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Osmotic adjustment, symplast volume, and nonstomatally mediated water stress inhibition of photosynthesis in wheat.

At low water potential (psi(w)), dehydration reduces the symplast volume of leaf tissue. The effect of this reduction on photosynthetic capacity was investigated. The influence of osmotic adjustment on this relationship was also examined. To examine these relationships, comparative studies were undertaken on two wheat cultivars, one that osmotically adjusts in response to water deficits (;Condor'), and one that lacks this capacity (;Capelle Desprez'). During a 9-day stress cycle, when water was withheld from plants grown in a growth chamber, the relative water content of leaves declined by 30% in both cultivars. Leaf osmotic potential (psi(s)) declined to a greater degree in Condor plants. Measuring psi(s) at full turgor indicated that osmotic adjustment occurred in stressed Condor, but not in Capelle plants. Two methods were used to examine the degree of symplast (i.e. protoplast) volume reduction in tissue rapidly equilibrated to increasingly low psi(w). Both techniques gave similar results. With well-watered plants, symplast volume reduction from the maximum (found at high psi(w) for each cultivar) was the same for Condor and Capelle. After a stress cycle, volume was maintained to a greater degree at low psi(w) in Condor leaf tissue than in Capelle. Nonstomatally controlled photosynthesis was inhibited to the same degree at low psi(w) in leaf tissue prepared from well-watered Condor and Capelle plants. However, photosynthetic capacity was maintained to a greater degree at low psi(w) in tissue prepared from stressed Condor plants than in tissue from stressed Capelle plants. Net CO(2) uptake in attached leaves was monitored using an infrared gas analyzer. These studies indicated that in water stressed plants, photosynthesis was 106.5% higher in Condor than Capelle at ambient [CO(2)] and 21.8% higher at elevated external [CO(2)]. The results presented in this report were interpreted as consistent with the hypothesis that there is a causal association between protoplast (and presumably chloroplast) volume reduction at low psi(w) and low psi(w) inhibition of photosynthesis. Also, the data indicate that osmotic adjustment allows for maintenance of relatively greater volume at low psi(w), thus reducing low psi(w) inhibition of chloroplast photosynthetic potential.

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Regulation of photosynthesis in nitrogen deficient wheat seedlings.

Nitrogen effects on the regulation of photosynthesis in wheat (Triticum aestivum L., cv Remia) seedlings were examined. Ribulose 1,5-bisphosphate carboxylase/oxygenase was rapidly extracted and tested for initial activity and for activity after incubation in presence of CO(2) and Mg(2+). Freeze clamped leaf segments were extracted for determinations of foliar steady state levels of ribulose 1,5-bisphosphate, triose phosphate, 3-phosphoglycerate, ATP, and ADP. Nitrogen deficient leaves showed increased ATP/ADP and triose phosphate/3-phosphoglycerate ratios suggesting increased assimilatory power. Ribulose 1,5-bisphosphate levels were decreased due to reduced pentose phosphate reductive cycle activity. Nevertheless, photosynthesis appeared to be limited by ribulose 1,5-bisphosphate carboxylase/oxygenase, independent of nitrogen nutrition. Its degree of activation was increased in nitrogen deficient plants and provided for maximum photosynthesis at decreased enzyme protein levels. It is suggested that ribulose 1,5-bisphosphate carboxylase/oxygenase activity is regulated according to the amount of assimilatory power.

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Depression of Photosynthesis, Growth, and Yield in Field-Grown Green Pepper (Capsicum annuum L.) Exposed to Acidic Fog and Ambient Ozone.

