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The regulation by glutamine of the leaf transcript level corresponding to the Arabidopsis thaliana (L.) Heynh. nitrate reductase gene nia2 was examined using a novel approach: we took advantage of the ability of a ferredoxin-dependent glutamate synthase-deficient gluS mutant of A. thaliana to accumulate glutamine in the leaves when illuminated under conditions that favour photorespiration. The accumulation of glutamine in gluS mutant leaves and the concomitant decline in the leaf glutamate pool were not correlated with a reduction in the foliar nia2 transcript level. This result indicates that glutamine may not exert a negative control of the leaf nia2 transcript pool. The pattern of diurnal nia2 mRNA oscillation did not change upon illumination of the gluS mutant in air, although the leaf glutamine level remained high during the diurnal cycle. The amplitude of the diurnal fluctuation in nia2 transcript abundance, therefore, does not seem to depend on the size of the leaf glutamine pool (which normally fluctuates in opposite phase). This result also appears to argue against a role of glutamine as an effective repressor of nia2 transcript accumulation. The application of a solution containing 100 mM glutamine to the roots of A. thaliana resulted in an increase in the leaf glutamine level and in a decrease in the leaf nia2 transcript level. Net CO2 uptake and chlorophyll fluorescence quenching by attached leaves of A. thaliana were determined as a control of the physiological status of the plants and remained unaffected by the glutamine treatment. However, there was a decrease in the foliar nitrate level. The negative effect on the nia2 transcript pool exerted by exogeneous glutamine may, therefore, be explained as a result of the down-regulation of nitrate-uptake permeases in the roots by glutamine.

The regulation by photorespiration of the transcript level corresponding to plastidic glutamine synthetase (GS-2) was investigated in the leaves of Arabidopsis thaliana (L.) Heynh.. Photorespiration was suppressed by growing the plants in an atmosphere containing 300 Pa CO2. Suppression of photorespiration was demonstrated by the ability of the conditionally lethal serine hydroxymethyltransferase (SHMT)-deficient stm mutant of A. thaliana to grown normally under these conditions. In contrast to previous studies with bean or pea that were performed at very high CO2 partial pressure (2-4 kPa; Edwards and Coruzzi, 1989, Plant Cell 1:241-248; Cock et al., 1991, Plant Mol Biol 17: 761-771), suppression of photorespiration during growth of A. thaliana in an atmosphere with 300 Pa CO2 had no effect of the leaf GS-2 transcript level. In the short term, neither suppression of photorespiration induced by the transfer of air-grown A. thaliana plants into a CO2-enriched atmosphere, nor an increase in the rate of photorespiration achieved by the transfer of high-CO2-grown A. thaliana plants into air resulted in a change in the GS-2 mRNA level. The absence of photorespiratory ammonium release in leaves of the stm mutant had no effect on the GS-2 transcript level. Overall, our data argue against a control by photorespiration of the A. thaliana leaf GS-2 mRNA pool. In contrast, regulation of the leaf SHMT mRNA level may involve a negative feedback effect of at least one metabolite derived from the glycine/serine conversion during photorespiration, as indicated by the overexpression of SHMT transcripts in the leaves of the stm mutant.

Attached leaves of sunflower (Helianthus annuus L.) were exposed to (14)CO2 during steady-state photosynthesis for 2 to 30 min in 345 μl/l CO2 and 21% O2 at 29° C and a light intensity of 1300 μE m(-2)s(-1). Glycolic acid was extracted with water and diethyl ether, and was determined in the aqueous residue by high-pressure liquid column chromatography. The relative specific radioactivity of the glycolic acid synthesized during photosynthesis reached about 100% after 30 min of photosynthesis and was almost equal to that of the CO2 evolved during photorespiration, their ratio at all times being nearly one. These results provide strong in-vivo evidence that the glycolic acid is the substrate for CO2 evolved by sunflower leaves in light.

