17.23‰ indoors), which may indicate DIC supply limited carbon uptake in seawater.