The relationship among physiological, injury, growth, and yield responses was examined in field-grown green pepper (Capsicum annuum L. ;California Wonder') subjected to two airborne environmental stresses. The primary objectives were to determine if the stresses could cause alterations in the plant responses, and to determine if any stress induced alterations in physiological or injury responses were correlated with effects on growth or yield. Responses were monitored in green pepper exposed to simulated acidic fog alone, or in combination with ambient concentrations of ozone in open-top field chambers. Both highly acidic fog and ambient ozone depressed green pepper growth and yield responses via the inhibition of photosynthesis. Applications of highly acidic fog (i.e. two exposures of pH 1.68 fog per week for 11 weeks) caused a significant depression of net photosynthesis, reduction in leaf buffering capacity, and an extensive amount of leaf injury. These alterations closely paralleled decreases in growth and yield on a percentage basis. In contrast, ambient ozone had similar impacts on net photosynthesis, growth and yield, but enhanced leaf buffering capacity, and caused no visible injury. The pollutant-specific differences in plant response are discussed with respect to whole-plant carbon metabolism and physiological compensation.

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Determination of the Rate Limiting Step for Photosynthesis in a Nearly Isonuclear Rapeseed (Brassica napus L.) Biotype Resistant to Atrazine.

Plant biotypes that are resistant to S-triazines under most conditions often grow less vigorously and have lower quantum yields and lower maximum rates of photosynthesis. The photosynthetic reactions responsible for these effects were identified in whole leaves and thylakoids of nearly isonuclear lines of oilseed rape (Brassica napus L.). The lower quantum yield was a result of poor efficiency in the use of separated charge at the photosystem II reaction center. Charge separation occurred normally, but over 30% of the charges recombined instead of being used for oxygen evolution and for reduction capacity in photosystem I. The lower maximum rate of photosynthesis in the resistant biotype was set by the transfer of electrons between the primary, Q(A), and secondary, Q(B), acceptors of photosystem II. This charge transfer reaction became rate limiting in resistant biotypes. The decreased quantum yield and decreased maximum rate of photosynthesis are both believed to be consequences of changes in the 32 kilodalton herbicide binding protein. As such, it is likely that these traits will not be genetically separable.

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Mild water stress effects on carbon-reduction-cycle intermediates, ribulose bisphosphate carboxylase activity, and spatial homogeneity of photosynthesis in intact leaves.

We have examined the effect of mild water stress on photosynthetic chloroplast reactions of intact Phaseolus vulgaris leaves by measuring two parameters of ribulose bisphosphate (RuBP) carboxylase activity and the pool sizes of RuBP, 3-phosphoglycerate (PGA), triose phosphates, hexose monophosphates, and ATP. We also tested for patchy stomatal closure by feeding (14)CO(2). The k(cat) of RuBP carboxylase (moles CO(2) fixed per mole enzyme per second) which could be measured after incubating the enzyme with CO(2) and Mg(2+) was unchanged by water stress. The ratio of activity before and after incubation with CO(2) and Mg(2+) (the carbamylation state) was slightly reduced by severe stress but not by mild stress. Likewise, the concentration of RuBP was slightly reduced by severe stress but not by mild stress. The concentration of PGA was markedly reduced by both mild and severe water stress. The concentration of triose phosphates did not decline as much as PGA. We found that photosynthesis in water stressed leaves occurred in patches. The patchiness of photosynthesis during water stress may lead to an underestimation of the effect of stomatal closure. We conclude that reductions in whole leaf photosynthesis caused by mild water stress are primarily the result of stomatal closure and that there is no indication of damage to chloroplast reactions.

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Low oxygen inhibition of photosynthesis is caused by inhibition of starch synthesis.

Photosynthesis of C(3) plants is occasionally inhibited upon switching from normal to low partial pressure of O(2). Leaves of Solanum tuberosum exhibited this effect reproducibly under saturating light and 700 microbars of CO(2). We determined the partitioning of recent photosynthate between starch and sucrose and measured the concentration of hexose monophosphates in the stroma and cytosol after nonaqueous fractionation. The reduction in the rate of photosynthesis upon switching to low partial pressure of O(2) was caused by reduced starch synthesis. The concentration of hexose monophosphates in the stroma fell and the glucose 6-phosphate to fructose 6-phosphate to fructose 6-phosphate ratio fell from 2.7 to 1.3, indicating an inhibition of phosphoglucoisomerase as described by K-J Dietz ([1985] Biochim Biophys Acta 839: 240-248). The concentration of hexose monophosphates in the cytosol increased, ruling out a sucrose synthesis limitation by reduced transport from the chloroplast as the explanation for low O(2) inhibition of photosynthesis.

