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Effect of low root temperature on net photosynthesis, stomatal conductance and carbohydrate concentration in Engelmann spruce (Picea engelmannii Parry ex Engelm.) seedlings.

The effect of low root temperature on net photosynthesis, stomatal conductance and carbohydrate concentration in potted Engelmann spruce (Picea engelmannii Parry ex Engelm.) seedlings was examined under controlled growth-chamber conditions. Root temperature had no effect on net photosynthesis between 10 and 20 degrees C, however, conductance and photosynthesis declined sharply below 8 degrees C. Net photosynthesis and stomatal conductance decreased to 50 and 34% of the initial values after 7 days at a root temperature of 0.7 degrees C. Low root temperature also caused a decrease in photosynthetic utilization of internal CO(2), carboxylation efficiency and apparent quantum yield, and it was concluded that the decrease in photosynthetic rate was caused primarily by nonstomatal limitations. Root chilling caused a small increase in starch content in needles and stems and induced the hydrolysis of starch to glucose in roots. The increase in glucose concentration may enable continued root growth at low temperatures.

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On estimating canopy photosynthesis and stomatal conductance in a deciduous forest with clumped foliage.

The foliage in a fully-leafed deciduous forest canopy is clumped. Consequently, theory indicates that the probability of beam penetration will be estimated more accurately with a model based on the negative binomial distribution than with a model based on the Poisson distribution, incorporating an assumption of a spherical leaf inclination angle distribution. Flux densities of photosynthetically active radiation (PAR) were measured in and above a deciduous forest canopy and were computed with the canopy radiative transfer models based on the negative binomial and Poisson distributions. These radiation values were used to compute canopy photosynthesis and stomatal conductance. Canopy photosynthesis and stomatal conductance, based on the negative binomial model, overestimated values computed from measured PAR profiles by 8 and 9%. respectively. The canopy photosynthesis and stomatal conductance values computed with the spherical Poisson model under-estimated measured values by 17 and 10%, respectively. Thus, the negative binomial radiative transfer model improves estimates of canopy photosynthesis and, to a lesser extent, stomatal conductance, inside a deciduous forest.

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Effects of season, needle age and elevated atmospheric CO(2) on photosynthesis in Scots pine (Pinus sylvestris).

Five-year-old Scots pine (Pinus sylvestris L.) seedlings were grown in open-top chambers at ambient and elevated (ambient + 400 &mgr;mol mol(-1)) CO(2) concentrations. Net photosynthesis (A), specific leaf area (SLA) and concentrations of nitrogen (N), carbon (C), soluble sugars, starch and chlorophyll were measured in current-year and 1-year-old needles during the second year of CO(2) enrichment. The elevated CO(2) treatment stimulated photosynthetic rates when measured at the growth CO(2) concentration, but decreased photosynthetic capacity compared with the ambient CO(2) treatment. Acclimation to elevated CO(2) involved decreases in carboxylation efficiency and RuBP regeneration capacity. Compared with the ambient CO(2) treatment, elevated CO(2) reduced light-saturated photosynthesis (when measured at 350 &mgr;mol mol(-1) in both treatments) by 18 and 23% (averaged over the growing season) in current-year and 1-year-old needles, respectively. We observed significant interactive effects of CO(2) treatment, needle age and time during the growing season on photosynthesis. Large seasonal variations in photosynthetic parameters were attributed to changes in needle chemistry, needle structure and feedbacks governed by whole-plant growth dynamics. Down-regulation of photosynthesis was probably a result of reduced N concentration on an area basis, although a downward shift in the relationship between photosynthetic parameters and N was also observed.

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Leaf phosphorus and nitrogen concentrations and net photosynthesis in Eucalyptus seedlings.