Using an open gas-exchange system, apparent photosynthesis, true photosynthesis (TPS), photorespiration (PR) and dark respiration of sunflower (Helianthus annuus L.) leaves were determined at three temperatures and between 50 and 400 μl/l external CO2. The ratio of PR/TPS and the solubility ratio of O2/CO2 in the intercellular spaces both decreased with increasing CO2. The rate of PR was not affected by the CO2 concentration in the leaves and was independent of the solubility ratio of oxygen and CO2 in the leaf cell. At photosynthesis-limiting concentrations of CO2, the ratio of PR/TPS significantly increased from 18 to 30°C and the rate of PR increased from 4.3 mg CO2 dm(-2) h(-1) at 18°C to 8.6 mg CO2 dm(-2) h(-1) at 30°C. The specific activity of photorespired CO2 was CO2-dependent but temperature-independent, and the carbon traversing the glycolate pathway appeared to be derived both from recently fixed assimilate and from older reserve materials. It is concluded that PR as a percentage of TPS is affected by the concentrations of O2 and CO2 around the photosynthesizing cells, but the rate of PR may also be controlled by other factors.

Chloroplasts with high rates of photosynthetic O2 evolution (up to 120 μmol O2· (mg Chl)(-1)·h(-1) compared with 130 μmol O2· (mg Chl)(-1)·h(-1) of whole cells) were isolated from Chlamydomonas reinhardtii cells grown in high and low CO2 concentrations using autolysine-digitonin treatment. At 25° C and pH=7.8, no O2 uptake could be observed in the dark by high- and low-CO2 adapted chloroplasts. Light saturation of photosynthetic net oxygen evolution was reached at 800 μmol photons·m(-2)·s(-1) for high- and low-CO2 adapted chloroplasts, a value which was almost identical to that observed for whole cells. Dissolved inorganic carbon (DIC) saturation of photosynthesis was reached between 200-300 μM for low-CO2 adapted chloroplasts, whereas high-CO2 adapted chloroplasts were not saturated even at 700 μM DIC. The concentrations of DIC required to reach half-saturated rates of net O2 evolution (Km(DIC)) was 31.1 and 156 μM DIC for low- and high-CO2 adapted chloroplasts, respectively. These results demonstrate that the CO2 concentration provided during growth influenced the photosynthetic characteristics at the whole cell as well as at the chloroplast level.

The (12)CO2- and (14)CO2-exchange of illuminated corn leaf discs were measured at normal (21%) and low (1%) oxygen. After periods of exposure to (14)CO2 or to (14)CO2 followed by (12)CO2, the discs were killed and the specific activities of some metabolites were determined. At both O2 concentrations the specific activity of 3-PGA increased and decreased rapidly during the first 5 min of (14)CO2-feeding or (12)CO2-flushing but did not equilibrate with that of the CO2 in the assimilation chamber even after 15 min. The specific activity of aspartic acid also showed bimodal kinetics during both feeding and flushing. The specific activities of 3-phosphoglyceric acid (3-PGA), aspartic acid and alanine were higher at 1% O2 than at 21% O2, but glycine and serine were lower in specific activity at 1% O2. The results are in agreement with the proposed initial fixation of CO2 into C4-dicarboxylic acids and subsequent transfer of this carbon to 3-PGA. Indirect evidence supports the idea that at 21% O2, CO2 was produced by the corn leaf discs in the light and was refixed into C4-dicarboxylic acids. At 1% O2, the photorespiratory process could also have been active although the flux of carbon through the glycolate pathway was probably smaller than at 21% O2.