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Salinity effects on photosynthesis in isolated mesophyll cells of cowpea leaves.

Mesophyll cells from leaves of cowpea (Vigna unquiculata [L.] Walp.) plants grown under saline conditions were isolated and used for the determination of photosynthetic CO(2) fixation. Maximal CO(2) fixation rate was obtained when the osmotic potential of both cell isolation and CO(2) fixation assay media were close to leaf osmotic potential, yielding a zero turgor pressure. Hypotonic and hypertonic media decreased the rate of photosynthesis regardless of the salinity level during plant growth. No decrease in photosynthesis was obtained for NaCl concentrations up to 87 moles per cubic meter in the plant growing media and only a 30% decrease was found at 130 moles per cubic meter when the osmotic potential of cell isolation and CO(2) fixation media were optimal. The inhibition was reversible when stress was relieved. At 173 moles per cubic meter NaCl, photosynthesis was severely and irreversibly inhibited. This inhibition was attributed to toxic effects caused by high Cl(-) and Na(+) accumulation in the leaves. Uptake of sorbitol by intact cells was insignificant, and therefore not associated with cell volume changes. The light response curve of cells from low salinity grown plants was similar to the controls. Cells from plants grown at 173 moles per cubic meter NaCl were light saturated at a lower radiant flux density than were cells from lower salinity levels.

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Stromal Phosphate Concentration Is Low during Feedback Limited Photosynthesis.

It has been hypothesized that photosynthesis can be feedback limited when the phosphate concentration cannot be both low enough to allow starch and sucrose synthesis at the required rate and high enough for ATP synthesis at the required rate. We have measured the concentration of phosphate in the stroma and cytosol of leaves held under feedback conditions. We used non-aqueous fractionation techniques with freeze-clamped leaves of Phaseolus vulgaris plants grown on reduced phosphate nutrition. Feedback was induced by holding leaves in low O(2) or high CO(2) partial pressure. We found 7 millimolar phosphate in the stroma of leaves in normal oxygen but just 2.7 millimolar phosphate in leaves held in low oxygen. Because 1 to 2 millimolar phosphate in the stroma may be metabolically inactive, we estimate that in low oxygen, the metabolically active pool of phosphate is between negligible and 1.7 millimolar. We conclude that halfway between these extremes, 0.85 millimolar is a good estimate of the phosphate concentration in the stroma of feedback-limited leaves and that the true concentration could be even lower. The stromal phosphate concentration was also low when leaves were held in high CO(2), which also induces feedback-limited photosynthesis, indicating that the effect is related to feedback limitation, not to low oxygen per se. We conclude that the concentration of phosphate in the stroma is usually in excess and that it is sequestered to regulate photosynthesis, especially starch synthesis. The capacity for this regulation is limited by the coupling factor requirement for phosphate.

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Carbonic anhydrase activity in leaves and its role in the first step of c(4) photosynthesis.

In C(4) plants carbonic anhydrase catalyzes the critical first step of C(4) photosynthesis, the hydration of CO(2) to bicarbonate. The maximum activity of this enzyme in C(4) leaf extracts, measured by H(+) production with saturating CO(2) and extrapolated to 25 degrees C, was found to be 3,000 to 10,000 times the maximum photosynthesis rate for these leaves. Similar activities were found in C(3) leaf extracts. However, the calculated effective activity of this enzyme at in vivo CO(2) concentrations was apparently just sufficient to prevent the rate of conversion of CO(2) to HCO(3) (-) from limiting C(4) photosynthesis. This conclusion was supported by the mass spectrometric determination of leaf carbonic anhydrase activities.