Seedlings of Eucalyptus grandis Hill ex Maiden, E. pilularis Smith and E. gummifera (Sol. ex Gaertner) Hochr. were grown in solution culture with 100 micromol phosphorus (P). After eight weeks, half of the seedlings were transferred to solution cultures containing 1 micromol P. After a further four weeks, growth, net photosynthesis and foliar P and nitrogen (N) concentrations were measured. The seeds of E. grandis came from a relatively fertile site and those of the other two species from phosphorus-deficient sites. Growth and net photosynthesis did not change in E. pilularis subjected to the low-P treatment, whereas in E. grandis, and to a limited extent in E. gummifera, the low-P treatment resulted in an increase in net photosynthesis that was associated with higher foliar N concentrations (especially protein-N), possibly as a result of nitrogen being retranslocated from the roots. In response to the low-P treatment, leaf phosphorus concentration was reduced by 50-60% in E. grandis and E. pilularis and by 20-30% in E. gummifera. Of the chemical fractions examined, the greatest decrease occurred in the inorganic-P pool. The data suggest that photosynthesis is not limited by leaf phosphorus concentrations typical of those found in Eucalyptus seedlings growing on phosphorus-deficient sites.

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Isolation of regulatory mutants in photosynthesis gene expression in Rhodobacter sphaeroides 2.4.1 and partial complementation of a PrrB mutant by the HupT histidine-kinase.

The photosynthetic bacterium Rhodobacter sphaeroides responds to the transition from aerobiosis to anaerobic photosynthesis by increasing the expression of the photosynthesis genes. Mutants have been isolated based on their inability, following such a transition, to increase transcription of the puc operon encoding the apoproteins of the light-harvesting complex II. Mutant D5, a representative of one mutant class, described here, although remaining photosynthetically competent, produced only low levels of the photosynthetic spectral complexes. Complementation analysis revealed that either the gene for the photosynthesis response regulator prrA or the gene encoding its cognate sensor kinase, prrB, was capable of rescuing this mutant. However, partial complementation of this mutant was achieved by placing in trans additional copies of other defined genes from the cosmid library of R. sphaeroides. We describe this effect in detail, attributable to the hupT gene, which has been proposed to encode a histidine-kinase for the hydrogen uptake system in Rhodobacter capsulatus. The effect of HupT on the expression of the photosynthesis genes was mediated through PrrA and independent of a functioning hydrogen uptake system. Thus, we raise the possibility that HupT can participate in phosphorylation of the heterologous response regulator PrrA by so-called cross-talk and therefore partially compensate for the defect in the mutant described. The observation of cross-talk, together with the complementation analysis, allowed us to assign the original mutation to the prrB gene; this was confirmed by DNA sequencing. Analysis of cross-talk in the wild-type, prrB and prrA genetic backgrounds suggested that besides kinase activity, PrrB may possess phosphatase activity toward PrrA. We also report the cloning, organization and structure of some of the hup genes from R. sphaeroides and construction of a Hup- strain.

Amino Acid Sequence↗

Mining thermophile photosynthesis genes: a synthetic operon expressing Chloroflexota species reaction center genes in Rhodobacter sphaeroides.

Photosynthesis is the foundation of the vast majority of life systems, and therefore the most important bioenergetic process on earth, and the greatest diversity in photosynthetic systems are found in microorganisms. However, understanding of the biophysical and biochemical processes that transduce light to chemical energy has derived from the relatively small subset of proteins from microbes that are amenable to cultivation, in contrast to the huge number of microbial DNA sequences encoding proteins that catalyze the initial photochemical reactions that has been deposited in databases, such as from metagenomics. We describe the use of a Rhodobacter sphaeroides laboratory strain for expression of heterologous photosynthesis genes to demonstrate the feasibility of mining this resource, focusing on hot spring Chloroflexota gene sequences. Using a synthetic operon of genes, we produced a photochemically active complex of reaction center proteins in our biological system. We also present bioinformatic analyses of anoxygenic type II reaction center sequences from metagenomic samples collected from hot (42-90° C) springs available through the JGI IMG database, to generate a resource of diverse sequences that potentially are adapted to photosynthesis at such temperatures. These data provide a view into the natural diversity of anoxygenic photosynthesis, through a lens focused on high-temperature environments. The approach we took to express such genes can be applied for potential biotechnology purposes as well as for studies of fundamental catalytic properties of these heretofore inaccessible protein complexes.