The O2- and CO2-exchange of detached, illuminated leaves from 18 species of plants grown in the Botanical Garden at the University of Frankfurt was measured at 22° C in an open gas stream (400 ppm CO2 supplied to the assimilation chamber) at both 2% O2 and 21% O2.At a low partial pressure of oxygen (2%) the ratios of CO2 uptake to O2 evolution were in all cases near unity, with a mean of 0.93±0.01. In normal atmosphere (21% O2) the CO2/O2 ratios were higher than those in 2% O2 and averaged 1.26±0.01.The results are interpreted as showing that oxygen is involved in the regulation of metabolism and may cause a shift in the direction of carbon flow in photosynthesizing tissue.

Sunflower (Helianthus annuus L.) leaf discs were exposed to (14)CO2 or (14)CO2 followed by (12)CO2 at 21% O2 and three different CO2 concentrations. After intervals of up to 15 min, the specific activity of some photosynthetic intermediates was determined. At all CO2 concentrations, the specific activity of 3-phosphoglyceric acid (3-PGA) increased most rapidly and after 15 min of (14)CO2 feeding was 92% (967 ppm CO2), 87% (400 ppm CO2) and 53% (115 ppm CO2) of CO2 supplied to the assimilation chamber. The specific activity of glycine, serine and the photorespiratory CO2 was similar at all CO2 concentrations, in aggreement with their proposed close metabolic relationship in the glycolate pathway. However, the kinetics of serine and glycine labelling suggested that serine was not totally derived from glycine. Because the specific activity of these glycolate-pathway intermediates was very differnet from that of 3-PGA at all CO2 concentrations, not all of the carbon traversing this pathway came directly from the Calvin cycle. The non-equilibration of the 3-PGA with the feeding gas reflects the recycling of C from the glycolate pathway into the photosynthetic reduction cycle. Measurements of the rates of CO2 evolution in the light and estimates of the C flux through the glycolate pathway suggest that the photorespiratory activity was high and similar at 115 ppm CO2 and 400 ppm CO2 but inhibited at 967 ppm CO2.

Sunflower (Helianthus annuus L.) leaf discs were exposed to (14)CO2 or (14)CO2 followed by (12)CO2 in an open gas-exchange system with incoming gas of approximately 400 ppm CO2 and either 21% or 1% O2. The (14)CO2 and (12)CO2 gas-exchange of the leaf discs were measured, and the specific activities of several metabolites were determined after different lengths of time. The rate of CO2 efflux by the leaf discs was ca. 20% of the net photosynthetic rate at 21% O2 but no CO2 efflux could be detected at 1% O2. At both O2 concentrations the specific activity of 3-phosphoglyceric acid (3-PGA) increased and decreased rapidly for the first 5 min, and then more slowly during (14)CO2 feeding and (12)CO2 flushing. At 21% O2, glycine, serine and alanine changed more slowly in specific activity than 3-PGA and at 1% O2 their specific activities were much lower than at 21% O2. The results at both O2 concentrations indicated that the glycolate pathway compounds were not derived solely from Calvin-cycle intermediates. At 1% O2 the flux of carbon from the immediate fixation products was inhibited and serine was at least partially produced from a precursor of higher specific activity than glycine, although the glycolate pathway may have been active even at 1% O2. The difference between the specific activities of 3-PGA and the feeding gases could be explained by the recycling of C from the glycolate pathway.

Carbon dioxide and oxygen gas exchange of illuminated Amaranthus and Phaseolus leaves was measured from 0-600 ppm of CO2 in an open system.At low oxygen concentration (2% O2) the ratio of CO2 uptake to O2 evolution came close to 1.At high oxygen partial pressure (42% O2) the O2 compensation point of an Amaranthus leaf was increased and oxygen evolution was depressed. Accordingly the CO2/O2 quotients were variable; the lowest value of 1,9 differed significantly from 1,0.The oxygen and carbon dioxide compensation points of a Phaseolus leaf were increased at high oxygen concentration (42% O2) and oxygen evolution as well as carbon dioxide uptake were reduced. Therefore the ratios CO2 over O2 varied and differed greatly from 1,0.It was concluded that the nature of photosynthates is regulated by the gas composition around the leaves.