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Cooperative regulation of cytoplasmic streaming and ca fluxes by pfr and photosynthesis in vallisneria mesophyll cells.

In mesophyll cells of Vallisneria gigantea Graebner, Ca(2+) regulates the induction and cessation of cytoplasmic streaming. Streaming is induced when the level of calcium in the cytoplasm is lowered through light-accelerated release of Ca(2+) from the cells (S Takagi, R Nagai [1988] Plant Physiol 88: 228-232). We have now initiated an investigation on the nature of the photoreceptor(s) that are involved in the regulation of Ca(2+) movements across the cell membrane and of streaming. Streaming is induced only when phytochrome exists in the phytochrome-far redabsorbing form (Pfr)-and photosynthesis is allowed to take place for at least 4 minutes. The former effect is typically photoreversible by red and far-red light, and phytochrome is spectro-photometrically detectable in the crude extract from the leaves. The latter effect is assessed in terms of the wavelength dependency and the effects of diuron and atrazine, two inhibitors of photosynthesis. A similar requirement for Pfr and photosynthesis is found to be associated with the acceleration of Ca(2+) efflux in the protoplasts. The results suggest that phytochrome and photosynthetic pigment(s) cooperatively regulate cytoplasmic streaming via modulation of the Ca(2+) transport in the cell membrane.

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Rapid Modulation of Spinach Leaf Nitrate Reductase Activity by Photosynthesis : I. Modulation in Vivo by CO(2) Availability.

It has been shown recently that in spinach leaves (Spinacia oleracea) net photosynthesis and nitrate reduction are closely linked: when net photosynthesis was low because of stomatal closure, rates of nitrate reduction decreased (WM Kaiser, J Förster [1989] Plant Physiol 91: 970-974). Here we present evidence that photosynthesis regulates nitrate reduction by modulating nitrate reductase activity (NRA, EC 1.6.6.1). When spinach leaves were exposed to low CO(2) in the light, extractable NRA declined rapidly with a half-time of 15 minutes. The inhibition was rapidly reversed when leaves were brought back to air. NRA was also inhibited when leaves were wilted in air; this inhibition was due to decreased CO(2) supply as a consequence of stomatal closure. The modulation of NRA was stable in vitro. It was not reversed by gel filtration. In contrast, the in vitro inhibition of nitrate reductase (NR) by classical inhibitors such as cyanide, hydroxylamin, or NADH disappeared after removal of free inhibitors by gel filtration. The negative modulation of NRA in -CO(2)-treated leaves became manifest as a decrease in total enzyme activity only in the presence of free Mg(2+) or Ca(2+). Mg(2+) concentrations required for observing half-maximal inhibition were about 1 millimolar. In the presence of EDTA, the enzyme activity was always high and rather independent of the activation status of the enzyme. NRA was also independent of the pH in the range from pH 7 to pH 8, at saturating substrate and Mg(2+) concentrations. The apparent substrate affinities of NR were hardly affected by the in vivo modulation of NR. Only V(max) changed.

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Reduction of ribulose-1,5-bisphosphate carboxylase/oxygenase content by antisense RNA reduces photosynthesis in transgenic tobacco plants.