Chloroflexota↗

NADP-Malate Dehydrogenase in the C4 Plant Flaveria bidentis (Cosense Suppression of Activity in Mesophyll and Bundle-Sheath Cells and Consequences for Photosynthesis).

Flaveria bidentis, a C4 dicot, was transformed with sorghum (a monocot) cDNA clones encoding NADP-malate dehydrogenase (NADP-MDH; EC 1.1.1.82) driven by the cauliflower mosaic virus 35S promoter. Although these constructs were designed for over-expression, many transformants contained between 5 and 50% of normal NADP-MDH activity, presumably by cosense suppression of the native gene. The activities of a range of other photosynthetic enzymes were unaffected. Rates of photosynthesis in plants with less than about 10% of normal activity were reduced at high light and at high [CO2], but were unaffected at low light or at [CO2] below about 150 [mu]L L-1. The large decrease in maximum activity of NADP-MDH was accompanied by an increase in the activation state of the enzyme. However, the activation state was unaffected in plants with 50% of normal activity. Metabolic flux control analysis of plants with a range of activities demonstrates that this enzyme is not important in regulating the steady-state flux through C4 photosynthesis in F. bidentis. Cosense suppression of gene expression was similarly effective in both the mesophyll and bundle-sheath cells. Photosynthesis of plants with very low activity of NADP-MDH in the bundle-sheath cells was only slightly inhibited, suggesting that the presence of the enzyme in this compartment is not essential for supporting maximum rates of photosynthesis.

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Does Decrease in Ribulose-1,5-Bisphosphate Carboxylase by Antisense RbcS Lead to a Higher N-Use Efficiency of Photosynthesis under Conditions of Saturating CO2 and Light in Rice Plants?

Rice (Oryza sativa L.) plants with decreased ribulose-1,5-bisphosphate carboxylase (Rubisco) were obtained by transformation with the rice rbcS antisense gene under the control of the rice rbcS promoter. The primary transformants were screened for the Rubisco to leaf N ratio, and the transformant with 65% wild-type Rubisco was selected as a plant set with optimal Rubisco content at saturating CO2 partial pressures for photosynthesis under conditions of high irradiance and 25[deg]C. This optimal Rubisco content was estimated from the amounts and kinetic constants of Rubisco and the gas-exchange data. The R1 selfed progeny of the selected transformant were grown hydroponically with different N concentrations. Rubisco content in the R1 population was distributed into two groups: 56 plants had about 65% wild-type Rubisco, whereas 23 plants were very similar to the wild type. Although the plants with decreased Rubisco showed 20% lower rates of light-saturated photosynthesis in normal air (36 Pa CO2), they had 5 to 15% higher rates of photosynthesis in elevated partial pressures of CO2, (100-115 Pa CO2) than the wild-type plants for a given leaf N content. We conclude that the rice plants with 65% wild-type Rubisco show a higher N-use efficiency of photosynthesis under conditions of saturating CO2 and high irradiance.

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Photosynthesis and transpiration of monterey pine seedlings as a function of soil water suction and soil temperature.

Rates of photosynthesis, respiration, and transpiration of Monterey pine (Pinus radiata D. Don) were measured under controlled conditions of soil water suction and soil temperature. Air temperature, relative humidity, light intensity, and air movement were maintained constant. Rates of net photosynthesis, respiration, and transpiration decreased with increasing soil water suction. The decrease in the rates of net photosynthesis and transpiration as a function of the soil temperature at low soil water suctions may be attributed to changes in the viscosity of water. At soil water suctions larger than 0.70 bars rates of transpiration and net photosynthesis may be affected in the same proportion by changes in stomatal apertures.

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Photosynthesis: action spectra for leaves in normal and low oxygen.

The action spectrum of apparent photosynthesis for attached radish (Raphanus sativus L. var. Early Scarlet Globe) and corn (Zea mays L. var. Pride V.) leaves was measured at 300 mul/l CO(2) and both 21% and 2% O(2). The spectra were measured at light intensities where apparent photosynthesis was proportional to intensity. For radish, a high compensation point plant, oxygen had an inhibiting effect on photosynthesis at all wavelengths from 402 to 694 mmu. If a constant rate of photosynthesis at 21% O(2) for the different wavelengths was chosen, then the percent increase in net CO(2) fixation at 2% O(2) was constant. For corn, a low compensation point plant, no inhibitory effect of oxygen concentration from 2% to 21% O(2) was found over the visible spectrum. The CO(2) compensation point for light intensities greater than the light compensation point was found to be constant and independent of wavelength for both radish and corn leaves. For radish, the lowering of the oxygen concentration from 21% to 2% at these intensities was found to reduce the CO(2) compensation point by the same amount for the wavelengths studied.

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Differing sensitivity of photosynthesis to low leaf water potentials in corn and soybean.

Rates of net photosynthesis were studied in soil-grown corn (Zea mays) and soybean (Glycine max) plants having various leaf water potentials. Soybean was unaffected by desiccation until leaf water potentials were below -11 bars. Rates of photosynthesis in corn were inhibited whenever leaf water potentials dropped below -3.5 bars.The differences in photosynthetic behavior could be attributed solely to differences in stomatal behavior down to leaf water potentials of -16 bars in soybean and -10 bars in corn. Below these potentials, other factors in addition to stomatal closure caused inhibition, although their effect was relatively small.Corn, which has the C(4)-dicarboxylic acid pathway for carbon fixation, generally had a higher rate of photosynthesis than soybean during desiccation. Nevertheless, since inhibition of photosynthesis began at higher potentials than in soybean, and since corn was less able to withstand severe desiccation without tissue death, it was concluded that the C(4) pathway confers no particular ability to withstand low leaf water potentials.

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Plants under Climatic Stress: I. Low Temperature, High Light Effects on Photosynthesis.

Photosynthetic rates of both C(4)- and C(3)-pathway plants grown at 25 C were measured before and during a period of chilling stress at 10 C, and then again at 25 C following various periods at 10 C. When temperatures are first lowered photosynthetic rates drop immediately, then undergo a further reduction which is quite rapid in species such as Sorghum, maize, and Pennisetum; slower in soybean; and very slow in Paspalum and ryegrass. Visible light causes progressive permanent damage to the photosynthetic capacity of leaves during this period of lowered photosynthesis. The extent of damage increases with light intensity and the length of time leaves are held at 10 C but varies greatly between species, being roughly correlated with the extent to which chilling initially and subsequently lowers photosynthesis. Three days of chilling (10 C) at 170 w.m(-2) reduces the photosynthetic capacity of youngest-mature Paspalum leaves only 30 to 40% while Sorghum leaves are essentially inoperative when returned to 25 C after the same stress. Root temperature has a substantial rapid effect on photosynthesis of soybean and little immediate effect on Sorghum. Photosynthesis of stress-intolerant species (Sorghum) is reduced only slightly more than that of semitolerant species (Paspalum) when temperatures are lowered at mid-photo-period, but to a far greater extent if temperatures are reduced at the commencement of a photoperiod.

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Effects of light intensity and oxygen on photosynthesis and translocation in sugar beet.