A complementary DNA for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was cloned from tobacco (Nicotiana tabacum) and fused in the antisense orientation to the cauliflower mosaic virus 35S promoter. This antisense gene was introduced into the tobacco genome, and the resulting transgenic plants were analyzed to assess the effect of the antisense RNA on Rubisco activity and photosynthesis. The mean content of extractable Rubisco activity from the leaves of 10 antisense plants was 18% of the mean level of activity of control plants. The soluble protein content of the leaves of anti-small subunit plants was reduced by the amount equivalent to the reduction in Rubisco. There was little change in phosphoribulokinase activity, electron transport, and chlorophyll content, indicating that the loss of Rubisco did not affect these other components of photosynthesis. However, there was a significant reduction in carbonic anhydrase activity. The rate of CO(2) assimilation measured at 1000 micromoles quanta per square meter per second, 350 microbars CO(2), and 25 degrees C was reduced by 63% (mean value) in the antisense plants and was limited by Rubisco activity over a wide range of intercellular CO(2) partial pressures (p(i)). In control leaves, Rubisco activity only limited the rate of CO(2) assimilation below a p(i) of 400 microbars. Despite the decrease in photosynthesis, there was no reduction in stomatal conductance in the antisense plants, and the stomata still responded to changes in p(i). The unchanged conductance and lower CO(2) assimilation resulted in a higher p(i), which was reflected in greater carbon isotope discrimination in the leaves of the antisense plants. These results suggest that stomatal function is independent of total leaf Rubisco activity.

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Theoretical Considerations when Estimating the Mesophyll Conductance to CO(2) Flux by Analysis of the Response of Photosynthesis to CO(2).

The conductance for CO(2) diffusion in the mesophyll of leaves can limit photosynthesis. We have studied two methods for determining the mesophyll conductance to CO(2) diffusion in leaves. We generated an ideal set of photosynthesis rates over a range of partial pressures of CO(2) in the stroma and studied the effect of altering the mesophyll diffusion conductance on the measured response of photosynthesis to intercellular CO(2) partial pressure. We used the ideal data set to test the sensitivity of the two methods to small errors in the parameters used to determine mesophyll conductance. The two methods were also used to determine mesophyll conductance of several leaves using measured rather than ideal data sets. It is concluded that both methods can be used to determine mesophyll conductance and each method has particular strengths. We believe both methods will prove useful in the future.

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Dark Respiration Protects Photosynthesis Against Photoinhibition in Mesophyll Protoplasts of Pea (Pisum sativum).

The optimal light intensity required for photosynthesis by mesophyll protoplasts of pea (Pisum sativum) is about 1250 microeinsteins per square meter per second. On exposure to supra-optimal light intensity (2500 microeinsteins per square meter per second) for 10 min, the protoplasts lost 30 to 40% of their photosynthetic capacity. Illumination with normal light intensity (1250 microeinsteins per square meter per second) for 10 min enhanced the rate of dark respiration in protoplasts. On the other hand, when protoplasts were exposed to photoinhibitory light, their dark respiration also was markedly reduced along with photosynthesis. The extent of photoinhibition was increased when protoplasts were incubated with even low concentrations of classic respiratory inhibitors: 1 micromolar antimycin A, 1 micromolar sodium azide, and 1 microgram per milliliter oligomycin. At these concentrations, the test inhibitors had very little or no effect directly on the process of photosynthetic oxygen evolution. The promotion of photoinhibition by inhibitors of oxidative electron transport (antimycin A, sodium azide) and phosphorylation (oligomycin) was much more pronounced than that by inhibitors of glycolysis and tricarboxylic acid cycle (sodium fluoride and sodium malonate, respectively). We suggest that the oxidative electron transport and phosphorylation in mitochondria play an important role in protecting the protoplasts against photoinhibition of photosynthesis. Our results also demonstrate that protoplasts offer an additional experimental system for studies on photoinhibition.

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Regulation of photosynthesis by end-product accumulation in leaves of plants storing starch, sucrose, and hexose sugars.