The mass transfer rate of (14)C-sucrose translocation from sugar beet (Beta vulgaris, L.) leaves was measured over a range of net photosynthesis rates from 0 to 60 milligrams of CO(2) decimeters(-2) hour(-1) under varying conditions of light intensity, CO(2) concentration, and O(2) concentration. The resulting rate of translocation of labeled photosynthate into total sink tissue was a linear function (slope = 0.18) of the net photosynthesis rate of the source leaf regardless of light intensity (2000, 3700, or 7200 foot-candles), O(2) concentration (21% or 1% O(2)), or CO(2) concentration (900 microliters/liter of CO(2) to compensation concentration). These data support the theory that the mass transfer rate of translocation under conditions of sufficient sink demand is limited by the net photosynthesis rate or more specifically by sucrose synthesis and this limitation is independent of light intensity per se. The rate of translocation was not saturated even at net photosynthesis rates four times greater than the rate occurring at 300 microliters/liter of CO(2), 21% O(2), and saturating light intensity.

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Limitation of acetylene reduction (nitrogen fixation) by photosynthesis in soybean having low water potentials.

The role of photosynthesis and transpiration in the desiccation-induced inhibition of acetylene reduction (nitrogen fixation) was investigated in soybean (Glycine max [L.] Merr. var. Beeson) using an apparatus that permitted simultaneous measurements of acetylene reduction, net photosynthesis, and transpiration. The inhibition of acetylene reduction caused by low water potentials and their aftereffects could be reproduced by depriving shoots of atmospheric CO(2) even though the soil remained at water potentials that should have favored rapid acetylene reduction. The inhibition of acetylene reduction at low water potentials could be partially reversed by exposing the shoots to high CO(2) concentrations. When transpiration was varied independently of photosynthesis and dark respiration in plants having high water potentials, no effects on acetylene reduction could be observed. There was no correlation between transpiration and acetylene reduction in the CO(2) experiments. Therefore, the correlation that was observed between transpiration and acetylene reduction during desiccation was fortuitous. We conclude that the inhibition of shoot photosynthesis accounted for the inhibition of nodule acetylene reduction at low water potentials.

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Oxygen Inhibition of Photosynthesis: I. Temperature Dependence and Relation to O(2)/CO(2) Solubility Ratio.

The magnitude of the percentage inhibition of photosynthesis by atmospheric levels of O(2) in the C(3) species Solanum tuberosum L., Medicago sativa L., Phaseolus vulgaris L., Glycine max L., and Triticum aestivum L. increases in a similar manner with an increase in the apparent solubility ratio of O(2)/CO(2) in the leaf over a range of solubility ratios from 25 to 45. The solubility ratio is based on calculated levels of O(2) and CO(2) in the intercellular spaces of leaves as derived from whole leaf measurements of photosynthesis and transpiration. The solubility ratio of O(2)/CO(2) can be increased by increased leaf temperature under constant atmospheric levels of O(2) and CO(2) (since O(2) is relatively more soluble than CO(2) with increasing temperature); by increasing the relative levels of O(2)/CO(2) in the atmosphere at a given leaf temperature, or by increased stomatal resistance. If the solubility ratio of O(2)/CO(2) is kept constant, as leaf temperature is increased, by varying the levels of O(2) or CO(2) in the atmosphere, then the percentage inhibition of photosynthesis by O(2) is similar. The decreased solubility of CO(2) relative to O(2) (decreased CO(2)/O(2) ratio) may be partly responsible for the increased percentage inhibition of photosynthesis by O(2) under atmospheric conditions with increasing temperature.

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Oxaloacetate as the Source of Carbon Dioxide for Photosynthesis in Bundle Sheath Cells of the C(4) Species Panicum maximum.

3-Mercaptopicolinic acid (3-MPA), an inhibitor of phosphoenolpyruvate carboxykinase, was employed to study the role of organic acid decarboxylation during C(4) photosynthesis. Treatment of detached Panicum maximum leaves with 5 mm 3-MPA inhibited photosynthesis 70 to 75%. Oxygen was found to have no effect on the degree of inhibition. The postillumination (14)CO(2) burst associated with P. maximum photosynthesis was almost abolished by 5 mm 3-MPA. The turnover rates of malate and aspartate during C(4) photosynthesis were severely reduced as well as the rates of formation of C(3) cycle intermediates in P. maximum leaves treated with 3-MPA. These results are interpreted as direct evidence for the fixation of CO(2), arising from the decarboxylation of oxaloacetate, by the C(3) cycle in bundle sheath cells of P. maximum leaves.

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Effects of Sink Removal on Photosynthesis and Senescence in Leaves of Soybean (Glycine max L.) Plants.

Photosynthetic rate, ribulose 1,5-bisphosphate carboxylase activity, specific leaf weight, and leaf concentrations of carbohydrates, proteins, chlorophyll, and inorganic phosphate were determined periodically from midbloom until maturity in leaves of soybean plants (Glycine max L., var. Hodgson) from which reproductive and vegetative sinks had been removed 32 hours before measurement, or continuously since midbloom.Leaf photosynthesis, measured in the top of the canopy, was partially inhibited by both sink removal treatments. This inhibition was of constant magnitude from midbloom until maturity.Leaf photosynthesis in the top of the canopy declined from midbloom until maturity in the control as well as in the desinked plants. The decline in photosynthesis was gradual at first, but later became more abrupt. The photosynthetic decline was equally evident in the yellowing leaves of control plants and in the dark green leaves of the continuously desinked plants.Neither the inhibition of photosynthesis by sink removal nor the decline in photosynthetic rate with time was clearly related to any of the measured traits.

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Photosynthesis of Grass Species Differing in Carbon Dioxide Fixation Pathways: IV. ANALYSIS OF REDUCED OXYGEN RESPONSE IN PANICUM MILIOIDES AND PANICUM SCHENCKII.

Reduced photorespiration has been reported in Panicum milioides on the basis of lower CO(2) compensation concentrations than in C(3) species, lower CO(2) evolution in the light, and less response of apparent photosynthesis to O(2) concentration. The lower response to O(2) in P. milioides could be due to reduced O(2) competition with CO(2) for reaction with ribulose 1,5-bisphosphate, to a reduced loss of CO(2), or to an initial fixation of CO(2) by phosphoenolpyruvate carboxylase. Experiments were carried out with Panicum maximum Jacq., a C(4) species having no apparent photorespiration; tall fescue (Festuca arundinacea Schreb.), a C(3) species; P. milioides Nees ex Trin.; and Panicum schenckii Hack. The latter two species are closely related and have low photorespiration rates. CO(2) exchange was measured at five CO(2) concentrations ranging from 0 to 260 microliters per liter at both 2 and 21% O(2). Mesophyll conductance or carboxylation efficiency was estimated by plotting substomatal CO(2) concentrations against apparent photosynthesis. In the C(4) species P. maximum, mesophyll conductance was 0.96 centimeters per second and was unaffected by O(2) concentration. At 21% O(2) mesophyll conductance of tall fescue was decreased 32% below the value at 2% O(2). Decreases in mesophyll conductance at 21% O(2) for P. milioides and P. schenckii were similar to that for tall fescue. On the other hand, loss of CO(2) in CO(2)-free air, estimated by extrapolating the CO(2) response curve to zero CO(2), was increased from 1.8 to 6.5 milligrams per square decimeter per hour in tall fescue as O(2) was raised from 2-21%. Loss of CO(2) was less than 1 milligram per square decimeter per hour for P. milioides and P. schenckii and was unaffected by O(2). The results suggest that the reduced O(2) response in P. milioides and P. schenckii is due to a lower loss of CO(2) in the light rather than less inhibition of carboxylation by O(2), since the decrease in carboxylation efficiency at 21% O(2) was similar for P. milioides, P. schenckii, and tall fescue. The inhibition of apparent photosynthesis by 21% O(2) in these three species at low light intensities was similar at 31 to 36% which also indicates similar O(2) effects on carboxylation. Apparent photosynthesis at high light intensity was inhibited less by 21% O(2) in P. milioides (16.8%) and P. schenckii (23.8%) than in tall fescue (28.4%). This lower inhibition in the Panicum species may have been due to a higher degree of recycling of photorespired CO(2) in these species than in tall fescue.

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