In the present study, leaves of different plant species were girdled by the hot wax collar method to prevent export of assimilates. Photosynthetic activity of girdled and control leaves was evaluated 3 to 7 days later by two methods: (a) carbon exchange rate (CER) of attached leaves was determined under ambient CO(2) concentrations using a closed gas system, and (b) maximum photosynthetic capacity (A(max)) was determined under 3% CO(2) with a leaf disc O(2) electrode. Starch, hexoses, and sucrose were determined enzymically. Typical starch storers like soybean (Glycine max L.) (up to 87.5 milligrams of starch per square decimeter in girdled leaves), cotton (Gossypium hirsutum L.), and cucumber (Cucumis sativus L.) responded to 7 days of girdling by increased (80-100%) stomatal resistance (r(s)) and decreased A(max) (>50%). On the other hand, spinach (Spinacia oleracea L.), a typical sucrose storer (up to 160 milligrams of sucrose per square decimeter in girdled leaves), showed only a slight reduction in CER and almost no change in A(max). Intermediate plants like tomato (Lycopersicon esculentum Mill.), sunflower (Helianthus annuus L.), broad bean (Vicia faba L.), bean (Phaseolus vulgaris L.), and pea (Pisum sativum L.), which upon girdling store both starch and sucrose, responded to the girdle by a considerable reduction in CER but only moderate inhibition of A(max), indicating that the observed reduction in CER was primarily a stomatal response. Both the wild-type tobacco (Nicotiana sylvestris) (which upon girdling stored starch and hexoses) and the starchless mutant (which stored only hexoses, up to 90 milligrams per square decimeter) showed 90 to 100% inhibition of CER and approximately 50% inhibition of A(max). In general, excised leaves (6 days) behaved like girdled leaves of the respective species, showing 50% reduction of A(max) in wild-type and starchless N. sylvestris but only slight decline of A(max) in spinach. The results of the present study demonstrate the possibility of the occurrence of end-product inhibition of photosynthesis in a large number of crop plants. The long-term inhibition of photosynthesis in girdled leaves is not confined to stomatal responses since the A(max) declined up to 50%. The inhibition of A(max) by girdling was strongest in starch storers, but starch itself cannot be directly responsible, because the starchless mutant of N. sylvestris was also strongly inhibited. Similarly, the inhibition cannot be attributed to hexose sugars either, because soybean, cotton, and cucumber are among the plants most strongly inhibited although they do not maintain a large hexose pool. Spinach, a sucrose storer, showed the least inhibition in both girdled and excised leaf systems, which indicates that sucrose is probably not directly responsible for the end-product inhibition of photosynthesis. The occurrence of strong end-product inhibition appears to be correlated with high acid-invertase activity in fully expanded leaves. The inhibition may be related to the nature of soluble sugar metabolism in the extrachloroplastic compartment and may be caused by a metabolite that has different rates of accumulation and turnover in sucrose storers and other plants.

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Stomatal and nonstomatal limitations to net photosynthesis in seedlings of woody angiosperms.

Comparative responses of net photosynthesis (A) to water stress in woody species from a variety of habitats were studied to assess the relationship between photosynthetic attributes and drought tolerance. Stomatal and nonstomatal limitations to A were compared in three-month-old white oak (Quercus alba L.), post oak (Quercus stellata Wangenh.), sugar maple (Acer saccharum Marsh.), and black walnut (Juglans nigra L.) seedlings during a drying cycle. Relative stomatal limitation of photosynthesis (I) was less than 50% in all species except for Q. stellata seedlings subjected to severe water stress. No significant changes in I were observed in Q. alba and J. nigra before, during, and after drought. In A. saccharum, I was generally low and decreased significantly under water stress. Under well-watered conditions, A was highest in Q. stellata, intermediate in Q. alba, and lower in A. saccharum and J. nigra. High A in well-watered Q. stellata was associated with high stomatal conductance and carboxylation efficiency, whereas low A was associated with low stomatal conductance and carboxylation efficiency in A. saccharum and low stomatal conductance, low carboxylation efficiency, and high CO(2) compensation point in J. nigra. Under severe water stress, A, carboxylation efficiency, and stomatal conductance decreased substantially in all species; however, Q. stellata had the highest carboxylation efficiency and lowest CO(2) compensation point under these conditions. After 5 days at high soil moisture after drought, stomatal and mesophyll components of A in A. saccharum and J. nigra had not recovered to predrought levels, whereas they had completely recovered in Q. stellata and Q. alba. The photosynthetic apparatus, especially mesophyll components, of drought-tolerant Quercus species showed either less inhibition under water stress, superior recovery to predrought capacity, or both. Exposure of the leaves to (14)CO(2) indicated apparent asymmetric stomatal closure for mildly water-stressed seedlings, but not for leaves of well-watered, severely stressed, or rehydrated plants. These results suggest that patchy stomatal closure under mild water stress might be important for water stress-induced inhibition of photosynthesis, but not under the more severe water stress imposed in this study.

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Probing corticular photosynthesis through in vivo chlorophyll fluorescence measurements: evidence that high internal CO levels suppress electron flow and increase the risk of photoinhibition.

Twigs of many woody plants possess chlorenchyma under a well-developed periderm which lacks stomata and impedes both gas diffusion and light penetration. The so-called corticular photosynthesis, occurring in the shade and under extremely high CO(2) concentrations, was probed in this study through in vivo chlorophyll fluorescence measurements. Field comparisons between twigs and corresponding leaves in five species indicated that both the dark- and light-adapted PSII photochemical efficiencies are considerably lower in twigs at all incident photon fluence rates, in spite of the significant attenuation of solar radiation by the periderm. Light saturation curves for linear electron transport rates (corrected according to the actual light intensities reaching twig chlorenchyma) were compatible with a shade-acclimated photosynthetic machinery, showing very low maximum electron transport rates (at approximately 5% of the corresponding leaf values) and threshold irradiances for light saturation. However, removing periderms from twig segments (i.e. relieving the twig interior form the high CO(2) partial pressures) considerably improved the light-adapted (but not the dark-adapted) PSII photochemical efficiency, allowing the assumption that the high internal CO(2) levels may interfere with the smooth functioning of photosynthesis. Indeed, laboratory experiments with twig segments equilibrated under various CO(2) levels (0.036-20%), resulted in a progressive decrease of light-adapted PSII photochemical yield, with the values obtained at 20% CO(2) being similar to those obtained with intact twigs in the field. Further experiments indicated that high CO(2) combined with high light suppressed the development of a photoprotective non-photochemical quenching through a reduction of its fast relaxing component, accompanied by a higher risk of photoinhibition. It is suggested that high internal CO(2) concentrations in twigs impede photosynthesis possibly through acidification of protoplasm and impairment of the pH-dependent high energy state quenching followed by reduction in the efficiency of heat dissipation.

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Monitoring of oxygenic and anoxygenic photosynthesis in a unicyanobacterial biofilm, grown in benthic gradient chamber.

In order to assess the role of cyanobacteria in the formation and dynamics of microenvironments in microbial mats, we studied an experimental biofilm of a benthic, halotolerant strain, belonging to the Halothece cluster of cyanobacteria. The 12-week-old biofilm developed in a sand core incubated in a benthic gradient chamber under opposing oxygen and sulfide vertical concentration gradients. At the biofilm surface, and as a response to high light irradiances, specific accumulation of myxoxanthophyll was detected in the cells, consistent with the typical vertical distribution of sun versus shade species in nature. The oxygen turn-over in terms of gross photosynthesis and net productivity rates was comparable to oxygen dynamics in natural microbial mats. Sulfide blocked O(2) production at low irradiances in deep biofilm layers but the dynamics of H(2)S and pH demonstrated that sulfide removal by anoxygenic photosynthesis was taking place. At higher irradiances, as soon as H(2)S was depleted, the cells switched to oxygenic photosynthesis as has been postulated for natural communities. The similarities between this experimental biofilm and natural benthic microbial mats demonstrate the central role of cyanobacteria in shaping microenvironmental gradients and processes in other complex microbial communities.

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